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Verfassungsblog: On Matters Constitutional

This chapter examines the steps which take place after legislation has been passed. It also looks at the principles and rules that exist to ensure the legality and legitimacy of administrative action implementing EU law. It begins with an overview of the key institutions and agencies of the EU and what they do. It then discusses the applicable law which is key to developing notions of accountability and the protection of rights in this field.

UV chromatographic data in combination with multivariate data analysis (MVDA) has been extensively used for bioprocess monitoring. However, they are usually attributed to shifts along the retention time and require preprocessing. Misaligned UV chromatographic data result in inconsistent MVDA models. Numerous preprocessing techniques are available, each varying in the number of meta-parameters to optimize, complexity and computational time. Therefore, we aimed at developing a generic workflow to screen for preprocessing techniques. We chose four datasets with increasing complexity containing UV chromatographic data from reverse-phase and size exclusion chromatography HPLC. We aligned all four datasets using three preprocessing techniques, namely icoshift, PAFFT and RAFFT algorithms. We chose several statistical tools to validate the performance of the preprocessing techniques and to screen for meta-parameters. We validated the performance of the preprocessing techniques in terms of data preservation, complexity and computational time, and identified the optimal ranges of meta-parameters for each dataset. Finally, we established principal component analysis (PCA) models to evaluate the chosen alignment technique. Summarizing, in this study a generic workflow has been developed to validate alignment of chromatographic data using statistical tools.

One of the European Union’s most ambitious policy projects to date is the ‘economic and monetary union whose currency is the euro’ (EMU, Article 3(4) TEU). The EMU’s two polices—the economic union and the monetary union—are an unequal set of twins. On one hand, the monetary union’s central elements are well developed: as an element of substance, the introduction of the euro as a single currency; as an institutional achievement, the creation of the European System of Central Banks (ESCB) together with the European Central Bank (ECB) on the EU level as a highly independent body having the power to adopt a diverse range of measures. Additionally, the Treaties contain specific provisions on the goals and principles of monetary policy.

Any attempt to survey the sectoral administrative law of the European Union begs a number of fundamental questions. First, how do we understand ‘administrative law’, especially in relation to a supranational structure and system, such as that of the Union? Secondly, what exactly might we understand by ‘sectoral’ or ‘specialized’ administrative law, again especially in the context of the EU? Thirdly, just what is the difference between European Union law as such, and its administrative law. These distinctions, and the definitions attempted, are not addressed here just as scholarly abstractions. We take the view that understanding them and differentiating between these terms will make clear from the outset what this book attempts to cover, especially with respect to the concepts of both general EU administrative law and specialized EU administrative law employed here (which may or may not be similar to conceptualizations found in national contexts where these terms are employed).

In the context of the developing European Economic and Monetary Union (EMU), questions of political and judicial means of control of the bodies central to shaping this policy are of great relevance. This paper looks at how responsibility towards political and judicial means of control can be ensured in reality and what the case law of the CJEU can teach about the accountability standards of a structurally independent executive body such as the ECB. In the inverse, it also looks at the more general lessons the CJEU's Gauweiler case has for today's understanding of the EMU as central part of EU public law.

We report on a label-free, noninvasive method for determination of spore inoculum quality of Penicillium chrysogenum prior to cultivation/germination. Raman microspectroscopy providing direct, molecule-specific information was used to extract information on the viability state of spores sampled directly from the spore inoculum. Based on the recorded Raman spectra, a supervised classification method was established for classification between living and dead spores and thus determining spore inoculum quality for optimized process control. A fast and simple sample preparation method consisting of one single dilution step was employed to eliminate interfering signals from the matrix and to achieve isolation of single spores on the sample carrier (CaF2). Aiming to avoid any influence of the killing procedure in the Raman spectrum of the spore, spores were considered naturally dead after more than one year of storage time. Fluorescence staining was used as reference method. A partial least squares discriminant analysis classifier was trained with Raman spectra of 258 living and dead spores (178 spectra for calibration, 80 spectra for validation). The classifier showed good performance when being applied to a 1 mu L droplet taken from a 1:1 mixture of living and dead spores. Of 135 recorded spectra, 51% were assigned to living spores while 49% were identified as dead spores by the classifier. The results obtained in this work are a fundamental step towards developing an automated, label-free, and noninvasive screening method for assessing spore inoculum quality.

Horseradish peroxidase (HRP), a versatile heme-containing glycoprotein frequently used in medical diagnostics, is primarily isolated from plant. This process involves filtration, salt precipitation and several chromatography steps, which is expensive, renders low yields and gives isoenzyme mixtures. Although single isoenzymes can be recombinantly produced in Pichia pastoris, they get hyper-glycosylated in the yeast rendering the downstream process cumbersome and thus not competitive. In this study, we analyzed the purification of three HRP isoenzymes differing in the number of N-glycosylation sites recombinantly produced in P. pastoris. We wanted to 1) determine potential correlations between the mode of protein production and the resulting product quality and downstream process, 2) investigate correlations between the number of N-glycosylation sites of HRP and the mode of purification, and 3) find the optimal purification strategy for the recombinantly produced HRP isoenzymes. Thus, we applied different cultivation strategies and tested downstream processes employing both particlebased resins as well as monolithic supports. We showed that the mode of cultivation affected product purity but not the subsequent downstream process. Purification of each of the three HRP isoenzymes using monolithic supports was successful and independent of the number of N-glycosylation sites, whereas purification by particle-based resins was highly affected by glycosylation. Summarizing, we demonstrated a novel feasibility of using monoliths operated in flow through mode for the purification of comparatively small biomolecules.

This paper shows the ease of application and usefulness of mid-IR measurements for the investigation of orthogonal cell states on the example of the analysis of Pichia pastoris cells. A rapid method for the discrimination of entire yeast cells grown under carbon and nitrogen-limited conditions based on the direct acquisition of mid-IR spectra and partial least squares discriminant analysis (PLS-DA) is described. The obtained PLS-DA model was extensively validated employing two different validation strategies: (i) statistical validation employing a method based on permutation testing and (ii) external validation splitting the available data into two independent sub-sets. The Variable Importance in Projection scores of the PLS-DA model provided deeper insight into the differences between the two investigated states. Hence, we demonstrate the feasibility of a method which uses IR spectra from intact cells that may be employed in a second step as an in-line tool in process development and process control along Quality by Design principles.