Influence of handgrip strength in oxidative stress in children

El ejercicio físico puede producir estrés oxidativo en el individuo lo que pueden condicionar el riesgo cardiovascular en niños y adolescentes. Este estudio pretende analizar el estrés oxidativo según la fuerza muscular isométrica de las extremidades superiores en la edad pediátrica. Se estudiaron 70 niños sanos con edades entre 10 y 14 años, y se analizaron en saliva los lipoperóxidos (LPO), el glutatión reducido (GSH), la ratio GSH/LPO y la catalasa, como marcadores de estrés oxidativo. La muestra se dividió en dos grupos según una condición física superior o inferior medida a través de dinamometría manual (TKK 5110); se diferenció la serie en sujetos prepuberales y puberales. Se encontraron niveles significativamente inferiores de GSH y GSH/LPO en el grupo de niños con fuerza superior, y en los puberales con la mismas características; estos resultados podrían indicar la existencia de un mayor estrés oxidativo en esta situación. En conclusión, los mayores niveles de fuerza músculo esquelética, medida a través de dinamometría manual, posiblemente pueden estar asociados a un mayor estrés oxidativo en niños púberes con condición física musculoesquelética superior.Physical exercise can produce oxidative stress, this situation could contribute cardiovascular risk in children and adolescents. The following study tries to evaluate the oxidative stress produced according to the handgrip strength in infancy. 70 healthy male subjects, ages 10 to 14 years, were studied. In the saliva samples, lipoperoxides (LPO), reduced glutathione (GSH), glutathione/lipoperoxides ratio and catalase were analyzed as biomarkers of oxidative stress. Children were divided into two groups according to their handgrip strength (high or low) measured by handgrip strength (TKK 5110). The groups were also divided into prepubertal and pubertal subjects. We found GSH’s significantly low levels and of GSH/LPO in pubertal group with high handgrip strength. We could possibly conclude that the higher levels of handgrip strength could be associated with more oxidative stress in pubertal group with high fitness

Influencia de la fuerza muscular isométrica de las extremidades superiores en el estrés oxidativo en niños. (Influence of handgrip strength in oxidative stress in children).

<b>Resumen</b><p align="justify">El ejercicio físico puede producir estrés oxidativo en el individuo lo que pueden condicionar el riesgo cardiovascular en niños y adolescentes. Este estudio pretende analizar el estrés oxidativo según la fuerza muscular isométrica de las extremidades superiores en la edad pediátrica. Se estudiaron 70 niños sanos con edades entre 10 y 14 años, y se analizaron en saliva los lipoperóxidos (LPO), el glutatión reducido (GSH), la ratio GSH/LPO y la catalasa, como marcadores de estrés oxidativo. La muestra se dividió en dos grupos según una condición física superior o inferior medida a través de dinamometría manual (TKK 5110); se diferenció la serie en sujetos prepuberales y puberales. Se encontraron niveles significativamente inferiores de GSH y GSH/LPO en el grupo de niños con fuerza superior, y en los puberales con la mismas características; estos resultados podrían indicar la existencia de un mayor estrés oxidativo en esta situación. En conclusión, los mayores niveles de fuerza músculo esquelética, medida a través de dinamometría manual, posiblemente pueden estar asociados a un mayor estrés oxidativo en niños púberes con condición física musculoesquelética superior.</p><b>Abstract</b><p align="justify">Physical exercise can produce oxidative stress, this situation could contribute cardiovascular risk in children and adolescents. The following study tries to evaluate the oxidative stress produced according to the handgrip strength in infancy. 70 healthy male subjects, ages 10 to 14 years, were studied. In the saliva samples, lipoperoxides (LPO), reduced glutathione (GSH), glutathione/lipoperoxides ratio and catalase were analyzed as biomarkers of oxidative stress. Children were divided into two groups according to their handgrip strength (high or low) measured by handgrip strength (TKK 5110). The groups were also divided into prepubertal and pubertal subjects. We found GSH’s significantly low levels and of GSH/LPO in pubertal group with high handgrip strength. We could possibly conclude that the higher levels of handgrip strength could be associated with more oxidative stress in pubertal group with high fitness.</p>doi:10.5232/ricyde2011.0220

Development of a thermosensitive hydrogel based on Polaxamer 407 and gellan gum with inclusion complexes (Sulfobutylated-β-cyclodextrin-Farnesol) as a local drug delivery system

This work proposes the development of a thermosensitive local drug release system based on Polaxamer 407, also known as Pluronic® F-127 (PF-127), Gellan Gum (GG) and the inclusion complex Sulfobutylated-β-cyclodextrin (CD) with Farnesol (FOH). Rheological properties of the hydrogels and their degradation were studied. According to the rheological results, a solution of 20% w/v of PF-127 forms a strong gel with a gelling temperature of about 25 °C (storage modulus of 15,000 Pa). The addition of the GG increased the storage modulus (optimal concentration of 0.5 % w/v) twofold without modifying the gelling temperature. Moreover, including 0.5% w/v of GG also increased 6 times the degradation time of the hydrogel. Regarding the inclusion complex, the addition of free CD decreased the viscosity and the gel strength since polymer chains were included in CD cavity without affecting the gelling temperature. Contrarily, the inclusion complex CD-FOH did not significantly modify any property of the formulation because the FOH was hosted in the CD. Furthermore, a mathematical model was developed to adjust the degradation time. This model highlights that the addition of the GG decreases the number of released chains from the polymeric network (which coincides with an increase in the storage modulus) and that the free CD reduces the degradation rate, protecting the polymeric chains. Finally, FOH release was quantified with a specific device, that was designed and printed for this type of system, observing a sustainable drug release (similar to FOH aqueous solubility, 8 μM) dependent on polymer degradation

