Mannheim University of Applied Sciences

About: Mannheim University of Applied Sciences is a education organization based out in Mannheim, Germany. It is known for research contribution in the topics: Gene & Mass spectrometry imaging. The organization has 593 authors who have published 779 publications receiving 19248 citations. The organization is also known as: FH Mannheim & Mannheim College.

Fast Nanoliter‐Scale Cell Assays Using Droplet Microarray–Mass Spectrometry Imaging

Abstract: In pharmaceutical research and development, cell-based assays are primarily used with readout that rely on fluorescence-based and other label-dependent techniques for analysis of different cellular processes Superhydrophobic-hydrophilic droplet microarrays (DMA) and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) have recently emerged as key technologies for miniaturized high-throughput cell assays and for label-free molecular high-content drug profiling, respectively Here, nanoliter-scale cell assays are integrated on DMAs with MALDI-MS imaging (MALDI-MSI) approaches to a droplet microarray-mass spectrometry imaging (DMA-MSI) platform Using A549 lung cancer cells, concentration-response profiling of a pharmaceutical compound, the fatty acid synthase inhibitor GSK2194069, are demonstrated Direct cell culture on DMAs enables combination of microscopy and high speed, high molecular content analysis using MALDI-MSI Miniaturization of array spots down to 05 mm confining 40 nL droplets allows for MALDI imaging analysis of as few as ten cells per spot Partial automation ensures a fast sample preparation workflow Taken together, the integrated DMA-MSI platform that combines MALDI-MSI, as a label-free analytical readout, with the miniaturized droplet microarray platform is a valuable complement to high throughput cell-based assays technologies

Sharing Proofs of Retrievability across Tenants

Abstract: Proofs of Retrievability (POR) are cryptographic proofs which provide assurance to a single tenant (who creates tags using his secret material) that his files can be retrieved in their entirety. However, POR schemes completely ignore storage-efficiency concepts, such as multi-tenancy and data deduplication, which are being widely utilized by existing cloud storage providers. Namely, in deduplicated storage systems, existing POR schemes would incur an additional overhead for storing tenants' tags which grows linearly with the number of users deduplicating the same file. This overhead clearly reduces the (economic) incentives of cloud providers to integrate existing POR/PDP solutions in their offerings. In this paper, we propose a novel storage-efficient POR, dubbed SPORT, which transparently supports multi-tenancy and data deduplication. More specifically, SPORT enables tenants to securely share the same POR tags in order to verify the integrity of their deduplicated files. By doing so, SPORT considerably reduces the storage overhead borne by cloud providers when storing the tags of different tenants deduplicating the same content. We show that SPORT resists against malicious tenants/cloud providers (and against collusion among a subset of the tenants and the cloud). Finally, we implement a prototype based on SPORT, and evaluate its performance in a realistic cloud setting. Our evaluation results show that our proposal incurs tolerable computational overhead on the tenants and the cloud provider.

[Recommended actions: Reinforcing clinicians' resilience and supporting second victims during the COVID-19 pandemic to maintain capacity in the healthcare system]

Abstract: The term second victim describes a person involved in patient care who, due to an extraordinary patient care situation, also becomes traumatized. This phenomenon is largely unknown to the general public, although it is widespread, and is being exacerbated by the COVID-19 pandemic. Pronounced psychological strain among clinicians entails the risk of increasing pressure on the healthcare system even further. The ensuing threat to the safety of both patients and staff needs to be taken seriously. The second victim phenomenon is extensively researched and requires a two-pronged strategy. Second victims need fast, personal and confidential support within a comprehensive, easily accessible, stratified system and reinforcing clinicians' resilience is crucial. Leadership and appropriate crisis communication can sustainably support clinicians' resilience, and thus their ability to function effectively in the long term. Consequently, management can make both a short-term as well as a sustainable contribution to patient safety and therefore increasing the chances of survival for many patients during and after the COVID-19 pandemic.

