Surface receptors that mediate the interaction of Borrelia burgdorferi with macrophages: death and inflammation.

182 p.La enfermedad de Lyme es causada por la transmisión de la bacteria llamada Borrelia burgdorferi. Esta tesis tiene como objetivo el estudio de la respuesta inmune de los macrófagos a B. burgdorferi, centrándonos en los elementos involucrados en su actividad fagocítica y la posterior respuesta inflamatoria mediada por estas células, a fin de comprender más profundamente la interacción de este tipo de células y la bacteria. Mediante enfoques analíticos de gran escala, como la transcriptómica y la proteómica, hemos estudiado en macrófagos murinos y humanos, las rutas de señalización implicadas en la fagocitosis de este patógeno, y hemos descrito un conjunto de receptores de membranas responsables de su ingestión y eliminación. Nuestro estudio ha permitido el descubrimiento y descripción de los receptores CD180 y CD64 como elementos moduladores y mediadores de la respuesta inmune a B. burgdorferi, así como del consiguiente proceso inflamatorio. También hemos esclarecido la importancia y el comportamiento de diferentes factores de señalización, como SHIP1, FcR¿, Syk y MyD88, revelando la existencia de una compleja conexión entre las familias de los receptores que intervienen en el proceso de fagocitosis de este patógeno. Este conocimiento conducirá a una mejor comprensión del rol de los macrófagos en respuesta a B. burgdorferi, arrojando luz sobre los síntomas inflamatorios observados durante la borreolisis de Lyme y revelando nuevas dianas terapéuticas para un tratamiento más efectivo de la enfermedad.CICbioGUN

Surface receptors that mediate the interaction of Borrelia burgdorferi with macrophages: death and inflammation.

182 p.La enfermedad de Lyme es causada por la transmisión de la bacteria llamada Borrelia burgdorferi. Esta tesis tiene como objetivo el estudio de la respuesta inmune de los macrófagos a B. burgdorferi, centrándonos en los elementos involucrados en su actividad fagocítica y la posterior respuesta inflamatoria mediada por estas células, a fin de comprender más profundamente la interacción de este tipo de células y la bacteria. Mediante enfoques analíticos de gran escala, como la transcriptómica y la proteómica, hemos estudiado en macrófagos murinos y humanos, las rutas de señalización implicadas en la fagocitosis de este patógeno, y hemos descrito un conjunto de receptores de membranas responsables de su ingestión y eliminación. Nuestro estudio ha permitido el descubrimiento y descripción de los receptores CD180 y CD64 como elementos moduladores y mediadores de la respuesta inmune a B. burgdorferi, así como del consiguiente proceso inflamatorio. También hemos esclarecido la importancia y el comportamiento de diferentes factores de señalización, como SHIP1, FcR¿, Syk y MyD88, revelando la existencia de una compleja conexión entre las familias de los receptores que intervienen en el proceso de fagocitosis de este patógeno. Este conocimiento conducirá a una mejor comprensión del rol de los macrófagos en respuesta a B. burgdorferi, arrojando luz sobre los síntomas inflamatorios observados durante la borreolisis de Lyme y revelando nuevas dianas terapéuticas para un tratamiento más efectivo de la enfermedad.CICbioGUN

Cueva del sol, a solar marker in the sierra de la plata (Tarifa, Cádiz)

