Mycolactone-dependent depletion of endothelial cell thrombomodulin is strongly associated with fibrin deposition in Buruli ulcer lesions

A well-known histopathological feature of diseased skin in Buruli ulcer (BU) is coagulative necrosis caused by the Mycobacterium ulcerans macrolide exotoxin mycolactone. Since the underlying mechanism is not known, we have investigated the effect of mycolactone on endothelial cells, focussing on the expression of surface anticoagulant molecules involved in the protein C anticoagulant pathway. Congenital deficiencies in this natural anticoagulant pathway are known to induce thrombotic complications such as purpura fulimans and spontaneous necrosis. Mycolactone profoundly decreased thrombomodulin (TM) expression on the surface of human dermal microvascular endothelial cells (HDMVEC) at doses as low as 2ng/ml and as early as 8hrs after exposure. TM activates protein C by altering thrombin's substrate specificity, and exposure of HDMVEC to mycolactone for 24 hours resulted in an almost complete loss of the cells' ability to produce activated protein C. Loss of TM was shown to be due to a previously described mechanism involving mycolactone-dependent blockade of Sec61 translocation that results in proteasome-dependent degradation of newly synthesised ER-transiting proteins. Indeed, depletion from cells determined by live-cell imaging of cells stably expressing a recombinant TM-GFP fusion protein occurred at the known turnover rate. In order to determine the relevance of these findings to BU disease, immunohistochemistry of punch biopsies from 40 BU lesions (31 ulcers, nine plaques) was performed. TM abundance was profoundly reduced in the subcutis of 78% of biopsies. Furthermore, it was confirmed that fibrin deposition is a common feature of BU lesions, particularly in the necrotic areas. These findings indicate that there is decreased ability to control thrombin generation in BU skin. Mycolactone's effects on normal endothelial cell function, including its ability to activate the protein C anticoagulant pathway are strongly associated with this. Fibrin-driven tisischemia could contribute to the development of the tissue necrosis seen in BU lesions

Transcriptomic Analyses of Inner Ear Sensory Epithelia in Zebrafish

Analysis of gene expression has the potential to assist in the understanding of multiple cellular processes including proliferation, cell-fate specification, senesence, and activity in both healthy and disease states. Zebrafish model has been increasingly used to understand the process of hearing and the development of the vertebrate auditory system. Within the zebrafish inner ear, there are three otolith organs, each containing a sensory macula of hair cells. The saccular macula is primarily involved in hearing, the utricular macula is primarily involved in balance and the function of the lagenar macula is not completely understood. The goal of this study is to understand the transcriptional differences in the sensory macula associated with different otolith organs with the intention of understanding the genetic mechanisms responsible for the distinct role each organ plays in sensory perception. The sensory maculae of the saccule, utricle, and lagena were dissected out of adult Et(krt4:GFP) zebrafish expressing green fluorescent protein in hair cells for transcriptional analysis. The total RNAs of the maculae were isolated and analyzed by RNA GeneChip microarray. Several of the differentially expressed genes are known to be involved in deafness, otolith development and balance. Gene expression among these otolith organs was very well conserved with less than 10% of genes showing differential expression. Data from this study will help to elucidate which genes are involved in hearing and balance. Furthermore, the findings of this study will assist in the development of the zebrafish model for human hearing and balance disorders. Anat Rec, 303:527-543, 2020. © 2019 American Association for Anatomy

Is coracohumeral distance associated with pain-function, and shoulder range of movement, in chronic anterior shoulder pain?

