Diagnosis and Stratification of Pseudomonas aeruginosa Infected Patients by Immunochemical Quantitative Determination of Pyocyanin From Clinical Bacterial Isolates

Pseudomonas aeruginosa; Monoclonal antibody; PyocyaninPseudomonas aeruginosa; Anticuerpo monoclonal; PiocianinaPseudomonas aeruginosa; Anticòs monoclonal; PiocianinaThe development of a highly sensitive, specific, and reliable immunochemical assay to detect pyocyanin (PYO), one of the most important virulence factors (VFs) of Pseudomonas aeruginosa, is here reported. The assay uses a high-affinity monoclonal antibody (mAb; C.9.1.9.1.1.2.2.) raised against 1-hydroxyphenazine (1-OHphz) hapten derivatives (PC1; a 1:1 mixture of 9-hydroxy- and 6-hydroxy-phenazine-2-carobxylic acids). Selective screening using PYO and 1-OHphz on several cloning cycles allowed the selection of a clone able to detect PYO at low concentration levels. The microplate-based ELISA developed is able to achieve a limit of detection (LoD) of 0.07 nM, which is much lower than the concentrations reported to be found in clinical samples (130 μM in sputa and 2.8 μM in ear secretions). The ELISA has allowed the investigation of the release kinetics of PYO and 1-OHphz (the main metabolite of PYO) of clinical isolates obtained from P. aeruginosa-infected patients and cultured in Mueller-Hinton medium. Significant differences have been found between clinical isolates obtained from patients with an acute or a chronic infection (~6,000 nM vs. ~8 nM of PYO content, respectively) corroborated by the analysis of PYO/1-OHphz levels released by 37 clinical isolates obtained from infected patients at different stages. In all cases, the levels of 1-OHphz were much lower than those of PYO (at the highest levels 6,000 nM vs. 300 nM for PYO vs. 1-OHphz, respectively). The results found point to a real potential of PYO as a biomarker of P. aeruginosa infection and the possibility to use such VF also as a biomarker for patient stratification[2] and for an effective management of these kinds of infections.This work has been funded by the Spanish Government to M-PM through the Ministry of Science and Innovation (SAF2015-67476-R, RTI2018-096278-B-C21, PI, M-PM) and by Fundació Marató de TV3 (201825-30-31, PI, M-PM). The Nb4D group is a consolidated research group (Grup de Recerca) of the Generalitat de Catalunya and has support from the Departament d’Universitats, Recerca i Societat de la Informació de la Generalitat de Catalunya (expedient: 2017 SGR 1441). CIBER Actions are financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund (ERDF). Moreover, BR-U has an FI fellowship from the AGAUR (Agència de Gestió d’Ajuts Universitaris I de Recerca) of the Government of Catalonia (Generalitat de Catalunya) (2019FI_B00289). El Fons social Europeu Inverteix en el teu futur

An Immunochemical Approach to Detect the Quorum Sensing-Regulated Virulence Factor 2-Heptyl-4-Quinoline N-Oxide (HQNO) Produced by Pseudomonas aeruginosa Clinical Isolates