Validation of the Women’s Views of Birth Labor Satisfaction Questionnaire (WOMBLSQ4) in the Spanish Population

The satisfaction of women with the birth experience has implications for the health and wellness of the women themselves and also of their newborn baby. The objectives of this study were to determine the factor structure of the Women’s Views of Birth Labor Satisfaction Questionnaire (WOMBLSQ4) questionnaire on satisfaction with the attention received during birth delivery in Spanish women and to compare the level of satisfaction of pregnant women during the birth process with that in other studies that validated this instrument. A cross-sectional study using a self-completed questionnaire of 385 Spanish-speaking puerperal women who gave birth in the Public University Hospitals of Granada (Spain) was conducted. An exploratory factor analysis of the WOMBLSQ4 questionnaire was performed to identify the best fit model. Those items that showed commonalities higher than 0.50 were kept in the questionnaire. Using the principal components method, nine factors with eigenvalues greater than one were extracted after merging pain-related factors into a single item. These factors explain 90% of the global variance, indicating the high internal consistency of the full scale. In the model resulting from the WOMBLSQ4 questionnaire, its nine dimensions measure the levels of satisfaction of puerperal women with childbirth care. Average scores somewhat higher than those of the original questionnaire and close to those achieved in the study carried out in Madrid (Spain) were obtained. In clinical practice, this scale may be relevant for measuring the levels of satisfaction during childbirth of Spanish-speaking women

A personalized intervention to prevent depression in primary care: cost-effectiveness study nested into a clustered randomized trial

Abstract Background: Depression is viewed as a major and increasing public health issue, as it causes high distress in the people experiencing it and considerable financial costs to society. Efforts are being made to reduce this burden by preventing depression. A critical component of this strategy is the ability to assess the individual level and profile of risk for the development of major depression. This paper presents the cost-effectiveness of a personalized intervention based on the risk of developing depression carried out in primary care, compared with usual care. Methods: Cost-effectiveness analyses are nested within a multicentre, clustered, randomized controlled trial of a personalized intervention to prevent depression. The study was carried out in 70 primary care centres from seven cities in Spain. Two general practitioners (GPs) were randomly sampled from those prepared to participate in each centre (i.e. 140 GPs), and 3326 participants consented and were eligible to participate. The intervention included the GP communicating to the patient his/her individual risk for depression and personal risk factors and the construction by both GPs and patients of a psychosocial programme tailored to prevent depression. In addition, GPs carried out measures to activate and empower the patients, who also received a leaflet about preventing depression. GPs were trained in a 10- to 15-h workshop. Costs were measured from a societal and National Health care perspective. Qualityadjustedlife years were assessed using the EuroQOL five dimensions questionnaire. The time horizon was 18 months.This work was supported by grants from the Spanish Ministry of Health, the Institute of Health Carlos III (ISCIII) and the European Regional Development Fund (ERDF) ’A way to build Europe’(grant references PS09/02272, PS09/02147, PS09/01095, PS09/00849 and PS09/00461); the Andalusian Council of Health (grant reference PI-0569-2010); the Spanish Network of Primary Care Research ’redIAPP’ (RD06/0018, RD12/0005/0001); the ’Aragón group’ (RD06/0018/0020, RD12/0005/0006); the ’Bizkaya group’ (RD06/0018/0018, RD12/0005/0010); the Castilla-León Group (RD06/0018/0027); the Mental Health (SJD) Barcelona Group (RD06/0018/0017, RD12/0005/0008); and the Mental-Health, Services and Primary Care (SAMSERAP) MálagaGroup (RD06/0018/0039, RD12/0005/0005)

Influencia de las variables de la etapa intestinal en la digestibilidad de macronutrientes en productos de bollería

