Search

There is a growing concern about the presence of pharmaceuticals on all trophic levels of the 3 aquatic environment. The presence of pharmaceuticals in the marine environment, which is the 4 final receiver of the continental water including treated wastewater, has been much less 5 investigated than in freshwater. Marine mammals are very suitable sentinel species of the 6 marine environment because they often feed at high trophic levels, have unique fat stores and 7 long lifespan. Small delphinids in particular serve as excellent sentinel species for contamination 8 in the marine environment worldwide. To the best of our knowledge, no pharmaceuticals have 9 been detected or reported in dolphins so far. 10 In the present study, muscle, liver and blubbler samples from four common dolphins (Delphinus 11 delphis) and six striped dolphins (Stenella coeruleoalba) stranded along the Basque Coast were 12 collected. A total of 95 pharmaceuticals based on detectability and predicted ability to 13 bioaccumulate in fish were included in the analysis. 14 At least one pharmaceutical was found in 70% of the individuals. Only three of the 95 monitored 15 pharmaceuticals (orphenadrine, pizotifen, and promethazine) were detected in dolphin ́s tissues 16 (liver or blubber) at very low concentrations (<1ng/g). 17 In this study we also highlight the gap in the knowledge regarding the study organisms and 18 marine environments with respect to pharmaceutical pollution, which demands further 19 research to understand the threat pharmaceuticals might pose for these apex predators.

318 p., El objetivo de esta tesis es comparar la efectividad, seguridad y eficiencia de los nuevos anticoagulantes orales (dabigatran, apixaban y rivaroxaban- NACO) frente a acenocumarol en los pacientes con fibrilación auricular que comienzan tratamiento con alguno de estos fármacos. Se seleccionan 10.843 nuevos usuarios de estos fármacos entre 2013 y 2016 en la CAPV y seguidos hasta diciembre de 2019. Se realizan análisis de efectividad comparada y análisis de supervivencia ajustada mediante técnicas de propensity score, ponderación por el inverso de la probabilidad de tratamiento y gradient boosting machine.Los NACO presentan una efectividad similar a acenocumarol en cuanto al riesgo de aparición de ictus oeventos tromboembólicos; el riesgo de hemorragias severas es menor con el empleo de un NACO, aunque los resultados no sean totalmente concluyentes.El uso de los NACO comparado con acenocumarol es coste-efectivo para una disponibilidad a pagar de 20.000 ¿/Año de Vida Ganado. Esta eficiencia se confirma empleando las distintas técnicas de ajuste mencionadas.El elevado tamaño muestral, una media de seguimiento de los pacientes superior a los 4 años, y la utilización de técnicas adecuadas para el análisis de datos de vida real (RWD) son aspectos novedosos que sobre este tema aporta esta tesis.

330 p., En la presente memoria para optar al grado de doctor se presenta un resumen de los resultados y las conclusiones más importantes extraídas de los estudios llevados a cabo con el fin de elucidar el efecto que tiene, por un lado, la temperatura y, por otro, el enriquecimiento con compuestos y mezclas de compuestos con potencial capacidad antioxidante sobre el proceso de oxidación de aceites ricos en grupos acilo poliinsaturados, como son el aceite de girasol y de nuez, sometidos a condiciones de almacenaje acelerado. Para ello, muestras de los aceites sin enriquecer y enriquecidas en compuestos con potencial capacidad antioxidante se sometieron a condiciones de almacenaje acelerado y el seguimiento de su proceso de oxidación se llevó a cabo mediante Resonancia Magnética Nuclear de Protón tanto a través de la degradación de los grupos acilo poliinsaturados del aceite, como de la formación y evolución de la concentración de un gran número de compuestos de oxidación también denominados oxilipinas. De forma general y resumida, se ha puesto de manifiesto que el aumento de la temperatura de almacenaje de 40 ºC a 70 ºC acelera el proceso de oxidación del aceite, aunque no modifica la secuencia de la formación de las distintas oxilipinas. Sin embargo, el enriquecimiento de estos aceites, bien con alfa-tocoferol o con acetato de hidroxitirosilo de forma independiente o con mezclas binarias de ambos, modifica el proceso de oxidación de manera bien distinta. Mientras que el alfa-tocoferol ejerce un claro efecto prooxidante acelerando el proceso de oxidación en la primera etapa y ralentizándolo en la segunda, el acetato de hidroxitirosilo retrasa e inhibe el proceso de oxidación en ambas etapas. Las mezclas de ambos compuestos ejercen un efecto complejo, predominando de forma notable el efecto del alfa-tocoferol sobre el del acetato de hidroxitirosilo en la primera etapa, debido a que aparentemente este último regenera a aquel a costa de su propia degradación, y quedando reducido

246 p., To date, there are no studies on the food environment (FE) in European universities, that include both external and personal domains. This cross-sectional observational study was designed to assess the FE through both dimensions at two European public universities, the University of the Basque Country ¿ UPV/EHU (in northern Spain) and OsloMet ¿ Oslo Metropolitan University (Norway). Food availability, product properties, prices, and marketing, as well as accessibility, food acquisition behaviours, choice determinants and opinions on the food availability on-campus were considered. The results showed that the supply of commercial foods of low nutritional quality was greater at the UPV/EHU in comparison with OsloMet; however, overall no differences were found in the percentage of ultra-processed products. Food purchasing behaviours in both universities were related to their food supply. The top determinants of food choice were those related to taste and cost. Interventions that improve food prices, availability of products of high nutritional quality and information on labelling would be well-received in these communities. These findings are important steps for understanding baseline campus environments and for guiding intervention programme developers and policymakers in their efforts to create campus FEs fully supportive of healthy eating.

405 p., La suplementación génica consiste en la administración de ácidos nucleicos (ADN o ARNm) para inducir la producción de una proteína terapéutica. En la actualidad, el uso del ARNm se ha extendido debido su alta eficacia, perfil de seguridad y versatilidad para una rápida producción de proteínas. Las partículas sólidas lipídicas (SLNs) son consideradas uno de los sistemas no virales lipídicos más eficaces para la administración de ácidos nucleicos. La terapia génica se ha propuesto como una alternativa eficaz para el tratamiento de enfermedades oculares y, en concreto, para la inflamación corneal. Los corticoides son el principal tratamiento para la inflamación corneal, pero su eficacia es limitada y presentan múltiples efectos adversos. La administración de la interleuquina 10 (IL-10) antiinflamatoria a nivel ocular se ha propuesto como una alternativa eficaz. Sin embargo, su escasa biodisponibilidad ocular y su corta vida media limitan su uso. El objetivo de la presente tesis doctoral fue el desarrollo y evaluación de sistemasde liberación de ácidos nucleicos (ADNp o ARNm) basados en SLNs para el tratamiento de la inflamación corneal mediante la administración tópica ocular. Para ello, tras la caracterización fisicoquímica y evaluación in vitro de las formulaciones en diferentes líneas celulares, se evaluó su capacidad para transfectar e inducir la expresión de IL-10 in vivo en la córnea de los ratones tras su administración en forma de colirio. Las formulaciones fueron capaces de producir IL-10 en la córnea. Este trabajo muestra el potencial de la suplementación génica no viral para el tratamiento de la inflamación corneal.