Pseudomonas aeruginosa; Quorum sensing; VirulencePseudomonas aeruginosa; Detecció de quòrum; VirulènciaPseudomonas aeruginosa; Detección de quórum; VirulenciaUnderstanding quorum sensing (QS) and its role in the development of pathogenesis may provide new avenues for diagnosing, surveillance, and treatment of infectious diseases. For this purpose, the availability of reliable and efficient analytical diagnostic tools suitable to specifically detect and quantify these essential QS small molecules and QS regulated virulence factors is crucial. Here, we reported the development and evaluation of antibodies and an enzyme-linked immunosorbent assay (ELISA) for HQNO (2-heptyl-4-quinoline N-oxide), a QS product of the PqsR system, which has been found to act as a major virulence factor that interferes with the growth of other microorganisms. Despite the nonimmunogenic character of HQNO, the antibodies produced showed high avidity and the microplate-based ELISA developed could detect HQNO in the low nM range. Hence, a limit of detection (LOD) of 0.60 ± 0.13 nM had been reached in Müeller Hinton (MH) broth, which was below previously reported levels using sophisticated equipment based on liquid chromatography coupled to mass spectrometry. The HQNO profile of release of different Pseudomonas aeruginosa clinical isolates analyzed using this ELISA showed significant differences depending on whether the clinical isolates belonged to patients with acute or chronic infections. These data point to the possibility of using HQNO as a specific biomarker to diagnose P. aeruginosa infections and for patient surveillance. Considering the role of HQNO in inhibiting the growth of coinfecting bacteria, the present ELISA will allow the investigation of these complex bacterial interactions underlying infections. IMPORTANCE Bacteria use quorum sensing (QS) as a communication mechanism that releases small signaling molecules which allow synchronizing a series of activities involved in the pathogenesis, such as the biosynthesis of virulence factors or the regulation of growth of other bacterial species. HQNO is a metabolite of the Pseudomonas aeruginosa-specific QS signaling molecule PQS (Pseudomonas quinolone signal). In this work, the development of highly specific antibodies and an immunochemical diagnostic technology (ELISA) for the detection and quantification of HQNO was reported. The ELISA allowed profiling of the release of HQNO by clinical bacterial isolates, showing its potential value for diagnosing and surveillance of P. aeruginosa infections. Moreover, the antibodies and the ELISA reported here may contribute to the knowledge of other underlying conditions related to the pathology, such as the role of the interactions with other bacteria of a particular microbiota environment.This work has been funded by the Ministry of Science and Innovation (SAF2015-67476-R and RTI2018-096278-B-C21) and Fundación Marató de TV3 (TV32018-201825-30-31). The Nb4D group is a consolidated research group (Grup de Recerca) of the Generalitat de Catalunya and has support from the Departament d’Universitats, Recerca i Societat de la Informació de la Generalitat de Catalunya (expedient: 2017 SGR 1441). CIBER-BBN is an initiative funded by the Spanish National Plan for Scientific and Technical Research and Innovation from 2013 to 2016, Iniciativa Ingenio 2010, Consolider Program, and CIBER Actions were financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund. Enrique J. Montagut and Juan Raya wish to thank the FPI fellowship (BES-2016-076496 and PRE2019-087542, respectively) from the Spanish Ministry of Science and Innovation. The Custom Antibody Service (CAbS) is acknowledged for its assistance and support in the production of HQNO antibodies

The teleconnection of the tropical Atlantic to Indo-Pacific sea surface temperatures on inter-annual to centennial time scales: a review of recent findings

In this paper, the teleconnections from the tropical Atlantic to the Indo-Pacific region from inter-annual to centennial time scales will be reviewed. Identified teleconnections and hypotheses on mechanisms at work are reviewed and further explored in a century-long pacemaker coupled ocean-atmosphere simulation ensemble. There is a substantial impact of the tropical Atlantic on the Pacific region at inter-annual time scales. An Atlantic Niño (Niña) event leads to rising (sinking) motion in the Atlantic region, which is compensated by sinking (rising) motion in the central-western Pacific. The sinking (rising) motion in the central-western Pacific induces easterly (westerly) surface wind anomalies just to the west, which alter the thermocline. These perturbations propagate eastward as upwelling (downwelling) Kelvin-waves, where they increase the probability for a La Niña (El Niño) event. Moreover, tropical North Atlantic sea surface temperature anomalies are also able to lead La Niña/El Niño development. At multidecadal time scales, a positive (negative) Atlantic Multidecadal Oscillation leads to a cooling (warming) of the eastern Pacific and a warming (cooling) of the western Pacific and Indian Ocean regions. The physical mechanism for this impact is similar to that at inter-annual time scales. At centennial time scales, the Atlantic warming induces a substantial reduction of the eastern Pacific warming even under CO2 increase and to a strong subsurface cooling