[ES] La insuficiencia pancreática exocrina (IPE) es una afección grave asociada a diversas enfermedades caracterizadas por la incapacidad del páncreas para secretar la cantidad suficiente de enzimas, ocasionando una maldigestión y malabsorción de nutrientes, especialmente de los lípidos. El tratamiento consiste en una terapia de suplementación enzimática (TSE), administrando al paciente pancreatina encapsulada (Kreon®). En la actualidad, su dosificación se establece en función de la edad y el peso de cada individuo, además de su ingesta diaria de grasa. Sin embargo, no se tienen en cuenta factores intrínsecos al alimento (origen y tipo de grasa, estructura matricial, etc.) y extrínsecos (pH intestinal y concentración biliar del individuo, etc.) que pueden afectar a la eficiencia del suplemento y por tanto a la digestibilidad de las grasas. En el presente estudio se ha empleado un modelo de digestión gastrointestinal in vitro para analizar la eficacia de distintas dosis del suplemento enzimático Kreon® (0-4000 UL/ g grasa), así como la influencia de las condiciones fisiológicas de la etapa intestinal (pH 6 y 7; concentración biliar 1 y 10 mM), en la digestión de lípidos y glúcidos de diferentes productos de bollería industrial (donut, gofre, croissant y magdalena), acompañados o no de leche. Los resultados muestran que el aumento de la dosis de Kreon® aumentó la digestibilidad de las grasas hasta alcanzar un valor máximo a partir del cual un aumento de la dosis no se tradujo en una mayor lipólisis. En cuanto al efecto del pH y la concentración biliar, en general una mayor concentración de sales biliares favoreció una mayor digestibilidad de las grasas. La glucólisis aumentó en presencia de Kreon® no siendo significativa la dosis de encima (1000-4000), mientras que el pH y la concentración biliar contribuyeron a la glucólisis dependiendo del tipo de producto.[EN] Exocrine pancreatic insufficiency (IPE) is a serious disease associated with different illnesses characterized by the pancreas¿s inability to secrete enough enzymes, causing maldigestión and malabsorption of nutrients, especially lipids. The treatment is an enzymatic supplementation therapy (TSE), administering pancreatin in the form of capsules (Kreon®) to the patient. At present, dosage is established based on age and weigh of each person, too their daily fat intake. However, food¿s intrinsic factors (fat¿s origin and type, matrix structure, etc.) and extrinsic factors (type of processing, intestinal pH and biliary concentration, etc.) that can affect the supplement¿s efficiency and therefore, fats digestibility. In this study, an in vitro gastrointestinal digestion model was used to analyze the efficacy of different doses of the enzyme supplement (Kreon®) (0-4000UL/g fat), as well as, the influence of different physiological conditions during the intestinal stage (pH 6 or 7, bile concentration 1 or 10 mM) in the digestion of lipids and sugars of different industrial bakery products (donut, waffle, croissant and muffin). Results show that increasing the dose of Kreon® increased the digestibility of the fats until a maximum value from which an increase of the dose did not translate into a greater lipolysis. As to how pH and biliary concentration affect, in general a higher concentration of bile salts favored a higher digestibility of fats. Glycolysis increased in the presence of Kreon® regardless the dose (1000-4000), whereas pH and biliary concentration contributed to glycolysis depending on the type of product.Dios Pérez, ID. (2017). Influencia de las variables de la etapa intestinal en la digestibilidad de macronutrientes en productos de bollería. http://hdl.handle.net/10251/87753.TFG

Desarrollo de sistemas inteligentes para tratamientos contra el cáncer colorrectal

