Factors affecting the performance of P22 ELISA for the diagnosis of caprine tuberculosis in milk samples

Caprine tuberculosis (TB) is a zoonosis caused by members of the Mycobacterium tuberculosis complex (MTBC). Caprine TB eradication programmes are based mainly on intradermal tuberculin tests and slaughterhouse surveillance. However, the use of serological test has been extended as a potential diagnostic tool in goats through the use of serum, plasma, or even milk samples. Milk production and the antibodies (Ab) present in milk can vary depending on several circumstances. In the present study, different factors that may affect the performance of humoral TB diagnosis were analysed using goat milk samples: 1) lactation stage, 2) a recent previous skin test (booster effect) and 3) the effect of freeze-thaw cycles on milk samples preserved with azidiol. TB-infected animals (n = 44) were selected to evaluate the evolution of the Ab levels during the 6-month lactation period, along with its potential effect on the P22 ELISA results. In general, no significant changes (p = 0.079) were observed throughout the study as regards Ab levels in milk samples between consecutive analysis although the reactivity to P22 ELISA decreased when samplings were performed at the last two months of the lactation. Regarding the booster effect, the quantitative results showed a significant variation (p < 0.001) for both milk and serum samples when serological tests were carried out 15 days after the skin test. Finally, there were no significant differences (p = 0.99) in the P22 ELISA results when using milk samples preserved with azidiol that had undergone freeze-thaw cycles.This study was funded by the Spanish Ministry of Science and Innovation through the project “Análisis del proceso de erradicación de la tuberculosis caprina a largo plazo y desarrollo de pruebas de diagnóstico y medidas de control para su mejora (GoaTBfree-UCM, reference PID2019-105155RB-C31) and the Spanish Ministry of Agriculture, Fisheries and Food. JO was supported by an FPU (Formación de Profesorado Universitario) contract-fellowship provided by the Spanish Ministerio de Ciencia, Innovación y Universidades (FPU18/05197).S

Pulmonary delivery of cationic gold nanoparticles boost antigen-specific CD4+ T Cell Proliferation

To address how surface charge affects the fate of potential nanocarriers in the lung, gold nanoparticles (AuNPs) coated with polyvinyl alcohol containing either positively (NH2) or negatively (COOH) charged functional groups were intra-nasally instilled in mice, and their uptake by antigen presenting cell populations (APC) in broncho-alveolar lavage (BAL) fluid, trachea, and lung parenchyma, as well as trafficking to the lung draining lymph nodes (LDLNs) was assessed by flow cytometry. Furthermore, CD4+ T cell proliferation in LDLNs was investigated following instillation. All APC subpopulations preferentially captured positively-charged AuNPs compared to their negatively-charged counterparts. Uptake of AuNPs up-regulated expression of co-stimulatory molecules on all APC populations. Furthermore, positively-charged AuNPs induced enhanced OVA-specific CD4+ T cell stimulation in LDLNs compared to negatively-charged AuNPs, or polymer alone. Our findings demonstrate surface charge as a key parameter determining particle uptake by APC, and down-stream immune responses depend on the presence of particle core-bound polymer

REGULATION OF BETA-ADRENOCEPTORS ACTIVITYUSING SYNTHETIC LIGHT-REGULATED MOLECULES

Beta-adrenoceptors (ß-AR) are prototypical G proteincoupled receptors and important pharmacological targets for many diseases. Indeed, a number of approved drugs target these receptors due to their key role on many physiological functions. Among other examples, we encounter ß1-AR antagonists (ß- Blockers), which constitute the first-line therapy for the treatment of heart diseases, and ß2-AR agonists, which act as bronchodilators for the treatment of breathing pathologies. Considering the relevance of these receptors, achieving a reversible and localised control of their activity would provide a powerful tool, both for its research applications and its clinical potential. In this context, photopharmacology arises as a potent approach. Photopharmacology is an emerging field based on the use of synthetic light-regulated molecules to allow reversible spatiotemporal control of target receptors in native tissues. These ligands have the potential to provide a precise and controllable therapeutic action with increased efficacy and reduced side effects. Moreover, the fine regulation on demand of the receptor activation state is of great interest for their study in non-modified cells, tissues and organisms. The present project provides the first proof of concept for beta-adrenoceptor photopharmacology. We first designed and synthesised libraries of lightregulated compounds in order to regulate ß-AR activity with spatiotemporal precision. Subsequent testing highlighted the successful development of compounds with promising pharmacological properties which can be reversibly and irreversibly controlled by light. The discovered molecules enable a fine control of ß-AR in their native environment that will certainly open the door to innovative research procedures and may inspire future personalized therapies targeting these receptors