Los estudios 5 y 6 pertenecientes al capítulo 4 están sujetos a confidencialidad por la autora., Before conducting a child-centered study, the selection of the methodology to use is of special importance since it should be adapted to their cognitive, physical, and social stage of development. On this regard, researchers are force to reconsider the existing methodologies on consumer science, mainly design for adults, or to develop new ones.It is thought that the measure of emotions could contribute the understanding of consumer,s preferences and food choices. Since emotions have a multicomponent character, the application of methodologies that measure each component of the emotion are necessary to reach a holistic perspective. This dissertation aimed to design new child-friendly methodologies appropriate to measure food-evoked emotions holistically. To achieve this goal an emoji-based tool was developed and its applicability in food studies was validated to examine the cognitive inherent of emotion. A combined methodology was also designed to apply automatic facial coding and the measure of skin conductance response (SCR) to disentangle the behavioural and physiological components of emotion. On this regard, the applicability of facial coding on the evaluation of spontaneous facial expressions elicited by food stimuli was examined. After, a methodological approach was developed to overcome some limitations frequently associated with the use of facial coding and SCR and their applicability in a broad range of sensory tasks was tested.

Gene therapeutics are promising for treating diseases at the genetic level, with some already validated for clinical use. Recently, nanostructures have emerged for the targeted delivery of genetic material. Nanomaterials, exhibiting advantageous properties such as a high surface-to-volume ratio, biocompatibility, facile functionalization, substantial loading capacity, and tunable physicochemical characteristics, are recognized as non-viral vectors in gene therapy applications. Despite progress, current non-viral vectors exhibit notably low gene delivery efficiency. Progress in nanotechnology is essential to overcome extracellular and intracellular barriers in gene delivery. Specific nanostructures such as carbon nanotubes (CNTs), carbon quantum dots (CQDs), nanodiamonds (NDs), and similar carbon-based structures can accommodate diverse genetic materials such as plasmid DNA (pDNA), messenger RNA (mRNA), small interference RNA (siRNA), micro RNA (miRNA), and antisense oligonucleotides (AONs). To address challenges such as high toxicity and low transfection efficiency, advancements in the features of carbon-based nanostructures (CBNs) are imperative. This overview delves into three types of CBNs employed as vectors in drug/gene delivery systems, encompassing their synthesis methods, properties, and biomedical applications. Ultimately, we present insights into the opportunities and challenges within the captivating realm of gene delivery using CBNs.

Opuntia ficus-indica fruits have been widely used due to their nutritional composition and beneficial effects on health, particularly against chronic diseases such as diabetes, obesity, cardiovascular diseases and cancer, among others. In recent years, prickly pear peel and pulp extracts have been characterised, and a high number of bioactive compounds have been identified. This study aimed to analyse the triglyceride-lowering effect of prickly pear peel and pulp extracts obtained from fruits of three varieties (Pelota, Sanguinos, and Colorada) in 3T3-L1 maturing and mature adipocytes. At a concentration of 50 µg/mL, peel extracts from Colorada reduced triglyceride accumulation in pre-adipocytes and mature adipocytes. Additionally, at 25 µg/mL, Pelota peel extract decreased triglyceride content in mature adipocytes. Moreover, maturing pre-adipocytes treated with 50 and 25 µg/mL of Sanguinos pulp extract showed a reduction of triglyceride accumulation. In addition, the lipid-lowering effect of the main individual betalain and phenolic compounds standards were assayed. Piscidic acid and isorhamnetin glycoside (IG2), found in Colorada peel extract, were identified as the bioactive compounds that could contribute more notably to the triglyceride-lowering effect of the extract. Thus, the betalain and phenolic-rich extracts from Opuntia ficus indica fruits may serve as an effective tool in obesity management.

Marine algae are valuable sources of bioactive compounds that have the potential to be used in the management of various pathologies. Despite the increasing prevalence of NAFLD, the absence of an approved effective pharmacological treatment with demonstrable effectiveness persists. In this context, the aim of the present study is to assess the effect of Gracilaria vermiculophylla red seaweed dietary supplementation on hepatic lipid accumulation, as well as on oxidative stress, inflammation and fibrosis- related markers on obese fa/fa Zucker rats fed with a standard diet, supplemented or not with 2.5% or 5% dehydrated Gracilaria vermiculophylla. After a six-week supplementation with the macroalga, no significant reduction in hepatic total lipid content or hepatic triglyceride content was observed. However, both doses were able to diminish hepatic NEFA concentration by reducing de novo lipogenesis and increasing mitochondrial biogenesis. Moreover, supplementation with the dose of 2.5% improved some oxidative stress and inflammation-related markers. Supplementation with the dose of 5% did not exert these clear beneficial effects. Thus, this study demonstrates that while Gracilaria vermiculophylla may not mitigate hepatic steatosis, it could exert protective effects on the liver by reducing NEFA content and enhancing oxidative stress and inflammation parameters.

(1) Background: Food insecurity (FI) among university students has received less attention in Europe than in other regions before and during the COVID-19 pandemic. (2) Methods: A cross-sectional study was conducted between January and March 2022 using an online questionnaire (n = 924). The questionnaire addressed food security status; demographic, socioeconomic, and educational variables; academic performance; food consumption; and social support networks. The validated Food Insecurity Experience Scale was used to measure food security. Binary logistic regressions adjusted by age and gender were applied to identify FI-related factors. (3) Results: Just over 17% of the students were living with some level of FI, nearly one in three students reported having consumed few kinds of food, and 3.9% spent an entire day without eating due to a lack of resources. Food insecurity was associated with a higher likelihood of negative academic performance, decreased food consumption, and a lower likelihood of having a large support network, when compared to food-secure respondents. (4) Conclusions: The findings suggest that FI negatively impacts students’ academic performance and food consumption. Future public health programs should be prioritized to prevent students from experiencing hunger due to financial or resource constraints.

The only treatment for celiac disease (CD) is a strict and lifelong gluten-free diet (GFD), which must be safe and nutritionally balanced. Avoiding gluten brings difficulties with following the diet and can affect the social life of people with CD. The Zeliakide Project is a nutrition education program aimed at increasing the knowledge of the general population about healthy diets, CD and GFD, and, therefore, to improve the social inclusion and quality of life of people with CD. It is a one-month intervention program, two-armed cluster, non-randomised and controlled trial, conducted among 10–12-year-old children. Pre- and post-intervention evaluation and 1 month follow-up will be carried out to assess the effectiveness of the program. It is based on competencies and their respective learning outcomes. The teaching methodology chosen is a STEAM methodology: inquiry-based learning (IBL). A teaching unit has been created to develop the project, which, in the future, will be useful for the self-application of the program. This study will provide a valid and useful tool to achieve changes in the diet at the school level and will help to promote the social inclusion of people with CD. Moreover, it will enforce the STEAM competences of children.

This study investigated antibiotic utilization in artisanal dairies and residue occurrence throughout the raw milk cheese production chain using commercial testing (Charm KIS and Eclipse Farm3G) and UHPLC-QqQ-MS/MS and LC-QqQ-MS/MS. The cross-sectional survey results revealed gaps in the producers’ knowledge of antibiotic use. Commercial testing detected antibiotic levels close to the LOD in 12.5 % of the samples, mainly in raw milk and whey, with 10.0 % testing positive, specifically in fresh and ripened cheeses, indicating that antibiotics are concentrated during cheese-making. Chromatographically, several antibiotics were identified in the faeces of healthy animals, with chlortetracycline (15.7 ± 34.5 µg/kg) and sulfamethazine (7.69 ± 16.5 µg/kg) predominating. However, only tylosin was identified in raw milk (3.28 ± 7.44 µg/kg) and whey (2.91 ± 6.55 µg/kg), and none were found in fresh or ripened cheeses. The discrepancy between commercial and analytical approaches is attributed to compounds or metabolites not covered chromatographically.