En el ámbito geográfico que nos ocupa, entornos de la Laguna de la Janda y Campo de Gibraltar, existen unos trescientos abrigos con arte rupestre escasamente estudiados. En veintidós de ellos está representada la figura soliforme en sus paredes rocosas. La denominada Cueva del Sol es uno de los abrigos que forman parte de este conjunto. Sus especiales características, tanto geográficas como iconográficas, y los reveladores resultados de observaciones astronómicas apuntan a la existencia de un marcador solar del solsticio de invierno, y convierten a esta cavidad en una de las más excepcionales del conjunto y, tal vez, de la península ibérica. Los datos obtenidos hacen que este abrigo pueda llevarnos a determinar algunos aspectos concretos sobre los cultos aso - ciados al comportamiento solar y su función como regulador de las actividades productivas y, por ende, acercarnos a la cosmovisión de los hacedores de estas pinturas. Para ello es condición sine qua non un análisis individual y profundo, realizado a partir de un enfoque teórico-metodológico basado sobre todo en la Arqueoastronomía, la Arqueología del Paisaje, la Semiótica o la Etnografía, entre otras disciplinas.In the geographical area under study, surround - ings of the Laguna de la Janda and the Campo de Gibraltar , there are over three hundred rock-shelters with cave paint - ings, which have barely been studied. The sun-shaped figure is depicted on the rocky walls of twenty-two of these rock- shelters. The Cueva del Sol is one of the rock-shelters that forms part of this set. Due to both its special geographical and iconographic characteristics as well as the revealing results of astronomic observation that confirmed the existence of a winter solstice marker (sun dagger), this rock-shelter is one of the most exceptional of the set and maybe of the ibe - rian peninsula. The characteristics of this rock-shelter enable us to determine some specific aspects on worship associated to the behaviour of the sun and its role in regulating produc - tion activities thus bringing us closer to the worldview of the creators of these paintings. For this purpose, a detailed and individual analysis from a theoretical and methodological ap - proach, mainly based on Archaeoastronomy, Landscape Archaeology, Semiotics and Ethnography, amongst other disci - plines, is a conditio sine qua non

Structural basis for selective recognition of acyl chains by the membrane-associated acyltransferase PatA

The biosynthesis of phospholipids and glycolipids are critical pathways for virtually all cell membranes. PatA is an essential membrane associated acyltransferase involved in the biosynthesis of mycobacterial phosphatidyl-myo-inositol mannosides (PIMs). The enzyme transfers a palmitoyl moiety from palmitoyl-CoA to the 6-position of the mannose ring linked to 2-position of inositol in PIM1/PIM2. We report here the crystal structures of PatA from Mycobacterium smegmatis in the presence of its naturally occurring acyl donor palmitate and a nonhydrolyzable palmitoyl-CoA analog. The structures reveal an alpha/beta architecture, with the acyl chain deeply buried into a hydrophobic pocket that runs perpendicular to a long groove where the active site is located. Enzyme catalysis is mediated by an unprecedented charge relay system, which markedly diverges from the canonical HX4D motif. Our studies establish the mechanistic basis of substrate/membrane recognition and catalysis for an important family of acyltransferases, providing exciting possibilities for inhibitor design.This work was supported by the European Commission Contract HEALTH-F3-2011-260872, the Spanish Ministry of Economy and Competitiveness Contract BIO2013-49022-C2-2-R, and the Basque Government (to M.E.G.); Slovak Research and Development Agency Contract No. DO7RP-0015-11 (to K.M.) and the NIH/NIAID grant AI064798 (to M.J.). D.A.-J. acknowledges the support from Fundacion Biofisica Bizkaia. We gratefully acknowledge Sonia Lopez-Fernandez (Unit of Biophysics, CSIC, UPV/EHU, Spain), Drs E. Ogando and T. Mercero (Scientific Computing Service UPV/EHU, Spain) for technical assistance. We thank the Swiss Light Source (SLS), and the Diamond Light Source (DLS) for granting access to synchrotron radiation facilities and their staff for the onsite assistance. We specially thank the BioStruct-X project to support access to structural biology facilities. We also acknowledge all members of the Structural Glycobiology Group (Spain) for valuable scientific discussions. The following reagent was obtained through BEI Resources, NIAID, NIH: Mycobacterium tuberculosis, Strain H37Rv, Purified Phosphatidylinositol Mannosides 1 and 2 (PIM1,2), NR-14846

Borrelia burgdorferi infection induces long-term memory-like responses in macrophages with tissue-wide consequences in the heart