In vitro and in vivo regulation of ß-Adrenoceptors signaling using synthetic light-regulated molecules

Beta-adrenoceptors (ß-AR) are prototypical G protein-coupled receptors (GPCR) and important pharmacological targets for numerous diseases. Indeed, a number of approved drugs target ß-AR, which are key regulators of many physiological functions. Among other examples, ß1-AR antagonists (known as ß-Blockers) are first-line therapies for the treatment of heart failure, and ß2-AR agonists, which act as bronchodilators, are widely used for the treatment of breathing pathologies. Considering the medical relevance of these receptors, achieving a reversible and localized control of their activity would provide a powerful research and clinical tool. GPCR signaling is currently recognized as a multidimensional process governed by molecular, spatial and temporal components. Uncovering the role of each of these dimensions is crucial to improve our knowledge on cell communication, to understand how different pathways give rise to cellular and physiological effects, and to know how can we interact with biological systems with precision using drugs. Photopharmacology is an emerging field in which light-sensitive molecules are used to control the function of a given target protein in native tissues. The modulation of the target activity is achieved by small, drug-like, photoregulated ligands. By the use of light, both spatial and temporal control of the compound activity can be achieved in unprecedented manners compared to conventional pharmacology. These ligands have the potential to provide highly precise and controllable therapeutic actions that may result in increased efficacies and reduced side effects. Importantly, photopharmacology may allow to gain mechanistic insight on the interplay between the activation time and the receptor location during signaling processes in non-modified cells, tissues and whole organisms. Our research focused on the generation of new molecular tools for beta-adrenoceptors photopharmacology will be presented in this communication. First, several libraries of light-sensitive compounds with the aim to regulate ß-AR activity with spatiotemporal precision were designed and synthesized. Subsequent testing in cell preparations demonstrated the successful development of compounds with promising pharmacological properties, which can be reversibly and irreversibly controlled by light. Among those, several hit compounds were identified as ligands for beta-1 and beta-2 adrenoceptors with low nanomolar activities. These libraries compounds were found to be active enough to become useful photopharmacological tools, so we also performed in vivo experiments to determine their research potential in physiological environments. Indeed, the discovered molecules enabled a fine control of ß-AR in their native environment. We believe that the results of these studies will certainly open the door to innovative research procedures and may inspire future therapies targeting ß-AR

The templated growth of a chiral transition metal chalcogenide

We demonstrate that an intrinsically chiral, high Miller index surface of an achiral metal can be used to template the enantioselective growth of chiral transition metal chalcogenide films. Specifically, Cu(643)R can be used as a template for the enantioselective growth of a chiral copper telluride alloy surface. Beyond a critical alloy thickness the chiral influence of the Cu(643)R surface diminishes and an achiral surface forms. Our work demonstrates a new method of producing chiral transition metal chalcogenide surfaces, with potential applications in the study of structurally chiral topological insulators

Toward Multimodal Analytics in Ubiquitous Learning Environments

While Ubiquitous Learning Environments (ULEs) have shown several benefits for learning, they pose challenges for orchestration. Teachers need to be aware of the learning process, which is difficult to achieve when it occurs across a heterogeneous set of spaces, resources and devices. In addition, ULEs can benefit from multimodal analyses due to the heterogeneity of the data sources available (e.g., logs, geolocation, sensor information, learning artifacts). In previous works, we proposed an orchestration system with some analytics features that can gather multimodal datasets during the learning process. Based on this experience, in this paper we describe the technological support provided by the system to collect data from multiple spaces and sources as well as the structure of the generated dataset. We also reflect about the challenges of multimodal learning analytics (MMLA) in ULEs, and we pose some ideas about how the system could better support MMLA in the future to mitigate those challenges

NG2 antigen is involved in leukemia invasiveness and central nervous system infiltration in MLL-rearranged infant B-ALL

