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The implementation of single-use technologies offers several major advantages, e.g. prevention of cross-contamination, especially when spore-forming microorganisms are present. This study investigated the application of a single-use bioreactor in batch fermentation of filamentous fungus Penicillium sp. (IBWF 040-09) from the Institute of Biotechnology and Drug Research (IBWF), which is capable of intracellular production of a protease inhibitor against parasitic proteases as a secondary metabolite. Several modifications to the SU bioreactor were suggested in this study to allow the fermentation in which the fungus forms pellets. Simultaneously, fermentations in conventional glass bioreactor were also conducted as reference. Although there are significant differences in the construction material and gassing system, the similarity of the two types of bioreactors in terms of fungal metabolic activity and the reproducibility of fermentations could be demonstrated using statistic methods. Under the selected cultivation conditions, growth rate, yield coefficient, substrate uptake rate, and formation of intracellular protease-inhibiting substance in the single-use bioreactor were similar to those in the glass bioreactor.
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%.
Productive biofilms are gaining growing interest in research due to their potential of producing valuable compounds and bioactive substances such as antibiotics. This is supported by recent developments in biofilm photobioreactors that established the controlled phototrophic cultivation of algae and cyanobacteria. Cultivation of biofilms can be challenging due to the need of surfaces for biofilm adhesion. The total production of biomass, and thus production of e.g. bioactive substances, within the bioreactor volume highly depends on the available cultivation surface. To achieve an enlargement of surface area for biofilm photobioreactors, biocarriers can be implemented in the cultivation. Thereby, material properties and design of the biocarriers are important for initial biofilm formation and growth of cyanobacteria. In this study, special biocarriers were designed and additively manufactured to investigate different polymeric materials and surface designs regarding biofilm adhesion of the terrestrial cyanobacterium Nostoc flagelliforme (CCAP 1453/33). Properties of 3D-printed materials were characterized by determination of wettability, surface roughness, and density. To evaluate the influence of wettability on biofilm formation, material properties were specifically modified by gas-phase fluorination and biofilm formation was analyzed on biocarriers with basic and optimized geometry in shaking flask cultivation. We found that different polymeric materials revealed no significant differences in wettability and with identical surface design no significant effect on biomass adhesion was observed. However, materials treated with fluorination as well as optimized biocarrier design showed improved wettability and an increase in biomass adhesion per biocarrier surface.
The purpose of this article is to evaluate optimal expected utility risk measures (OEU) in a risk-constrained portfolio optimization context where the expected portfolio return is maximized. We compare the portfolio optimization with OEU constraint to a portfolio selection model using value at risk as constraint. The former is a coherent risk measure for utility functions with constant relative risk aversion and allows individual specifications to the investor’s risk attitude and time preference. In a case study with three indices, we investigate how these theoretical differences influence the performance of the portfolio selection strategies. A copula approach with univariate ARMA-GARCH models is used in a rolling forecast to simulate monthly future returns and calculate the derived measures for the optimization. The results of this study illustrate that both optimization strategies perform considerably better than an equally weighted portfolio and a buy and hold portfolio. Moreover, our results illustrate that portfolio optimization with OEU constraint experiences individualized effects, e.g., less risk-averse investors lose more portfolio value in the financial crises but outperform their more risk-averse counterparts in bull markets.