Lyme carditis is an extracutaneous manifestation of Lyme disease characterized by episodes of atrioventricular block of varying degrees and additional, less reported cardiomyopathies. The molecular changes associated with the response to Borrelia burgdorferi over the course of infection are poorly understood. Here, we identify broad transcriptomic and proteomic changes in the heart during infection that reveal a profound down-regulation of mitochondrial components. We also describe the long-term functional modulation of macrophages exposed to live bacteria, characterized by an augmented glycolytic output, increased spirochetal binding and internalization, and reduced inflammatory responses. In vitro, glycolysis inhibition reduces the production of tumor necrosis factor (TNF) by memory macrophages, whereas in vivo, it produces the reversion of the memory phenotype, the recovery of tissue mitochondrial components, and decreased inflammation and spirochetal burdens. These results show that B. burgdorferi induces long-term, memory-like responses in macrophages with tissue-wide consequences that are amenable to be manipulated in vivo.Supported by grants from the Spanish Ministry of Science, Innovation and Universities (MCIU) co-financed with FEDER funds (SAF2015-65327-R and RTI2018-096494-B-100 to JA; BFU2016-76872-R to EB, AGL2017-86757-R to LA, SAF2017-87301-R to MLMC, SAF2015-64111-R to AP, SAF2015-73549-JIN to HR), Instituto de Salud Carlos III (PIE13/0004 to AP), the Basque Government Department of Health (2015111117 to LA), the Basque Foundation for Innovation and Health Research (BIOEF), through the EiTB Maratoia grant BIO15/CA/016/BS to MLMC, the regional Government of Andalusia co-funded by CEC and FEDER funds (Proyectos de Excelencia P12-CTS-2232) and Fundación Domingo Martínez (to AP). LA is supported by the Ramon y Cajal program (RYC-2013-13666). DB, MMR and TMM are recipients of MCIU FPI fellowships. ACG and AP are recipients of fellowships form the Basque Government. APC is a recipient of a fellowship from the University of the Basque Country. We thank the MCIU for the Severo Ochoa Excellence accreditation (SEV-2016-0644), the Basque Department of Industry, Tourism and Trade (Etortek and Elkartek programs), the Innovation Technology Department of the Bizkaia Province and the CIBERehd network. DB and JA are supported by a grant from the Jesús de Gangoiti Barrera Foundation

DD04107-Derived neuronal exocytosis inhibitor peptides: Evidences for synaptotagmin-1 as a putative target

15 pags, 8 figs, 3 tabs. -- Supplementary data to this article can be found online at https://doi.org/10.1016/j.bioorg.2021.105231.The analgesic peptide DD04107 (Pal-EEMQRR-NH2) and its acetylated analogue inhibit α-calcitonin gene-related peptide (α-CGRP) exocytotic release from primary sensory neurons. Examining the crystal structure of the SNARE-Synaptotagmin-1(Syt1) complex, we hypothesized that these peptides could inhibit neuronal exocytosis by binding to Syt1, hampering at least partially its interaction with the SNARE complex. To address this hypothesis, we first interrogate the role of individual side-chains on the inhibition of α-CGRP release, finding that E1, M3, Q4 and R6 residues were crucial for activity. CD and NMR conformational analysis showed that linear peptides have tendency to adopt α-helical conformations, but the results with cyclic analogues indicated that this secondary structure is not needed for activity. Isothermal titration calorimetry (ITC) measurements demonstrate a direct interaction of some of these peptides with Syt1-C2B domain, but not with Syt7-C2B region, indicating selectivity. As expected for a compound able to inhibit α-CGRP release, cyclic peptide derivative Pal-E-cyclo[EMQK]R-NH2 showed potent in vivo analgesic activity, in a model of inflammatory pain. Molecular dynamics simulations provided a model consistent with KD values for the interaction of peptides with Syt1-C2B domain, and with their biological activity. Altogether, these results identify Syt1 as a potential new analgesic target.This work was supported by the Spanish Ministerio de Economía y Competitividad (MINECO-FEDER), RTI2018-097189-C2 and CTQ2017-84371-P), and the Spanish National Research Council (CSIC, 201880E109, 201980E030). The NMR experiments were performed in the “Manuel Rico” NMR laboratory, LMR, CSIC, a node of the Spanish Large-Scale National Facility ICTS R-LRB. We thank Prof. Josep Rizo and R. Voleti (Dept. Biophysics, Biochemistry and Pharmacology, UT Southwestern Medical Center, Dallas, USA) for providing the clones required for expressing Syt1 and Syt7 proteins. SG-R and AB belong to the Instituto de Investigación Sanitaria del Principado de Asturias (ISPA).Peer reviewe

Exposure to secondhand and thirdhand smoke in private vehicles: Measurements in air and dust samples

