Finance 1978-79

IPEDS Finance annual report contains the following information: • Revenues by source (e.g., tuition and fees, government, private gifts) • Expenses by function (e.g., instruction, research, plant maintenance and operation) • Scholarships, physical plant assets and indebtedness • Assets, liabilities and net assets • Different formats are used based on the institution’s accounting standard

Chronic kidney disease induces systemic microvascular functional impairment

Chronic kidney disease in young patients is associated with a high mortality rate due to cardiac events, which is in stark contrast to age-matched healthy controls. The mechanisms leading to terminal heart failure in chronic kidney disease are, however, still poorly understood. Two mouse models of experimental uremia mimicking chronic kidney disease were employed. The data acquired were used for extensive correlation analyses, using serum urea levels and microvessel density in cremaster muscle (derived from identical animals in a related study) as reference parameters. Functional parameters were measured in vivo in the cremaster muscle as a model skeletal muscle by intravital microscopy techniques. The arterial-venous oxygen saturation difference in uremic animals was found to be significantly diminished in a dose-dependent manner. Similarly, in uremic animals, a reduced capability to adjust vessel tone was detected. Despite elevated IL-6 serum levels, no reduction in leukocyte rolling velocity was shown, which is in line with previous results showing an impaired neutrophil recruitment. Post mortem analyses of microvessel density in myocardium showed a significant correlation with microvascular rarefaction in skeletal muscle detected in vivo, implying that uremia has a systemic impact. Aortic cross sections of uremic mice, however, showed normal morphology and no differences to controls, suggesting that microvascular rarefaction precedes macrovascular pathology. These results indicated chronic hypoxic conditions in the respective tissues. Therefore, the expression of genes involved in vessel growth, integrity, and remodeling was determined in cardiac muscle. While VEGF-A, Ang-1, and MMP-2 remained unaffected by uremia, HIF 1alpha, VEGFR-2, Tie-1, Tie-2, Ang-2, and MMP-9 displayed significant downregulation. Thus, VEGF and Ang/Tie-2 signaling was disrupted in uremia, while HIF 1alpha levels were depleted, suggesting a dysregulated angiogenic response. In fact, uremia resulted in a reversed relationship of hypoxia and angiogenesis. This study shows that uremia leads, in a dose-dependent manner, to a profound systemic dysregulation of the sensitive interplay between physiological and cellular factors needed to maintain homeostasis, and this process may already be far advanced before becoming clinically apparent. A regulatory network model of feedback loops is presented which may explain the progression of disease. Upon continuous damage, the regulatory network may decompensate, resulting in a sudden onset of progressively amplifying dysregulatory circles, leading to severe complications which may ultimately result in cardiac death. Therefore, microvascular rarefaction is not only an early systemic marker of cardiovascular impairment in chronic kidney disease but also a promising target for future therapeutic approaches.In der