Mixed-lineage leukemia (MLL)-rearranged (MLLr) infant B-cell acute lymphoblastic leukemia (iMLLr-B-ALL) has a dismal prognosis and is associated with a pro-B/mixed phenotype, therapy refractoriness and frequent central nervous system (CNS) disease/relapse. Neuron-glial antigen 2 (NG2) is specifically expressed in MLLr leukemias and is used in leukemia immunophenotyping because of its predictive value for MLLr acute leukemias. NG2 is involved in melanoma metastasis and brain development; however, its role in MLL-mediated leukemogenesis remains elusive. Here we evaluated whether NG2 distinguishes leukemia-initiating/propagating cells (L-ICs) and/or CNS-infiltrating cells (CNS-ICs) in iMLLr-B-ALL. Clinical data from the Interfant cohort of iMLLr-B-ALL demonstrated that high NG2 expression associates with lower event-free survival, higher number of circulating blasts and more frequent CNS disease/relapse. Serial xenotransplantation of primary MLL-AF4 + leukemias indicated that NG2 is a malleable marker that does not enrich for L-IC or CNS-IC in iMLLr-B-All. However, NG2 expression was highly upregulated in blasts infiltrating extramedullar hematopoietic sites and CNS, and specific blockage of NG2 resulted in almost complete loss of engraftment. Indeed, gene expression profiling of primary blasts and primografts revealed a migratory signature of NG2 + blasts. This study provides new insights on the biology of NG2 in iMLLr-B-ALL and suggests NG2 as a potential therapeutic target to reduce the risk of CNS disease/relapse and to provide safer CNS-directed therapies for iMLLr-B-ALL

Two-dimensional multi-component photometric decomposition of CALIFA galaxies

We present a two-dimensional multi-component photometric decomposition of 404 galaxies from the Calar Alto Legacy Integral Field Area Data Release 3 (CALIFA-DR3). They represent all possible galaxies with no clear signs of interaction and not strongly inclined in the final CALIFA data release. Galaxies are modelled in the g, r, and i Sloan Digital Sky Survey (SDSS) images including, when appropriate, a nuclear point source, bulge, bar, and an exponential or broken disc component. We use a human-supervised approach to determine the optimal number of structures to be included in the fit. The dataset, including the photometric parameters of the CALIFA sample, is released together with statistical errors and a visual analysis of the quality of each fit. The analysis of the photometric components reveals a clear segregation of the structural composition of galaxies with stellar mass. At high masses (log(M⋆/M⊙) > 11), the galaxy population is dominated by galaxies modelled with a single Sérsic or a bulge+disc with a bulge-to-total (B/T) luminosity ratio B/T > 0.2. At intermediate masses (9.5 < log(M⋆/M⊙) < 11), galaxies described with bulge+disc but B/T < 0.2 are preponderant, whereas, at the low mass end (log(M⋆/M⊙)< 9.5), the prevailing population is constituted by galaxies modelled with either pure discs or nuclear point sources+discs (i.e., no discernible bulge). We obtain that 57% of the volume corrected sample of disc galaxies in the CALIFA sample host a bar. This bar fraction shows a significant drop with increasing galaxy mass in the range 9.5 < log(M⋆/M⊙) < 11.5. The analyses of the extended multi-component radial profile result in a volume-corrected distribution of 62%, 28%, and 10% for the so-called Type I (pure exponential), Type II (down-bending), and Type III (up-bending) disc profiles, respectively. These fractions are in discordance with previous findings. We argue that the different methodologies used to detect the breaks are the main cause for these differences.PostprintPeer reviewe

Comprehensive characterization of a novel, oncogenic and targetable SEPTIN6::ABL2 fusion in T-ALL

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Nanoparticle colloidal stability in cell culture media and impact on cellular interactions

Nanomaterials are finding increasing use for biomedical applications such as imaging, diagnostics, and drug delivery. While it is well understood that nanoparticle (NP) physico-chemical properties can dictate biological responses and interactions, it has been difficult to outline a unifying framework to directly link NP properties to expected in vitro and in vivo outcomes. When introduced to complex biological media containing electrolytes, proteins, lipids, etc., nanoparticles (NPs) are subjected to a range of forces which determine their behavior in this environment. One aspect of NP behavior in biological systems that is often understated or overlooked is aggregation. NP aggregation will significantly alter in vitro behavior (dosimetry, NP uptake, cytotoxicity), as well as in vivo fate (pharmacokinetics, toxicity, biodistribution). Thus, understanding the factors driving NP colloidal stability and aggregation is paramount. Furthermore, studying biological interactions with NPs at the nanoscale level requires an interdisciplinary effort with a robust understanding of multiple characterization techniques. This review examines the factors that determine NP colloidal stability, the various efforts to stabilize NP in biological media, the methods to characterize NP colloidal stability in situ, and provides a discussion regarding NP interactions with cell