Satisfaction with Orthopedic Treatments

To determine the effectiveness and satisfaction with orthopodologic treatments in users of the University Clinic of podiatry at the University of A Coruña, according to various parameters. After approval from the ethics committee of the University of A Coruña, an observational retrospective study (n = 125). We analyzed the effectiveness and satisfaction with the orthopodologic treatments depending on the reason for consultation, diagnosis, treatment and goals of treatment. We performed a descriptive analysis of all variables collected. The most frequent reason for consultation was for pain of the hindfoot (58.2%).The most frequent diagnosis was plantar fasciitis, followed by metatarsalgia (29.7% vs. 18.6%). The orthotic treatment corrective was the most used (68.5%) with pronation control (52.3%). The majority of patients reported improvement in pain, and a high degree of satisfaction with the treatment used. The profile of the patient who consults the Podiatry clinic for a orthopodologic treatment is that of a man over the age of 50, who consulted for pain of the hindfoot. The most frequent diagnosis is plantar fasciitis and the treatment carried out the corrective for pronation control. The majority of patients used the brace between 4 and 8 hours a day, with a high satisfaction with the treatment and improvement in the evolution of the pain. The degree of satisfaction was significantly associated with age, younger patients more satisfied. The improvement of pain was significantly associated with age, younger patients who show improvement

Meta-DiSc 2.0:a web application for meta-analysis of diagnostic test accuracy data

BACKGROUND: Diagnostic evidence of the accuracy of a test for identifying a target condition of interest can be estimated using systematic approaches following standardized methodologies. Statistical methods for the meta-analysis of diagnostic test accuracy (DTA) studies are relatively complex, presenting a challenge for reviewers without extensive statistical expertise. In 2006, we developed Meta-DiSc, a free user-friendly software to perform test accuracy meta-analysis. This statistical program is now widely used for performing DTA meta-analyses. We aimed to build a new version of the Meta-DiSc software to include statistical methods based on hierarchical models and an enhanced web-based interface to improve user experience. RESULTS: In this article, we present the updated version, Meta-DiSc 2.0, a web-based application developed using the R Shiny package. This new version implements recommended state-of-the-art statistical models to overcome the limitations of the statistical approaches included in the previous version. Meta-DiSc 2.0 performs statistical analyses of DTA reviews using a bivariate random effects model. The application offers a thorough analysis of heterogeneity, calculating logit variance estimates of sensitivity and specificity, the bivariate I-squared, the area of the 95% prediction ellipse, and the median odds ratios for sensitivity and specificity, and facilitating subgroup and meta-regression analyses. Furthermore, univariate random effects models can be applied to meta-analyses with few studies or with non-convergent bivariate models. The application interface has an intuitive design set out in four main menus: file upload; graphical description (forest and ROC plane plots); meta-analysis (pooling of sensitivity and specificity, estimation of likelihood ratios and diagnostic odds ratio, sROC curve); and summary of findings (impact of test through downstream consequences in a hypothetical population with a given prevalence). All computational algorithms have been validated in several real datasets by comparing results obtained with STATA/SAS and MetaDTA packages. CONCLUSION: We have developed and validated an updated version of the Meta-DiSc software that is more accessible and statistically sound. The web application is freely available at www.metadisc.es

Nanoscale Phase Separation in Colossal Magnetoresistance Materials: A Lesson for the Cuprates?

A recent vast experimental and theoretical effort in manganites has shown that the colossal magnetoresistance effect can be understood based on the competition of charge-ordered and ferromagnetic phases. The general aspects of the theoretical description appear to be valid for any compound with intrinsic phase competition. In high temperature superconductors, recent experiments have shown the existence of intrinsic inhomogeneities in many materials, revealing a phenomenology quite similar to that of manganese oxides. Here, the results for manganites are briefly reviewed with emphasis on the general aspects. In addition, theoretical speculations are formulated in the context of Cu-oxides by mere analogy with manganites. This includes a tentative explanation of the spin-glass regime as a mixture of antiferromagnetic and superconducting islands, the rationalization of the pseudogap temperature T* as a Griffiths temperature where clusters start forming upon cooling, the prediction of "colossal" effects in cuprates, and the observation that quenched disorder may be far more relevant in Cu-oxides than previously anticipated.Comment: 25 pages, 14 figures, post-muSR2002 Superconductivity Worksho

