A prospective study to analyse the concentration of octenidine in hand wounds after disinfection by LC-MS/MS.

Toxic reactions can appear after pressurised flushing of soft tissue with octenidine (OCT) containing disinfectants. Their use for surgical disinfection could complicate the diagnosis of possible contamination. In patients with open lacerations of their hand's subcutaneous tissue samples were taken before and after surgical disinfection with Octenisept® and analysed by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). In 16 out of 20 tissue samples, OCT was detected after disinfection (lower limit of quantification (LLOQ)=10 pg/mL/mg). The concentration of OCT was below the LLOQ, estimation of mean of 0.6 pg/mL/mg (0.22-0.98 pg/mL/mg, 95%-CI) before disinfection and mean of 179.4 pg/mL/mg (13.35-432.0 pg/mL/mg, 95%-CI) after disinfection. This study shows that the disinfection of open wounds with Octenisept® leads to a quantifiable concentration of OCT in open wounds. In cases of suspected OCT-mediated toxic reaction, the use of antiseptics containing OCT should be avoided

Conception d’un outil d’analyse du comportement de systèmes biologiques

Les innovations variétales sont depuis toujours un élément clé de la production en agriculture. Evaluer les comportements de nouvelles variétés face à une diversité de milieux (sol, climat, manière de produire) est alors un enjeu important pour les acteurs, depuis ceux qui sélectionnent ces nouvelles variétés jusqu’à ceux qui les cultivent ou en utilisent les produits. Pour concevoir un outil qui aide les acteurs à réaliser cette évaluation, agronomes et ergonomes ont collaboré pour articuler trois dimensions dans le travail de conception : (i) la cristallisation dans l’outil d’un modèle de l’activité, (ii) la plasticité de l’outil, et (iii) le développement conjoint de l’outil et de l’activité. Pour produire des spécifications en relation avec ces trois dimensions, nous analysons respectivement, (i) les invariants du système d’activité de l’évaluation variétale, (ii) la diversité les actions conduites par les acteurs de l’évaluation, (iii) les contradictions qui apparaissent dans le système d’activité. Notre analyse valorise des travaux d’historiens et s’appuie surtout sur des entretiens conduits auprès de 21 acteurs de l’évaluation variétale. Elle nous permet de mettre en avant le rôle central joué par l’outil qu’est le réseau d’expérimentation et de d’étudier la façon dont les acteurs mettent à profit l’indétermination des outils de l’activité pour les constituer en instruments. Elle montre que l’objet de l’activité a été historiquement constitué autour de l’évaluation à la stabilité du comportement d’une variété et que cette dernière continue à orienter les actions des acteurs. Elle permet d’identifier 11 actions d’évaluation variétale différentes. Elle montre les contradictions qui émergent aujourd’hui au sein de l’objet compte tenu des nouveaux enjeux auxquels doivent faire face les acteurs. Elle débouche sur des spécifications pour un prototype d’outil suffisamment ouvert pour s’adapter à la diversité des actions et susceptible d’aider les acteurs à lever les contradictions identifiées.New cultivars have always been a key element in the development of cropping systems. The assessment of new cultivar performance in a diverse range of environments (soil, climate, production methods) is a key issue for the actors along the chain which links breeders to growers and food suppliers. As agronomists and ergonomists, we worked together to design a tool aimed at supporting these actors in assessing cultivar performance in order to link three aspects of the design