[ES] El cáncer colorrectal (CCR) es uno de los diferentes tipos de enfermedades que engloba el concepto de cáncer, ocupando el tercer lugar en incidencia a nivel mundial y es el segundo en mortalidad. La detección temprana es crucial para la supervivencia, y el tratamiento varía desde la cirugía en etapas tempranas hasta opciones más complejas en etapas avanzadas. A pesar de los avances, la investigación es esencial para reducir los efectos secundarios de los tratamientos y optimizar su selectividad, mejorando así la calidad de vida de los pacientes. La baja biodisponibilidad y falta de especificidad de los medicamentos antineoplásicos convencionales han llevado al interés creciente en el uso de nanomedicina en quimioterapia. Por ello, se han desarrollado sistemas de administración de fármacos que mejoran la biodistribución, actividad terapéutica y selectividad de los tratamientos. El objetivo central de esta tesis ha sido mejorar la eficacia de las terapias existentes y reducir los efectos secundarios, aprovechando las diferencias metabólicas entre células normales y cancerígenas. Con este fin, se llevaron a cabo tres estrategias. La primera estrategia desarrollada se enfocó en inhibir la proliferación celular al actuar simultáneamente sobre dos rutas metabólicas de las células cancerígenas. Para lo cual se utilizaron dos compuestos: pregnenolona (P5) y farnesol (FOH). P5 impide que las células tengan la energía necesaria para proliferar al inhibir la acción de la enzima G6PD en la ruta pentosa fosfato (PPP), mientras que, FOH impide la formación de membranas en las nuevas células inhibiendo la síntesis de fosfatidilcolina. Sin embargo, como ocurre con muchos de los fármacos anticancerígenos, estos compuestos tienen carácter hidrofóbico. Por lo que, se propuso el uso de ciclodextrinas (CD), oligosacáridos de alta solubilidad y estructura cónica hidrófoba, capaces de formar complejos de inclusión con moléculas huésped y mejorar las propiedades fisicoquímicas y farmacológicas de dichos compuestos. En primer lugar, se caracterizó la formación de los complejos de inclusión formados entre las moléculas de FOH y P5 con una ciclodextrina, SBE-β-CD. Las razones para usar esta CD es que tiene una elevada solubilidad acuosa y la ventaja de estar aprobada por la FDA. Los resultados de caracterización revelaron que la temperatura afectaba significativamente el proceso de complejación y que era instantáneo, obteniendo complejos con una estabilidad mínima de 2 meses. A continuación, se evaluó la toxicidad in vitro de P5 y FOH, tanto en su forma libre como en complejo (SBE-β-CD – P5 y SBE-β-CD – FOH), determinándose en cada caso, la concentración necesaria para inhibir un 50 % la proliferación celular (IC50). Dicho estudio evidenció que, en el caso de SBE-β-CD, las concentraciones superiores a 1 mM eran desfavorables para la viabilidad celular, señalando la necesidad de trabajar con niveles inferiores, para que la ciclodextrina no afecte a la proliferación celular. Respecto a la pregnenolona (P5), se presentaron ciertos problemas, como la necesidad de concentraciones elevadas de SBE-β-CD – P5 para alcanzar la IC50 o la precipitación de P5 cuando se utilizaba en su forma libre. Por estas razones se descartó la pregnenolona como posible tratamiento anticancerígeno y se suspendió su investigación. Por el contrario, los resultados obtenidos con el farnesol, tanto en su forma libre como en el complejo con SBE-β- CD, mostraron una IC50 prometedora, destacando mayor selectividad en la viabilidad de las líneas cancerígenas de colon, hígado y mama, en comparación con los fibroblastos normales. Los análisis del ciclo celular revelaron que el mecanismo de acción de FOH consistía en inhibir la síntesis de fosfatidilcolina (PC), este hecho se corroboró con ensayos de viabilidad celular. A su vez, la visualización microscópica destacó cambios morfológicos, como pérdida de adherencia y reducción celular, confirmando la eficacia del tratamiento. La investigación de esta estrategia finalizó con la validación in vitro de la eficacia terapéutica de SBE-β-CD – FOH, mediante ensayos con cocultivos. Los resultados demostraron su capacidad para reducir significativamente la viabilidad en líneas cancerígenas de colon e hígado, preservando la viabilidad en células normales. Además, se exploró la posibilidad de reducir costos al verificar que la toxicidad de farnesol no dependía significativamente del tipo de isómero utilizado. En conjunto, los resultados sugieren que los complejos de inclusión con FOH tienen un potencial terapéutico significativo en líneas cancerígenas. Sin embargo, con el propósito de dirigir de forma más específica los fármacos hacia las células cancerígenas, se llevó a cabo una segunda estrategia basada en encapsular los compuestos en exosomas. En primer lugar, se aislaron los exosomas utilizando diferentes métodos y se caracterizaron. Inicialmente se utilizó el producto comercial ExoQuick-TC® ULTRA para aislar exosomas, pero las muestras presentaban interferencias en la cuantificación y determinación de proteínas, lo que llevó a descartar este método. En contraste, el método de ultracentrifugación resultó efectivo, permitiendo el aislamiento de exosomas cuya forma esferoide se confirmó mediante microscopía electrónica de transmisión (TEM). Además, el análisis de Dispersión Dinámica de Luz (DLS) reveló que tenían diámetros inferiores a 200 nm. Y el ensayo de Western Blot (WB) confirmó la presencia de proteínas CD9 y HSP-70 confirmando que se trataba de exosomas. Posteriormente, los exosomas fueron cargados con los compuestos hidrófobos utilizando diversas técnicas. No obstante, debido al descarte de P5, se decidió trabajar con un compuesto ampliamente estudiado, Paclitaxel (PTX). La cuantificación de fármacos reveló que la extrusión era el método más eficaz, ya que se alcanzaban concentraciones de FOH y PTX más altas. Con estos exosomas se realizó una evaluación de la toxicidad in vitro y los resultados indicaron que los exosomas sin carga no afectaban significativamente a la viabilidad celular después de 48 horas, mientras que la viabilidad se redujo con los exosomas cargados con FOH y PTX. Los exosomas procedentes de la línea cancerígena cargados con FOH redujeron en mayor medida la viabilidad celular que los procedentes de la línea normal. Además, se produjo el mismo efecto con los cargados con PTX, aunque en este caso la reducción de la viabilidad fue mucho mayor. Al comparar estos resultados con los de SBE-β-CD – FOH, se observó que los exosomas redujeron la viabilidad