Cell wound repair in Drosophila occurs through three distinct phases of membrane and cytoskeletal remodeling

Single-cell wound repair in Drosophila involves mechanistically distinct expansion, contraction, and closure phases

Suppression of charge-ordering and appearance of magnetoresistance in a spin-cluster glass manganite La0.3Ca0.7Mn0.8Cr0.2O3

The magnetic properties of electron-doped manganite La0.3Ca0.7MnO3 and La0.3Ca0.7Mn0.8Cr0.2O3 polycrystalline samples prepared by sol-gel technique have been investigated between 5 and 300 K in magnetic fields ranging from 0 to 5 T. The transition at 260 K, attributed to charge ordering in La0.3Ca0.7MnO3, is completely suppressed in the Cr-substituted sample while the onset of a magnetic remanence followed by the appearance of a magnetic irreversibility at lower temperatures is observed in both samples. These features indicate that ferromagnetic clusters coexist with either an antiferromagnetic phase for La0.3Ca0.7MnO3 or a spin-cluster glass phase for La0.3Ca0.7Mn0.8Cr0.2O3 at the lowest temperatures. The exponential temperature dependence of the resistivity for the Cr-substituted sample is consistent with the small polaron hopping model for 120 K < T < 300 K, while the data are better described by Mott's hopping mechanism for T < 120 K. Whereas the parent compound La0.3Ca0.7MnO3 is known to show no magnetoresistance, a large negative magnetoresistance is observed in the La0.3Ca0.7Mn0.8Cr0.2O3 sample below 120 K. The appearance of the CMR is attributed to spin dependent hopping between spin clusters and/or between ferromagnetic domains

Adjuvant dabrafenib and trametinib for patients with resected BRAF-mutated melanoma: DESCRIBE-AD real-world retrospective observational study

BRAF and MEK inhibitor, dabrafenib plus trametinib, adjuvant therapy is effective for high-risk resected melanoma patients with BRAF-V600 mutations. However, real-world evidence is limited. We aimed to determine the feasibility of this therapy in routine clinical practice. DESCRIBE-AD, a retrospective observational study, collected real-world data from 25 hospitals in Spain. Histologically confirmed and resected BRAF-mutated melanoma patients aged & GE;18 years who were previously treated with dabrafenib plus trametinib adjuvant therapy, were included. The primary objectives were treatment discontinuation rate and time to discontinuation. The secondary objectives included safety and efficacy. From October 2020 to March 2021, 65 patients were included. Dabrafenib and trametinib discontinuation rate due to treatment-related adverse events (TRAEs) of any grade was 9%. Other reasons for discontinuation included patients' decisions (6%), physician decisions (6%), unrelated adverse events (3%), disease progression (5%), and others (5%). The median time to treatment discontinuation was 9 months [95% confidence interval (CI), 5-11]. G3-4 TRAEs occurred in 21.5% of patients, the most common being pyrexia (3%), asthenia (3%), and diarrhoea (3%). Unscheduled hospitalisations and clinical tests occurred in 6 and 22% of patients, respectively. After 20-month median follow-up (95% CI, 18-22), 9% of patients had exitus due to disease progression, with a 12-month relapse-free survival and overall survival rates of 95.3% and 100%, respectively. Dabrafenib and trametinib adjuvant therapy proved effective for melanoma patients in a real-world setting, with a manageable toxicity profile. Toxicity frequencies were low leading to low incidence of unscheduled medical visits, tests, and treatment discontinuations

Environmental justice and transformations to sustainability

Global carbon emissions continue to rise,1 rates of global biodiversity loss continue to increase,2 and social and economic inequalities continue to widen.3 Significant global social movements such as Fridays for Future are declaring this situation an “emergency,” regarding it as a crime against humanity in which political and business leaders stand accused of ignoring the plight of current and future vulnerable people. This association between environmental crises and social injustice is now widely accepted. Many feel that time is running out for incremental approaches to prove effective and that there is an inescapable need for a radical, transformative change that combines sustainability and justice