Evaluation of the use of goat’s milk-derived exosomes as theragnostic platform in oncology

Exosomes are biologically active nanovesicles from endosomal origin and lipid bilayer involved in cell communication, secreted by different types of cells and present in a wide variety of biological fluids. Due to their size, liposome-like characteristics and their strong relation with oncology, new applications for therapy and diagnosis are being explored in the field of medical imaging. During this thesis, the isolation, characterization and functionalization with fluorescent dyes has been performed to examine the possibility of using goat’s milk derived exosomes as theragnostic platform. With this purpose, the size, shape and charge of exosomes have been characterized, and the conjugation with two different fluorophores has proved to unalter their physicochemical properties. The biodistribution of functionalized exosomes, their toxicity and in vitro accumulation have been analyzed to evaluate that these nanovesicles can be used not only for diagnosis, but also for therapy and drug delivery in the future.Ingeniería Biomédica (Plan 2010

The contribution of microfluidics to the fight against tuberculosis

The high mortality associated with tuberculosis brings forward the urgency of developing new therapies and strategies against the disease. With the advance of drug-resistant strains, traditional techniques have proven insufficient to manage the disease appropriately. Microfluidic devices have characteristics that can enhance treatment prescription and significantly advance our knowledge about the disease and its interaction within the human body. In addition, microfluidic systems provide advantages in terms of time and costs, which are particularly important in countries with low income and resources. This review will highlight how microdevices can help bridge the gaps in disease management, including their use for drug testing and development, drug susceptibility, basic research, and novel approaches to anti-TB vaccines and organ-on-chip studies.This project has received funding from the Innovative Medicines Initiative 2 Joint Undertaking (JU) under grant agreement No. 853989. The JU receives support from the European Union's Horizon 2020 research and innovation programme and EFPIA and Global Alliance for TB Drug Development non-profit organisation, Bill & Melinda Gates Foundation and University of Dundee. This work was partially funded by Ministerio de Ciencia, Innovación y Universidades, Agencia Estatal de Investigación, under grant PID2019-109820RB-I00, MCIN/AEI/10.13039/501100011033/, co-finance by European Regional Development Fund (ERDF), "A way of making Europe

Development of an Inverted Epifluorescence Microscope for Long-Term Monitoring of Bacteria in Multiplexed Microfluidic Devices

Developing more efficient methods for antibiotic susceptibility testing is a pressing issue in novel drug development as bacterial resistance to antibiotics becomes increasingly common. Microfluidic devices have been demonstrated to be powerful platforms that allow researchers to perform multiplexed antibiotic testing. However, the level of multiplexing within microdevices is limited, evidencing the need of creating simple, low-cost and high-resolution imaging systems that can be integrated in antibiotic development pipelines. This paper describes the design and development of an epifluorescence inverted microscope that enables long-term monitoring of bacteria inside multiplexed microfluidic devices. The goal of this work is to provide a simple microscope powerful enough to allow single-cell analysis of bacteria at a reduced cost. This facilitates increasing the number of microscopes that are simultaneously used for antibiotic testing. We prove that the designed system is able to accurately detect fluorescent beads of 100 nm, demonstrating comparable features to high-end commercial microscopes and effectively achieving the resolution required for single-cell analysis of bacteria. The proposed microscope could thus increase the efficiency in antibiotic testing while reducing cost, size, weight, and power requirements, contributing to the successful development of new antibiotic drugs.This work was partially funded by project TEC2016-78052-R from the Spanish Ministry of Economy and PID2019-109820RB-I00 from the Spanish Ministry of Science and Innovation. The research leading to these results received funding from the Innovative Medicines Innitiative 2 Joint Undertaking (JU) under grant agreement No 853989. The JU receives support from the European Union’s Horizon 2020 research and innovation programme and EFPIA and Global Alliance for TB Drug Development non profit organisation, Bill & Melinda Gates Foundation and University of Dundee

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

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

Single Plane Illumination Microscopy for Microfluidic Device Imaging

Three-dimensional imaging of live processes at a cellular level is a challenging task. It requires high-speed acquisition capabilities, low phototoxicity, and low mechanical disturbances. Three-dimensional imaging in microfluidic devices poses additional challenges as a deep penetration of the light source is required, along with a stationary setting, so the flows are not perturbed. Different types of fluorescence microscopy techniques have been used to address these limitations; particularly, confocal microscopy and light sheet fluorescence microscopy (LSFM). This manuscript proposes a novel architecture of a type of LSFM, single-plane illumination microscopy (SPIM). This custom-made microscope includes two mirror galvanometers to scan the sample vertically and reduce shadowing artifacts while avoiding unnecessary movement. In addition, two electro-tunable lenses fine-tune the focus position and reduce the scattering caused by the microfluidic devices. The microscope has been fully set up and characterized, achieving a resolution of 1.50 μm in the x-y plane and 7.93 μm in the z-direction. The proposed architecture has risen to the challenges posed when imaging microfluidic devices and live processes, as it can successfully acquire 3D volumetric images together with time-lapse recordings, and it is thus a suitable microscopic technique for live tracking miniaturized tissue and disease models

Tuberculosis: integrated studies for a complex disease 2050

Tuberculosis (TB) has been a disease for centuries with various challenges [1]. Like other places where challenges and opportunities come together, TB challenges were the inspiration for the scientific community to mobilize different groups for the purpose of interest. For example, with the emergence of drug resistance, there has been a huge volume of research on the discovery of new medicines and drug delivery methods and the repurposing of old drugs [2, 3]. Moreover, to enhance the capacity to detect TB cases, studies have sought diagnostics and biomarkers, with much hope recently expressed in the direction of point-of-care tests [4]. Despite all such efforts as being highlighted in 50 Chapters of this volume, we are still writing about TB and thinking about how to fight this old disease–implying that the problem of TB might be complex, so calling the need for an integrated science to deal with multiple dimensions in a simultaneous and effective manner. We are not the first one; there have been proposed integrated platform for TB research, integrated prevention services, integrated models for drug screening, integrated imaging protocol, integrated understanding of the disease pathogenesis, integrated control models, integrated mapping of the genome of the pathogen, etc. [5–12], to name some. These integrated jobs date back decades ago. So, a question arises: why is there a disease named TB yet? It might be due to the fact that this integration has happened to a scale that is not global, and so TB remains to be a problem, especially in resource-limited settings. Hope Tuberculosis: Integrated Studies for a Complex Disease helps to globalize the integrated science of TB.info:eu-repo/semantics/publishedVersio