de las células cancerosas de manera similar, pero afectando en menor medida a las células normales. Por lo que los exosomas, se presentan como una estrategia prometedora para el transporte selectivo en el tratamiento del cáncer colorrectal. Los resultados obtenidos sugieren que los exosomas pueden ofrecer ventajas significativas en términos de selectividad y eficacia en comparación con otras formas de administración de fármacos. Para finalizar esta tesis, se buscó mejorar el tratamiento local de CCR, llevándose a cabo una tercera estrategia que implica el desarrollo de una terapia local con hidrogeles termosensibles cargados con FOH. En una primera etapa, se determinó la composición óptima de hidrogeles compuestos por Pluronic F-127 (PF-127) y Goma Gellan (GG), a través de pruebas reológicas y evaluación de la degradación del gel. Se concluyó que la concentración óptima de PF-127 para obtener geles termosensibles estables era del 20 % w/v. Esta concentración permitió trabajar con soluciones líquidas a temperaturas inferiores a la ambiente y garantizó la formación de geles estables una vez alcanzada la temperatura de gelificación. La adición de GG al gel favoreció la formación de interacciones poliméricas, resultando en un aumento del módulo de almacenamiento, la viscosidad y la velocidad de degradación. GG demostró ser eficaz para controlar la sensibilidad a la temperatura, y una concentración del 0.50 % w/v duplicó la velocidad de degradación sin afectar significativamente la temperatura de gelificación. Posteriormente, se estudió el impacto de añadir FOH, tanto como fármaco libre o en forma de complejo de inclusión, en la reología y degradación del gel, utilizando geles con un 20 % de FP-127 y 0.5 % de GG. La principal conclusión fue que concentraciones elevadas de FOH libre afectaban negativamente la reología del gel debido a la formación de una emulsión. Por otro lado, la presencia de ciclodextrina (CD) producía una disminución del módulo de almacenamiento y la viscosidad, pero una prolongación del tiempo de degradación, esto se debe a que las cadenas hidrófobas de PF-127 formaban un complejo con la CD. Finalmente, se llevó a cabo un estudio de la cinética de liberación de FOH, permitiendo prever los parámetros que controlan la transferencia de materia tanto en el interior como en el exterior del hidrogel. Los estudios de liberación del fármaco indicaron que era sostenida y proporcional a la degradación del gel. Sin embargo, aunque la liberación de FOH libre resultó en concentraciones superiores a su solubilidad, se recomendaba el uso del complejo para lograr una solubilidad total del fármaco liberado con el fin de que pueda internalizar en las células. Estos resultados respaldan la posibilidad de emplear dicho hidrogeles como sistemas de liberación controlada para el tratamiento oncológico localizado. Además, se destaca el potencial de estos hidrogeles para ser utilizados en la simulación de tumores como se detalla en el Anexo I. Los resultados obtenidos de las tres estrategias presentaron avances significativos en la mejora del tratamiento del CCR. La estrategia de utilizar farnesol en forma de complejo con la ciclodextrina demostró eficacia terapéutica, los exosomas cargados con FOH y PTX exhibieron prometedores niveles de selectividad y los hidrogeles termosensibles ofrecen una terapia local controlada con liberación sostenida de FOH. Estos enfoques, basados en la nanotecnología y la terapia local, abren nuevas perspectivas para optimizar los tratamientos actuales contra el CCR. Todos estos estudios establecen una sólida base para futuras investigaciones, como el desarrollo de hidrogeles que permitan la liberación de exosomas cargados con farnesol y Paclitaxel, de tal forma que actúen únicamente contra células cancerígenas. Además, se podrían perfeccionar los hidrogeles, con el fin de que tengan una doble acción: liberación local de fármacos y regeneración del tejido dañado. [EN] Colorectal cancer (CRC) is one of the types of diseases that includes the concept of cancer, ranking third globally in incidence and second in mortality. Early detection is crucial for survival, the treatment varies from surgery in early stages to more complex options in advanced stages. Despite advances, research is essential to reduce the side effects of treatments and optimize their selectivity, thereby improving the quality of life for patients. The low bioavailability and lack of specificity of conventional antineoplastics have led to increasing interest in the use of nanomedicine in chemotherapy. For his reason, drug delivery systems (DDS) have been developed to enhance drug biodistribution, therapeutic activity, and treatment selectivity. The thesis aims has been to improve the effectiveness of current therapies and minimize side effects, taking advantage of the metabolic distinctions between normal and cancer cells. Three strategies were employed to achieve this goal. The first strategy focused on inhibiting cellular proliferation by simultaneously acting on different metabolic pathways of cancer cells. For this purpose, two compounds were used: pregnenolone (P5) and farnesol (FOH). P5 prevents cells from having the necessary energy to proliferate by inhibiting the action of the G6PD enzyme in the pentose phosphate pathway (PPP), while FOH prevents the formation of membranes in the new cells by inhibiting the synthesis of phosphatidylcholine. However, similar to many anticancer drugs, these compounds are hydrophobic. Therefore, the use of cyclodextrins (CD) was proposed. CD are oligosaccharides with high solubility and a hydrophobic conical structure, capable of forming inclusion complexes with guest molecules, thus improving their physicochemical and pharmacological properties. Firstly, the inclusion complex was characterized using SBE-β-CD and FOH/P5 molecules. SBE-β-CD was selected because it is an FDA-approved cyclodextrin and it has high aqueous solubility. Characterization results revealed that temperature significantly affected the complexation process, which was instantaneous, producing complexes with a minimum stability of 2 months. Next, the in vitro toxicity of P5 and FOH, both in their free form and in complex (SBE-β-CD – P5 and SBE-β-CD – FOH), was evaluated, determining the half-maximal inhibitory concentration (IC50) in each case. The study revealed that concentrations of SBE-β-CD above 1 mM affected adversely at cell viability, emphasizing the need to work with lower levels. Regarding pregnenolone (P5), certain issues arose, such as the