Effect of post-induction substrate oscillation on recombinant alkaline phosphatase production expressed in Escherichia coli

Microorganisms are exposed to fast changes in microenvironment in large scale bioreactors. Because of their fast response to the changes, overall performance of biological system in small scale differs from large scale. Hence the variations in the environment that microorganisms are living in are mimicked in small scale. For this purpose one way is to feed substrate into the bioreactor in an oscillatory profile. In this work two different types of triangular oscillatory feeding profiles were applied as the post induction feeding strategy in intracellular recombinant alkaline phosphatase production expressed in Escherichia coli to find out if this biological system behaves in inhomogeneous environment differently. On line and offline measurements provide evaluation of product quality and quantity. Then the results of the experiments were compared with those of the control run at which constant feeding rate was executed. The results showed that oscillatory feeding at which cells were not starved led to higher yield of protein per substrate (0.027 C-mol/C-mol) and higher activity per protein (0.79 U/mg) when compared to a constant feeding rate (0.011 C-mol/C-mol and 0.11 U/mg).     

AnDiaTec Salmonella sp. PCR-ELISA for analysis of food samples

A commercially available PCR kit (AnDiaTec Salmonella sp. PCR-ELISA) was developed and evaluated for the detection of Salmonella sp. in food samples. The test is based on PCR amplification and hybridization of the amplified DNA to a microtiter plate followed by the detection of PCR product in the manner of an enzyme-linked immunosorbent assay test. The sensitivity and specificity were evaluated first with Salmonella pure cultures and artificially contaminated food samples, including food types for which an inhibition of the PCR reaction was expected. Both experiments proved a very good sensitivity, specificity, and reliability of the test with a very broad range of food types. In a second evaluation study, more than 1,100 food samples of different types were tested in parallel with the PCR method and with the International Standardization Organization 6579 bacteriological reference method. The results of this evaluation study and the results from other experiments on dilutions of artificially contaminated food samples led to the establishment of a positive-negative cutoff value (optical density at 450 nm of more than 0.9) with respect to the conventional bacteriological method. Using this positive-negative cutoff, 98% agreement to the bacteriological method was obtained.     

Investigation on electrostatic assist and gravure process parameters on solid mottle reduction for shrink films

Gravure is a high throughput printing process, normally associated with speed, quality, and long print runs. It is widely used for printing on shrink films and other substrates. The shrink films, in particular, polyvinyl chloride (PVC) and glycol-modified polyethylene terephthalate (PET-G), are two dominant substrates widely consumed and printed by gravure process. The PVC and PET-G offer different properties which greatly influence the printability. The surface energy of the substrate determines the adhesion and wettability of ink, while electrical properties such as surface and volume resistivity impact electrostatic assist (ESA) performance. The introduction of ESA in gravure further improved the print quality by eliminating dot skips with reduced impression pressure. However, print defects such as print mottle is inevitable. Print mottle occurs due to a discrepancy between substrate, ink, and process parameters which degrade the print quality. These complexities need to be addressed to deliver higher productivity with less print waste. Therefore, the study investigates the effect of process parameters, i.e., substrate type, line screen, air gap (distance between charge bar and impression roller), viscosity, voltage, and speed, and aims to quantify their effect numerically on defect minimization. The Design of Experiments (DOE) was generated for the above-mentioned parameters and analyzed to extract the best combination of process parameters. The optimized setting showed a reduction in solid mottle by 54% and 57% for PET-G and PVC, respectively.     

SARS-CoV-2 Spike Protein Binding of Glycated Serum Albumin—Its Potential Role in the Pathogenesis of the COVID-19 Clinical Syndromes and Bias towards Individuals with Pre-Diabetes/Type 2 Diabetes and Metabolic Diseases

The immune response to SARS-CoV-2 infection requires antibody recognition of the spike protein. In a study designed to examine the molecular features of anti-spike and anti-nucleocapsid antibodies, patient plasma proteins binding to pre-fusion stabilised complete spike and nucleocapsid proteins were isolated and analysed by matrix-assisted laser desorption ionisation–time of flight (MALDI-ToF) mass spectrometry. Amongst the immunoglobulins, a high affinity for human serum albumin was evident in the anti-spike preparations. Careful mass comparison revealed the preferential capture of advanced glycation end product (AGE) forms of glycated human serum albumin by the pre-fusion spike protein. The ability of bacteria and viruses to surround themselves with serum proteins is a recognised immune evasion and pathogenic process. The preference of SARS-CoV-2 for AGE forms of glycated serum albumin may in part explain the severity and pathology of acute respiratory distress and the bias towards the elderly and those with (pre)diabetic and atherosclerotic/metabolic disease.     

The live cell irradiation and observation setup at SNAKE

We describe a new setup at the ion microprobe SNAKE (Superconducting Nanoscope for Applied nuclear (Kern-) physics Experiments) at the Munich 14 MV Tandem accelerator that facilitates both living cell irradiation with sub micrometer resolution and online optical imaging of the cells before and after irradiation by state of the art phase contrast and fluorescence microscopy. The cells are kept at standard cell growth conditions at 37 °C in cell culture medium. After irradiation it is possible to switch from single ion irradiation conditions to cell observation within 0.5 s. First experiments were performed targeting substructures of a cell nucleus that were tagged by TexasRed labeled nucleotides incorporated in the cellular DNA by 55 MeV single carbon ion irradiation. In addition we show first online sequences of short time kinetics of Mdc1 protein accumulation in the vicinity of double strand breaks after carbon ion irradiation.     

Einfluss einer gezielt aufgeprägten Mikrogeometriestreuung auf das Anregungsverhalten von Kegelradverzahnungen

In the design process of transmissions, one major criterion is the resulting noise emission of the powertrain due to the gear excitation. Within the past years, a lot of investigations have shown that the noise emission can be correlated to the quasi-static transmission error. Therefore, the transmission error can be used as a characteristic value for quality assurance by experimental inspections as well as for a tooth contact analysis in the design process.The noise behavior of gearboxes is mainly caused by the excitation in the tooth mesh. The standardized design and calculation methods for gears concentrate on the reduction of the excitation level. But often the physical sound characteristics do not fit in with the human noise perception. Thus gear design rules are required that are able to rate the excitation according to the perception.The effect of the targeted topography scatter generally described is the reduction of the gear mesh amplitudes with an increase of the background noise. The objective of the report is to develop an understanding of the influence of different micro geometry scatters on the excitation behavior.Finally, it is the aim to design an aurally-accurate micro geometry scattering for the optimization of ground bevel gears in terms of tonality reduction and increased background noise. By means of a variant calculation and weighted target variables, psychoacoustic optimized micro geometry scattering is designed. At the same time, attention is also focused on a simple production of mixed topographies.