In: Chemie Ingenieur Technik, 109(7), 1151. ISSN 0009286X

Abstract: In dieser Arbeit wird die Möglichkeit aufgezeigt, mithilfe einer sehr empfindlichen Einzel‐Photonenmesstechnik Streulicht von Partikeln zu messen, das durch den Einsatz von Lasern mit extrem geringen Laserintensitäten erzeugt wurde. Die entstehenden Vorteile für nach diesem Prinzip zu bauende Geräte liegen im Bereich Explosionsschutz, Vermeidung von Produktschädigungen und Arbeitssicherheit. Es werden Messaufbauten zum Vermessen von Suspensionen und Aerosolen beschrieben. Laserleistungen kleiner 5 µW werden eingesetzt. Die Messungen beginnen ab einer Partikelgröße von 200 nm und werden in einer 90°‐Anordnung als worst case durchgeführt, da bei diesem Winkel ein Minimum an Streuung vorliegt.

Tobias Teumer, Thomas Hufnagel, Thomas Schäfer, Robert Schlarp‐Horvath, Heike P. Karbstein, Frank‐Jürgen Methner, Matthias Rädle

In: Proteomics Clin. Appl. 2018, 1800029

Abstract: PURPOSE: To facilitate the transition of MALDI-MS Imaging (MALDI-MSI) from basic science to clinical application, it is necessary to analyze formalin-fixed paraffin-embedded (FFPE) tissues. The aim is to improve in situ tryptic digestion for MALDI-MSI of FFPE samples and determine if similar results would be reproducible if obtained from different sites.
EXPERIMENTAL DESIGN: FFPE tissues (mouse intestine, human ovarian teratoma, tissue microarray of tumor entities sampled from three different sites) are prepared for MALDI-MSI. Samples are coated with trypsin using an automated sprayer then incubated using deliquescence to maintain a stable humid environment. After digestion, samples are sprayed with CHCA using the same spraying device and analyzed with a rapifleX MALDI Tissuetyper at 50 µm spatial resolution. Data are analyzed using flexImaging, SCiLS, and R.
RESULTS: Trypsin application and digestion are identified as sources of variation and loss of spatial resolution in the MALDI-MSI of FFPE samples. Using the described workflow, it is possible to discriminate discrete histological features in different tissues and enabled different sites to generate images of similar quality when assessed by spatial segmentation and PCA.
CONCLUSIONS AND CLINICAL RELEVANCE: Spatial resolution and site-to-site reproducibility can be maintained by adhering to a standardized MALDI-MSI workflow.

Ly A1, Longuespée R2, Casadonte R3, Wandernoth P3, Schwamborn K4, Bollwein C4, Marsching C5, Kriegsmann K6, Hopf C5, Weichert W4, Kriegsmann J3, Schirmacher P2, Kriegsmann M2, Deininger SO1.

1Bruker Daltonik GmbH, Bremen, Germany.
2Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany.
3Proteopath GmbH, Trier, Germany.
4Institute of Pathology, Technical University of Munich, Munich, Germany.
5Center for Biomedical Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Mannheim, Germany.
6Department of Hematology, Oncology and Rheumatology, University Hospital Heidelberg, Heidelberg, Germany.

In: Journal of Sensors and Sensor Systems, 7, 543-549, 2018

Abstract: This paper presents an explosion-proof two-channel Raman photometer designed for chemical process monitoring in hazardous explosive atmospheres. Due to its design, alignment of components is simplified and economic in comparison to spectrometer systems. Raman spectrometers have the potential of becoming an increasingly important tool in process analysis technologies as part of molecular-specific concentration monitoring. However, in addition to the required laser power, which restricts use in potentially explosive atmospheres, the financial hurdle is also high. Within the scope of a proof of concept, it is shown that photometric measurements of Raman scattering are possible. The use of highly sensitive detectors allows the required excitation power to be reduced to levels compliant for operation in potentially explosive atmospheres. The addition of an embedded platform enables stable use as a self-sufficient sensor, since it carries out all calculations internally.

