45 research outputs found
Cytotoxicity and toxicokinetics of new psychoactive substances
To expand the knowledge about toxicokinetic and cytotoxic properties of new psychoactive substances (NPS), different in vitro and in vivo tests were (further) developed and applied on structurally different compounds. First, in vitro and in vivo models were used to determine the metabolic stability (in vitro half-life) and the metabolic fate to elucidate targets for urine-based screenings. Investigations on isozymes, which contribute to their metabolism, and on the plasma protein binding were conducted to identify potential interactions with other NPS or drugs (of abuse). Finally, an existing cytotoxicity high-content screening assay (HCSA) using HepG2 cells was optimized to specify the cytotoxic potential of NPS. Synthetic cannabinoids showed a more extensive metabolism compared to synthetic opioids, which was in accordance with the number of involved isozymes. Since plasma protein binding values exceeded 70% in all cases, relevant drug-drug interactions cannot be excluded. Based on the optimized HCSA, a cytotoxic potential was assigned for seven NPS. Therefore, for two NPS a previously reported in vivo hepatotoxicity can be confirmed in vitro. Furthermore, metabolism-based effects on their cytotoxicity were observed for two NPS.Zur Erforschung der toxikokinetischen und cytotoxischen Eigenschaften von neuen psychoaktiven Substanzen (NPS) wurden verschiedene in vitro und in vivo Tests (weiter-)entwickelt und an strukturell unterschiedlichen Verbindungen angewendet. Mittels Stoffwechselversuchen wurden unter anderem analytische Zielmoleküle für Urinuntersuchungen identifiziert. Um mögliche Interaktionen mit anderen NPS oder Drogen zu untersuchen, wurden die am Metabolismus beteiligte Isoenzyme sowie die Plasmaproteinbindung bestimmt. Im letzten Schritt ging es um die Optimierung eines bestehenden Hochdurchsatzverfahrens zur Bestimmung der zytotoxischen Eigenschaften von NPS an HepG2 Zellen. Synthetische Cannabinoide zeigten einen ausgeprägteren Metabolismus als synthetische Opioide, welches sich auch in der Anzahl der daran beteiligten Isoenzyme widerspiegelte. Die Plasmaproteinbindung der untersuchten NPS lag über 70%, weshalb Wechselwirkungen mit anderen Drogen nicht ausgeschlossen werden können. Mittels des optimierten Zytotoxizitäts-Assays konnten sieben NPS als potenziell zytotoxisch eingestuften werden. In zwei Fällen konnte somit eine zuvor beschriebene in vivo Hepatotoxizität in vitro bestätigt werden. Außerdem wurden Metabolismus-basierte Effekte an der Zytotoxizität zweier NPS beobachtet
RIO Country Report 2015: Cyprus
The 2015 series of RIO Country Reports analyse and assess the policy and the national research and innovation system developments in relation to national policy priorities and the EU policy agenda with special focus on ERA and Innovation Union. The executive summaries of these reports put forward the main challenges of the research and innovation systems.JRC.J.6-Innovation Systems Analysi
Recent trends in drugs of abuse metabolism studies for mass spectrometry-based analytical screening procedures
The still increasing number of drugs of abuse, particularly the so-called new psychoactive substances (NPS), poses an analytical challenge for clinical and forensic toxicologists but also for doping control. NPS usually belong to various classes such as synthetic cannabinoids, phenethylamines, opioids, or benzodiazepines. Like other xenobiotics, NPS undergo absorption, distribution, metabolism, and excretion processes after consumption, but only very limited data concerning their toxicokinetics and safety properties is available once they appear on the market. The inclusion of metabolites in mass spectral libraries is often crucial for the detection of NPS especially in urine screening approaches. Authentic human samples may represent the gold standard for identification of metabolites but are often not available and clinical studies cannot be performed due to ethical concerns. However, numerous alternative in vitro and in vivo models are available. This trends article will give an overview on selected models, discuss current studies, and highlight recent developments
Investigation on the Future of Enterprise Architecture in Dynamic Environments
En la economía actual, el cambio constante se ha convertido en la nueva normalidad. Las consecuencias de este desarrollo son vívidamente visibles. La dinámica en los entornos corporativos está aumentando y las empresas que no se adapten a las condiciones cambiantes serán menos exitosas y finalmente acabarán en cierre.