Background: This study aimed to estimate airborne nicotine concentrations and nicotine, cotinine, and tobaccospecific nitrosamines (TSNAs) in settled dust from private cars in Spain and the UK. Methods: We measured vapor-phase nicotine concentrations in a convenience sample of 45 private cars from Spain (N = 30) and the UK (N = 15) in 2017-2018. We recruited non-smoking drivers (n = 20), smoking drivers who do not smoke inside the car (n = 15), and smoking drivers who smoke inside (n = 10). Nicotine, cotinine, and three TSNAs (NNK, NNN, NNA) were also measured in settled dust in a random subsample (n = 20). We computed medians and interquartile ranges (IQR) of secondhand smoke (SHS) and thirdhand smoke (THS) compounds according to the drivers' profile. Results: 24-h samples yielded median airborne nicotine concentrations below the limit of quantification (LOQ) (IQR

Primary and Secondary Immunodeficiency Diseases in Oncohaematology: Warning Signs, Diagnosis, and Management

Background: Immunodeficiencies (ID), in particular primary immunodeficiencies (PID), are often associated with haematological manifestations, such as peripheral cytopenias or lymphoproliferative syndromes. Early diagnosis and management have significant prognostic implications. Secondary immunodeficiencies (SID) may also be induced by oncohaematological diseases and their treatments. Haematologists and oncologists must therefore be aware of the association between blood disorders and cancer and ID, and be prepared to offer their patients appropriate treatment without delay. Our aim was to define the warning signs of primary and secondary IDs in paediatric and adult patients with oncohaematological manifestations.Methods: A multidisciplinary group of six experts (2 haematologists, 2 immunologists, and 2 paediatricians specializing in ID) conducted a literature review and prepared a document based on agreements reached an in-person meeting. An external group of 44 IDs specialists from all over Spain assessed the document and were consulted regarding their level of agreement.Results: This document identifies the haematological and extra-haematological diseases that should prompt a suspicion of PIDs in adults and children, in both primary care and haematology and oncology departments. Cytopenia and certain lymphoproliferative disorders are key diagnostic pointers. The diagnosis must be based on a detailed clinical history, physical exploration, complete blood count and standard laboratory tests. The immunological and haematological tests included in the diagnostic process will depend on the care level. Patients who are candidates for immunoglobulin replacement therapy must be carefully selected, and treatment should be offered as soon as possible to avoid the development of complications. Finally, this document recommends procedures for monitoring these patients.Conclusions: This document combines scientific evidence with the opinion of a broad panel of experts, and emphasizes the importance of an early diagnosis and treatment to avoid complications. The resulting document is a useful tool for primary care physicians and specialists who see both adult and paediatric patients with oncohaematological diseases

Burden of disease attributable to risk factors in European countries: a scoping literature review

Objectives: Within the framework of the burden of disease (BoD) approach, disease, and injury burden estimates attributable to risk factors are a useful guide for policy formulation and priority setting in disease prevention. Considering the important differences in methods, and their impact on burden estimates, we conducted a scoping literature review to: (1) map the BoD assessments including risk factors performed across Europe, and (2) identify the methodological choices in comparative risk assessment (CRA) and risk assessment methods. Methods: We searched multiple literature databases, including grey literature websites, and targeted public health agencies' websites. Results: A total of 113 studies were included in the synthesis and further divided into independent BoD assessments (54 studies) and studies linked to the Global Burden of Disease (59 papers). Our results showed that the methods used to perform CRA varied substantially across independent European BoD studies. While there were some methodological choices that were more common than others, we did not observe patterns in terms of country, year, or risk factor. Each methodological choice can affect the comparability of estimates between and within countries and/or risk factors since they might significantly influence the quantification of the attributable burden. From our analysis, we observed that the use of CRA was less common for some types of risk factors and outcomes. These included environmental and occupational risk factors, which are more likely to use bottom-up approaches for health outcomes where disease envelopes may not be available. Conclusions: Our review also highlighted misreporting, the lack of uncertainty analysis, and the under-investigation of causal relationships in BoD studies. Development and use of guidelines for performing and reporting BoD studies will help understand differences, and avoid misinterpretations thus improving comparability among estimates.info:eu-repo/semantics/publishedVersio