Multi-pixel photon counters (MPPCs) with large detection areas of 1350 µm2 are implemented as detectors. As a result, the sensitivity of the sensor is strongly increased. This gain in sensitivity is primarily achieved through two characteristics: first, the operating principle “avalanche breakdown” to detect single photons is used; second, the size of the image projected onto the MPPC is much bigger than the pixel area in competing Raman-Spectrometers resulting in higher photon flux. This combination enables reduction of the required excitation power to levels compliant for operation in potentially explosive atmospheres. All presented experiments are performed with strongly attenuated laser power of 35 mW. These include the monitoring of the analytes ethanol and hydrogen peroxide as well as the reversible binding of CO2 to amine. Accordingly, the described embedded sensor is ideally suited as a process analytical technology (PAT) tool for applications in environments with limitations on power input.

Marcel Nachtmann1, Shaun Paul Keck1, Frank Braun1, Hanns Simon Eckhardt2, Christoph Mattolat2, Norbert Gretz3, Stephan Scholl4, and Matthias Rädle1

1Institute for Process Control and Innovative Energy Conversion, Mannheim University of Applied Sciences, Mannheim, 68163, Germany
2tec5 AG, Oberursel/Ts, 61440, Germany
3Medical Research Center, Medical Faculty Mannheim, Heidelberg University, Mannheim, 68167, Germany
4Institute for Chemical and Thermal Process Engineering, Technical University Braunschweig, Braunschweig, 38106, Germany

Prof. Dr. Rüdiger Rudolf über die Entwicklung von „Organoiden“ auf Basis komplexer 3D-Zellkulturen für das Screening neuer bioaktiver Wirkstoffe für Hautpflegeprodukte und die Biomedizin.

Organoide – Wirkstoffscreening treibt Entwicklung neuer Technologien voran
Bislang beruhte das Screening von Wirkstoffen und Arzneimitteln meist entweder auf zweidimensionalen (2D) Zellkulturen oder auf Tierversuchen. Beiden Ansätzen wohnen jedoch bedeutende Nachteile inne. Zum einen sind 2D-Zellkulturen zwar preisgünstig in der Herstellung und Durchführung der Versuche, sie sind für hohe Durchsatzraten geeignet und sie erlauben die Verwendung von Zellen menschlichen Ursprungs, flache Petrischalen stellen jedoch aufgrund ihrer Steifigkeit und der Beschränkung der Kontaktbildung auf eine Seite der Zelloberfläche eine höchst unnatürliche Umgebung für das Screening dar. Darüber hinaus gestaltet sich die Durchdringung der Zellen mit Wirkstoffen ebenso wie die Verteilung von Abfallprodukten, Sauerstoff und Nährstoffen in solchen 2D-Kulturen völlig anders als in den dreidimensionalen Anordnungen echten menschlichen Gewebes.

Bitte hier Blickwinkel Structures Oktober 2018 weiterlesen.

In: International Journal of Molecular Sciences, Vol. 19, Issue 7

Abstract: Vertebrate neuromuscular junctions (NMJs) have been conceived as tripartite synapses composed of motor neuron, Schwann cell, and muscle fiber. Recent work has shown the presence of sympathetic neurons in the immediate vicinity of NMJs and experimental and clinical findings suggest that this plays an eminent role in adult NMJ biology. The present study examined the postnatal development and distribution of sympathetic innervation in different muscles using immunofluorescence, confocal microscopy, and Western blot. This demonstrates the proximity of sympathetic neurons in diaphragm, extensor digitorum longus, tibialis anterior, soleus, and levator auris longus muscles. In extensor digitorum longus muscle, sympathetic innervation of NMJs was quantified from perinatal to adult stage and found to increase up to two months of age. In diaphragm muscle, an extensive network of sympathetic neurons was prominent along the characteristic central synapse band. In summary, these data demonstrate that an elaborate sympathetic innervation is present in several mouse skeletal muscles and that this is often next to NMJs. Although the presence of sympathetic neurons at the perisynaptic region of NMJs increased during postnatal development, many synapses were already close to sympathetic neurons at birth. Potential implications of these findings for treatment of neuromuscular diseases are discussed.