need for elevated concentrations of SBE-β-CD – P5 to achieve IC50 or the precipitation of P5 when used in its free form. For these reasons, pregnenolone was ruled out as a potential anticancer treatment, and its investigation was suspended. In contrast, results with farnesol, both in its free form and in complex with SBE-β-CD, showed a promising IC50, highlighting greater selectivity in the viability of colon, liver, and breast cancer cell lines compared to normal fibroblasts. Cell cycle analysis revealed that the mechanism of action of FOH consisted of inhibiting the synthesis of phosphatidylcholine (PC), supported by cell viability assays. Microscopic visualization highlighted morphological changes, such as loss of adherence and cell reduction, confirming the effectiveness of the treatment. This strategy culminated with the in vitro validation of the therapeutic efficacy of SBE-β-CD – FOH, through co-culture assays. The results demonstrated its ability to significantly reduce viability in colon and liver cancer cell lines while preserving viability in normal cells. Furthermore, cost reduction possibilities were explored by confirming that farnesol toxicity did not significantly depend on the type of isomer used. Overall, the results suggest that inclusion complexes with FOH have significant therapeutic potential in cancer cell lines. However, with the aim of more specifically targeting drugs to cancer cells, a second strategy was implemented, involving encapsulating compounds in exosomes. Initially, exosomes were isolated using different methods and characterized. The initial attempt to use the commercial product ExoQuick-TC® ULTRA to isolate exosomes presented interferences in protein determination, leading to the dismissal of this method. In contrast, the ultracentrifugation method was effective, allowing the isolation of exosomes with a spheroid shape confirmed by transmission electron microscopy (TEM). Additionally, Dynamic Light Scattering (DLS) analysis revealed diameters below 200 nm. Western Blot (WB) assays confirmed the presence of CD9 and HSP-70 proteins, confirming their identity as exosomes. Subsequently, the exosomes were loaded with hydrophobic compounds using various techniques. However, due to the discarding of P5, it was decided to work with a widely studied compound, Paclitaxel (PTX). Drug quantification revealed that extrusion was the most effective method, achieving the highest concentrations. In vitro toxicity assessment with these exosomes indicated that unloaded exosomes did not significantly affect cell viability after 48 hours, while viability decreased with exosomes loaded with FOH and PTX. Exosomes from the cancer cell line loaded with FOH reduced cell viability to a greater extent than those from the normal cell line. Additionally, the same effect occurred with those loaded with PTX, although the reduction in viability was much greater. Comparing the results with SBE-β-CD – FOH, it was observed that exosomes reduced cancer cell viability similarly, with a lesser impact on normal cells. Therefore, exosomes emerge as a promising strategy for selective transport in colorectal cancer treatment. The results suggest that exosomes may offer significant advantages in terms of selectivity and efficacy compared to other drug administration forms. To conclude this thesis, a third strategy was implemented to improve local CRC treatment, involving the development of a local therapy with thermosensitive hydrogels loaded with FOH. In the first stage, the optimal composition of hydrogel, using Pluronic F-127 (PF-127) and Gellan Gum (GG),was determined through rheological tests and evaluation of gel degradation. The optimal concentration of PF-127 for obtaining stable thermosensitive gels was identified as 20%. This concentration allowed working with liquid solutions at temperatures below room temperature and ensured the formation of stable gels once the gelation temperature was reached. The addition of GG to the gel favoured the formation of polymeric interactions, resulting in an increase in storage modulus, viscosity, and degradation rate. GG proved effective in controlling temperature sensitivity, and a concentration of 0.50% doubled the degradation rate without significantly affecting the gelation temperature. Subsequently, the impact of adding farnesol on the rheology and degradation of the gel was studied. Using gels with 20% FP-127 and 0.5% GG, the impact of incorporating farnesol into the gel, both as a free drug and in the form of an inclusion complex, was studied. The main conclusion was that high concentrations of free FOH negatively affected the gel rheology due to the formation of an emulsion. On the other hand, the presence of cyclodextrin (CD) resulted in a decrease in storage modulus and viscosity but an extension of the degradation time because the hydrophobic chains of PF-127 formed a complex with CD. Finally, a study of the release kinetics of farnesol was conducted, allowing the anticipation of parameters controlling material transfer both inside and outside the hydrogel. Drug administration studies indicated sustained release proportional to gel degradation. However, although the release of free FOH resulted in concentrations exceeding its solubility, the use of the complex was recommended to achieve complete solubility of the released drug, allowing it to be internalized into cells. These results confirm the viability of hydrogels as controlled release systems for localized oncological treatment. Additionally, the potential of these hydrogels for simultaneous use in tissue regeneration is highlighted, as detailed in Anexo I. These findings establish a solid foundation for future research and clinical applications. The results obtained from the three strategies presented significant advances in improving CRC treatment. The strategy of using farnesol in complex form with cyclodextrin demonstrated therapeutic efficacy, loaded exosomes exhibited promising levels of selectivity, and thermosensitive hydrogels offer controlled local therapy with sustained release of farnesol. These approaches, based on nanotechnology and local therapy, present new perspectives for optimizing current CRC treatments. All these studies establish a solid foundation for future research, such as the development of hydrogels that allow the release of exosomes loaded with farnesol and Paclitaxel, acting only against cancer cells. Additionally, hydrogels could be refined to have a dual action: local drug release and regeneration of damaged tissue

