A Reference Architecture for a Workflow Management System Front End Designed for Augmented Reality Headsets

A well-known approach to managing and controlling workflows in organizations is the workflow management system (WFMS). Recently, approaches utilizing augmented reality headsets as WFMS front ends have been discussed, promising higher efficiency, effectiveness, and usability for certain application scenarios. However, existing design-oriented approaches lack actionable guidance for implementation. A well-known approach to address such knowledge gaps is a reference architecture, which inter alia reduces development times and risks and facilitates collaboration between developers. Based on an existing tentative design theory for an augmented reality-based WFMS front end, we contribute a reference architecture containing an extended design theory, user interface design, and models for use cases, components, classes, and sequence flows in the unified modeling language. The reference architecture was successfully operationalized in a prototype and positively evaluated via a survey of potential users

Evaluación de riesgos ecotoxicológicos derivados del empleo del hongo entomopatógeno Metarhizium spp. para el control de plagas

La agricultura ha provisto de alimentos al ser humano desde su origen, pero en el último siglo, la producción agraria se ha intensificado gracias, entre otros, al empleo de insecticidas. Sin embargo, esta intensificación no ha estado exenta de problemas de contaminación por residuos en los alimentos o de efectos negativos sobre la biodiversidad y el medioambiente. La legislación agraria actual promueve una agricultura acorde con la creciente preocupación de los consumidores por la salud y por el medioambiente. En este escenario, el desarrollo de estrategias ambientalmente sostenibles y seguras para los consumidores se sitúa en primera línea en los programas de investigación para el control de plagas. Los ascomicetos mitospóricos entomopatógenos (AMEs) y en particular el género Metarhizium, cumplen los requisitos de seguridad para el ser humano y el medio ambiente y además han mostrado un gran éxito en el control de plagas de insectos debido a su modo de acción por contacto, su presencia natural en diversos ecosistemas y su capacidad de secretar compuestos con actividad insecticida. No obstante, la implantación en el mercado de los micoinsecticidas es lenta, y tropieza con una barrera fundamental que es la escasa información sobre el destino de algunos metabolitos secundarios en la cadena alimentaria y su riesgo para la salud humana y animal, información clave para abordar su registro. Por tanto, existe la necesidad de desarrollar y validar métodos analíticos con alta sensibilidad para su determinación a bajas concentraciones en diferentes matrices biológicas. La destruxina A es uno de los principales metabolitos secundarios producidos por el AME Metarhizium spp., pero la falta de estudios sobre su producción por parte del hongo es probablemente el mayor obstáculo para el registro de nuevas cepas de esta especie fúngica. El objetivo principal de esta tesis ha sido desarrollar nuevas herramientas para la detección y cuantificación de destruxinas, así como investigar el destino de la destruxina A en la cadena trófica. En el capítulo II de esta Tesis, se ha determinado la producción de destruxinas por parte de cuatro cepas de Metarhizium (BIPESCO5, EAMa 01/58-Su, ARSEF 23 y ART 2825) con un método mejorado de cromatografía líquida de ultra alto rendimiento en tándem con espectrometría de masas (UHPLC-MS / MS) en cuatro medios de cultivo (CM, MM, CN2, OSM) que representan diferentes condiciones de estrés. Cada 3 días durante 18 días se tomaron muestras para análisis que permitieron detectar 15 destruxinas, siendo las destruxinas A y B las más abundantes. Además, se detectaron diferencias significativas entre las cepas en la producción de destruxinas, que a su vez fue altamente dependiente del medio de cultivo. En el capítulo III la colonización endofítica y la producción de destruxina A en plantas de patata se monitorizaron a las 24, 48, 72, 96 y 120 h después de la inoculación con dos cepas de Metarhizium brunneum Petch. (BIPESCO5 y EAMa 01/58-Su), lo que puso de manifiesto que la concentración de destruxina A en los tejidos vegetales es muy baja en comparación con los niveles de colonización. Aunque se observó una colonización similar para ambas cepas, hubo diferencias a lo largo de la planta, con valores más altos en las hojas a 96 h para EAMa 01/58-Su (83.3 %) y BIPESCO5 (81.6 %), y más bajos en tubérculo y raíz a las 72, 96 y 120 h después de la inoculación para ambas cepas (10.0-13.3 %). Para la cepa EAMa 01/58-Su, la destruxina A se cuantificó a las 24 h en el tubérculo y la raíz (2.0 ± 1.4 y 2.49 ± 1.7 μg / kg, respectivamente) y a las 96 h con igual concentración también en tubérculo y raíz (2.5 ± 1.7 μg /kg); para