Tatjana Straka1,2,3, Veronica Vita4, Kaltrina Prokshi1,2, Sarah Janice Hörner1,2, Muzamil Majid Khan1,2,3, Marco Pirazzini4, Marion Patrick Ivey Williams1, Mathias Hafner1, Tania Zaglia4,5,6 and Rüdiger Rudolf1,2,3
1 Institute of Molecular and Cell Biology, Mannheim University of Applied Sciences, 68163 Mannheim, Germany
2 Interdisciplinary Center for Neurosciences, Heidelberg University, 69120 Heidelberg, Germany
3 Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, 76344 Eggenstein-Leopoldshafen, Germany
4 Venetian Institute of Molecular Medicine, 35129 Padua, Italy
5 Department of Biomedical Sciences, University of Padua, 35131 Padua, Italy
6 Department of Cardiac Thoracic and Vascular Sciences, University of Padua, 35128 Padua, Italy

In:Frontiers in Molecular Neuroscience - Front. Mol. Neurosci. 2018, 11, 206

Abstract: Alzheimer's disease (AD) is characterized by progressive neurodegeneration and a concomitant loss of synapses and cognitive abilities. Recently, we have proposed that an alteration of neuronal membrane lipid microdomains increases neuronal resistance toward amyloid-β stress in cultured neurons and protects from neurodegeneration in a mouse model of AD. Lipid microdomains are highly enriched in a specific subclass of glycosphingolipids, termed gangliosides. The enzyme glucosylceramide synthase (GCS) catalyzes the rate-limiting step in the biosynthesis of these gangliosides. The present work now demonstrates that genetic GCS deletion in subsets of adult forebrain neurons significantly improves the spatial memory and counteracts the loss of dendritic spines in the hippocampal dentate gyrus of 5x familial AD mice (5xFAD//Ugcgf/f//Thy1-CreERT2//EYFP mice), when compared to 5xFAD//Ugcgf/f littermates (5xFAD mice). Aberrantly activated glial cells and their expression of pro-inflammatory cytokines have emerged as the major culprits for synaptic loss in AD. Typically, astrocytic activation is accompanied by a thickening of astrocytic processes, which impairs astrocytic support for neuronal synapses. In contrast to 5xFAD mice, 5xFAD//Ugcgf/f//Thy1-CreERT2//EYFP display a less pronounced thickening of astrocytic processes and a lower expression of tumor necrosis factor-α and interleukin 1-α in the hippocampus. Thus, this work further emphasizes that GCS inhibition may constitute a potential therapeutic target against AD.

Herzer S1,2, Hagan C1,3, von Gerichten J1,4, Dieterle V1,2, Munteanu B5, Sandhoff R1,4, Hopf C5, Nordström V1,2.

1Division of Cellular and Molecular Pathology, German Cancer Research Center, Heidelberg, Germany.
2Interdisciplinary Center for Neurosciences, Heidelberg University, Heidelberg, Germany.
3Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO, United States.
4Lipid Pathobiochemistry Group, German Cancer Research Center, Heidelberg, Germany.
5Center for Mass Spectrometry (CeMOS), University of Heidelberg and Mannheim University of Applied Sciences, Mannheim, Germany.

In: Bioconjugate Chem., 2018, 29 (5), pp 1525–1533

Abstract: Gold nanoparticles (AuNPs) have widely been used for 70 years in cancer treatment, but only in the last 15 years has the focus been on specific AuNPs with homogeneous size and shape for various areas in science. They constitute a perfect platform for multifunctionalization and therefore enable the enhancement of target affinity. Here we report on the development of tumor specific AuNPs as diagnostic tools intended for the detection of prostate cancer via fluorescence imaging and positron emission tomography (PET). The AuNPs were further evaluated in vitro and in vivo and exhibited favorable diagnostic properties concerning tumor cell uptake, biodistribution, clearance, and tumor retention.

