W196 and the ß -Hairpin Motif Modulate the Redox Switch of Conformation and the Biomolecular Interaction Network of the Apoptosis-Inducing Factor

The human apoptosis-inducing factor (hAIF) is a moonlight flavoprotein involved in mitochondrial respiratory complex assembly and caspase-independent programmed cell death. These functions might be modulated by its redox-linked structural transition that enables hAIF to act as a NAD(H/+) redox sensor. Upon reduction with NADH, hAIF undergoes a conformational reorganization in two specific insertions - the flexible regulatory C-loop and the 190-202 ß-harpin - promoting protein dimerization and the stabilization of a long-life charge transfer complex (CTC) that modulates its monomer-dimer equilibrium and its protein interaction network in healthy mitochondria. In this regard, here, we investigated the precise function of the ß-hairpin in the AIF conformation landscape related to its redox mechanism, by analyzing the role played by W196, a key residue in the interaction of this motif with the regulatory C-loop. Mutations at W196 decrease the compactness and stability of the oxidized hAIF, indicating that the ß-hairpin and C-loop coupling contribute to protein stability. Kinetic studies complemented with computational simulations reveal that W196 and the ß-hairpin conformation modulate the low efficiency of hAIF as NADH oxidoreductase, contributing to configure its active site in a noncompetent geometry for hydride transfer and to stabilize the CTC state by enhancing the affinity for NAD+. Finally, the ß-hairpin motif contributes to define the conformation of AIF's interaction surfaces with its physiological partners. These findings improve our understanding on the molecular basis of hAIF''s cellular activities, a crucial aspect for clarifying its associated pathological mechanisms and developing new molecular therapies

Comparison of constant load exercise intensity for verification of maximal oxygen uptake following a graded exercise test in older adults

Maximal oxygen uptake (VO2max) declines with advancing age and is a predictor of morbidity and mortality risk. The purpose here was to assess the utility of constant load tests performed either above or below peak work rate obtained from a graded exercise test for verification of VO2max in older adults. Twenty-two healthy older adults (9M, 13F, 67 ± 6 years, BMI: 26.3 ± 5.1 kg·m−2) participated in the study. Participants were asked to complete two experimental trials in a randomized, counterbalanced cross-over design. Both trials (cycle ergometer) consisted of (1) an identical graded exercise test (ramp) and (2) a constant load test at either 85% (CL85; n = 22) or 110% (CL110; n = 20) of the peak work rate achieved during the associated ramp (performed 10-min post ramp). No significant differences were observed for peak VO2 (L·min−1) between CL85 (1.86 ± 0.72; p = 0.679) or CL110 (1.79 ± 0.73; p = 0.200) and the associated ramp (Ramp85, 1.85 ± 0.73; Ramp110, 1.85 ± 0.57). Using the study participant\u27s mean coefficient of variation in peak VO2 between the two identical ramp tests (2.9%) to compare individual differences between constant load tests and the associated ramp revealed 19/22 (86%) of participants achieved a peak VO2 during CL85 that was similar or higher versus the ramp, while only 13/20 (65%) of participants achieved a peak VO2 during CL110 that was similar or higher versus the ramp. These data indicate that if a verification of VO2max is warranted when testing older adults, a constant load effort at 85% of ramp peak power may be more likely to verify VO2max as compared to an effort at 110% of ramp peak power

Digital Inclusion Across the Americas and Caribbean

This research brings together digital inequality scholars from across the Americas and Caribbean to examine efforts to tackle digital inequality in Uruguay, Chile, Peru, Brazil, Mexico, Cuba, Jamaica, the United States, and Canada. As the case studies show, governmental policy has an important role to play in reducing digital disparities, particularly for potential users in rural or remote areas, as well as populations with great economic disparities. We find that public policy can effectively reduce access gaps when it combines the trifecta of network, device, and skill provision, especially through educational institutions. We also note, that urban populations have benefitted from digital inclusion strategies to a greater degree. This underscores that, no matter the national context, rural-urban digital inequality (and often associated economic inequality) is resistant to change. Even when access is provided, potential users may not find it affordable, lack skills, and/or see no benefit in adoption. We see the greatest potential for future digital inclusion in two related approaches: 1) initiatives that connect with hard-to-reach, remote, and rural communities outside urban cores and 2) initiatives that learn from communities about how best to provide digital resources while respecting their diversely situated contexts, while meeting social, economic and political needs

SARS-CoV-2 testing and infection control strategies in European paediatric emergency departments during the first wave of the pandemic.