BIPESCO5, solamente se cuantificó destruxina A en el tubérculo a las 24 h y en la raíz a las 48 h (6.8 ± 4.8 y 2.1 ± 1.4 μg / kg, respectivamente). En el capítulo IV se investiga por primera vez la dinámica de crecimiento de las cepas BIPESCO5 y EAMa 01/58-Su de M. brunneum y la secreción de destruxina A durante el proceso de infección de larvas del insecto modelo Galleria mellonella L. (Lepidoptera; Pyralidae). Se observó que la secreción de destruxina A fue paralela a la evolución de la cantidad de ADN fúngico en el interior del insecto para la cepa EAMa 01/58-Su, no así para BIPESCO5. Las cepas EAMa 01/58-Su y BIPESCO5 secretaron destruxina A desde los días 2 al 6 y desde el día 2 hasta el día 5 después del tratamiento, respectivamente. Para EAMa 01/58-Su y BIPESCO5, la máxima cantidad de destruxina A producida en el insecto hospedante fue de 0.369 y 0.06 μg/larva a los 4 días del tratamiento, respectivamente, y a lo largo del proceso patogénico, la producción fue de 0.6 y 0.09 μg / larva, respectivamente. En el capítulo V se realizaron bioensayos presa-depredador para evaluar el comportamiento y la supervivencia de las larvas del depredador generalista Chrysoperla carnea (Stephens) (Neuroptera; Chrysopidae) al alimentarse con larvas insecto polífago Spodoptera littoralis (Boisd.) (Lepidoptera; Noctuidae) inoculadas con las cepas BIPESCO5 y EAMa 01/58-Su. Además, se llevaron a cabo estudios ecotoxicológicos para supervisar el destino de la destruxina A en el sistema presa-depredador. La concentración máxima de destruxina A producida por la cepa BIPESCO5 fue el día 4 con un valor de 0.000054 μg/insecto (aproximadamente 0.014 μg/g) y para la cepa EAMa 01/58-Su el día 5 con 0.00012 μg/insecto (aproximadamente 0.031 μg/g), mientras que el metabolito no se detectó en larvas de C. carnea. El porcentaje de crisopas que se alimentó de larvas de S. littoralis 24 horas después de la infección fue de 96.6, 75.0 y 65.0 % para el control, EAMa 01/58-Su y BIPESCO5, respectivamente, mientras que 5 días después de la infección fue 38.3 % para control y 33.3 % en los tratamientos con las cepas EAMa 01/58-Su y BIPESCO5. La cantidad de larvas de S. littoralis consumidas por C. carnea 24 h después de la infección fue 5.6, 2.2 y 2.3 para las tratadas con el control, EAMa 01/58-Su y BIPESCO5 respectivamente, mientras que 5 días después de la infección consumió una sola larva per cápita. Esto puso de manifiesto que los tratamientos de M. brunneum contra las larvas de S. littoralis fueron seguros para C. carnea debido tanto a la ausencia de mortalidad relacionada con los hongos en el depredador como a la falta de movimiento de la destruxina A de la presa al depredador. Es importante resaltar que en los capítulos IV y V, para ambas cepas, la mortalidad de las larvas debido a otras causas fue mucho mayor que la mortalidad con crecimiento fúngico. Sin embargo, la secreción de destruxina A fue mayor para EAMa 01/58-Su que para BIPESCO5, lo que sugiere que la destruxina A podría ser un factor de virulencia de la primera, mientras que la segunda podría requerir la participación de otros factores además de destruxina A durante el proceso de infección. Los resultados obtenidos proporcionan métodos analíticos valiosos para llevar a cabo evaluaciones de riesgo sobre el empleo de AME, así como resultados que indican que su empleo supone un bajo nivel de riesgo para la salud humana, animal y medio ambiental.Agriculture produces the vast majority of the world’s food supply, and in last century the global food production has grown at a huge rate mainly from the increased yields resulting from greater inputs of insecticides and other technologies. Meanwhile overuse or improper use of insecticides and other agrochemicals has raised issues about related environment and health costs, with current legislation promoting sustainable agriculture, in which scenario, the development of environmentally sustainable strategies is mandatory for research programs regarding pest control. The entomopathogenic mitosporic ascomycetes (EMAs) and in particular the genus Metarhizium have shown great success in the control of insect pests due to their contact mode of action, natural presence in the ecosystems and their ability to secrete compounds with insecticidal activity, and even, they comply with the security requirements for human health and environment, whereas information about the fate of their secondary metabolites in the food chain and their risk to human and animal health is still scarce. There is a need to develop and validate analytical methods with high sensitivity for metabolite determination at low concentrations in different biological matrices. Destruxin A is one of the major secondary metabolite produced by the genus Metarhizium spp., but the lack of studies concerning destruxin A production is most likely the biggest obstacle for registration of new fungal strains. The main goal