process: (i) the crystallisation in the tool of an activity model, (ii) the flexibility of the tool, (iii) the joint development of the tool and the activity. To produce specifications based on these aspects, we analysed respectively (i) the invariants of the activity system of cultivar assessment, (ii) the diversity of actions performed by actors involved in cultivar assessment, (iii) the contradictions which arise within the activity system. Our analysis calls on existing historical studies but is based mainly on interviews with 21 people involved in cultivar assessment and gave us an insight into the actions of assessment activity. This analysis enables us to point out that the network of field trials is a key tool for the activity of cultivar assessment and to study how actors take advantage of the vagueness of the various tools to transform them into instruments. It also shows that, historically, the object of the activity has been built around assessing the stability of cultivar performance and that stability still gives direction to the actions undertaken by actors today. Eleven different actions for evaluating cultivars were identified. Our analysis highlights the contradictions which emerge today within the object of the activity system due to the new issues that actors have to face. It led us to define specifications for a prototype tool which is flexible enough to adapt to the diversity of actions and which could be used to enable actors to overcome some of the contradictions identified.Las innovaciones varietales han sido, desde siempre, un elemento clave de la producción en la agricultura. La evaluación de los comportamientos de las nuevas variedades frente a una diversidad de entornos (suelo, clima, forma de producir), es entonces un desafío importante para los actores, desde aquellos que seleccionan estas nuevas variedades hasta quienes las cultivan o utilizan los productos. Para diseñar una herramienta que ayude a los actores a realizar dicha evaluación, han colaborado agrónomos y ergónomos para articular tres dimensiones en el trabajo de concepción : (i) la cristalización en la herramienta de un modelo de la actividad (ii) la plasticidad de la herramienta, y (iii) el desarrollo conjunto de la herramienta y de la actividad. Para producir las especificaciones con relación a estas tres dimensiones, analizamos respectivamente, (i) las invariantes del sistema de actividad de la evaluación varietal, (ii) la diversidad de las acciones realizadas por los actores de la evaluación, (iii) las contradicciones que aparecen en el sistema de actividad. Nuestro análisis valoriza el trabajo de los historiadores y, sobre todo, se apoya en las entrevistas realizadas a 21 actores de la evaluación varietal. Nos permite poner de relieve el rol central jugado por la herramienta que es la red de experimentación, así como estudiar la forma en que los actores aprovechan de la indeterminación de las herramientas de la actividad para convertirlas en instrumentos. El análisis muestra que el objeto de la actividad ha estado históricamente constituido alrededor de la evaluación de la estabilidad del comportamiento de una variedad, y que esta última continúa a orientar las acciones de los actores. Asimismo, permite identificar 11 acciones diferentes de evaluación varietal y muestra las contradicciones que emergen en torno del objeto, habida cuenta de los desafíos que deben enfrentar los actores. Finalmente, el análisis desemboca en especifaciones para un prototipo de herramienta suficientemente abierto para adaptarse a la diversidad de acciones y susceptible de ayudar a los actores a remontar las contracciones identificadas

