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

Magneto-mechanical system to reproduce and quantify complex strain patterns in biological materials

Biological cells and tissues are continuously subjected to mechanical stress and strain cues from their surrounding substrate. How these forces modulate cell and tissue behavior is a major question in mechanobiology. To conduct studies under controlled varying physiological strain scenarios, a new virtually-assisted experimental system is proposed allowing for non-invasive and real-time control of complex deformation modes within the substrates. This approach is based on the use of extremely soft magneto-active polymers, which mimic the stiffness of biological materials. Thus, the system enables the untethered control of biological substrates providing reversible mechanical changes and controlling heterogeneous patterns. Motivated on a deep magneto-mechanical characterization across scales, a multi-physics and multi-scale in silico framework was developed to guide the experimental stimulation setup. The versatility and viability of the system have been demonstrated through its ability to reproduce complex mechanical scenarios simulating local strain patterns in brain tissue during a head impact, and its capability to transmit physiologically relevant mechanical forces to dermal fibroblasts. The proposed framework opens the way to understanding the mechanobiological processes that occur during complex and dynamic deformation states, e.g., in traumatic brain injury, pathological skin scarring or fibrotic heart remodeling during myocardial infarction.The authors thank Denis Wirtz (Johns Hopkins University) and Jean-Christophe Olivo-Marin (Institute Pasteur) for relevant discussion. The authors acknowledge support from the European Research Council (ERC) under the European Union's Horizon 2020 Research and Innovation Program (Grant agreement No. 947723, project: 4D-BIOMAP), and from Programa de Apoyo a la Realizacion de Proyectos Interdiscisplinares de I+D para Jovenes Investigadores de la Universidad Carlos III de Madrid and Comunidad de Madrid (project: BIOMASKIN). MAMM and CGC acknowledges support from the Ministerio de Ciencia, Innovacion y Universidades, Spain (FPU19/03874 and FPU20/01459) and DGG acknowledges support from the Talent Attraction grant (CM 2018 - 2018-T2/IND-9992) from the Comunidad de Madrid

Tango-Therapy Intervention for Older Adults with Cognitive Impairment Living in Nursing Homes: Effects on Quality of Life, Physical Abilities and Gait

Cognitive impairment in older adults is associated with poor gait performance, physical decline, falls and poor quality of life. This paper analyzes the feasibility and efficacy of tango-based intervention in older people living in nursing homes with and without cognitive impairment. A multicenter study, with pre- and post-test, was carried out. Intervention attendance, well-being, physical abilities (short physical performance battery), walking performance, functional capacities (Katz Index) and quality of life (quality of life in Alzheimer’s disease) were assessed. Fifty-four participants (84.9 ± 6.7 years, mini mental state examination 14.5 ± 7.4) completed the protocol. Intervention attendance was 92%, and the mean subjective well-being after each session was 4.5 ± 0.5 (on a five-point scale). A statistically significant improvement was found in the quality of life (p = 0.030). Non-statistically significant changes were found in walking performance (p = 0.159), physical abilities (p = 0.876) and in functional capacities (p = 0.253). This study shows feasibility and suggests evidence for the effects of tango therapy on well-being and quality of life. Further studies are necessary to contrast these findings and to support the role of tango interventions as a holistic approach to prevent functional decline in older people with cognitive impairment

Deletion of PTEN in microglia ameliorates chronic neuroinflammation following repetitive mTBI

Traumatic brain injury is a leading cause of morbidity and mortality in adults and children in developed nations. Following the primary injury, microglia, the resident innate immune cells of the CNS, initiate several inflammatory signaling cascades and pathophysiological responses that may persist chronically; chronic neuroinflammation following TBI has been closely linked to the development of neurodegeneration and neurological dysfunction. Phosphoinositide 3-kinases (PI3Ks) are a family of lipid kinases that have been shown to regulate several key mechanisms in the inflammatory response to TBI. Increasing evidence has shown that the modulation of the PI3K/AKT signaling pathway has the potential to influence the cellular response to inflammatory stimuli. However, directly targeting PI3K signaling poses several challenges due to its regulatory role in several cell survival pathways. We have previously identified that the phosphatase and tensin homolog deleted on chromosome 10 (PTEN), the major negative regulator of PI3K/AKT signaling, is dysregulated following exposure to repetitive mild traumatic brain injury (r-mTBI). Moreover, this dysregulated PI3K/AKT signaling was correlated with chronic microglial-mediated neuroinflammation. Therefore, we interrogated microglial-specific PTEN as a therapeutic target in TBI by generating a microglial-specific, Tamoxifen inducible conditional PTEN knockout model using a CX3CR1 Cre recombinase mouse line PTENfl/fl/CX3CR1+/CreERT2 (mcg-PTENcKO), and exposed them to our 20-hit r-mTBI paradigm. Animals were treated with tamoxifen at 76 days post-last injury, and the effects of microglia PTEN deletion on immune-inflammatory responses were assessed at 90-days post last injury. We observed that the deletion of microglial PTEN ameliorated the proinflammatory response to repetitive brain trauma, not only reducing chronic microglial activation and proinflammatory cytokine production but also rescuing TBI-induced reactive astrogliosis, demonstrating that these effects extended beyond microglia alone. Additionally, we observed that the pharmacological inhibition of PTEN with BpV(HOpic) ameliorated the LPS-induced activation of microglial NFκB signaling in vitro. Together, these data provide support for the role of PTEN as a regulator of chronic neuroinflammation following repetitive mild TBI

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

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