Hydrochar derived from Argan nut shell (ANS) was synthesized and applied to remove bisphenol A (BPA) and diuron. The results indicated that the hydrochar prepared at 200 °C (HTC@ANS-200) possessed a higher specific surface area (42 m2/g) than hydrochar (HTC@ANS-180) prepared at 180 °C (17 m2/g). The hydrochars exhibited spherical particles, which are rich in functional groups. The HTC@ANS-200 exhibited high adsorption efficiency, of about 92% of the BPA removal and 95% of diuron removal. The maximum Langmuir adsorption capacities of HTC@ANS-200 at room temperature were 1162.79 mg/for Bisphenol A and 833.33 mg/g for diuron (higher than most reported adsorbents). The adsorption process was spontaneous (− ΔG°) and exothermic (− ΔH°). Excellent reusability was reclaimed after five cycles, the removal efficiency showed a weak decrease of 4% for BPA and 1% for diuron. The analysis of Fourier transforms infrared spectrometry demonstrated that the aromatic C=C and OH played major roles in the adsorption mechanisms of BPA and diuron in this study. The high adsorption capacity was attributed to the beneficial porosity (The pore size of HTC@ANS-200 bigger than the size of BPA and diuron molecule) and surface functional groups. BPA and diuron adsorption occurred also via multiple adsorption mechanisms, including pore filling, π–π interactions, and hydrogen bonding interactions on HTC@ANS-200.
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.
A comprehensive overview is provided evaluating direct real-world CO2 emissions of both diesel and petrol cars newly registered in Europe between 1995 and 2015. Before 2011, European diesel cars emitted less CO2 per kilometre than petrol cars, but since then there is no appreciable difference in per-km CO2 emissions between diesel and petrol cars. Real-world CO2 emissions of diesel cars have not declined appreciably since 2001, while the CO2 emissions of petrol cars have been stagnant since 2012. When adding black carbon related CO2-equivalents, such as from diesel cars without particulate filters, diesel cars were discovered to have had much higher climate relevant emissions until the year 2001 when compared to petrol cars. From 2001 to 2015 CO2-equivalent emissions from new diesel cars and petrol cars were hardly distinguishable. Lifetime use phase CO2-equivalent emissions of all European passenger vehicles were modelled for 1995–2015 based on three scenarios: the historic case, another scenario freezing percentages of diesel cars at the low levels from the early 1990s (thus avoiding the observed “boom” in new diesel registrations), and an advanced mitigation scenario based on high proportions of petrol hybrid cars and cars burning gaseous fuels. The difference in CO2-equivalent emissions between the historical case and the scenario avoiding the diesel car boom is only 0.4%. The advanced mitigation scenario would have been able to achieve a 3.4% reduction in total CO2-equivalent emissions over the same time frame. The European diesel car boom appears to have been ineffective at reducing climate-warming emissions from the European transport sector.
Electric drive systems are increasingly used in automobiles. However, the combination of comfort, dynamics and safety requirements places high demands on the torque accuracy. The complex interplay of battery, inverter and electrical machine causes a lot of system uncertainties based on parameter fluctuations and measurement errors that influence the system performance. In this paper these influences on the closed loop torque control are analyzed and quantified using a variance based sensitivity analysis. The method enables to connect the variance of the torque accuracy with the parameter uncertainties causing this variance. Moreover, it quantifies the influences of the parameters independent of the complexity of the analyzed system. In addition, two methods to ensure convergence of the estimated variance based sensitivity measures are proposed. The results of the analysis are presented for 19 static working points of an battery electric drive system.
Passenger cars in Europe have become both heavier and more powerful over the past decades. This trend has increased vehicle utility but it might have also offset technical improvements in powertrain efficiency. Here, we analyze efficiency trade-offs and CO2 emissions for three popular compact cars in Germany. We find that mass, power, and front area of model variants has increased by 66%, 147%, and 22%, respectively between 1980 and 2018. In the same period, fuel consumption decreased 14% for gasoline models but it increased 9% for diesel models. However, if vehicle mass, power, and front area had remained at 1980 levels, technical efficiency improvements would have decreased the fuel consumption of gasoline and diesel models by 23% and 24%, respectively. The related efficiency trade-offs amount to 24 g CO2/km or 13% of the current fuel consumption for gasoline models and 40 g CO2/km or 25% of the current fuel consumption for diesel models. These findings suggest that about half of the technical efficiency improvements in gasoline models and all of the technical efficiency improvements in diesel models are offset through other vehicle attributes. By accounting for the observed efficiency trade-offs, climate policy could become more effective.