223 p., In recent years, organ transplantation technology has made significant improvements regarding long-term survival, substantially decreasing the risk of graft rejection. However, with each passing year,the gap between available donors and the number of patients needing an organ transplant is steadilyincreasing. Consequently, there is a growing demand to create engineered tissues to minimize the needfor organ transplantation. However, the rejection of engineered tissue implants has become a criticalchallenge today. Primarily, rejection occurs due to the crosstalk between immune cells, such asmacrophages, and the engineered tissue structure. Therefore, it is important to be able to influence theimmune cell milieu of the implantation site to achieve graft integration. With this perspective in mind,this doctoral thesis aimed to provide two different strategies: The first one is the use of human hairfollicle derived mesenchymal stromal cells (HF-MSCs) for immunomodulation. For that, we isolate andcharacterize MSCs from human hair follicles and then, we analyzed the immunomodulatory capacity ofthe cells by means of the inhibition of peripheral mononuclear cells (PBMCs), inducing T regulatoryphenotype and skewing macrophages towards a tissue remodeling phenotype (aka M2). The secondstrategy relied on the application of tissue-engineered blood vessels to interact with different macrophagephenotypes and thus promoting a tissue remodeling response around the implant. The effects of bloodvessels on undifferentiated (M0), pro-inflammatory (M1) and regenerative (M2 and M1M2) macrophagephenotypical change were assessed by single cell analysis, specifically using dimensionality reductionand clustering algorithms for a thorough characterization of resulted macrophage phenotype. Therefore,we could conclude that both strategies could have an important developmental impact towards theimplementation of immunomodulatory techniques and could have important consequences for tissue engineeri

199 p., En la presente tesis doctoral se abordan aspectos relacionados con la biocompatibilidad y bioseguridad de los sistemas de microencapsulación celular para la liberación sostenida de moléculas terapéuticas.En una primera parte, se analizó el comportamiento de una línea de células madre mesenquimales D1 (D1-MSC), genéticamente modificadas para secretar eritropoyetina (EPO), dentro de dichos sistemas. Para ello, se realizaron pruebas de viabilidad y metabolismo celular, proliferación y secreción de la molécula activa, durante 45 días, tanto in vitro como in vivo, en un modelo de ratón. El estudio incluyó 4 grupos de microcápsulas con diferentes cargas celulares, de forma que fue posible evaluar la influencia de dicho parámetro en el comportamiento final de los sistemas. Estas células poseen características de inmunomodulación muy adecuadas para mitigar la respuesta a cuerpo extraño que suele observarse con este tipo de implantes.En una segunda parte, se desarrolló un completo, pero al mismo tiempo sencillo, modelo matemático para el cálculo de la dosis administrada mediante sistemas de micropartículas esféricas, donde el principio activo se encuentra encapsulado en el interior de esferas de hidrogel, o cuando la molécula terapéutica es sintetizada por células inmovilizadas dentro de dichas esferas. El modelo nos da una relación entre un volumen, una cantidad de esferas y una cantidad de células o de principio activo, permitiendo una dosificación exacta y fiable, medida en unidades volumétricas. La validación de las expresiones matemáticas se llevó a cabo con el ejemplo de la tecnología de microencapsulación celular para la liberación sostenida de principios activos.

244 p., El término iPPSD (inactivating PTH/PTHrP signalling disorder) hace referencia a un grupo de enfermedades (epi)genéticas raras caracterizadas todas ellas por una disfunción en la cascada de señalización de PTH/PTHrP. En la presente tesis se ha trabajado con una cohorte de pacientes para identificar alteraciones en los genes asociados a iPPSD, pero también para identificar nuevos mecanismos causales mediante la tecnología de secuenciación masiva (NGS). Recientemente, se han propuesto unos criterios para proceder al diagnóstico clínico de iPPSD. En nuestra investigación también hemos intentado validar esa clasificación mediante estudios de correlación genotipo-fenotipo.Una vez identificadas las alteraciones (epi)genéticas, si las variantes no han sido descritas previamente, es necesario realizar estudios funcionales para validar la patogenicidad de dicha variante. Por tanto, es interesante poder acceder fácilmente a un buen tipo celular para utilizarlo como modelo de la enfermedad (pudiendo hacer modificaciones en su genoma). Con ese fin, se han conseguido aislar células madre mesenquimales derivadas de orina (hUSC), cuya obtención es sencilla, económica y no invasiva. Su caracterización nos ha permitido determinar que son un tipo celular adecuado para su aislamiento, mantenimiento y transporte. Finalmente, se ha analizado en las hUSC la eficiencia de transfección de los vectores no virales conocidos como niosomas catiónicos. Estos vectores son prometedores para el transporte de material genético al interior celular, ya que su fabricación es simple y económica, no presentan problemas de inmunogenicidad y tienen muy buenas características físico-químicas. Se han dado unos primeros pasos a la hora de desarrollar una estrategia de transfección en hUSCs mediante niosomas catiónicos, que podrá ser utilizada con diferentes fines, como por ejemplo terapia génica en enfermedades monogénicas.

In the present study, a novel composite bone cement based on calcium sulfate hemihydrate (CSH) and Mg, Sr-containing bioactive glass (BG) as solid phase, and solution of chitosan as liquid phase were developed. The phase composition, morphology, setting time, injectability, viscosity, and cellular responses of the composites with various contents of BG (0, 10, 20, and 30 wt.%) were investigated. The pure calcium sulfate cement was set at approximately 180 min, whereas the setting time was drastically decreased to 6 min by replacing 30 wt.% glass powder for CSH in the cement solid phase. BG changed the microscopic morphology of the set cement and decreased the size and compaction of the precipitated gypsum phase. Replacing the CSH phase with BG increased injection force of the produced cement; however, all the cements were injected at a nearly constant force, lower than 20 N. The viscosity measurements in oscillatory mode determined the shear-thinning behavior of the pastes. Although the viscosity of the pastes increased with increasing BG content, it was influenced by the frequency extent. Pure calcium sulfate cement exhibited some transient cytotoxicity on human-derived bone mesenchymal stem cells and it was compensated by introducing BG phase. Moreover, BG improved the cell proliferation and mineralization of extracellular matrix as shown by calcein measurements. The results indicate the injectable composite cement comprising 70 wt.% CSH and 30 wt.% Mg, Sr-doped BG has better setting, mechanical and cellular behaviors and hence, is a potential candidate for bone repair, however more animal and human clinical evaluations are essential.

Background: Lenvatinib, a tyrosine kinase inhibitor (TKI) approved for the treatment of progressive and radioactive iodine (RAI)-refractory differentiated thyroid cancer (DTC), is associated with significant adverse effects that can be partially mitigated through the development of novel drug formulations. The utilization of nanoparticles presents a viable option, as it allows for targeted drug delivery, reducing certain side effects and enhancing the overall quality of life for patients. This study aimed to produce and assess, both in vitro and in vivo, the cytotoxicity, biodistribution, and therapeutic efficacy of lenvatinib-loaded PLGA nanoparticles (NPs), both with and without decoration using antibody conjugation (cetuximab), as a novel therapeutic approach for managing aggressive thyroid tumors. Methods: Poly(lactic-co-glycolic acid) nanoparticles (NPs), decorated with or without anti-EGFR, were employed as a lenvatinib delivery system. These NPs were characterized for size distribution, surface morphology, surface charge, and drug encapsulation efficiency. Cytotoxicity was evaluated through MTT assays using two cellular models, one representing normal thyroid cells (Nthy-ori 3-1) and the other representing anaplastic thyroid cells (CAL-62). Additionally, an in vivo xenograft mouse model was established to investigate biodistribution and therapeutic efficacy following intragastric administration. Results: The NPs demonstrated success in terms of particle size, polydispersity index (PDI), zeta potential, morphology, encapsulation efficiency, and cetuximab distribution across the surface. In vitro analysis revealed cytotoxicity in both cellular models with both formulations, but only the decorated NPs achieved an ID50 value in CAL-62 cells. Biodistribution analysis following intragastric administration in xenografted thyroid mice demonstrated good stability in terms of intestinal barrier function and tumor accumulation. Both formulations were generally well tole