Designing a tool to analyse the performance of biological systems

Les innovations variétales sont depuis toujours un élément clé de la production en agriculture. Evaluer les comportements de nouvelles variétés face à une diversité de milieux (sol, climat, manière de produire) est alors un enjeu important pour les acteurs, depuis ceux qui sélectionnent ces nouvelles variétés jusqu’à ceux qui les cultivent ou en utilisent les produits. Pour concevoir un outil qui aide les acteurs à réaliser cette évaluation, agronomes et ergonomes ont collaboré pour articuler trois dimensions dans le travail de conception  : (i) la cristallisation dans l’outil d’un modèle de l’activité, (ii) la plasticité de l’outil, et (iii) le développement conjoint de l’outil et de l’activité. Pour produire des spécifications en relation avec ces trois dimensions, nous analysons respectivement, (i) les invariants du système d’activité de l’évaluation variétale, (ii) la diversité les actions conduites par les acteurs de l’évaluation, (iii) les contradictions qui apparaissent dans le système d’activité. Notre analyse valorise des travaux d’historiens et s’appuie surtout sur des entretiens conduits auprès de 21 acteurs de l’évaluation variétale. Elle nous permet de mettre en avant le rôle central joué par l’outil qu’est le réseau d’expérimentation et de d’étudier la façon dont les acteurs mettent à profit l’indétermination des outils de l’activité pour les constituer en instruments. Elle montre que l’objet de l’activité a été historiquement constitué autour de l’évaluation à la stabilité du comportement d’une variété et que cette dernière continue à orienter les actions des acteurs. Elle permet d’identifier 11 actions d’évaluation variétale différentes. Elle montre les contradictions qui émergent aujourd’hui au sein de l’objet compte tenu des nouveaux enjeux auxquels doivent faire face les acteurs. Elle débouche sur des spécifications pour un prototype d’outil suffisamment ouvert pour s’adapter à la diversité des actions et susceptible d’aider les acteurs à lever les contradictions identifiées.New cultivars have always been a key element in the development of cropping systems. The assessment of new cultivar performance in a diverse range of environments (soil, climate, production methods) is a key issue for the actors along the chain which links breeders to growers and food suppliers. As agronomists and ergonomists, we worked together to design a tool aimed at supporting these actors in assessing cultivar performance in order to link three aspects of the design process: (i) the crystallisation in the tool of an activity model, (ii) the flexibility of the tool, (iii) the joint development of the tool and the activity. To produce specifications based on these aspects, we analysed respectively (i) the invariants of the activity system of cultivar assessment, (ii) the diversity of actions performed by actors involved in cultivar assessment, (iii) the contradictions which arise within the activity system. Our analysis calls on existing historical studies but is based mainly on interviews with 21 people involved in cultivar assessment and gave us an insight into the actions of assessment activity. This analysis enables us to point out that the network of field trials is a key tool for the activity of cultivar assessment and to study how actors take advantage of the vagueness of the various tools to transform them into instruments. It also shows that, historically, the object of the activity has been built around assessing the stability of cultivar performance and that stability still gives direction to the actions undertaken by actors today. Eleven different actions for evaluating cultivars were identified. Our analysis highlights the contradictions which emerge today within the object of the activity system due to the new issues that actors have to face. It led us to define specifications for a prototype tool which is flexible enough to adapt to the diversity of actions and which could be used to enable actors to overcome some of the contradictions identified.Las innovaciones varietales han sido, desde siempre, un elemento clave de la producción en la agricultura. La evaluación de los comportamientos de las nuevas variedades frente a una diversidad de entornos (suelo, clima, forma de producir), es entonces un desafío importante para los actores, desde aquellos que seleccionan estas nuevas variedades hasta quienes las cultivan o utilizan los productos. Para diseñar una herramienta que ayude a los actores a realizar dicha evaluación, han colaborado agrónomos y ergónomos para articular tres dimensiones en el trabajo de concepción : (i) la cristalización en la herramienta de un modelo de la actividad (ii) la plasticidad de la herramienta, y (iii) el desarrollo conjunto de la herramienta y de la actividad. Para producir las especificaciones con relación a estas tres dimensiones, analizamos respectivamente, (i) las invariantes del sistema de actividad de la evaluación varietal, (ii) la diversidad de las acciones realizadas por los actores de la evaluación, (iii) las contradicciones que aparecen en el sistema de actividad. Nuestro análisis valoriza el trabajo de los historiadores y, sobre todo, se apoya en las entrevistas realizadas a 21 actores de la evaluación varietal. Nos permite poner de relieve el rol central jugado por la herramienta que es la red de experimentación, así como estudiar la forma en que los actores aprovechan de la indeterminación de las herramientas de la actividad para convertirlas en instrumentos. El análisis muestra que el objeto de la actividad ha estado históricamente constituido alrededor de la evaluación de la estabilidad del comportamiento de una variedad, y que esta última continúa a orientar las acciones de los actores. Asimismo, permite identificar 11 acciones diferentes de evaluación varietal y muestra las contradicciones que emergen en torno del objeto, habida cuenta de los desafíos que deben enfrentar los actores. Finalmente, el análisis desemboca en especifaciones para un prototipo de herramienta suficientemente abierto para adaptarse a la diversidad de acciones y susceptible de ayudar a los actores a remontar las contracciones identificadas

Specific GAG ratios in the diagnosis of mucopolysaccharidoses.