Marc Pretze*† , Andreas Hien‡, Matthias Rädle‡, Ralf Schirrmacher§ , Carmen Wängler∥, and Björn Wängler*†
†Molecular Imaging & Radiochemistry, Department of Clinical Radiology and Nuclear Medicine and ∥Biomedical Chemistry, Medical Faculty Mannheim of Heidelberg University, Mannheim 68167, Germany
‡ Institute of Process Control and Innovative Energy Conversion, Mannheim University of Applied Sciences, Mannheim 68163, Germany
§ Oncologic Imaging, Department of Oncology, University of Alberta, Edmonton 6820, Alberta, Canada

In: Scientific Reports 8, Article number: 313 (2018)

Abstract: Multimodal imaging combines complementary platforms for spatially resolved tissue analysis that are poised for application in life science and personalized medicine. Unlike established clinical in vivo multimodality imaging, automated workflows for in-depth multimodal molecular ex vivo tissue analysis that combine the speed and ease of spectroscopic imaging with molecular details provided by mass spectrometry imaging (MSI) are lagging behind. Here, we present an integrated approach that utilizes non-destructive Fourier transform infrared (FTIR) microscopy and matrix assisted laser desorption/ionization (MALDI) MSI for analysing single-slide tissue specimen. We show that FTIR microscopy can automatically guide high-resolution MSI data acquisition and interpretation without requiring prior histopathological tissue annotation, thus circumventing potential human-annotation-bias while achieving >90% reductions of data load and acquisition time. We apply FTIR imaging as an upstream modality to improve accuracy of tissue-morphology detection and to retrieve diagnostic molecular signatures in an automated, unbiased and spatially aware manner. We show the general applicability of multimodal FTIR-guided MALDI-MSI by demonstrating precise tumor localization in mouse brain bearing glioma xenografts and in human primary gastrointestinal stromal tumors. Finally, the presented multimodal tissue analysis method allows for morphology-sensitive lipid signature retrieval from brains of mice suffering from lipidosis caused by Niemann-Pick type C disease.

Jan-Hinrich Rabe1,2, Denis A. Sammour1,2, Sandra Schulz1,2, Bogdan Munteanu1,2, Martina
Ott3, Katharina Ochs3,5, Peter Hohenberger4, Alexander Marx4, Michael Platten3,4,
Christiane A. Opitz5,6, Daniel S. Ory7 & Carsten Hopf1,2

1Center for Applied Research in Applied Biomedical Mass Spectrometry (ABIMAS), Mannheim University of Applied Sciences, Mannheim, Germany
2Institute of Medical Technology, Heidelberg University and Mannheim University of Applied Sciences, Mannheim, Germany
3German Cancer Consortium (DKTK) CCU Neuroimmunology and Brain Tumor Immunology, German Cancer Research Center (DKFZ), Heidelberg, Germany
4University Medical Center Mannheim of Heidelberg University, Mannheim, Germany
5Brain Cancer Metabolism Group, German Cancer Research Center (DKFZ), Heidelberg, Germany
6Department of Neurology and National Center of Tumor Diseases, University Hospital Heidelberg, Heidelberg, Germany
7Diabetic Cardiovascular Disease Center and Department of Medicine, Washington University School of Medicine

In: Journal of Cellular Biotechnology 3 (2017) 41–50

Abstract: In recent years, many different methods were introduced for generation of 3D cell culture. However, many currently available three-dimensional techniques are not suitable for certain cell lines and sometimes showed a lack of reproducibility. Therefore, specific protocols for cell lines are needed. In this work, we demonstrate different methods of generating 3D cell culture for SCC4 tongue cancer cell line and discuss their applicability. Using three different methods, tumor spheroids were generated from SCC4 cells and cultured for 20 days. To investigate the influence of initial seeding density on spheroid morphology and size during long term culture, the same set of different cell numbers was used for each method. Using phase-contrast microscopy, spheroids were monitored until day 20 and their sizes were determined. We observed that spheroids were formed within 24 hours regardless of the method and initial cell density. Further, in all groups the spheroid size was maximal at day 2, followed by a decline until day 20. Spheroids remained stable until day 20 independent of initial seeding concentration in all groups. We have generated long-term culture spheroids of SCC4 cells. The size of the spheroids can be influenced by varying the initial cell seeding density until day 20. This may be useful if different sizes of spheroids are required, e.g. in hypoxia research.