Wines covered by PDO or PGI quality labels have specific sensory characteristics. According to EU regulations, product characteristics (including the sensory description) must be defined in the PDO technical specification and should be verified for their compliance. There exist internationally harmonized quantitative descriptive sensory methods applied to products such as virgin olive oil, with well-defined attributes described in the method itself. Currently, in the case of wine, there is no harmonized international sensory descriptive method that allows comparison of results between different PDOs or laboratories. In this work, a qualitative and quantitative olfactory profile for a broad variety of wines (11 PDO and 37 wine types) and their corresponding reference standards are proposed. The sensory profile obtained can be used both to verify compliance with the technical specifications of the product and to categorize wines by type or region, thus becoming a powerful tool for the wine sector.

Consumption of meat from suckling lambs is typical in some regions. However, sensory differences between meat from suckling and non-suckling lambs are barely described in previous studies. The objectives of the present study were (a) to develop a method to describe the sensory characteristics of lamb meat, including the development of sensory references for odor, flavor, and texture attributes; and (b) to study the sensory differences between lamb meat commercialized as “suckling lamb” and that commercialized without this designation. Twenty-three sensory attributes were selected, and their corresponding sensory references were developed. A detailed procedure to evaluate the samples was also set up. This methodology was used to characterize samples (n = 48) from a survey of lamb meat from different types of stores (n = 23). Half of the samples were commercialized as sucking lamb and the other half (older lambs) without this indication. Samples were bought in two seasons (May and December) to consider possible seasonal effects. Samples were evaluated in triplicate by 12 trained assessors. Data were analyzed by ANOVA (p ≤ .05). Apart from how long juiciness was maintained (“maintained juiciness”), all the selected attributes were appropriate to discriminate between samples. Lamb meat sold as “suckling” did not differ from the other lamb samples in odor and flavor, but there were clear differences in texture attributes: meat sold as suckling lamb was tenderer and juicier, with higher crumbliness, and with lower fibrousness, chewiness, and residue than non-suckling lamb meat. Several sensory characteristics, mainly related to odor and flavor, varied according to the season.

Insulin resistance and type 2 diabetes are obesity-related health alterations, featuring an ever-increasing prevalence. Besides inadequate feeding patterns, gut microbiota alterations stand out as potential contributors to these metabolic disturbances. The aim of this study was to investigate whether the administration of a probiotic (Lactobacillus rhamnosus GG) effectively prevents diet-induced insulin resistance in rats and to compare these potential effects with those exerted by its heat-inactivated parabiotic. For this purpose, 34 male Wistar rats were fed a standard or a high-fat high-fructose diet, alone or supplemented with viable or heat-inactivated Lactobacillus rhamnosus GG. The body and white adipose tissue weight increases, induced by the obesogenic diet, were prevented by probiotic and parabiotic administration. The trend towards higher basal glucose levels and significantly higher serum insulin concentration observed in the non-treated animals fed with the obesogenic diet were effectively reverted by both treatments. Similar results were also found for serum adiponectin and leptin, whose levels were brought back by the probiotic and parabiotic administration to values similar to those of the control animals. Noteworthily, parabiotic administration significantly reduced skeletal muscle triglyceride content and activated CPT-1b compared to the non-treated animals. Finally, both treatments enhanced Akt and AS160 phosphorylation in the skeletal muscle compared to the non-treated animals; however, only parabiotic administration increased GLUT-4 protein expression in this tissue. These results suggest that heat-inactivated Lactobacillus rhamnosus GG seem to be more effective than its probiotic of origin in preventing high-fat high-fructose diet-induced insulin resistance in rats.

Background The COVID-19 pandemic was a health emergency requiring rapid scal resource mobilisation to support national responses. The use of e ective health nancing mechanisms and policies, or lack thereof, a ected the impact of the pandemic on the population, particularly vulnerable groups and individuals. We provide an overview and illustrative examples of health nancing policies adopted in 15 countries during the pandemic, develop a framework for resilient health nancing, and use this pandemic to argue a case to move towards universal health coverage (UHC). Methods In this case study, we examined the national health nancing policy responses of 15 countries, which were purposefully selected countries to represent all WHO regions and have a range of income levels, UHC index scores, and health system typologies. We did a systematic literature review of peer-reviewed articles, policy documents, technical reports, and publicly available data on policy measures undertaken in response to the pandemic and complemented the data obtained with 61 in-depth interviews with health systems and health nancing experts. We did a thematic analysis of our data and organised key themes into a conceptual framework for resilient health nancing. Findings Resilient health nancing for health emergencies is characterised by two main phases: (1) absorb and recover, where health systems are required to absorb the initial and subsequent shocks brought about by the pandemic and restabilise from them; and (2) sustain, where health systems need to expand and maintain scal space for health to move towards UHC while building on resilient health nancing structures that can better prepare health systems for future health emergencies. We observed that ve key nancing policies were implemented across the countries— namely, use of extra-budgetary funds for a swift initial response, repurposing of existing funds, e cient fund disbursement mechanisms to ensure rapid channelisation to the intended personne

Rhabdomyosarcoma is a rare cancer arising in skeletal muscle that typically impacts children and young adults. It is a worldwide challenge in child health as treatment outcomes for metastatic and recurrent disease still pose a major concern for both basic and clinical scientists. The treatment strategies for rhabdomyosarcoma include multi-agent chemotherapies after surgical resection with or without ionization radiotherapy. In this comprehensive review, we first provide a detailed clinical understanding of rhabdomyosarcoma including its classification and subtypes, diagnosis, and treatment strategies. Later, we focus on chemotherapy strategies for this childhood sarcoma and discuss the impact of three mechanisms that are involved in the chemotherapy response including apoptosis, macro-autophagy, and the unfolded protein response. Finally, we discuss in vivo mouse and zebrafish models and in vitro three-dimensional bioengineering models of rhabdomyosarcoma to screen future therapeutic approaches and promote muscle regeneration.

Collagen is a cornerstone protein for tissue engineering and 3D bioprinting due to its outstanding biocompatibility, low immunogenicity, and natural abundance in human tissues. Nonetheless, it still poses some important challenges, such as complicated and limited extraction processes, usually accompanied by batchto-batch reproducibility and influence of factors, such as temperature, pH, and ionic strength. In this work, we evaluated the suitability and performance of new, fibrillar type I collagen as standardized and reproducible collagen source for 3D printing and bioprinting. The acidic, native fibrous collagen formulation (5% w/w) performed remarkably during 3D printing, which was possible to print constructs of up to 27 layers without collapsing. On the other hand, the fibrous collagen mass has been modified to provide a fast, reliable, and easily neutralizable process. The neutralization with TRIS-HCl enabled the inclusion of cells without hindering printability. The cell-laden constructs were printed under mild conditions (50–80 kPa, pneumatic 3D printing), providing remarkable cellular viability (>90%) as well as a stable platform for cell growth and proliferation in vitro. Therefore, the native, type I collagen masses characterized in this work offer a reproducible and reliable source of collagen for 3D printing and bioprinting purposes.