Background: The aim of this study was twofold: (i) to assess the intrarater reliability of coracohumeral distance; (ii) to investigate the level of association between coracohumeral distance measured by ultrasonography, and pain-disability and shoulder range of movement, in patients suffering from chronic anterior shoulder pain. Methods: An observational, cross sectional study was carried out. A convenience sample comprised of 87 patients with chronic anterior shoulder pain was assessed from 3 primary care centres. Main outcomes as pain and function were measured through the shoulder pain and disability index. Furthermore, shoulder range of movement-free of pain in shoulder elevation, as well as coracohumeral distance at both 0 and 60 degrees, were collected. Results: Absence of any correlation was found between coracohumeral distance and shoulder pain and disability index at both 0 and 60 degrees of shoulder elevation. Furthermore, absence of any correlation was found between coracohumeral distance measurements and active shoulder range of movement -free of pain. Conclusions: There was poor association between coracohumeral distance and shoulder pain and function, as well as with shoulder range of movement, in patients with chronic anterior shoulder pain. Hence, clinicians should consider, not only increasing this space, but also other possibilities in their therapies, when patients with anterior shoulder pain are treated

Nonlinear dynamics and chaos in an optomechanical beam

[EN] Optical nonlinearities, such as thermo-optic mechanisms and free-carrier dispersion, are often considered unwelcome effects in silicon-based resonators and, more specifically, optomechanical cavities, since they affect, for instance, the relative detuning between an optical resonance and the excitation laser. Here, we exploit these nonlinearities and their intercoupling with the mechanical degrees of freedom of a silicon optomechanical nanobeam to unveil a rich set of fundamentally different complex dynamics. By smoothly changing the parameters of the excitation laser we demonstrate accurate control to activate two-and four-dimensional limit cycles, a period-doubling route and a six-dimensional chaos. In addition, by scanning the laser parameters in opposite senses we demonstrate bistability and hysteresis between two-and four-dimensional limit cycles, between different coherent mechanical states and between four-dimensional limit cycles and chaos. Our findings open new routes towards exploiting silicon-based optomechanical photonic crystals as a versatile building block to be used in neurocomputational networks and for chaos-based applications.This work was supported by the European Comission project PHENOMEN (H2020-EU-713450), the Spanish Severo Ochoa Excellence program and the MINECO project PHENTOM (FIS2015-70862-P). DNU, PDG and MFC gratefully acknowledge the support of a Ramon y Cajal postdoctoral fellowship (RYC-2014-15392), a Beatriu de Pinos postdoctoral fellowship (BP-DGR 2015 (B) and a Severo Ochoa studentship, respectively. We would like to acknowledge Jose C. Sabina de Lis, J.M. Plata Suarez, A. Trifonova and C. Masoller for fruitful discussions.Navarro-Urrios, D.; Capuj, NE.; Colombano, MF.; García, PD.; Sledzinska, M.; Alzina, F.; Griol Barres, A.... (2017). Nonlinear dynamics and chaos in an optomechanical beam. Nature Communications. 