Between February and May 2020, during the first wave of the COVID-19 pandemic, paediatric emergency departments in 12 European countries were prospectively surveyed on their implementation of SARS-CoV-2 disease (COVID-19) testing and infection control strategies. All participating departments (23) implemented standardised case definitions, testing guidelines, early triage and infection control strategies early in the outbreak. Patient testing criteria initially focused on suspect cases and later began to include screening, mainly for hospital admissions. Long turnaround times for test results likely put additional strain on healthcare resources.Conclusion: Shortening turnaround times for SARS-CoV-2 tests should be a priority. Specific paediatric testing criteria are needed. What is Known: • WHO and public health authorities issued case definitions, testing and infection control recommendations for COVID-19 in January. • SARS-CoV-2 testing was made available across Europe in February. What is New: • Paediatric emergency departments implemented COVID-19-specific procedures rapidly, including case definitions, testing guidelines and early triage. • A third of surveyed departments waited more than 24 h for SARS-CoV-2 test to be reported, resulting in additional strain on resources

Analyzing multitarget activity landscapes using protein-ligand interaction fingerprints: interaction cliffs.

This is the original submitted version, before peer review. The final peer-reviewed version is available from ACS at http://pubs.acs.org/doi/abs/10.1021/ci500721x.Activity landscape modeling is mostly a descriptive technique that allows rationalizing continuous and discontinuous SARs. Nevertheless, the interpretation of some landscape features, especially of activity cliffs, is not straightforward. As the nature of activity cliffs depends on the ligand and the target, information regarding both should be included in the analysis. A specific way to include this information is using protein-ligand interaction fingerprints (IFPs). In this paper we report the activity landscape modeling of 507 ligand-kinase complexes (from the KLIFS database) including IFP, which facilitates the analysis and interpretation of activity cliffs. Here we introduce the structure-activity-interaction similarity (SAIS) maps that incorporate information on ligand-target contact similarity. We also introduce the concept of interaction cliffs defined as ligand-target complexes with high structural and interaction similarity but have a large potency difference of the ligands. Moreover, the information retrieved regarding the specific interaction allowed the identification of activity cliff hot spots, which help to rationalize activity cliffs from the target point of view. In general, the information provided by IFPs provides a structure-based understanding of some activity landscape features. This paper shows examples of analyses that can be carried out when IFPs are added to the activity landscape model.M-L is very grateful to CONACyT (No. 217442/312933) and the Cambridge Overseas Trust for funding. AB thanks Unilever for funding and the European Research Council for a Starting Grant (ERC-2013- StG-336159 MIXTURE). J.L.M-F. is grateful to the School of Chemistry, Department of Pharmacy of the National Autonomous University of Mexico (UNAM) for support. This work was supported by a scholarship from the Secretariat of Public Education and the Mexican government

Near-infrared photometry and spectroscopy of the low Galactic latitude globular cluster 2MASS-GC03

We present deep near-infrared photometry and spectroscopy of the globular cluster 2MASS-GC03 projected in the Galactic disk using MMIRS on the Clay telescope (Las Campanas Observatory) and VISTA Variables in the Via Lactea survey (VVV) data. Most probable cluster member candidates were identified from near-infrared photometry. Out of ten candidates that were followed-up spectroscopically, five have properties of cluster members, from which we calculate = -0.9 +/- 0.2 and a radial velocity of v_r > = -78 +- 12km/s. A distance of 10.8kpc is estimated from 3 likely RRLyrae members. Given that the cluster is currently at a distance of 4.2kpc from the Galactic center, the cluster's long survival time of an estimated 11.3 +/- 1.2 Gyr strengthens the case for its globular-cluster nature. The cluster has a hint of elongation in the direction of the Galactic center.Peer reviewedFinal Published versio

Acupuncture and rehabilitation of the painful shoulder: study protocol of an ongoing multicentre randomised controlled clinical trial [ISRCTN28687220]

BACKGROUND: Although the painful shoulder is one of the most common dysfunctions of the locomotor apparatus, and is frequently treated both at primary healthcare centres and by specialists, little evidence has been reported to support or refute the effectiveness of the treatments most commonly applied. According to the bibliography reviewed, physiotherapy, which is the most common action taken to alleviate this problem, has not yet been proven to be effective, because of the small size of sample groups and the lack of methodological rigor in the papers published on the subject. No reviews have been made to assess the effectiveness of acupuncture in treating this complaint, but in recent years controlled randomised studies have been made and these demonstrate an increasing use of acupuncture to treat pathologies of the soft tissues of the shoulder. In this study, we seek to evaluate the effectiveness of physiotherapy applied jointly with acupuncture, compared with physiotherapy applied with a TENS-placebo, in the treatment of painful shoulder caused by subacromial syndrome (rotator cuff tendinitis and subacromial bursitis). METHODS/DESIGN: Randomised controlled multicentre study with blind evaluation by an independent observer and blind, independent analysis. A study will be made of 465 patients referred to the rehabilitation services at participating healthcare centres, belonging to the regional public health systems of Andalusia and Murcia, these patients presenting symptoms of painful shoulder and a diagnosis of subacromial syndrome (rotator cuff tendinitis and subacromial bursitis). The patients will be randomised into two groups: 1) experimental (acupuncture + physiotherapy); 2) control (TENS-placebo + physiotherapy); the administration of rescue medication will also be allowed. The treatment period will have a duration of three weeks. The main result variable will be the change produced on Constant's Shoulder Function Assessment (SFA) Scale; as secondary variables, we will record the changes in diurnal pain intensity on a visual analogue scale (VAS), nocturnal pain intensity on the VAS, doses of non-steroid anti-inflammatory drugs (NSAIDs) taken during the study period, credibility scale for the treatment, degree of improvement perceived by the patient and degree of improvement perceived by the evaluator. A follow up examination will be made at 3, 6 and 12 months after the study period has ended. Two types of population will be considered for analysis: per protocol and per intention to treat. DISCUSSION: The discussion will take into account the limitations of the study, together with considerations such as the choice of a simple, safe method to treat this shoulder complaint, the choice of the control group, and the blinding of the patients, evaluators and those responsible for carrying out the final analysis