Gene therapy and optogenetics are becoming promising tools for treating several nervous system pathologies. Currently, most of these approaches use viral vectors to transport the genetic material inside the cells, but viruses present some potential risks, such as marked immunogenicity, insertional mutagenesis, and limited insert gene size. In this framework, non-viral nanoparticles, such as niosomes, are emerging as possible alternative tools to deliver genetic material, avoiding the aforementioned problems. To determine their suitability as vectors for optogenetic therapies in this work, we tested three different niosome formulations combined with three optogenetic plasmids in rat cortical neurons in vitro. All niosomes tested successfully expressed optogenetic channels, which were dependent on the ratio of niosome to plasmid, with higher concentrations yielding higher expression rates. However, we found changes in the dendritic morphology and electrophysiological properties of transfected cells, especially when we used higher concentrations of niosomes. Our results highlight the potential use of niosomes for optogenetic applications and suggest that special care must be taken to achieve an optimal balance of niosomes and nucleic acids to achieve the therapeutic effects envisioned by these technologies.

Molecularly imprinted polymers (MIPs) are synthetic polymers with specific binding sites that present high affinity and spatial and chemical complementarities to a targeted analyte. They mimic the molecular recognition seen naturally in the antibody/antigen complementarity. Because of their specificity, MIPs can be included in sensors as a recognition element coupled to a transducer part that converts the interaction of MIP/analyte into a quantifiable signal. Such sensors have important applications in the biomedical field in diagnosis and drug discovery, and are a necessary complement of tissue engineering for analyzing the functionalities of the engineered tissues. Therefore, in this review, we provide an overview of MIP sensors that have been used for the detection of skeletal- and cardiac-muscle-related analytes. We organized this review by targeted analytes in alphabetical order. Thus, after an introduction to the fabrication of MIPs, we highlight different types of MIP sensors with an emphasis on recent works and show their great diversity, their fabrication, their linear range for a given analyte, their limit of detection (LOD), specificity, and reproducibility. We conclude the review with future developments and perspectives.

Substrate reduction therapy (SRT) has been proposed as a new gene therapy for Fabry disease (FD) to prevent the formation of globotriaosylceramide (Gb3). Nanomedicines containing different siRNA targeted to Gb3 synthase (Gb3S) were designed. Formulation factors, such as the composition, solid lipid nanoparticles (SLNs) preparation method and the incorporation of different ligands, such as gold nanoparticles (GNs), protamine (P) and polysaccharides, were evaluated. The new siRNA–golden LNPs were efficiently internalized in an FD cell model (IMFE-1), with GNs detected in the cytoplasm and in the nucleus. Silencing efficacy (measured by RT-qPCR) depended on the final composition and method of preparation, with silencing rates up to 90% (expressed as the reduction in Gb3S-mRNA). GNs conferred a higher system efficacy and stability without compromising cell viability and hemocompatibility. Immunocytochemistry assays confirmed Gb3S silencing for at least 15 days with the most effective formulations. Overall, these results highlight the potential of the new siRNA–golden LNP system as a promising nanomedicine to address FD by specific SRT.

272 p., La bioimpresión 3D es una emergente tecnología de fabricación aditiva que ha demostrado tener una ventaja respecto a las técnicas convencionales debido a su automatización, rapidez y reproducibilidad. En este sentido, podría ser una herramienta eficaz para la fabricación de tejidos como el cartílago y el hueso, tejidos con una gran demanda debido a la alta incidencia de lesiones osteocondrales en la población adulta y anciana. Una de las claves para obtener una buena bioimpresión 3D es la biotinta. Para ello, en esta tesis, se han desarrollado diferentes biotintas a base de polímeros naturales como la nanocelulosa (NC) y el alginato (Alg) debido a sus buenas propiedades tanto mecánicas como biológicas. Está biotinta ha demostrado tener las propiedades reológicas adecuadas para fabricar estructuras 3D estables y con una similitud al diseño asistido por ordenador. Posteriormente, a esta biotinta base se le añadió ácido hialurónico (HA), un polímero que se encuentra de forma natural en la matriz extracelular de diversos tejidos humanos. El añadir HA ha demostrado mejorar las propiedades mecánicas de la biotinta base NC-Alg. Además, se ha establecido el autoclave de ciclo corto como método de esterilización para ambas biotintas debido a que ha demostrado ser la técnica menos destructiva al compararla con la esterilización por autoclave de ciclo largo y las radiaciones ionizantes. Por último, la biotinta NC-Alg-HA haaumentado la proliferación, viabilidad y actividad metabólica de las células mesenquimales D1 respecto a la biotinta NC-Alg, haciéndola una biotinta eficaz para su uso en la regeneración de diversos tejidos. Con el objetivo de enfocar la biotinta base NC-Alg a la regeneración de cartílago, se le ha añadido dos componentes que se encuentran en la matriz cartilaginosa como el sulfato de condroitina (CS) y el sulfato de dermatán (DS). Ambas biotintas (NC-Alg-CS) y (NC-Alg-DS) mejoraron tanto las propiedades reológicas como las biológicas en comparación con la ti

In the last few years, the importance of neoantigens in the development of personalized antitumor vaccines has increased remarkably. In order to study whether bioinformatic tools are effective in detecting neoantigens that generate an immune response, DNA samples from patients with cutaneous melanoma in different stages were obtained, resulting in a total of 6048 potential neoantigens gathered. Thereafter, the immunological responses generated by some of those neoantigens ex vivo were tested, using a vaccine designed by a new optimization approach and encapsulated in nanoparticles. Our bioinformatic analysis indicated that no differences were found between the number of neoantigens and that of non-mutated sequences detected as potential binders by IEDB tools. However, those tools were able to highlight neoantigens over non-mutated peptides in HLA-II recognition (p-value 0.03). However, neither HLA-I binding affinity (p-value 0.08) nor Class I immunogenicity values (p-value 0.96) indicated significant differences for the latter parameters. Subsequently, the new vaccine, using aggregative functions and combinatorial optimization, was designed. The six best neoantigens were selected and formulated into two nanoparticles, with which the immune response ex vivo was evaluated, demonstrating a specific activation of the immune response. This study reinforces the use of bioinformatic tools in vaccine development, as their usefulness is proven both in silico and ex vivo.

The genus Opuntia spp. includes plants capable of growing in arid, temperate and tropical climates. The vast majority of wild species grow in Mexico, but O. ficus-indica (prickly pear or nopal) is cultivated around the world and it is one of the most studied. This review shows the currently available knowledge concerning the action of O. ficus-indica and other Opuntia species (Opuntia vulgaris, Opuntia robusta, Opuntia streptacantha, Opuntia microdasys, Opuntia dillenii and Opuntia dejecta) on liver health. The available data demonstrate the positive effects of extracts, vinegar, juices or seed oil of the Opuntia genus on the alterations induced in the liver by inadequate feeding patterns or the administration of chemicals. In this regard, the potential beneficial effects of nopal are related to the attenuation of triglyceride accumulation, oxidative stress and/or inflammation. Nevertheless, there is no information concerning the bioactive compound’s characterisation in most of these studies; consequently, it is not possible to link the therapeutic effects of these plants to the presence of specific compounds in the nopal extracts. Therefore, further research is needed to confirm if the positive effects observed in animal models are also found in humans, in order to determine whether Opuntia can represent an effective tool to prevent and/or manage hepatic alterations.