Mientras el desarrollo y la mejora de las capacidades de adaptación para tener éxito en los entornos dinámicos requieren el trabajo conjunto de muchas partes dentro de la empresa, la Arquitectura Empresarial (Enterprise Architecture - EA) puede suponer una parte vital al habilitar y guiar a distintos elementos organizacionales para ser más efectivos en entornos dinámicos. Sin embargo, para poder hacerlo, la EA necesita transformarse a sí misma. Esta tesis ofrece resultados que describen cómo la EA puede ser efectiva en entornos dinámicos. Los resultados se han estructurado de acuerdo con las siguientes cuatro áreas.
Primero, se presenta una revisión del estado del arte sobre EA, en el que se describe el desarrollo de la disciplina a lo largo de las últimas tres décadas. Desde el análisis, es evidente que el enfoque de la investigación de EA se ha movido desde la comprensión y la definición de la EA hacia gestionar eficazmente la disciplina en entornos empresariales complejos. Las partes posteriores de esta tesis ponen énfasis en la gestión efectiva de la EA también al proporcionar enfoques de EA para circunstancias específicas, es decir, entornos con un mayor ritmo de cambio.
En segundo lugar, esta tesis ofrece una descripción formal de cómo los efectos del ritmo creciente de cambio influyen en la efectividad de la EA. El resultado primario de esta parte es un modelo, basado en la teoría de la complejidad, que resume las siguientes dependencias: El ritmo creciente del cambio conduce a una mayor complejidad dinámica para EA ya que existe la necesidad de administrar partes que están cambiando más y más rápido. Esta complejidad debe considerarse desde un punto de vista de negocio y tecnológico. En el modelo final, La complejidad dinámica de negocios y tecnológica se consideran como factores contextuales, los cuales influyen en el uso correcto de la EA y, en consecuencia, la efectividad de la disciplina.
Tercero, se presenta una colección de enfoques para mejorar la efectividad de la EA en ambientes dinámicos. Estos están estructurados en torno a cuatro dimensiones: la competencia EA, la cual considera quién en la organización está trabajando en EA; la metodología EA, que considera cómo se ejecuta EA en la organización; el contenido de EA, que considera la salida de EA; EA Tools que considera con qué EA está siendo creado y mantenido.
Cuarto, la parte final de esta tesis presenta los resultados en forma de arquitectura de referencia para EA en entornos dinámicos. Los enfoques de EA son nuevamente estructurados de acuerdo con las dimensiones descritas anteriormente. La arquitectura de referencia se describe en el nivel de los enfoques individuales, así como en el nivel de dimensión. En resumen, la competencia EA debe integrarse bien en la empresa. Además de esto, la metodología EA debe estar alineada con prácticas ágiles que permitan decisiones arquitectónicas rápidas. El contenido EA resultante debe ser adaptativo, lo que significa que la arquitectura se puede ajustar fácilmente en caso de que sea necesario. Por último, los arquitectos y otras partes interesadas de EA deberían recibir el soporte de las modernas herramientas de EA.
Esta tesis muestra que el objetivo subyacente de EA, en concreto, asegurar la alineación de diferentes facetas dentro de la empresa, incluso en las condiciones cambiantes de hoy en día, sigue siendo necesario. Sin embargo, los arquitectos trabajando en entornos dinámicos deberían revisar las dimensiones descritas (¿quién? - ¿cómo? - ¿qué? - ¿con qué?) en su práctica de la EA para seguir siendo efectivos.