VETTONIA PROJECT: A VIRTUAL ENVIRONMENT FOR THE EDUCATIONAL DISSEMINATION OF THE IRON AGE

The VETTONIA project aims to disseminate the rich heritage from the Iron Age of the western Iberian Peninsula and the archaeological investigations carried out on this topic in recent years. The project utilizes new technologies such as virtual tours, 3D models, and impressions to create interactive and stimulating ways to access the results of the most recent archaeological research. Using these resources, lectures and seminars are being given in various forums with diverse types of audiences to present the virtual tours and the rest of the dissemination initiatives. In addition, the project presents its different initiatives during the annual archaeological interventions developed in the oppidum of Ulaca (Solosancho, Ávila, Spain), with good reception by the attending public. The VETTONIA project represents a pioneering dissemination experience that takes advantage of the educational opportunities offered by new technologies. In the future, tools such as virtual tours to archaeological sites may prove essential in classroom teaching at different levels and could promote sustainable tourism in fragile natural environments such as those that constitute the major settlements of the Late Iron Age (ca. 400–50 BC)

Neutrophil Extracellular Traps Directly Induce Epithelial and Endothelial Cell Death: A Predominant Role of Histones

Neutrophils play an important role in innate immunity by defending the host organism against invading microorganisms. Antimicrobial activity of neutrophils is mediated by release of antimicrobial peptides, phagocytosis as well as formation of neutrophil extracellular traps (NET). These structures are composed of DNA, histones and granular proteins such as neutrophil elastase and myeloperoxidase. This study focused on the influence of NET on the host cell functions, particularly on human alveolar epithelial cells as the major cells responsible for gas exchange in the lung. Upon direct interaction with epithelial and endothelial cells, NET induced cytotoxic effects in a dose-dependent manner, and digestion of DNA in NET did not change NET-mediated cytotoxicity. Pre-incubation of NET with antibodies against histones, with polysialic acid or with myeloperoxidase inhibitor but not with elastase inhibitor reduced NET-mediated cytotoxicity, suggesting that histones and myeloperoxidase are responsible for NET-mediated cytotoxicity. Although activated protein C (APC) did decrease the histone-induced cytotoxicity in a purified system, it did not change NET-induced cytotoxicity, indicating that histone-dependent cytotoxicity of NET is protected against APC degradation. Moreover, in LPS-induced acute lung injury mouse model, NET formation was documented in the lung tissue as well as in the bronchoalveolar lavage fluid. These data reveal the important role of protein components in NET, particularly histones, which may lead to host cell cytotoxicity and may be involved in lung tissue destruction

An Off-Target Nucleostemin RNAi Inhibits Growth in Human Glioblastoma-Derived Cancer Stem Cells

Glioblastomas (GBM) may contain a variable proportion of active cancer stem cells (CSCs) capable of self-renewal, of aggregating into CD133+ neurospheres, and to develop intracranial tumors that phenocopy the original ones. We hypothesized that nucleostemin may contribute to cancer stem cell biology as these cells share characteristics with normal stem cells. Here we report that nucleostemin is expressed in GBM-CSCs isolated from patient samples, and that its expression, conversely to what it has been described for ordinary stem cells, does not disappear when cells are differentiated. The significance of nucleostemin expression in CSCs was addressed by targeting the corresponding mRNA using lentivirally transduced short hairpin RNA (shRNA). In doing so, we found an off-target nucleostemin RNAi (shRNA22) that abolishes proliferation and induces apoptosis in GBM-CSCs. Furthermore, in the presence of shRNA22, GBM-CSCs failed to form neurospheres in vitro or grow on soft agar. When these cells are xenotransplanted into the brains of nude rats, tumor development is significantly delayed. Attempts were made to identify the primary target/s of shRNA22, suggesting a transcription factor involved in one of the MAP-kinases signaling-pathways or multiple targets. The use of this shRNA may contribute to develop new therapeutic approaches for this incurable type of brain tumor