Polyphenol extraction from olive leaves to show Chemical Engineering students the importance of revaluating residues while improving their hands-on experience

[EN]The aim of this work was to try to implement a new laboratory class for Chemical Engineering students in which they could analyze how it is possible to revalue some organic residues. Concretely, the objective was to show them how to revalue olive leaves by extracting their polyphenols, since these leaves, being an abundant organic waste in Spain, were well-known by the students to whom this study was addressed. With the aim of improving student understanding of the basic Chemical Engineering concepts, two extraction techniques (solvent extraction process and ultrasound-assisted extraction) as well as their combination were investigated during the class. In addition, the effect that different pre-treatments had on polyphenol extraction efficiency was also studied to show students how important research is before fine-tuning an industrial process. To do this, students were divided into working groups to carry out polyphenol extraction under different experimental conditions. Ultimately, student groups compared and discussed the efficiency of the different employed extraction techniques. Likewise, students were surveyed to evaluate the suitability and training-usefulness of the proposed class. Obtained assignments revealed that it could be an interesting option to improve students’ hands-on experience while strengthening some theoretical concepts explained in the degree lectures

A Synergistic Approach Therapy for Colorectal Cancer Based on Exosomes and Exploitation of Metabolic Pathways

[EN]In order to reduce the side effects of traditional chemotherapy in the treatment of colorectal cancer (CRC), a newdrug delivery system has been developed in this work, based on exosomes that can host two drugs that act synergistically: farnesol (that stops the cell cycle) and paclitaxel (prevents microtubule system depolymerization). Firstly, exosomes were isolated from different cell cultures (from colorectal cancer and from f ibroblast as example of normal cell line) by different methods and characterized by western blot, TEM and DLS, and results showed that they express classical protein markers such as CD9 and HSP-70 and they showed spherical morphology with sizes from 93 nm to 129 nm depending on the source. These exosomes were loaded with both drugs and its effect was studied in vitro. The efficacy was studied by comparing the viability of cell cultures with a colorectal cancer cell line (HCT-116) and a normal cell line (fibroblast HS-5). Results showed that exosomes present a specific effect with more reduction in cell viability in tumour cultures than healthy ones. In summary, exosomes are presented in this work as a promising strategy for colorectal cancer treatmen

High School Students for Agricultural Science Research. Proceedings of the I Congress PIIISA