Mucopolysaccharidoses (MPS) screening is tedious and still performed by analysis of total glycosaminoglycans (GAG) using 1,9-dimethylmethylene blue (DMB) photometric assay, although false positive and negative tests have been reported. Analysis of differentiated GAGs have been pursued classically by gel electrophoresis or more recently by quantitative LC-MS assays. Secondary elevations of GAGs have been reported in urinary tract infections (UTI). In this manuscript, we describe the diagnostic accuracy of urinary GAG measurements by LC-MS for MPS typing in 68 untreated MPS and mucolipidosis (ML) patients, 183 controls and 153 UTI samples. We report age-dependent reference values and cut-offs for chondroitin sulfate (CS), dermatan sulfate (DS), heparan sulfate (HS) and keratan sulfate (KS) and specific GAG ratios. The use of HS/DS ratio in combination to GAG concentrations normalized to creatinine improves the diagnostic accuracy in MPS type I, II, VI and VII. In total 15 samples classified to the wrong MPS type could be correctly assigned using HS/DS ratio. Increased KS/HS ratio in addition to increased KS improves discrimination of MPS type IV by excluding false positives. Some samples of UTI patients showed elevation of specific GAGs, mainly CS, KS and KS/HS ratio and could be misclassified as MPS type IV. Finally, DMB photometric assay performed in MPS and ML samples reveal four false negative tests (sensitivity of 94%). In conclusion, specific GAG ratios in complement to quantitative GAG values obtained by LC-MS enhance discrimination of MPS types. Exclusion of patients with UTI improve diagnostic accuracy in MPS IV but not in other types

Improving Reconstituted HDL Composition for Efficient Post-Ischemic Reduction of Ischemia Reperfusion Injury

BACKGROUND: New evidence shows that high density lipoproteins (HDL) have protective effects beyond their role in reverse cholesterol transport. Reconstituted HDL (rHDL) offer an attractive means of clinically exploiting these novel effects including cardioprotection against ischemia reperfusion injury (IRI). However, basic rHDL composition is limited to apolipoprotein AI (apoAI) and phospholipids; addition of bioactive compound may enhance its beneficial effects. Objective The aim of this study was to investigate the role of rHDL in post-ischemic model, and to analyze the potential impact of sphingosine-1-phosphate (S1P) in rHDL formulations. Methods and RESULTS: The impact of HDL on IRI was investigated using complementary in vivo , ex vivo and in vitro IRI models. Acute post-ischemic treatment with native HDL significantly reduced infarct size and cell death in the ex vivo , isolated heart (Langendorff) model and the in vivo model (-48%, p<0.01). Treatment with rHDL of basic formulation (apoAI + phospholipids) had a non-significant impact on cell death in vitro and on the infarct size ex vivo and in vivo . In contrast, rHDL containing S1P had a highly significant, protective influence ex vivo , and in vivo (-50%, p<0.01). This impact was comparable with the effects observed with native HDL. Pro-survival signaling proteins, Akt, STAT3 and ERK1/2 were similarly activated by HDL and rHDL containing S1P both in vitro (isolated cardiomyocytes) and in vivo . CONCLUSION: HDL afford protection against IRI in a clinically relevant model (post-ischemia). rHDL is significantly protective if supplemented with S1P. The protective impact of HDL appears to target directly the cardiomyocyte

Population pharmacokinetic analyses of regorafenib and capecitabine in patients with locally advanced rectal cancer (SAKK 41/16 RECAP)