Con sus resultados, esta tesis presenta una guía para profesionales para que puedan tomar decisiones adecuadas y así optimizar la efectividad de la EA en entornos dinámicos. Al mismo tiempo, esta tesis contribuye al conocimiento académico sobre EA. Los modelos y enfoques presentados abordan la brecha con respecto al enfoque holístico actual de la EA en entornos dinámicos. Además, esta tesis señala diversas áreas que brindan oportunidades para futuras investigaciones. Se espera que estas inspirarán a investigadores a impulsar aún más la evolución de la EA desde el punto de vista académico.Administración y Dirección de Empresa
Do individuals care about fairness in burden sharing for climate change mitigation? Evidence from a lab experiment
One of the reasons for deadlock in global climate policy is countries' disagreement on how to share the mitigation burden. Normative theory suggests various fairness criteria for structuring burden sharing, most prominently, historical responsibility for emissions, economic capacity, and vulnerability to climate change. Governments have taken up these criteria in their rhetoric at UNFCCC negotiations. I examine whether normative criteria influence individual burden sharing preferences. This bottom-up perspective is important for two reasons. First, it is unknown if governments' fairness rhetoric matches citizens' actual preferences. Second, international climate agreements directly affect individuals through domestic policy measures (e.g. energy taxes), and therefore require domestic public support for successful implementation. I conducted two laboratory experiments where participants have to agree on how to share climate change mitigation costs in an ultimatum game. Treatment conditions include differences between proposer and responder in capacity, vulnerability (experiment 1), and historical emissions (experiment 2). Historical emissions are endogenously determined in a prior game. Capacity inequality strongly affects burden sharing, with richer players ending up paying more, and poorer players less. Vulnerability differences reduce the influence of fairness, leading to suggested cost distributions more unfavorable to vulnerable players. However, vulnerable responders still reject many "unfair” offers. Differences in historical responsibility result in cost distributions strongly correlated with players' relative contributions to climate change. The results suggest that more nuanced consideration of fairness criteria in burden sharing could make ambitious climate agreements more acceptable for reluctant countries and their citizens
Cytotoxicity, metabolism, and isozyme mapping of the synthetic cannabinoids JWH-200, A-796260, and 5F-EMB-PINACA studied by means of in vitro systems
Intake of synthetic cannabinoids (SC), one of the largest classes of new psychoactive substances, was reported to be associated with acute liver damage but information about their hepatotoxic potential is limited. The current study aimed to analyze the hepatotoxicity including the metabolism-related impact of JWH-200, A-796260, and 5F-EMB-PINACA in HepG2 cells allowing a tentative assessment of different SC subclasses. A formerly adopted high-content screening assay (HCSA) was optimized using a fully automated epifluorescence microscope. Metabolism-mediated effects in the HCSA were additionally investigated using the broad CYP inhibitor 1-aminobenzotriazole. Furthermore, phase I metabolites and isozymes involved were identified by in vitro assays and liquid chromatography–high-resolution tandem mass spectrometry. A strong cytotoxic potential was observed for the naphthoylindole SC JWH-200 and the tetramethylcyclopropanoylindole compound A-796260, whereas the indazole carboxamide SC 5F-EMB-PINACA showed moderate effects. Numerous metabolites, which can serve as analytical targets in urine screening procedures, were identified in pooled human liver microsomes. Most abundant metabolites of JWH-200 were formed by N-dealkylation, oxidative morpholine cleavage, and oxidative morpholine opening. In case of A-796260, most abundant metabolites included an oxidative morpholine cleavage, oxidative morpholine opening, hydroxylation, and dihydroxylation followed by dehydrogenation. Most abundant 5F-EMB-PINACA metabolites were generated by ester hydrolysis plus additional steps such as oxidative defluorination and hydroxylation. To conclude, the data showed that a hepatotoxicity of the investigated SC cannot be excluded, that metabolism seems to play a minor role in the observed effects, and that the extensive phase I metabolism is mediated by several isozymes making interaction unlikely
Toxicokinetics and analytical toxicology of the abused opioid U-48800 - in vitro metabolism, metabolic stability, isozyme mapping, and plasma protein binding
Due to the risk of new synthetic opioids (NSOs) for human health, the knowledge of their toxicokinetic characteristics is important for clinical and forensic toxicology. U‐48800 is an NSO structurally non‐related to classical opioids such as morphine or fentanyl and offered for abuse. As toxicokinetic data of U‐48800 is not currently available, the aims of this study were to identify the in vitro metabolites of U‐48800 in pooled human liver S9 fraction (pS9), to map the isozymes involved in the initial metabolic steps, and to determine further toxicokinetic data such as metabolic stability, including the in vitro half‐life (t1/2), and the intrinsic (CLint) and hepatic clearance (CLh). Furthermore, drug detectability studies in rat urine should be done using hyphenated mass spectrometry. In total, 13 phase I metabolites and one phase II metabolite were identified. N‐Dealkylation, hydroxylation, and their combinations were the predominant metabolic reactions. The isozymes CYP2C19 and CYP3A4 were mainly involved in these initial steps. CYP2C19 poor metabolizers may suffer from an increased U‐48800 toxicity. The in vitro t1/2 and CLint could be rated as moderate, compared to structural related compounds. After administration of an assumed consumer dose to rats, the unchanged parent compound was found only in very low abundance but three metabolites were detected additionally. Due to species differences, metabolites found in rats might be different from those in humans. However, phase I metabolites found in rat urine, the parent compound, and additionally the N‐demethyl metabolite should be used as main targets in toxicological urine screening approaches