pädiatrischen Nephrologie sind chronische Niereninsuffizienz und terminales Nierenversagen mit einer sehr hohen Mortalität verbunden. Auffallend dabei ist, dass jungen Patienten im Gegensatz zu gleichaltrigen gesunden Kontrollgruppen insbesondere an kardialen Ereignissen versterben. Der Pathomechanismus, der bei chronischer Niereninsuffizienz vor allem zum Herztod führt, ist dabei noch wenig verstanden. In dieser Arbeit wurden zwei Urämie-Mausmodelle zur chronischen Niereninsuffizienz für funktionelle Studien eingesetzt. Mit den gewonnenen Daten wurden umfassende Korrelationsanalysen durchgeführt, wobei Serumharnstoffwerte als Indikator für den Schweregrad der Niereninsuffizienz und die Dichte der Mikrovaskulatur des Musculus cremaster (als Modell für einen Skelettmuskel) derselben Tiere aus einer parallel durchgeführten Studie als Referenzen genutzt wurden. Die arterio-venöse Sauerstoffdifferenz war abhängig vom Urämiegrad der Tiere signifikant reduziert. Entsprechend war in urämischen Tieren auch die Fähigkeit zur Regulation des Gefäßtonus beeinträchtigt. Trotz erhöhter IL-6-Werte im Serum wurde keine Verminderung der Geschwindigkeit des Leukozytenrollens festgestellt. Dies entspricht früheren Studien, in denen bereits eine gestörte Rekrutierung von Neutrophilen beobachtet worden war. Post mortem-Analysen der Mikrovaskulaturdichte im Myokard zeigten eine signifikante Korrelation zur Mikrovaskulatur-Rarefizierung im Skelettmuskel in vivo, was auf einen systemischen Effekt der Urämie hindeutete. Aortenquerschnitte urämischer Versuchstiere zeigten jedoch eine normale Morphologie ohne Unterschiede zu Kontrolltieren. Dies legte nahe, dass eine Rarefizierung der Mikrovaskulatur der Pathologie großer Gefäße vorausgeht. Diese Ergebnisse wiesen auf eine chronische Hypoxie in den untersuchten Geweben hin. Daher wurde die Expression von solchen Genen im Herzmuskel untersucht, die Gefäßwachstum, -stabilität und -remodelling steuern. Während VEGF-A, Ang-1 und MMP-2 durch die Urämie unbeeinflusst blieben, waren die Expression von HIF 1alpha, VEGFR-2, Tie-1, Tie-2, Ang-2 und MMP-9 signifikant vermindert. Die VEGF- und Ang/Tie-2-Signalwege waren also bei Urämie gestört, während gleichzeitig der HIF 1alpha-Spiegel vermindert war, was eine Dysregulation der Angiogenese nahelegte. Tatsächlich bestand bei Urämie ein reziprokes Verhältnis zwischen Hypoxie und Angiogenese. Diese in vivo Studie ergab Hinweise darauf, dass Urämie abhängig von ihrem Schweregrad zu einer tiefgreifenden systemischen Dysregulation des empfindlichen Zusammenspiels physiologischer und zellulärer Faktoren führt, die zur Aufrechterhaltung einer zellulären Homöostase notwendig sind. Dieser Prozess kann bereits weit fortgeschritten sein, bevor er sich klinisch manifestiert. Der Krankheitsfortschritt kann durch ein Netzwerkmodell mit einer Vielzahl von Rückkopplungsschleifen erklärt werden. Sobald die Schädigung einen Schwellenwert überschreitet, kann dies zu einer Dekompensation durch sich zunehmend selbstverstärkende Rückkopplungsmechanismen führen, die schließlich in kardiale Komplikationen münden können. Die Rarefizierung der Mikrovaskulatur bei chronischer Niereninsuffizienz ist daher nicht nur ein früher klinischer Marker für drohende kardiale Ereignisse, sondern auch eine vielversprechende Zielstruktur für zukünftige therapeutische Ansätze