Aims: Locally advanced rectal cancer (LARC) is an area of unmet medical need with one third of patients dying from their disease. With response to neoadjuvant chemo-radiotherapy being a major prognostic factor, trial SAKK 41/16 assessed potential benefits of adding regorafenib to capecitabine-amplified neoadjuvant radiotherapy in LARC patients. Methods: Patients received regorafenib at three dose levels (40/80/120 mg once daily) combined with capecitabine 825 mg/m2 bidaily and local radiotherapy. We developed population pharmacokinetic models from plasma concentrations of capecitabine and its metabolites 5'-deoxy-5-fluorocytidine and 5'-deoxy-5-fluorouridine as well as regorafenib and its metabolites M-2 and M-5 as implemented into SAKK 41/16 to assess potential drug-drug interactions (DDI). After establishing parent-metabolite base models, drug exposure parameters were tested as covariates within the respective models to investigate for potential DDI. Simulation analyses were conducted to quantify their impact. Results: Plasma concentrations of capecitabine, regorafenib and metabolites were characterized by one and two compartment models and absorption was described by parallel first- and zero-order processes and transit compartments, respectively. Apparent capecitabine clearance was 286 L/h (relative standard error [RSE] 14.9%, interindividual variability [IIV] 40.1%) and was reduced by regorafenib cumulative area under the plasma concentration curve (median reduction of 45.6%) as exponential covariate (estimate -4.10 × 10-4 , RSE 17.8%). Apparent regorafenib clearance was 1.94 L/h (RSE 12.1%, IIV 38.1%). Simulation analyses revealed significantly negative associations between capecitabine clearance and regorafenib exposure. Conclusions: This work informs the clinical development of regorafenib and capecitabine combination treatment and underlines the importance of studying potential DDI with new anticancer drug combinations. Keywords: capecitabine; drug-drug interaction; population pharmacokinetics; rectal cancer; regorafenib

A local drug delivery system prolongs graft survival by dampening T cell infiltration and neutrophil extracellular trap formation in vascularized composite allografts.

INTRODUCTION The standard treatment for preventing rejection in vascularized composite allotransplantation (VCA) currently relies on systemic immunosuppression, which exposes the host to well-known side effects. Locally administered immunosuppression strategies have shown promising results to bypass this hurdle. Nevertheless, their progress has been slow, partially attributed to a limited understanding of the essential mechanisms underlying graft rejection. Recent discoveries highlight the crucial involvement of innate immune components, such as neutrophil extracellular traps (NETs), in organ transplantation. Here we aimed to prolong graft survival through a tacrolimus-based drug delivery system and to understand the role of NETs in VCA graft rejection. METHODS To prevent off-target toxicity and promote graft survival, we tested a locally administered tacrolimus-loaded on-demand drug delivery system (TGMS-TAC) in a multiple MHC-mismatched porcine VCA model. Off-target toxicity was assessed in tissue and blood. Graft rejection was evaluated macroscopically while the complement system, T cells, neutrophils and NETs were analyzed in graft tissues by immunofluorescence and/or western blot. Plasmatic levels of inflammatory cytokines were measured using a Luminex magnetic-bead porcine panel, and NETs were measured in plasma and tissue using DNA-MPO ELISA. Lastly, to evaluate the effect of tacrolimus on NET formation, NETs were induced in-vitro in porcine and human peripheral neutrophils following incubation with tacrolimus. RESULTS Repeated intra-graft administrations of TGMS-TAC minimized systemic toxicity and prolonged graft survival. Nevertheless, signs of rejection were observed at endpoint. Systemically, there were no increases in cytokine levels, complement anaphylatoxins, T-cell subpopulations, or neutrophils during rejection. Yet, tissue analysis showed local infiltration of T cells and neutrophils, together with neutrophil extracellular traps (NETs) in rejected grafts. Interestingly, intra-graft administration of tacrolimus contributed to a reduction in both T-cellular infiltration and NETs. In fact, in-vitro NETosis assessment showed a 62-84% reduction in NETs after stimulated neutrophils were treated with tacrolimus. CONCLUSION Our data indicate that the proposed local delivery of immunosuppression avoids off-target toxicity while prolonging graft survival in a multiple MHC-mismatch VCA model. Furthermore, NETs are found to play a role in graft rejection and could therefore be a potential innovative therapeutic target

