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Purification of mRNA with oligo(dT)-functionalized magnetic particles involves a series of magnetic separations for buffer exchange and washing. Magnetic particles interact and agglomerate with each other when a magnetic field is applied, which can result in a decreased total surface area and thus a decreased yield of mRNA. In addition, agglomeration may also be caused by mRNA loading on the magnetic particles. Therefore, it is of interest how the individual steps of magnetic separation and subsequent redispersion in the buffers used affect the particle size distribution. The lysis/binding buffer is the most important buffer for the separation of mRNA from the multicomponent suspension of cell lysate. Therefore, monodisperse magnetic particles loaded with mRNA were dispersed in the lysis/binding buffer and in the reference system deionized water, and the particle size distributions were measured. A concentration-dependent agglomeration tendency was observed in deionized water. In contrast, no significant agglomeration was detected in the lysis/binding buffer. With regard to magnetic particle recycling, the influence of different storage and drying processes on particle size distribution was investigated. Agglomeration occurred in all process alternatives. For de-agglomeration, ultrasonic treatment was examined. It represents a suitable method for reproducible restoration of the original particle size distribution.
Laboratory protocols using magnetic beads have gained importance in the purification of mRNA for vaccines. Here, the produced mRNA hybridizes specifically to oligo(dT)-functionalized magnetic beads after cell lysis. The mRNA-loaded magnetic beads can be selectively separated using a magnet. Subsequently, impurities are removed by washing steps and the mRNA is eluted. Magnetic separation is utilized in each step, using different buffers such as the lysis/binding buffer. To reduce the time required for purification of larger amounts of mRNA vaccine for clinical trials, high-gradient magnetic separation (HGMS) is suitable. Thereby, magnetic beads are selectively retained in a flow-through separation chamber. To meet the requirements of biopharmaceutical production, a disposable HGMS separation chamber with a certified material (United States Pharmacopeia Class VI) was developed which can be manufactured using 3D printing. Due to the special design, the filter matrix itself is not in contact with the product. The separation chamber was tested with suspensions of oligo(dT)-functionalized Dynabeads MyOne loaded with synthetic mRNA. At a concentration of cB = 1.6–2.1 g·L–1 in lysis/binding buffer, these 1 μm magnetic particles are retained to more than 99.39% at volumetric flows of up to 150 mL·min–1 with the developed SU-HGMS separation chamber. When using the separation chamber with volumetric flow rates below 50 mL·min–1, the retained particle mass is even more than 99.99%.
Modeling and executing knowledge-intensive processes (KiPs) are challenging with state-of-the-art approaches, and the specific demands of KiPs are the subject of ongoing research. In this context, little attention has been paid to the ontology-driven combination of data-centric and semantic business process modeling, which finds additional motivation by enabling the division of labor between humans and artificial intelligence. Such approaches have characteristics that could allow support for KiPs based on the inferencing capabilities of reasoners. We confirm this as we show that reasoners can infer the executability of tasks based on a currently researched ontology- and data-driven business process model (ODD-BP model). Further support for KiPs by the proposed inference mechanism results from its ability to infer the relevance of tasks, depending on the extent to which their execution would contribute to process progress. Besides these contributions along with the execution perspective (start-to-end direction), we will also show how our approach can help to reach specific process goals by inferring the relevance of process elements regarding their support to achieve such goals (end-to-start direction). The elements with the most valuable process progress can be identified in the intersection of both, the execution and goal perspective. This paper will introduce this new approach and verifies its practicability with an evaluation of a KiP in the field of emergency call centers.
Carbon footprinting of universities worldwide: Part I — objective comparison by standardized metrics
(2021)
Background: Universities, as innovation drivers in science and technology worldwide, should be leading the Great Transformation towards a carbon–neutral society and many have indeed picked up the challenge. However, only a small number of universities worldwide are collecting and publishing their carbon footprints, and some of them have defined zero emission targets. Unfortunately, there is limited consistency between the reported carbon footprints (CFs) because of different analysis methods, different impact measures, and different target definitions by the respective universities.
Results: Comprehensive CF data of 20 universities from around the globe were collected and analysed. Essential factors contributing to the university CF were identified. For the first time, CF data from universities were not only compared. The CF data were also evaluated, partly corrected, and augmented by missing contributions, to improve the consistency and comparability. The CF performance of each university in the respective year is thus homogenized, and measured by means of two metrics: CO2e emissions per capita and per m2 of constructed area. Both metrics vary by one order of magnitude across the different universities in this study. However, we identified ten universities reaching a per capita carbon footprint of lower than or close to 1.0 Mt (metric tons) CO2e/person and year (normalized by the number of people associated with the university), independent from the university’s size. In addition to the aforementioned two metrics, we suggested a new metric expressing the economic efficiency in terms of the CF per $ expenditures and year. We next aggregated the results for all three impact measures, arriving at an overall carbon performance for the respective universities, which we found to be independent of geographical latitude. Instead the per capita measure correlates with the national per capita CFs, and it reaches on average 23% of the national impacts per capita. The three top performing universities are located in Switzerland, Chile, and Germany.
Conclusion: The usual reporting of CO2 emissions is categorized into Scopes 1–3 following the GHG Protocol Corporate Accounting Standard which makes comparison across universities challenging. In this study, we attempted to standardize the CF metrics, allowing us to objectively compare the CF at several universities. From this study, we observed that, almost 30 years after the Earth Summit in Rio de Janeiro (1992), the results are still limited. Only one zero emission university was identified, and hence, the transformation should speed up globally.
Radar signal processing is a promising tool for vital sign monitoring. For contactless observation of breathing and heart rate a precise measurement of the distance between radar antenna and the patient’s skin is required. This results in the need to detect small movements in the range of 0.5 mm and below. Such small changes in distance are hard to be measured with a limited radar bandwidth when relying on the frequency based range detection alone. In order to enhance the relative distance resolution a precise measurement of the observed signal’s phase is required. Due to radar reflections from surfaces in close proximity to the main area of interest the desired signal of the radar reflection can get superposed. For superposing signals with little separation in frequency domain the main lobes of their discrete Fourier transform (DFT) merge into a single lobe, so that their peaks cannot be differentiated. This paper evaluates a method for reconstructing the phase and amplitude of such superimposed signals.