8. https://doi.org/10.1038/ncomms14965S8Strogatz, S. H. Nonlinear Dynamics and Chaos: With Applications to Physics, Biology, Chemistry, and Engineering Westview Press (2014).Lorenz, E. N. Deterministic nonperiodic ow. J. Atmos. Sci. 20, 130–141 (1963).Sparrow, C. The Lorenz Attractor: Bifurcations, Chaos and Strange Attractors Springer (1982).Aspelmeyer, M., Kippenberg, T. & Marquardt, F. Cavity optomechanics. Rev. Mod. Phys. 86, 1391 (2014).Kippenberg, T., Rokhsari, H., Carmon, T., Scherer, A. & Vahala, K. Analysis of radiation-pressure induced mechanical oscillation of an optical microcavity. Phys. Rev. Lett. 95, 033901 (2005).Marquardt, F., Harris, J. G. E. & Girvin, S. M. Dynamical multistability induced by radiation pressure in high-finesse micromechanical optical cavities. Phys. Rev. Lett. 96, 103901 (2006).Krause, A. G. et al. Nonlinear radiation pressure dynamics in an optomechanical crystal. Phys. Rev. Lett. 115, 233601 (2015).Metzger, C. et al. Self-induced oscillations in an optomechanical system driven by bolometric backaction. Phys. Rev. Lett. 101, 133903 (2008).Bakemeier, L., Alvermann, A. & Fehske, H. Route to chaos in optomechanics. Phys. Rev. Lett. 114, 013601 (2015).Sciamanna, M. & Shore, K. A. Physics and applications of laser diode chaos. Nat. Photon. 9, 151–162 (2015).Williams, C. R. et al. Experimental observations of group synchrony in a system of chaotic optoelectronic oscillators. Phys. Rev. Lett. 110, 064104 (2013).Sciamanna, M. Optomechanics: vibrations copying optical chaos. Nat. Photon. 10, 366–368 (2016).Carmon, T., Cross, M. C. & Vahala, K. J. Chaotic quivering of micron-scaled on-chip resonators excited by centrifugal optical pressure. Phys. Rev. Lett. 98, 167203 (2007).Carmon, T., Rokhsari, H., Yang, L., Kippenberg, T. J. & Vahala, K. J. Temporal behavior of radiation-pressure-induced vibrations of an optical microcavity phonon mode. Phys. Rev. Lett. 94, 223902 (2005).Monifi, F. et al. Optomechanically induced stochastic resonance and chaos transfer between optical fields. Nat. Photon. 10, 399–405 (2016).Wu, J. et al. Dynamical chaos in chip-scale optomechanical oscillators. Preprint at https://arxiv.org/abs/1608.05071 (2016).Navarro-Urrios, D., Tredicucci, A. & Sotomayor-Torres, C. M. Coherent phonon generation in optomechanical crystals. SPIE Newsroom, doi:10.1117/2.1201507.006036 (2015).Navarro-Urrios, D. et al. A self-stabilized coherent phonon source driven by optical forces. Sci. Rep. 5, 15733 (2015).Johnson, T. J., Borselli, M. & Painter, O. Self-induced optical modulation of the transmission through a high-Q silicon microdisk resonator. Opt. Express 14, 817–831 (2006).Navarro-Urrios, D. et al. Self-sustained coherent phonon generation in optomechanical cavities. J. Opt. 18, 094006 (2016).Kemiktarak, U., Durand, M., Metcalfe, M. & Lawall, J. Mode competition and anomalous cooling in a multimode phonon laser. Phys. Rev. Lett. 113, 030802 (2014).Rosenstein, M. T., Collins, J. J. & De Luca, C. J. A practical method for calculating largest Lyapunov exponents from small data sets. Physica D 65, 117–134 (1993).Sprott, J. C. Chaos and Time-Series Analysis Vol. 69, Citeseer (2003).Grassberger, P. & Procaccia, I. Characterization of strange attractors. Phys. Rev. Lett. 50, 346–349 (1983).Hoppensteadt, F. C. & Izhikevich, E. M. Synchronization of MEMS resonators and mechanical neurocomputing. IEEE Trans. Circuits Syst. I, Reg. Papers 48, 133–138 (2001).Pennec, Y. et al. Band gaps and cavity modes in dual phononic and photonic strip waveguides. AIP Adv. 1, 041901 (2011).Gomis-Bresco, J. et al. A one-dimensional optomechanical crystal with a complete phononic band gap. Nat. Commun. 5, 4452 (2014).Johnson, S. G. et al. Perturbation theory for Maxwells equations with shifting material boundaries. Phys. Rev. E 65, 066611 (2002).Chan, J., Safavi-Naeini, A. H., Hill, J. T., Meenehan, S. & Painter, O. Optimized optomechanical crystal cavity with acoustic radiation shield. Appl. Phys. Lett. 101, 081115 (2012).Pennec, Y. et al. Modeling light-sound interaction in nanoscale cavities and waveguides. Nanophotonics 3, 413–440 (2014)

Functional divergence in the role of N-linked glycosylation in smoothened signaling