In vitro metabolic fate of the synthetic cannabinoid receptor agonists QMPSB and QMPCB (SGT-11) including isozyme mapping and esterase activity
Quinolin-8-yl 4-methyl-3-(piperidine-1-sulfonyl)benzoate (QMPSB) and quinolin-8-yl 4-methyl-3-(piperidine-1-carbonyl)benzoate (QMPCB, SGT-11) are synthetic cannabinoid receptor agonists (SCRAs). Knowing their metabolic fate is crucial for the identification of toxicological screening targets and to predict possible drug interactions. The presented study aimed to identify the in vitro phase I/II metabolites of QMPSB and QMPCB and to study the contribution of different monooxygenases and human carboxylesterases by using pooled human liver S9 fraction (pHLS9), recombinant human monooxygenases, three recombinant human carboxylesterases, and pooled human liver microsomes. Analyses were carried out by liquid chromatography high-resolution tandem mass spectrometry. QMPSB and QMPCB showed ester hydrolysis, and hydroxy and carboxylic acid products were detected in both cases. Mono/dihydroxy metabolites were formed, as were corresponding glucuronides and sulfates. Most of the metabolites could be detected in positive ionization mode with the exception of some QMPSB metabolites, which could only be found in negative mode. Monooxygenase activity screening revealed that CYP2B6/CYP2C8/CYP2C9/CYP2C19/CYP3A4/CYP3A5 were involved in hydroxylations. Esterase screening showed the involvement of all investigated isoforms. Additionally, extensive non-enzymatic ester hydrolysis was observed. Considering the results of the in vitro experiments, inclusion of the ester hydrolysis products and their glucuronides and monohydroxy metabolites into toxicological screening procedures is recommended
Developing logistic software platforms: e-market place, a case study
This paper describes a framework for software development with emphasis on logistics platforms. Specifically, a case study is presented regarding the Logport program in Colombia’s Caribbean coast. The software development is based on the merging of modeling, design, and agile development techniques aimed towards software production. The agile methodologies discussed herein include Scrum, XP, and Crystal among others, and specific verification and validation aspects of CMMI 1.3 are evaluated. Furthermore, the integration of emerging technologies including elastic computing in cloud computing that allows scaled integration, security, load balancing, process speed, and high concurrency among other features are discussed. Finally, the proposed solutions cover actual and emergent needs in a B2B or B2C electronic commerce dynamic environment where suppliers and clients offer and demand transport, storage, and customs services. In this environment, their goal is to ensure there is added value to their logistics processes starting at the inputs and all the way to the outputs of the commercial business processes. Colombia’s Caribbean region is one such multimodal and multiport environment in which there is constant demand for low cost, time and storage optimization, and customs reliability for the businesspersons of this region, which serves as a Hub for the American Caribbean
Staphylococcal Superantigen (TSST-1) Mutant Analysis Reveals that T Cell Activation Is Required for Biological Effects in the Rabbit Including the Cytokine Storm
Staphylococcal superantigens (sAgs), such as toxic shock syndrome toxin 1 (TSST-1), induce massive cytokine production, which may result in toxic shock syndrome (TSS) and sepsis. Recently, we reported that in vitro studies in human peripheral blood mononuclear cells (PBMC) do not reflect the immunological situation of the host, because after exposure to superantigens (sAgs) in vivo, mononuclear cells (MNC) leave the circulation and migrate to organs, e.g., the spleen, liver and lung. Our experimental model of choice is the rabbit because it is comparable to humans in its sensitivity to sAg. T cell activation has been assessed by lymphocyte proliferation and IL-2 gene expression after in vivo challenge with TSST-1 and the mutant antigens; expression of the genes of proinflammatory cytokines were taken as indicators for the inflammatory reaction after the combined treatment with TSST-1 and LPS. The question as to whether the biological activities of TSST-1, e.g., lymphocyte extravasation, toxicity and increased sensitivity to LPS, are mediated by T cell activation or activation by MHC II-only, are unresolved and results are contradictory. We have addressed this question by studying these reactions in vivo, with two TSST-1 mutants: one mutated at the MHC binding site (G31R) with reduced MHC binding with residual activity still present, and the other at the T cell binding site (H135A) with no residual function detectable. Here, we report that the mutant G31R induced all the biological effects of the wild type sAg, while the mutant with non-functional TCR binding did not retain any of the toxic effects, proving the pivotal role of T cells in this system