The gluten-free diet (GFD) remains a complex paradigm in managing celiac disease (CeD) in children and adults, and there are many reasons why GFD adherence should be strict to improve outcomes. However, this is a challenging task for patients, since they need to have access to quality healthcare resources that facilitate optimal GFD adherence. Understanding the strengths and weaknesses of the GFD, tackling coexisting nutritional deficiencies, and dealing with complex situations, such as seronegative CeD or non-responsive CeD, all require the involvement of a multidisciplinary team. The short- and long-term follow-up of CeD patients should preferably be performed by a combined Gastroenterology and Nutrition service with well-defined quality standards and the multidisciplinary involvement of physicians, nurses, dietitians, and psychologists. Nutritional advice and counseling by an experienced dietitian can reduce the costs associated with long-term follow-up of CeD patients. Likewise, psychological interventions may be essential in specific scenarios where implementing and sustaining a lifelong GFD can cause a significant psychological burden for patients. This manuscript aims to provide guidelines to improve clinical practice in the follow-up and monitoring of CeD patients and provide information on the nutritional risks of an ill-advised GFD. Clinicians, biochemists, food technologists, dietitians, and psychologists with a global view of the disease have been involved in its writing.

227 p., The integration of novel compounds into formulations may open up new avenues and opportunities for improving the treatment of various diseases using a genetherapytechnique. To this purpose, my thesis aimed to study the impact of incorporation of biomaterials such sphingolipids or nanodiamonds as a helper component into a cationic niosome formulation for gene delivery, the formulations and related complexes were studied in terms of particle size,surface charge, dispersity, and morphology. Furthermore, biological research were conducted inboth in vitro and in vivo experimental trials to evaluate cellular uptake, intracellular trafficking pathways, and transfection efficiency.

The treatment for celiac disease (CD) involves a strict gluten-free diet, which can be challenging and lead to questions for patients. Pinpointing these uncertainties can enable the creation of efficient educational resources. In this study, a questionnaire was specifically designed to gain insights into the knowledge, concerns, and needs of individuals with CD and their supporters. The questionnaire was distributed through the Instagram social network and received adequate responses from 300 participants, 258 (86%) being female and 152 (50.7%) falling in the age range of 25–44 years. The concerns of individuals with celiac disease and celiac supporters were rated on a 1–4 scale, with a mean score of 3.5 indicating significant concern. A total of 255 (85%) of all participants expressed that their principal concern was the social limitations they faced, such as difficulties in eating out and sharing food with others. Every participant evaluated their overall disease knowledge, averaging at 2.92 out of 4, indicating a reasonable level of awareness. When asked if they believed that improving general knowledge about CD in the general population would enhance their quality of life, the vast majority responded affirmatively. This finding underscores the importance of not only educating individuals with CD but also reaching out to the wider population, especially those who have a direct impact on the daily lives of individuals with CD, such as family members, friends, and food service providers.

Drug adherence is a significant medical issue, often responsible for sub-optimal outcomes during the treatment of chronic diseases such as rheumatoid or psoriatic arthritis. Monoclonal antibodies (which are exclusively given parenterally) have been proven to be an effective treatment in these cases. The use of auto-injectors is an effective strategy to improve drug adherence in parenteral treatments since these pen-like devices offer less discomfort and increased user-friendliness over conventional syringe-based delivery. This study aims to investigate the feasibility of including a monoclonal antibody as a solid formulation inside an auto-injector pen. Specifically, the objective was to evaluate the drug stability after a concentration (to reduce the amount of solvent and space needed) and freeze-drying procedure. A preliminary screening of excipients to improve stability was also performed. The nano-DSC results showed that mannitol improved the stability of the concentrated, freeze-dried antibody in comparison to its counterpart without it. However, a small instability of the CH2 domain was still found for mannitol samples, which will warrant further investigation. The present results serve as a stepping stone towards advancing future drug delivery systems that will ultimately improve the patient experience and associated drug adherence.

Subunit vaccines typically require co-administration with an adjuvant to elicit protective immunity, adding development hurdles that can impede rapid pandemic responses. To circumvent the need for adjuvant in a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) subunit vaccine, we engineer a thermostable immunotargeting vaccine (ITV) that leverages the pan-HLA-DR monoclonal antibody 44H10 to deliver the viral spike protein receptor-binding domain (RBD) to antigen-presenting cells. X-ray crystallography shows that 44H10 binds to a conserved epitope on HLA-DR, providing the basis for its broad HLA-DR reactivity. Adjuvant-free ITV immunization in rabbits and ferrets induces robust anti-RBD antibody responses that neutralize SARS-CoV-2 variants of concern and protect recipients from SARS-CoV-2 challenge. We demonstrate that the modular nature of the ITV scaffold with respect to helper T cell epitopes and diverse RBD antigens facilitates broad sarbecovirus neutralization. Our findings support anti-HLA-DR immunotargeting as an effective means to induce strong antibody responses to subunit antigens without requiring an adjuvant.

The interest of extracellular vesicles (EVs) in cancer immunotherapy is increasing every day. EVs are lipid bilayer vesicles released by most cells, which contain the molecular signature of their parent cell. Melanoma-derived EVs present antigens specific to this aggressive type of cancer, but they also exert immunomodulatory and pro- metastatic activity. Until now, most reviews focus on the immunoevasive characteristics of tumour-derived EVs, but do not help to overcome the issues related to them. In this review, we describe isolation methods of EVs from melanoma patients and most interesting markers to oversee their effect if they are used as antigen carriers. We also discuss the methods developed so far to overcome the lack of immunogenicity of melanoma-derived EVs, which includes EV modification or adjuvant co-administration. In summary, we conclude that EVs can be an interesting antigen source for immunotherapy development once EV obtaining is optimised and the understanding of the mechanisms behind their multiple effects is further understood.

The formulation of hydrogels that meet the necessary flow characteristics for their extrusion-based 3D printing while providing good printability, resolution, accuracy and stability, requires long development processes. This work presents the technological development of a hydrogel-based ink of Laponite and alginate and evaluates its printing capacity. As a novelty, this article reports a standardizable protocol to quantitatively define the best printing parameters for the development of novel inks, providing new printability evaluation parameters such as the Printing Accuracy Escalation Index. As a result, this research develops a printable Laponite-Alginate hydrogel that presents printability characteristics. This ink is employed for the reproducible manufacture of 3D printed scaffolds with versatile and complex straight or curved printing patterns for a better adaptation to different final applications. Obtained constructs prove to be stable over time thanks to the optimization of a curing process. In addition, the study of the swelling and degradation behavior of the Laponite and alginate 3D printed scaffolds in different culture media allows the prediction of their behavior in future in vitro or in vivo developments. Finally, this study demonstrates the absence of cytotoxicity of the printed formulations, hence, setting the stage for their use in the field of biomedicine.

Non-alcoholic fatty liver disease (NAFLD) is considered the most common chronic liver alteration whose prevalence is increasing in Western countries. Microalgae and macroalgae have attracted great interest due to the high content in bioactive compounds with beneficial effects on health. The aim of the present study is to assess the potential interest of extracts rich in proteins obtained from the microalgae Chlorella vulgaris and Nannochloropsis gaditana and the macroalga Gracilaria vermiculophylla in the prevention of lipid accumulation in AML-12 hepatocytes. Toxicity was not observed at any of the tested doses. Both microalgae and the macroalga were effective in preventing triglyceride accumulation, with Nannochloropsis gaditana being the most effective one. Although the three algae extracts were able to increase different catabolic pathways involved in triglyceride metabolism, the mechanisms underlying the anti-steatotic effect were different in each algae extract. In conclusion, the present study demonstrates that Chlorella vulgaris, Nannochloropsis gaditana and Gracilaria vermiculophylla extracts are able to partially prevent the accumulation of triglycerides induced by palmitic acid in cultured hepatocytes, a model used to mimic the steatosis induced in liver by dietary patterns rich in saturated fat.