Autoantibodies neutralizing type I IFNs are present in ~4% of uninfected individuals over 70 years old and account for ~20% of COVID-19 deaths

Circulating autoantibodies (auto-Abs) neutralizing high concentrations (10 ng/mL, in plasma diluted 1 to 10) of IFN-α and/or -ω are found in about 10% of patients with critical COVID-19 pneumonia, but not in subjects with asymptomatic infections. We detect auto-Abs neutralizing 100-fold lower, more physiological, concentrations of IFN-α and/or -ω (100 pg/mL, in 1/10 dilutions of plasma) in 13.6% of 3,595 patients with critical COVID-19, including 21% of 374 patients &gt; 80 years, and 6.5% of 522 patients with severe COVID-19. These antibodies are also detected in 18% of the 1,124 deceased patients (aged 20 days-99 years; mean: 70 years). Moreover, another 1.3% of patients with critical COVID-19 and 0.9% of the deceased patients have auto-Abs neutralizing high concentrations of IFN-β. We also show, in a sample of 34,159 uninfected subjects from the general population, that auto-Abs neutralizing high concentrations of IFN-α and/or -ω are present in 0.18% of individuals between 18 and 69 years, 1.1% between 70 and 79 years, and 3.4% &gt;80 years. Moreover, the proportion of subjects carrying auto-Abs neutralizing lower concentrations is greater in a subsample of 10,778 uninfected individuals: 1% of individuals &lt;70 years, 2.3% between 70 and 80 years, and 6.3% &gt;80 years. By contrast, auto-Abs neutralizing IFN-β do not become more frequent with age. Auto-Abs neutralizing type I IFNs predate SARS-CoV-2 infection and sharply increase in prevalence after the age of 70 years. They account for about 20% of both critical COVID-19 cases in the over-80s, and total fatal COVID-19 cases

Autoantibodies neutralizing type I IFNs are present in ~4% of uninfected individuals over 70 years old and account for ~20% of COVID-19 deaths

Publisher Copyright: © 2021 The Authors, some rights reserved.Circulating autoantibodies (auto-Abs) neutralizing high concentrations (10 ng/ml; in plasma diluted 1:10) of IFN-alpha and/or IFN-omega are found in about 10% of patients with critical COVID-19 (coronavirus disease 2019) pneumonia but not in individuals with asymptomatic infections. We detect auto-Abs neutralizing 100-fold lower, more physiological, concentrations of IFN-alpha and/or IFN-omega (100 pg/ml; in 1:10 dilutions of plasma) in 13.6% of 3595 patients with critical COVID-19, including 21% of 374 patients >80 years, and 6.5% of 522 patients with severe COVID-19. These antibodies are also detected in 18% of the 1124 deceased patients (aged 20 days to 99 years; mean: 70 years). Moreover, another 1.3% of patients with critical COVID-19 and 0.9% of the deceased patients have auto-Abs neutralizing high concentrations of IFN-beta. We also show, in a sample of 34,159 uninfected individuals from the general population, that auto-Abs neutralizing high concentrations of IFN-alpha and/or IFN-omega are present in 0.18% of individuals between 18 and 69 years, 1.1% between 70 and 79 years, and 3.4% >80 years. Moreover, the proportion of individuals carrying auto-Abs neutralizing lower concentrations is greater in a subsample of 10,778 uninfected individuals: 1% of individuals 80 years. By contrast, auto-Abs neutralizing IFN-beta do not become more frequent with age. Auto-Abs neutralizing type I IFNs predate SARS-CoV-2 infection and sharply increase in prevalence after the age of 70 years. They account for about 20% of both critical COVID-19 cases in the over 80s and total fatal COVID-19 cases.Peer reviewe