The System Kato: Detecting Cases of Plagiarism for Answer-Set Programs

Plagiarism detection is a growing need among educational institutions and solutions for different purposes exist. An important field in this direction is detecting cases of source-code plagiarism. In this paper, we present the tool Kato for supporting the detection of this kind of plagiarism in the area of answer-set programming (ASP). Currently, the tool is implemented for DLV programs but it is designed to handle other logic-programming dialects as well. We review the basic features of Kato, introduce its theoretical underpinnings, and discuss an application of Kato for plagiarism detection in the context of courses on logic programming at the Vienna University of Technology

Sintering of ultrathin gold nanowires for transparent electronics

Ultrathin gold nanowires (AuNWs) with diameters below 2 nm and high aspect ratios are considered to be a promising base material for transparent electrodes. To achieve the conductivity expected for this system, oleylamine must be removed. Herein we present the first study on the conductivity, optical transmission, stability, and structure of AuNW networks before and after sintering with different techniques. Freshly prepared layers consisting of densely packed AuNW bundles were insulating and unstable, decomposing into gold spheres after a few days. Plasma treatments increased the conductivity and stability, coarsened the structure, and left the optical transmission virtually unchanged. Optimal conditions reduced sheet resistances to 50 Ω/sq

Multivalent bonds in self-assembled bundles of ultrathin gold nanowires

Ultrathin gold nanowires are unusual colloidal objects that assemble into bundles with line contacts between parallel wires. Each molecule in the contact line interacts with many ligand and solvent molecules. We used X-ray scattering and electron microscopy to study how these interactions control assembly

Cystic fibrosis sputum DNA has NETosis characteristics and neutrophil extracellular trap release is regulated by macrophage migration-inhibitory factor

Neutrophils are the main proinflammatory cell type in chronically infected lungs of cystic fibrosis (CF) patients; however, they fail to effectively clear the colonizing pathogens. Here, we investigated the molecular composition of non-mucoid and mucoid Pseudomonas aeruginosa-induced neutrophil extracellular traps (NETs) in vitro and compared them to the DNA-protein complexes present in the CF sputum. The protein composition of P. aeruginosa-induced NET fragments revealed that irrespective of the inducing stimuli, NET fragments were decorated with a conserved set of proteins. The DNA-protein complexes derived from CF sputum were consistent with NETosis and shared a similar protein signature, suggesting that the majority of the extracellular DNA was NET derived. The ability of polymorphonuclear leukocytes to produce NETs in response to P. aeruginosa was driven by macrophage migration-inhibitory factor (MIF) by promoting mitogen-activated protein kinase. Analysis of 132 CF patient samples revealed that elevated MIF protein levels correlated with poorer lung function. We suggest that targeting MIF by small molecular inhibitors might reduce the presence of extracellular DNA and serve as an adjunct to the use of antimicrobial drugs that could ultimately reduce bacterial fitness in the lungs during the later stages of CF disease

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

The response to unfolded protein is involved in osmotolerance of Pichia pastoris

Background The effect of osmolarity on cellular physiology has been subject of investigation in many different species. High osmolarity is of importance for biotechnological production processes, where high cell densities and product titers are aspired. Several studies indicated that increased osmolarity of the growth medium can have a beneficial effect on recombinant protein production in different host organisms. Thus, the effect of osmolarity on the cellular physiology of Pichia pastoris, a prominent host for recombinant protein production, was studied in carbon limited chemostat cultures at different osmolarities. Transcriptome and proteome analyses were applied to assess differences upon growth at different osmolarities in both, a wild type strain and an antibody fragment expressing strain. While our main intention was to analyze the effect of different osmolarities on P. pastoris in general, this was complemented by studying it in context with recombinant protein production. Results In contrast to the model yeast Saccharomyces cerevisiae, the main osmolyte in P. pastoris was arabitol rather than glycerol, demonstrating differences in osmotic stress response as well as energy metabolism. 2D Fluorescence Difference Gel electrophoresis and microarray analysis were applied and demonstrated that processes such as protein folding, ribosome biogenesis and cell wall organization were affected by increased osmolarity. These data indicated that upon increased osmolarity less adaptations on both the transcript and protein level occurred in a P. pastoris strain, secreting the Fab fragment, compared with the wild type strain. No transcriptional activation of the high osmolarity glycerol (HOG) pathway was observed at steady state conditions. Furthermore, no change of the specific productivity of recombinant Fab was observed at increased osmolarity. Conclusion These data point out that the physiological response to increased osmolarity is different to S. cerevisiae. Increased osmolarity resulted in an unfolded protein response (UPR) like response in P. pastoris and lead to pre-conditioning of the recombinant Fab producing strain of P. pastoris to growth at high osmolarity. The current data demonstrate a strong similarity of environmental stress response mechanisms and recombinant protein related stresses. Therefore, these results might be used in future strain and bioprocess engineering of this biotechnologically relevant yeast