of this research has been to develop new tools for destruxin detection and quantification and to investigate the fate of destruxin A in the trophic chain. In chapter II, destruxin production for Metarhizium strains BIPESCO5, EAMa 01/58- Su, ARSEF 23 and ART 2825 was determined with an improved method of ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), which has shown high precision in the detection and quantification of destruxins in four culture media (CM, MM, CN2, OSM) representing different stress conditions. Every 3 days samples were taken for analysis over 18 days that allowed detecting 15 destruxins, with destruxin A and B as the most abundant. However, significant differences among strains in destruxin production were detected, and for each strain, destruxin production was highly dependent on culture medium. In chapter III, endophytic colonisation and destruxin A production on potato plants were monitored at 24, 48, 72, 96 and 120 h after inoculation with Metarhizium brunneum strains (BIPESCO5 and EAMa 01/58-Su), which showed that the concentration of destruxin A in plant tissues was very low compared to the colonisation levels. Although a similar colonisation was observed for both strains, there were differences in percentages in different parts of the plants, with the higher values occurring in the leaves at 96 h for EAMa 01/58-Su (83.3 %) and BIPESCO5 (81.6 %), and the lower ones, 10.0-13.3 %, observed in tuber and root at 72, 96 and 120 h post-inoculation for both strains. For strain EAMa 01/58- Su, destruxin A was quantified at 24 h (2.49 ± 1.7 and 2.0 ± 1.4 μg/kg, respectively), and the same concentration was found in both tuber and root at 96 h (2.5 ± 1.7 μg/kg); for BIPESCO5, the concentrations differed in tuber at 24 h and in root at 48 h (6.8 ± 4.8 and 2.1 ± 1.4 μg/kg, respectively). In chapter IV, the dynamic of fungal growth and secretion of destruxin A by strains BIPESCO5 and EAMa 01/58-Su of Metarhizium brunneum Petch. during the infection process of larvae of the model insect Galleria mellonella L. (Lepidoptera; Pyralidae) was monitored for the first time. Data showed that destruxin A secretion was parallel to the fungal growth of EAMa 01/58-Su but not coupled with that for BIPESCO5. EAMa 01/58-Su and BIPESCO5 strains secreted destruxin A from days 2 to 6 and from day 2 to day 5 post treatment, respectively. For EAMa 01/58-Su and BIPESCO5, the maximum titer in the host on day 4 after treatment was 0.369 and 0.06 μg/larva, respectively, and throughout the pathogenic process, the production was 0.6 and 0.09 μg/larva, respectively. In chapter V, predator-prey bioassays were performed to evaluate the behavior and survival of larvae of the generalist predator Chrysoperla carnea (Stephens) (Neuroptera; Chrysopidae) when feeding on larvae of the polyphagous pest Spodoptera littoralis (Boisd.) (Lepidoptera; Noctuidae) challenged by M. brunneum BIPESCO5 and EAMa 01/58-Su strains. In addition, ecotoxicological studies based on HPLC-MS were performed to monitor the fate of destruxin A in the prey-predator system. The maximum concentration of destruxin A produced by the BIPESCO5 strain was on day 4 after treatment with a value of 0.000054 μg/insect (approx 0.014 μg/g), and for EAMa 01/58-Su was on day 5 with a value of 0.00012 μg/insect (approx 0.031 μg/g), whereas the metabolite was no detected in C. carnea larvae. The percentage of lacewings feeding on S. littoralis larvae 24 hour-post infection was 96.6, 75.0, and 65.0 % for the control, EAMa 01/58-Su, and BIPESCO5 treatments, respectively, whereas 5 days-post infection armyworm larvae were consumed by only 38.3 % of the control lacewings and 33.3 % of the EAMa 01/58-Su and BIPESCO5 treatment groups. C. carnea larvae feeding on 24 h-post infection armyworm larvae preyed 5.6, 2.2 and 2.3 larvae for the control, EAMa 01/58-Su and BIPESCO5 treatments, respectively, whereas those predator larvae feeding on 5 days-post infection armyworm larvae preyed on only one per capita larva. It showed that the M. brunneum treatments against S. littoralis larvae were safe for C. carnea due to both the lack of fungus-related mortality in the predator and the lack of movement of destruxin A from the prey to the predator. Notably in chapters IV and V, in both M. brunneum strains, mortality from other causes was higher than mortality with fungal outgrowth. However, destruxin A secretion was higher for EAMa 01/58-Su than for BIPESCO5. These results suggested that destruxin A could be a virulence factor for EAMa 01/58-Su strain, whereas for BIPESCO5, the virulence could require the involvement of other factors as well as destruxin A during the infection process. The results obtained provide valuable analytical methods for carrying out risk assessments on the use of EMAs. In addition, results indicate that their use poses a little potential hazard to human and animal health and the environment