Valeh Rustamova,b, Rüdiger Rudolfa,b, Vugar Yagubluc, Hella-Monika Kuhna, Mario Vitacolonnaa,1 and Mathias Hafnera,b,1

aInstitute of Molecular and Cell Biology, Faculty of Biotechnology, Mannheim University
of Applied Sciences, Mannheim, Germany
bInstitute of Medical Technology, Heidelberg University, Heidelberg, Germany
cDepartment of General, Visceral, Vascular, and Thoracic Surgery, Frankfurt Hoechst Hospital,
Frankfurt, Germany

In: Journal of Cellular Biotechnology 3 (2017) 61–80

Abstract: Major pathomechanisms underlying neurodegenerative diseases, such as Parkinson’s Disease, are still not well understood. Induced human pluripotent and rodent embryonic stem cells provide powerful disease models to address neurodegeneration-inducing pathomechanisms on a molecular and cellular level. Our aim is to establish a refined protocol to generate healthy and patient donor stem cell-derived dopaminergic neurons to investigate neurodegenerative events in vitro.METHODSHuman healthy donor- and patient-derived induced pluripotent stem cells were differentiated into stable dopaminergic progenitor cell lines and further differentiated into dopaminergic neurons. Induced pluripotent stem cells, neuronal progenitors and terminally differentiated neurons were characterized by confocal laser microscopy-based immunofluorescence analysis, live cell imaging demonstrating dopamine transporter-specific uptake of a fluorescent substrate and transcriptome analysis. Based on our immunofluorescence analysis, dopaminergic differentiation approaches predominantly yield dopaminergic neurons and GFAP-expressing glial cells. We detected a small partition of GABAergic neurons, yet neither serotonergic nor glutamatergic neurons. Dopaminergic neurons were successfully stained for pre- and postsynaptic and mitochondrial markers. Live cell imaging experiments verified dopamine transporter-dependent uptake of the fluorescent monoamine transporter substrate ASP+. Human stem cell-derived dopaminergic neurons are a suitable cellular system for fluorescence-based experimental approaches to address neurodegenerative events in vitro.

Yasmina Martí a,b,1, Elina Nürnberga,c,1, Sandra Horschitzb, Mathias Hafnerc, Patrick Schlossb, Andreas Meyer-Lindenbergb and Thorsten Laua

aCentral Institute of Mental Health, Hector Institute for Translational Brain Research, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany
bCentral Institute of Mental Health, Medical Faculty Mannheim, Heidelberg University, Department Psychiatry and Psychotherapy, Mannheim, Germany
cInstitute of Molecular and Cell Biology, Mannheim University of Applied Sciences, Mannheim, Germany

In: Journal of Cellular Biotechnology 3 (2017) 21–39

Abstract: Skin fulfils a plethora of eminent physiological functions ranging from physical barrier over immunity shield to the interface mediating social interaction. Prone to several acquired and inherited diseases, skin is therefore a major target of pharmaceutical and cosmetic research. The lack of similarity between human and animal skin and rising ethical concerns in the use of animal models have driven the search for novel realistic three-dimensional skin models. This review provides a survey of contemporary skin models and compares them in terms of applicability, reliability, cost and complexity.

Julia Klicksa, Elena von Molitora, Torsten Ertongur-Fauthb, Rüdiger Rudolfc and Mathias Hafnerc

aMannheim University of Applied Sciences, Institute of Molecular and Cell Biology, Mannheim, Germany
bBRAIN AG, Zwingenberg, Germany
cInstitute of Medical Technology, Heidelberg University, Heidelberg, Germany