On-line and real time cell counting and viability determination for animal cell process monitoring by in situ microscopy

Abstract: BackgroundTwo of the key parameters to be monitored during cellcultivation processes are cell concentration and viability.Until today, this is very often done off-line by sterilesampling and subsequent counting using a hemocyt-ometer or an electronic cell counter. Cell biology lacks ameasurable quantity by which single cells in suspensioncan be non-invasively diagnosed as dead or alive. How-ever, it would be of significance for process monitoringand in the light of initiatives like PAT if cell density aswell as viability could be determined directly and on-line.Optical measurement of cell density byin situ micro-scopy eliminates the need for sampling and allows forcontinuous monitoring of this key parameter; see e.g.[1,2]; Guez et al. [1] describe an in situ microscope(ISM) which does not use any moving mechanical partswithin or outside the fermentation vessel. It transmits inreal time images taken directly in the stirred suspensionwithin the bioreactor. Image data is processed and eval-uated to provide monitoring of cell-density and mor-phological parameters, e.g. cell size, by means ofassessing the obtained in situ cell-micrographs.Previously, we have extendedin situ microscopytowards viability assessment of suspended cells [3,5].Now, we present new findings on this topic and showthat in cultures of suspended cells, cell-death corre-sponds to measurable changes in morphometric para-meters as e.g. variance, contrast or entropy of thegreyvalues of in situ cell-micrographs. As an example,here we show viability determination via greyvaluedispersion.Material and methodsWe use a custom built high resolution ISM (HS Man-nheim) with water immersion objective, 40x magnifica-tion, numerical aperture 0.75 equipped with optical fiberillumination. Data acquisition is at 0.3– 15 frames persecond, frames have 1293x1040 pixels; primary dataanalysis results in cell micrographs (Figure 1a). We haveapplied the system to bench top and larger bioreactors(see e.g. [4]) and worked mainly with Jurkat, CHO andHybridoma cells.For the experiment presented in Figure 1a/b, cells arecultivated in a Biostat C30 (Sartorius BBI Systems) withthe ISM inserted in one of the existing probe ports. Pro-prietary hybridoma cells (InVivo BioTech Services) arecultured in serum free ISF-1 (InVivo BioTech Services;Biochrom AG) and monitored over the full length ofthe fermentation (not shown).For the experiment presented in Figure 1c, cells arecultivated in a custom built autoclavable steel bench topreactor(HSMannheim)with25mmporttoaccommo-date the ISM and a working volume of 0.7 L. To testreal time and viability determination capabilities of thesystem over a wide range of viabilities in a short time,cells were challenged with 3% Ethanol at 42 hours. Cellcounts (not shown) and viability were determined by insitu microscopy and, as reference, by means of a ViCellcell viability analyser (Beckman Coulter) and Flow Cyto-metry (Partec) using Annexin V / FITC and PI staining.Jurkat cells (DSMZ ACC 282) were cultured in 90%RPMI 1640 + 10% FBS.

Absorption and emission spectroscopic characterisation of 8-amino-riboflavin

Abstract: The flavin dye 8-amino-8-demethyl- d -riboflavin (AF) in the solvents water, DMSO, methanol, and chloroform/DMSO was studied by absorption and fluorescence spectroscopy. The first absorption band is red-shifted compared to riboflavin, and blue-shifted compared to roseoflavin (8-dimethylamino-8-demethyl-D-riboflavin). The fluorescence quantum yield of AF in the studied solvents varies between 20% and 50%. The fluorescence lifetimes were found to be in the 2–5 ns range. AF is well soluble in DMSO, weakly soluble in water and methanol, and practically insoluble in chloroform. The limited solubility causes AF aggregation, which was seen in differences between measured absorption spectra and fluorescence excitation spectra. Light scattering in the dye absorption region is discussed and approximate absorption cross-section spectra are determined from the combined measurement of transmission and fluorescence excitation spectra. The photo-stability of AF was studied by prolonged light exposure. The photo-degradation routes of AF are discussed.