The aim of this project was to make a comparison of the effects of antibody labeling chemistries on assays developed for the Gyrolab immunoassay platform. One of the labeling techniques was a heterogenous labeling technique targeting amino groups on the antibody. The other labeling technique was a site-specific labeling technique targeting the conserved Fc-glycan at the aspargine 297 residue on the IgG molecule. The site-specific labeling was performed using a kit from Genovis called GlyCLICK. The two labeling techniques were compared on four different assays developed for the Gyrolab platform. The assays tested in this project were two anti-drug antibody assays, a pharmacokinetics assay, a polyclonal antibody assay, and a monoclonal antibody assay. The drug tolerance was tested for the anti-drug antibody assays, resulting in better drug tolerance for reagents labeled with amino conjugation for the Humira assay with incubation overnight. A confirmatory analysis, testing the inhibition of negative control with addition of unlabeled drug in the Master Mix, was performed. This resulted in small differences in the inhibition between the different reagents, except for Keytruda on Gyrolab Bioaffy 200, for which the GlyCLICK labeled reagents led to a lower inhibition of the negative control. For all the assays the effects on signal to background ratio and limit of detection was investigated. The greatest advantages of GlyCLICK on the signal to background was observed for anti-drug antibody Keytruda assay and polyclonal antibody assay. For the polyclonal antibody assay, the results indicated potentially reduced need for the polishing step and for two wash solutions after addition of the detect reagent.

Cre-mediated site-specific recombination has emerged as an indispensable tool for the precise manipulation of genomes allowing lineage-tracing studies, temporal and spatial misexpressions, and in particular the generation of conditional knockout alleles. Previously, we and others showed that Cre and its ligand-inducible variant CreERT2 are also highly efficient in the developing and adult zebrafish. The number of Cre driver and effector lines is currently still limited in zebrafish. However, the recent advent of novel genome editing tools such as TALEN and CRISPR/Cas will significantly increase interest in the conditional Cre/lox-technology in this organism. The considerations of basic transgene design and subsequent transgenesis have been addressed elsewhere. Here we outline practical experimental steps for transient functionality tests of CreERT2 driver and effector constructs. In addition, we introduce detailed protocols to elicit CreERT2-mediated recombination in vivo at embryonic as well as adult stages.

Nitrogen (N) is a key nutrient input to crops and one of the major pollutants to the environment from agriculture in the United States. Recent developments in site-specific management (SSM) technology have the potential to reduce both N overapplication and underapplication and increase farmers' net returns. In Virginia, due to the high variability of within-field yield-limiting factors such as soil physical properties and fertility, the adoption of SSM is hindered by high gridsampling cost. Many Virginia corn-wheat-soybean farms have practiced generating yield maps using yield monitors for several years even though few variable applications based on yield maps were reported. It is unknown if the information generated by yield monitors under actual production situations can be used to direct N management for increased net returns in this area. The overall objective of the study is to analyze the economic and environmental impact of alternative management strategies for N in corn and wheat production based on site-specific information in eastern Virginia. Specifically, evaluations were made of three levels of site-specific information regarding crop N requirements combined with variable and uniform N application. The three levels of information are information about the yield potential of the predominant soil type within the field, information about yield potentials of all soils within the field (soil zones), information about yield potentials of smaller sub-field units which are aggregated into functional zones. Effects of information on expected net returns and net N (applied N that is not removed by the crop) were evaluated for corn-wheat-soybean fields in eastern Virginia. Ex post and ex ante evaluations of information were carried out. Historical weather data and farm-level yield data were used to generate yield sequences for individual fields. A Markov chain model was used to describe both temporal and spatial yield variation. Soil maps were used to divide a field into several soil management units. Cluster analysis was used to group subfield units into functional zones based on yield monitor data. Yield monitor data were used to evaluate ex post information and variable application values for 1995-1999, and ex ante information and variable application values for 1999. Ex post analysis results show that soil zone information increased N input but decreased net return, while functional zone information decreased N input and increased net returns. Variable application decreased N input compared with uniform application. Variable application based on soil zone information reduced net return due to cost of overapplication or underapplication. Variable application based on functional information increased net return. Ex ante results show that information on spatial variability was not able to increase farmers?net return due to the cost of variable N application and information. Variable rate application decreases N input relative to uniform application. However, imprecision in the spatial predictor makes the variable application unprofitable due to an imbalance between costs of under- and over-application of N. Sensitivity analysis showed that value of information was positive when temporal uncertainty was eliminated. The ex post results of this study suggest there is potential to improve efficiency of N use and farmers?net returns with site specific management techniques. The ex ante results suggest that site specific management improvements should be tested under conditions faced by farmers including imperfect information about temporal and spatial yield variability. / Ph. D.

京都大学 / 0048 / 新制・課程博士 / 博士(地域研究) / 甲第19833号 / 地博第189号 / 新制||地||66(附属図書館) / 32869 / 京都大学大学院アジア・アフリカ地域研究研究科東南アジア地域研究専攻 / (主査)教授 竹田 晋也, 教授 岩田 明久, 准教授 古澤 拓郎, 教授 神﨑 護 / 学位規則第4条第1項該当 / Doctor of Area Studies / Kyoto University / DGAM

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There is a new order on the farm, the animals are long since gone, new people have moved in. On a farm in Bergslagen, which is also my home, a barn (or lada in Swedish) stands empty. The original function of this barn, to house a small number of cows and their feed, has been lost. Since moving to the farm, we have asked ourselves how it should be used.  It is here the project I Want to Become a Barn plays out. In my degree project, I explore how to have a dia-logue with the building and how the barn can be approached by starting from its current condition.  The work process is guided by a care for that which has been, where the new has to adapt to the old. The history of the place, the ravages of time, and the forces of gravity are elementary starting points. The project proposes that the building can be made habitable and meet basic everyday needs. When does it stop being a stable for animals and when does it start to become a home for humans?

Diploma thesis is an environmentally and process oriented project which is based on the issue of the so-called bark beetle calamity. It deals with the personification of non-human life. It uses experience with materials, their handling and processing.

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