Characterising epithelial tissues using persistent entropy

In this paper, we apply persistent entropy, a novel topological statis- tic, for characterization of images of epithelial tissues. We have found out that persistent entropy is able to summarize topological and geomet- ric information encoded by -complexes and persistent homology. After using some statistical tests, we can guarantee the existence of signi cant di erences in the studied tissues.Ministerio de Economía y Competitividad MTM2015-67072-

Persistent entropy: a scale-invariant topological statistic for analyzing cell arrangements

In this work, we develop a method for detecting differences in the topological distribution of cells forming epithelial tissues. In particular, we extract topological information from their images using persistent homology and a summary statistic called persistent entropy. This method is scale invariant, robust to noise and sensitive to global topological features of the tissue. We have found significant differences between chick neuroepithelium and epithelium of Drosophila wing discs in both, larva and prepupal stages. Besides, we have tested our method, with good results, with images of mathematical tesselations that model biological tissues

Dual role of myosin II during Drosophila imaginal disc metamorphosis

The motor protein non-muscle myosin II is a major driver of the movements that sculpt three dimensional organs from two dimensional epithelia. The machinery of morphogenesis is well established but the logic of its control remains unclear in complex organs. Here we use live imaging and ex vivo culture to report a dual role of myosin II in regulating the development of the Drosophila wing. First, myosin II drives the contraction of a ring of cells that surround the squamous peripodial epithelium, providing the force to fold the whole disc through about 90°. Second, myosin II is needed to allow the squamous cells to expand and then retract at the end of eversion. The combination of genetics and live imaging allows us to describe and understand the tissue dynamics, and the logic of force generation needed to transform a relatively simple imaginal disc into a more complex and three-dimensional adult wing

Topological Progression in Proliferating Epithelia Is Driven by a Unique Variation in Polygon Distribution

Morphogenesis is consequence of lots of small coordinated variations that occur during development. In proliferating stages, tissue growth is coupled to changes in shape and organization. A number of studies have analyzed the topological properties of proliferating epithelia using the Drosophila wing disc as a model. These works are based in the existence of a fixed distribution of these epithelial cells according to their number of sides. Cell division, cell rearrangements or a combination of both mechanisms have been proposed to be responsible for this polygonal assembling. Here, we have used different system biology methods to compare images from two close proliferative stages that present high morphological similarity. This approach enables us to search for traces of epithelial organization. First, we show that geometrical and network characteristics of individual cells are mainly dependent on their number of sides. Second, we find a significant divergence between the distribution of polygons in epithelia from mid-third instar larva versus early prepupa. We show that this alteration propagates into changes in epithelial organization. Remarkably, only the variation in polygon distribution driven by morphogenesis leads to progression in epithelial organization. In addition, we identify the relevant features that characterize these rearrangements. Our results reveal signs of epithelial homogenization during the growing phase, before the planar cell polarity pathway leads to the hexagonal packing of the epithelium during pupal stages.España, Ministero de Ciencia BFU2011-2573

X-ray emission from a liquid curtain jet when irradiated by femtosecond laser pulses

Laser-based sources of ionizing radiation have attracted considerable attention in the last years for their broad potential applications. However, the stability and robustness of such sources are still issues that need to be addressed. Aiming to solve such problems, we propose a source that uses a liquid jet—rather than a solid—as a target for the production of X-rays. Liquid jets offer always a clean surface for every laser shot which represent a clear advantage over solids. In this work, we present an experimental characterization of the X-ray emission of such targets, and study the efficiency of the process when two temporally delayed pulses are used. According to the obtained results, the X-ray yield is comparable with commonly used targets.Ministerio de Economía y Competitividad FIS2016- 81056-

Nintedanib decreases muscle fibrosis and improves muscle function in a murine model of dystrophinopathy

Duchenne muscle dystrophy (DMD) is a genetic disorder characterized by progressive skeletal muscle weakness. Dystrophin deficiency induces instability of the sarcolemma during muscle contraction that leads to muscle necrosis and replacement of muscle by fibro-adipose tissue. Several therapies have been developed to counteract the fibrotic process. We report the effects of nintedanib, a tyrosine kinase inhibitor, in the mdx murine model of DMD. Nintedanib reduced proliferation and migration of human fibroblasts in vitro and decreased the expression of fibrotic genes such as COL1A1, COL3A1, FN1, TGFB1, and PDGFA. We treated seven mdx mice with 60 mg/kg/day nintedanib for 1 month. Electrophysiological studies showed an increase in the amplitude of the motor action potentials and an improvement of the morphology of motor unit potentials in the animals treated. Histological studies demonstrated a significant reduction of the fibrotic areas present in the skeletal muscles. Analysis of mRNA expression from muscles of treated mice showed a reduction in Col1a1, Col3a1, Tgfb1, and Pdgfa. Western blot showed a reduction in the expression of collagen I in skeletal muscles. In conclusion, nintedanib reduced the fibrotic process in a murine model of dystrophinopathy after 1 month of treatment, suggesting its potential use as a therapeutic drug in DMD patients.España, Ministerio de Economía y Competitividad BFU2016-74975-PEspaña, Instituto Ramón y Cajal PI13/0134