The G protein-coupled receptor (GPCR) Smoothened (Smo) is the requisite signal transducer of the evolutionarily conserved Hedgehog (Hh) pathway. Although aspects of Smo signaling are conserved from Drosophila to vertebrates, significant differences have evolved. These include changes in its active sub-cellular localization, and the ability of vertebrate Smo to induce distinct G protein-dependent and independent signals in response to ligand. Whereas the canonical Smo signal to Gli transcriptional effectors occurs in a G protein-independent manner, its non-canonical signal employs Gαi. Whether vertebrate Smo can selectively bias its signal between these routes is not yet known. N-linked glycosylation is a post-translational modification that can influence GPCR trafficking, ligand responsiveness and signal output. Smo proteins in Drosophila and vertebrate systems harbor N-linked glycans, but their role in Smo signaling has not been established. Herein, we present a comprehensive analysis of Drosophila and murine Smo glycosylation that supports a functional divergence in the contribution of N-linked glycans to signaling. Of the seven predicted glycan acceptor sites in Drosophila Smo, one is essential. Loss of N-glycosylation at this site disrupted Smo trafficking and attenuated its signaling capability. In stark contrast, we found that all four predicted N-glycosylation sites on murine Smo were dispensable for proper trafficking, agonist binding and canonical signal induction. However, the under-glycosylated protein was compromised in its ability to induce a non-canonical signal through Gαi, providing for the first time evidence that Smo can bias its signal and that a post-translational modification can impact this process. As such, we postulate a profound shift in N-glycan function from affecting Smo ER exit in flies to influencing its signal output in mice

Clinical features and health-related quality of life in adult patients with mucopolysaccharidosis IVA: the Spanish experience

Background: Mucopolysaccharidosis (MPS) IVA or Morquio A syndrome is a progressive and disabling disease characterized by a deficiency of the enzyme N-acetylgalactosamine-6-sulphate sulphatase. Its clinical presentation is very heterogeneous and poorly understood in adults. The aim of this study was to describe the clinical manifestations of MPS IVA in adult patients in Spain and to assess their health-related quality of life (HRQoL). Results: Thirty-three patients from nine reference centres participated in the study. The median age was 32 (interquartile range [IQR]: 20.5–40.5) years. The phenotype was classical in 54.5% of patients, intermediate in 33.3% of patients, and non-classical in 12.1% of patients. The most common clinical manifestation was bone dysplasia, with a median height of 118 (IQR: 106–136) cm. Other frequent clinical manifestations were hearing loss (75.7%), ligamentous laxity (72.7%), odontoid dysplasia (69.7%), limb deformities that required orthopaedic aids (mainly hip dysplasia and genu valgus) (63.6%), and corneal clouding (60.6%). In addition, 36.0% of patients had obstructive sleep apnoea/hypopnoea syndrome and 33.3% needed non-invasive ventilation. Cervical surgery and varisation osteotomy were the most common surgical interventions (36.4% each). Almost 80% of patients had mobility problems and 36.4% used a wheelchair at all times. Furthermore, 87.9% needed help with self-care, 33.3% were fully dependent, and 78.8% had some degree of pain. HRQoL according to the health assessment questionnaire was 1.43 (IQR: 1.03–2.00) in patients with the non-classical phenotype, but 2.5 (IQR: 1.68–3.00) in those with the classical phenotype. Seven patients were initiated on enzyme replacement therapy (ERT), but two of them were lost to follow-up. Lung function improved in four patients and slightly worsened in one patient. The distance achieved in the six-minute walk test increased in the four patients who could perform it. HRQoL was better in patients treated with elosulfase alfa, with a median (IQR) of 1.75 (1.25–2.34) versus 2.25 (1.62–3.00) in patients not treated with ERT. Conclusions: The study provides real-world data on patients with MPS IVA. Limited mobility, difficulties with self-care, dependence, and pain were common, together with poor HRQoL. The severity and heterogeneity of clinical manifestations require the combined efforts of multidisciplinary teams