Muñoz Dorado, José et al.-- Scientific editors: Alché Ramírez, Juan de Dios; Aranda, Elisabeth; Campos, Mercedes; Corpas, Francisco J.; García Romera, Inmaculada; Jiménez Zurdo, José I.; Martínez-Abarca, Francisco; Palma Martínez, José Manuel; Paredes, Daniel; Sampedro, Inmaculada.-- 46 páginas, con gráficos e imágenes.-- Proceedings of the I Congress PIIISA celebrado en la Estación Experimental del Zaidín, Granada, el 26 de abril de 2012.[ES] Lo que hay escrito a lo largo de estas páginas es el fruto de una andadura que comenzó en Noviembre de 2011: el proyecto PIIISA2012, al que algunos medios de comunicación calificaron como “Investigación con Acné”. El nombre era de lo más sugerente y apropiado, pero había que darle forma y resultado. Han sido casi seis meses de esfuerzo con mayúsculas. Esfuerzo sobre todo por parte de los jóvenes (alumnos de secundaria y Bachillerato), pero también de los investigadores a su cargo, así como de los profesores coordinadores de secundaria. Cada Proyecto ha tenido sus propias peculiaridades y dificultades, y como todo proyecto de Investigación, sus éxitos y sus fracasos; pero todos ellos han tenido un denominador común: la ilusión por conocer la ciencia y todos sus aspectos más escondidos. Los estudiantes han experimentado con la decepción, el acierto, la satisfacción final del trabajo acabado, la publicación y, lo más importante, la presentación a la sociedad que nos ha financiado y nos permite este lujo: el de INVESTIGAR. Estos alumnos “privilegiados” han entrado en contacto con la CIENCIA en mayúsculas desarrollando experimentos nunca realizados por otros. A su vez los investigadores han tenido que ser lo suficientemente flexibles y dinámicos para combinar el objetivo de estos proyectos (ilusionar a adolescentes por la ciencia) con la realidad y prioridades de unos alumnos de secundaria y bachillerato (el acné). A este último aspecto los coordinadores del proyecto estaban mucho más acostumbrados. El resultado lo podréis apreciar y disfrutar a lo largo de estas páginas, que seguro os sorprenderán. Que este libro sea no sólo un reflejo de lo que han sido estos seis meses sino el germen de un apasionante futuro.[EN] What is written along these pages is the fruit of a long journey that began back in November 2011: project PIIISA2012, which some media have described as “Research with Acne”. The name was indeed very appropriate, but it was necessary to actually put it into shape. We spent almost six months to do that devoting much of our time and efforts, as did the teen students involved in the project and their teachers and persons in charge. Each Project has had its own peculiarities and difficulties, and as in every research project, its successes and its failures. Nonetheless, each and every one of them had a common denominator: the excitement of science itself and of all its more hidden aspects. The students have experienced disappointment, success and the final satisfaction of the work well done, as well as its publication and dissemination to the general public responsible of our financing and of making research real and tangible. These “privileged” students have come into contact with the SCIENCE, developing experiments that had never been done before. Researchers, on their side, had to be the sufficiently flexible and dynamic to combine the aim of these projects (making science exciting and attractive to teenage students) with their everyday life and top priorities (acne). Needless to say that the coordinators of the projects were far more familiarized with this unsightly problem than with the more scientific aspects. The results of this endeavour will be thoroughly enjoyed along these pages and will surely surprise you. We hope that this book will not only be a reflection of what these past six months have been, but that it will also set the basis for an exciting scientific future.Biotic interactions in soil and rhizosphere: predation of Sinorhizobium meliloti by Myxococcus xanthus: this work was performed in the Microbiology and Symbiotic Systems Department at Estación Experimental del Zaidín (Consejo Superior de Investigaciones Científicas) within the frame of the Young Students in Science Program (PIIISA2012; http://emc2astronomy.blogspot.com/).It was mainly supported by research project MICINN Consolider‐Ingenio 2010 CSD2009‐00006 and Institutos de Educación Secundaria de Granada: Zaidín Vergeles, Angel Ganivet, Francisco Ayala, Mariana Pineda and Padre Manjón. We also appreciate the coordination of teachers Antonio Quesada, Francisco Javier López, Mª Concepción Martínez and Javier Cáceres.-- Does garlic‐generated nitric oxide (NO) affect the physiology of pepper fruits?: this work was supported by the ERDF‐cofinanced grant AGL2008‐00834 from the Ministry of Science and Innovation, Spain.-- Dry olive residue bioremediation by new saprobe fungi: this work was supported by the Ministerio de Economia y Competitividad (AGL2008‐00572).-- Flower development and pollen morphology in Cucurbitaceae: this work was supported by FEDER funds: The project MEyC BFU2011‐22779 and from “proyectos de excelencia”: (JA) P2010‐AGR6274, P2010‐CVI5767 and P2011‐CVI‐7487.-- Has cupper ion any effect on Sinorhizobium meliloti bacteria?: it was supported mainly by research project MICINN Consolider‐Ingenio 2010. CSD2009‐00006; Scientific research by Young Students in Science (PIIISA2012; http://emc2astronomy.blogspot.com/). Institutos de Educación Secundaria de Granada: Padre Manjón, Angel Ganivet y Fray Luis de Granada.Contiene: Biotic interactions in soil and rhizosphere: predation of Sinorhizobium meliloti by Myxococcus xanthus.Gabriela Tarifa Álvarez; Irene Marín Sánchez; Clara Pérez González; Ana Villar del Paso; Mª Ángeles Yélamos Lorente; Nazaret Ceballos Miján; Raúl Peña López; Manuel Jurado de Haro; José Muñoz Dorado; Juana Pérez; José I. Jiménez Zurdo. -- Determination of individualized breeding methods of Anthocoris nemoralis neonate nymphs. Nuria Sainz; Begoña Castillo; Carolina García; Francisco Sánchez; Ángela Lirola; Elio Gugliere; Mercedes Campos; Iván Batuecas; Daniel Paredes.-- Does garlic‐generated nitric oxide (NO) affect the physiology of pepper fruits?. Nuria García Carbonero; Carlos Nombela Durán; Inmaculada Caballero Guzmán; María del Mar Quesada Díaz; Pilar Montoza García; Alejandra Sánchez Alonso; María Jesús Campos; Carmelo Ruiz; Francisco J. Corpas; José Manuel Palma.-- Dry olive residue bioremediation by new saprobe fungi. María Naranjo Márquez; Ignacio Romero García; Belén Sola Bullejos; José Manuel Ruiz González; Álvaro Arregui Morales; Pablo Ortega Martín; Tomás Ortega Martín; Ramón de la Chica Mora; Carmen Fernández Lacal; David Jiménez Carretero; Elisabet Aranda; Inmaculada Sampedro; Inmaculada García‐Romera.-- Flower development and pollen morphology in Cucurbitaceae. David G. Caracuel; Antonio Jesús Castro; Ana María Cogolludo; Carlos Enríquez; José Carlos Morales; Irene Ruiz‐Gámez; María Victoria Ruiz‐Maldonado; Sergio Torreblanca; Guillermo Vicente; Agnieszka Zienkiewicz; Krzysztof Zienkiewicz; Juan de Dios Alché.-- Has cupper ion any effect on Sinorhizobium meliloti bacteria?. Blanca Barón Torcal; Ana Guardia Molina; Ángela Liébana Medina; Sara Miñán Polo; José Manuel Sanjuán Parra; Francisco Martínez‐Abarca.Peer reviewe