Rules of tissue packing involving different cell types: human muscle organization

Natural packed tissues are assembled as tessellations of polygonal cells. These include skeletal muscles and epithelial sheets. Skeletal muscles appear as a mosaic composed of two different types of cells: the “slow” and “fast” fibres. Their relative distribution is important for the muscle function but little is known about how the fibre arrangement is established and maintained. In this work we capture the organizational pattern in two different healthy muscles: biceps brachii and quadriceps. Here we show that the biceps brachii muscle presents a particular arrangement, based on the different sizes of slow and fast fibres. By contrast, in the quadriceps muscle an unbiased distribution exists. Our results indicate that the relative size of each cellular type imposes an intrinsic organization into natural tessellations. These findings establish a new framework for the analysis of any packed tissue where two or more cell types exist.España, Gobierno BFU2011-2573

Fundamental physical cellular constraints drive self-organization of tissues

Morphogenesis is driven by small cell shape changes that modulate tissue organization. Apical surfaces of proliferating epithelial sheets have been particularly well studied. Currently, it is accepted that a stereotyped distribution of cellular polygons is conserved in proliferating tissues among metazoans. In this work, we challenge these previous findings showing that diverse natural packed tissues have very different polygon distributions. We use Voronoi tessellations as a mathematical framework that predicts this diversity. We demonstrate that Voronoi tessellations and the very different tissues analysed share an overriding restriction: the frequency of polygon types correlates with the distribution of cell areas. By altering the balance of tensions and pressures within the packed tissues using disease, genetic or computer model perturbations, we show that as long as packed cells present a balance of forces within tissue, they will be under a physical constraint that limits its organization. Our discoveries establish a new framework to understand tissue architecture in development and disease. Synopsis Cell shapes in naturally packed tissues have different polygon distributions. Voronoi tessellations-based analysis suggests that polygon frequencies are restricted by the distribution of cell areas, and that this restriction emanates from the balance of forces within the tissue. Cell shapes in natural packed tissues present very different polygon distributions. These patterns can be reproduced by Voronoi tessellations. Natural tissues and Voronoi diagrams share some geometrical properties. There is a physical constraint that limits the organization of natural tissues. Unbalance of forces within the natural tissue breaks this restriction. Cell shapes in naturally packed tissues have different polygon distributions. Voronoi tessellations-based analysis suggests that polygon frequencies are restricted by the distribution of cell areas, and that this restriction emanates from the balance of forces within the tissue.Ministerio de Ciencia e Innovación BFU2011-2573

Neuromuscular disease classification system

Diagnosis of neuromuscular diseases is based on subjective visual assessment of biopsies from patients by the pathologist specialist. A system for objective analysis and classification of muscular dystrophies and neurogenic atrophies through muscle biopsy images of fluorescence microscopy is presented. The procedure starts with an accurate segmentation of the muscle fibers using mathematical morphology and a watershed transform. A feature extraction step is carried out in two parts: 24 features that pathologists take into account to diagnose the diseases and 58 structural features that the human eye cannot see, based on the assumption that the biopsy is considered as a graph, where the nodes are represented by each fiber, and two nodes are connected if two fibers are adjacent. A feature selection using sequential forward selection and sequential backward selection methods, a classification using a Fuzzy ARTMAP neural network, and a study of grading the severity are performed on these two sets of features. A database consisting of 91 images was used: 71 images for the training step and 20 as the test. A classification error of 0% was obtained. It is concluded that the addition of features undetectable by the human visual inspection improves the categorization of atrophic pattern

Scutoids are a geometrical solution to three-dimensional packing of epithelia

As animals develop, tissue bending contributes to shape the organs into complex three-dimensional structures. However, the architecture and packing of curved epithelia remains largely unknown. Here we show by means of mathematical modelling that cells in bent epithelia can undergo intercalations along the apico-basal axis. This phenomenon forces cells to have different neighbours in their basal and apical surfaces. As a consequence, epithelial cells adopt a novel shape that we term “scutoid”. The detailed analysis of diverse tissues confirms that generation of apico-basal intercalations between cells is a common feature during morphogenesis. Using biophysical arguments, we propose that scutoids make possible the minimization of the tissue energy and stabilize three-dimensional packing. Hence, we conclude that scutoids are one of nature's solutions to achieve epithelial bending. Our findings pave the way to understand the three-dimensional organization of epithelial organs.España Ministerio de Ciencia y Tecnología BFU2013-48988-C2-1-P and BFU2016-8079