This research aimed to evaluate the nutritional composition, in vitro digestibility, and gas production kinetics of 15 vegetable by-products generated by the agri-food industry compared with corn silage as a reference raw material. Nutritional characterization and in vitro ruminal fermentation tests were performed to determine in vitro organic matter digestibility and digestible energy values, short-chain fatty acids, and the gas production profile. Results indicate that vegetable by-products were more degradable, more extensively fermented, and fermented at a faster rate than corn silage. Going one step further in the valorization of these by-products in animal feed, the second part of the research aimed to compare the novel ration designed for calf fattening with a conventional one. An artificial rumen unit was used to obtain nutrient disappearance, rumen fermentation parameters, and gas production of rumen digesta. Very slight differences were observed between both experimental rations, with their composition being the main difference. Most of the unitary vegetable by-products and all mixes, as real examples of by-product generation in the agri-food industry, have higher digestibility and a greater nutritional value than corn silage. These by-products showed the potential to be used in ruminant-ensiled rations and could replace part of the ingredients in conventional diets.

Three dimensional (3D) bioprinting is a powerful tool, that was recently applied to tissue engineering. This technique allows the precise deposition of cells encapsulated in supportive bioinks to fabricate complex scaffolds, which are used to repair targeted tissues. Here, we review the recent developments in the application of 3D bioprinting to dental tissue engineering. These tissues, including teeth, periodontal ligament, alveolar bones, and dental pulp, present cell types and mechanical properties with great heterogeneity, which is challenging to reproduce in vitro. After highlighting the different bioprinting methods used in regenerative dentistry, we reviewed the great variety of bioink formulations and their effects on cells, which have been established to support the development of these tissues. We discussed the different advances achieved in the fabrication of each dental tissue to provide an overview of the current state of the methods. We conclude with the remaining challenges and future needs

BACKGROUNDDietary methyl donors might influence DNA methylation during carcinogenesis of colorectal cancer (CRC). However, whether the influence of methyl donor intake is modified by polymorphisms in such epigenetic regulators is still unclear.AIMTo improve the current understanding of the molecular basis of CRC.METHODSA literature search in the Medline database, Reference Citation Analysis (https:// www.referencecitationanalysis.com/), and manual reference screening were performed to identify observational studies published from inception to May 2022.RESULTSA total of fourteen case-control studies and five cohort studies were identified. These studies included information on dietary methyl donors, dietary components that potentially modulate the bioavailability of methyl groups, genetic variants of methyl metabolizing enzymes, and/or markers of CpG island methylator phenotype and/or microsatellite instability, and their possible interactions on CRC risk.CONCLUSIONSeveral studies have suggested interactions between methylenetetrahydrofolate reductase polymorphisms, methyl donor nutrients (such as folate) and alcohol on CRC risk. Moreover, vitamin B-6, niacin, and alcohol may affect CRC risk through not only genetic but also epigenetic regulation. Identification of specific mechanisms in these interactions associated with CRC may assist in developing targeted prevention strategies for individuals at the highest risk of developing CRC.

Research related to food insecurity (FI) among European university student populations is currently limited, especially the studies carried out during the COVID-19 pandemic. The present study aimed to assess the prevalence and identify possible predictors of FI among students from a Spanish public university, the University of the Basque Country UPV/EHU, during the COVID-19 pandemic. A cross-sectional observational study design was used, in which a total of 422 students completed an online survey. Results were weighted according to age and field of education. Binary logistic regressions adjusted by sex, age, and campus were applied to identify FI predictors. FI in 19.6, 2.6, and 0.7% of the population was mild, moderate, and severe, respectively. The three main predictors of FI were a decrease in the main source of income (OR, 2.80; 95% IC, 2.57–3.06), not receiving scholarships during the pandemic (OR, 2.32; 95% IC, 2.18–2.47), and living arrangements before the pandemic (not living with parents/relatives) (OR, 2.03, 95% IC, 1.89–2.18). This study found a high prevalence of FI among the students surveyed, and the strongest predictors of this FI were related to socioeconomic status. A robust and comprehensive policy response is recommended to mitigate FI in this population.

The influence of renewable packaging materials on the oxidative stability of sunflower oil was investigated to evaluate whether they could be used as alternatives to conventional plastics. Two renewable bottle materials, polylactic acid (PLA) and bio-polyethylene (Green-PE) were compared to conventional plastics consisting of virgin and recycled polyethylene terephthalate (PET, r-PET) and regular polyethylene (PE), in a storage study over a period of 56 days. The results showed that the progress of lipid oxidation in PLA was similar to PET and r-PET until day 28, while it was significantly increased in PE and Green-PE. Benzene was detected as the only migration compound in the oil stored in PET and r-PET, with concentrations of 0.153 ± 0.027 µg/g and 0.187 ± 0.024 µg/g after 56 days of storage. The study concluded that PLA could be used as an alternative packaging material for edible oils to replace PET.

Development of new medicinal products for particular therapeutic treatment or for better manipulations with better quality and less side effects are possible as a result of advanced inorganic and organic materials application, among which zeolites, due to their properties and versatility, have been gaining attention. This paper is an overview of the development in the use of zeolite materials and their composites and modifications as medicinal products for several purposes such as active agents, carriers, for topical treatments, oral formulations, anticancer, the composition of theragnostic systems, vaccines, parenteral dosage forms, tissue engineering, etc. The objective of this review is to explore the main properties of zeolites and associate them with their drug interaction, mainly addressing the advances and studies related to the use of zeolites for different types of treatments due to their zeolite characteristics such as molecule storage capacity, physical and chemical stability, cation exchange capacity, and possibility of functionalization. The use of computational tools to predict the drug—zeolite interaction is also explored. As conclusion was possible to realize the possibilities and versatility of zeolite applications as being able to act in several aspects of medicinal products.

El capítulo 2 está sujeto a confidencialidad por el autor. 335 p., In this doctoral thesis, we aimed to develop a complete biomaterial, following a circular economy thinking, which can tackle multiple stages of the wound healing process. In this sense, we designed a sponge-like scaffold composed of soy protein and ß-chitin with hemostatic properties, which can be combined with mesenchymal stromal cells (MSCs), for treating different wounds. Thanks to their anti-inflammatory, immunomodulatory and proregenerative properties, the MSCs have demonstrated to be very effective for treating multiple pathologies. However, despite the use of MSCs combined with biomaterials holds a promising potential, in this work we aimed to explore also the extracelular vesicle fraction of the MSCs secretome. Thus, this cell-free alternative presents the advantages of being; more easily handled, characterized, standardized, stored and potentially safer than MSCs, while exhibiting also therapeutic properties.

Opuntia cactus fruit (prickly pear flesh and agricultural residues such as peels and stalks) is an important source of bioactive compounds, including betalains and phenolic compounds. In this work, two double emulsion W1/O/W2 formulations (A and B) were designed to encapsulate green extracts rich in betalains and phenolic compounds obtained from Opuntia stricta var. dillenii (OPD) fruits with the aim of improving their stability and protecting them during the in vitro gastrointestinal digestion process. In addition, the characterization of the double emulsions was studied by microscopy and the evaluation of their physical and physico-chemical parameters. Formulation A, based on Tween 20, showed smaller droplets (1.75 µm) and a higher physical stability than Formulation B, which was achieved with sodium caseinate (29.03 µm). Regarding the encapsulation efficiency of the individual bioactives, betalains showed the highest values (73.7 ± 6.7 to 96.9 ± 3.3%), followed by flavonoids (68.2 ± 5.9 to 95.9 ± 7.7%) and piscidic acid (71 ± 1.3 to 70.2 ± 5.7%) depending on the formulation and the bioactive compound. In vitro digestive stability and bioaccessibility of the individual bioactives increased when extracts were encapsulated for both formulations (67.1 to 253.1%) in comparison with the non-encapsulated ones (30.1 to 64.3%), except for neobetanin. Both formulations could be considered as appropriate microcarrier systems for green OPD extracts, especially formulation A. Further studies need to be conducted about the incorporation of these formulations to develop healthier foods.