Consensus guidelines for the use and interpretation of angiogenesis assays

The formation of new blood vessels, or angiogenesis, is a complex process that plays important roles in growth and development, tissue and organ regeneration, as well as numerous pathological conditions. Angiogenesis undergoes multiple discrete steps that can be individually evaluated and quantified by a large number of bioassays. These independent assessments hold advantages but also have limitations. This article describes in vivo, ex vivo, and in vitro bioassays that are available for the evaluation of angiogenesis and highlights critical aspects that are relevant for their execution and proper interpretation. As such, this collaborative work is the first edition of consensus guidelines on angiogenesis bioassays to serve for current and future reference

Ultraschallfertigung zur Herstellung mikrofluidischer Systeme

The use of ultrasound for plasticizing materials, in particular thermoplastics, is the subject of research for over half a century [II.1]. Today, in the industry ultrasound machines are primarily used as an inexpensive and quick option for cleaning and bonding materials on a macroscale [II.2]. For this purpose, acoustic waveguides, so called sonotrodes, are used to transfer a high-frequency, mechanical vibration into the work piece making them moldable. The default machining area of a single ultrasonic machine has the size of a chip card.The range of application is extendable by usage of special molds, which enable the embossing of structures in the sub millimeter range [II.3]. The combination of this technique, the so-called ultrasonic hot embossing, with existing processing methods such as ultrasonic welding, -riveting, etc., allows the fabrication of a variety of microsystems. These ultrasonic assembly methods, with focus on ultrasonic hot embossing, were analyzed in this thesis and the practical implementation of the theoretical investigations were exemplified by the fabrication of different microsystems.For this purpose, the fundamentals of the ultrasonic manufacturing methods have been discussed first. The core areas were the distribution of the ultrasound through the wrought material, dimensioning of the mold with regard to the required functional and constructive geometries and the combination of different species of plastics. This allowed an estimation of the possibilities and limitations of the methods.One of the key aspects of the work was the construction of microfluidic devices. Therefore, a fluidic connection between the microsystems and devices above the micron range, such as pumps or gas cylinders, are required. In addition to commercially available connectors, own approaches with more exotic options have been tested. Connection components located planar and perpendicular to the system were developed and their advantages and disadvantages discussed.The evaluation of connectivity options resulted in a concept of modular micro-systems whose versatile and expandable kit was based on the ultrasonic fabrication methods. The aim was to provide the basis for an adaptable package of "disposable articles" in combination with a fast, flexible and cost-effective production process. The first elementary modules were systems that differ in their geometrical structure for the mixing and separation of fluids, which were subsequently extended by sensor units for the measurement of reaction temperatures and volumetric flow rates.Finally, two microsystems were reviewed outside the module kit that needed an alternative approach of manufacturing and assembly of the microstructure, necessitated by their requirements. A bioreactor for the growing of a yeast strain and the selective regulation of their gene expression. Here the typical approach of ultrasonic hot embossing the structure into a stack of foils hasn’t been used. Instead, the channel was prepared by ultrasonic hot embossing into a combination of a 0.125 mm foil and a 3 mm polymer plate. Secondly, a mea¬suring section, for the determination of diffusion constants with the smallest possible channel cross section at a maximum channel length was fabricated