Urinary tract infections (UTIs) are extremely common and a major driver for the use of antimicrobials. Calcium fosfomycin is an old antibiotic indicated for the treatment of UTIs; however, data about its urine pharmacokinetic profile are scarce. In this work, we have evaluated the pharmacokinetics of fosfomycin from urine concentrations after oral administration of calcium fosfomycin to healthy women. Moreover, we have assessed, by pharmacokinetic/pharmacodynamic (PK/PD) analysis and Monte Carlo simulations, its effectiveness considering the susceptibility profile of Escherichia coli, the main pathogen involved in UTIs. The accumulated fraction of fosfomycin excreted in urine was around 18%, consistent with its low oral bioavailability and its almost exclusively renal clearance by glomerular filtration as unchanged drug. PK/PD breakpoints resulted to be 8, 16, and 32 mg/L for a single dose of 500 mg, a single dose of 1000 mg, and 1000 mg q8h for 3 days, respectively. For empiric treatment, the estimated probability of treatment success was very high (>95%) with the three dose regimens, considering the susceptibility profile of E. coli reported by EUCAST. Our results show that oral calcium fosfomycin at a dose level of 1000 mg every 8 h provides urine concentrations sufficient to ensure efficacy for the treatment of UTIs in women.

Cheese microbiota contributes to various biochemical processes that lead to the formation of volatile compounds and the development of flavour during ripening. Nonetheless, the role of these microorganisms in volatile aroma compounds production is little understood. This work reports for the first time the dynamics and odour impact of volatile compounds, and their relationship to microbial shifts during the ripening of a raw ewe milk-derived cheese (Idiazabal). By means of SPME-GC-MS, 81 volatile compounds were identified, among which acids predominated, followed by esters, ketones and alcohols. The ripening time influenced the abundance of most volatile compounds, thus the moments of greatest abundance were determined (such as 30–60 days for acids). Through Odour Impact Ratio (OIR) values, esters and acids were reported as the predominant odour-active chemical families, while individually, ethyl hexanoate, ethyl 3-methyl butanoate, ethyl butanoate, butanoic acid or 3-methyl butanal were notable odorants, which would provide fruity, rancid, cheesy or malt odour notes. Using a bidirectional orthogonal partial least squares (O2PLS) approach with Spearman's correlations, 12 bacterial genera were reported as key bacteria for the volatile and aromatic composition of Idiazabal cheese, namely Psychrobacter, Enterococcus, Brevibacterium, Streptococcus, Leuconostoc, Chromohalobacter, Chryseobacterium, Carnobacterium, Lactococcus, Obesumbacterium, Stenotrophomonas and Flavobacterium. Non-starter lactic acid bacteria (NSLAB) were highly related to the formation of certain acids, esters and alcohols, such as 3-hexenoic acid, ethyl butanoate or 1-butanol. On the other hand, the starter LAB (SLAB) was related to particular ketones production, specifically 3-hydroxy-2-butanone; and environmental and/or non-desirable bacteria to certain ketones, hydrocarbons and sulphur compounds formation, such as 2-propanone, t-3-octene and dimethyl sulphone. Additionally, the SLAB Lactococcus an

Green chemistry has been a growing multidisciplinary field in recent years showing great promise in biomedical applications, especially for cancer therapy. Chitosan (CS) is an abundant biopolymer derived from chitin and is present in insects and fungi. This polysaccharide has favorable characteristics, including biocompatibility, biodegradability, and ease of modification by enzymes and chemicals. CS-based nanoparticles (CS-NPs) have shown potential in the treatment of cancer and other diseases, affording targeted delivery and overcoming drug resistance. The current review emphasizes on the application of CS-NPs for the delivery of a chemotherapeutic agent, doxorubicin (DOX), in cancer therapy as they promote internalization of DOX in cancer cells and prevent the activity of P-glycoprotein (P-gp) to reverse drug resistance. These nanoarchitectures can provide co-delivery of DOX with antitumor agents such as curcumin and cisplatin to induce synergistic cancer therapy. Furthermore, co-loading of DOX with siRNA, shRNA, and miRNA can suppress tumor progression and provide chemosensitivity. Various nanostructures, including lipid-, carbon-, polymeric- and metal-based nanoparticles, are modifiable with CS for DOX delivery, while functionalization of CS-NPs with ligands such as hyaluronic acid promotes selectivity toward tumor cells and prevents DOX resistance. The CS-NPs demonstrate high encapsulation efficiency and due to protonation of amine groups of CS, pH-sensitive release of DOX can occur. Furthermore, redox- and light-responsive CS-NPs have been prepared for DOX delivery in cancer treatment. Leveraging these characteristics and in view of the biocompatibility of CS-NPs, we expect to soon see significant progress towards clinical translation.

Oxidative stress and inflammation are well-known triggers of NAFLD onset and progression. The aim of this study is to compare the potential benefits of a viable probiotic (Lactobacillus rhamnosus GG) and its parabiotic (heat-inactivated) on oxidative stress, inflammation, DNA damage and cell death pathways in the liver of rats featuring diet-induced NAFLD. The consumption of the steatotic diet led to increased final body and liver weights, higher hepatic triacylglycerol content, altered serum transaminase levels and enhanced oxidative and inflammatory status. Administration of the probiotic and the parabiotic partially prevented the body weight increase induced by the steatotic diet, whereas the probiotic caused more effective decreasing hepatic triglyceride content. Sharp but nonstatistically significant decreases in serum transaminase levels were also observed for both treatments. The reduction in antioxidant enzyme activities found in the nontreated animals fed the steatotic diet was partially prevented by both treatments (GPx activity). Similarly, the reductions in nonenzymatic antioxidant protection (GSH content) and total antioxidant capacity (ORAC) found in the nontreated rats were restored by the administration of both treatments. These results show that both viable and heat-inactivated Lactobacillus rhamnosus GG administration partially prevent steatotic diet-induced liver oxidative stress and inflammation induced in rats.

Producers of milk and dairy products have been faced with the challenge of responding to European society’s demand for guaranteed animal welfare production. In recent years, measures have been taken to improve animal welfare conditions on farms and evaluation systems have been developed to certify them, such as the Welfare Quality® protocol. Among the markers used for this purpose, acute phase proteins stand out, with haptoglobin being one of the most relevant. However, the diagnostic power of these tools is limited and more sensitive and specific technologies are required to monitor animal health status. Different factors such as diet, stress, and diseases modify the metabolism of the animals, altering the composition of the milk in terms of oligosaccharides, proteins, and lipids. Thus, in order to study oxidative-stress-associated lipids, a collection of well-characterized milk samples, both by veterinary diagnosis and by content of the acute stress biomarker haptoglobin, was analyzed by mass spectrometry and artificial intelligence. Two lipid species (sphingomyelin and phosphatidylcholine) were identified as potential biomarkers of health status in dairy cows. Both lipids allow for the discrimination of milk from sick animals and also milk from those with stress. Moreover, lipidomics revealed specific lipid profiles depending on the origin of the samples and the degree of freedom of the animals on the farm. These data provide evidence for specific lipid changes in stressed animals and open up the possibility that haptoglobin could also affect lipid metabolism in cow’s milk.