cGMP-Signaltransduktion in vaskulären Prozessen der Niere(S. 252)
Jens Schlossmann, Elisabeth Schinner, Andrea Schramm
The second messenger cyclic guanosine monophosphate (cGMP) regulates various (patho-)physiological peripheral and central processes. Elucidation of cGMP-mediated mechanisms – the cGMP signalling – is essential to comprehend these diverse functions. Recently, some of these signalling pathways were established in the renovascular system. Furthermore, new drugs were approved which are promising for treatment of thrombotic and hypertensive diseases. Here, we provide an overview regarding cGMP signalling and its future therapeutical options in the renovascular system.
Export von Membranproteinen durch bakterielle Injektionsnadeln(S. 256)
Lea Krampen, Samuel Wagner
Many pathogenic bacteria use type III secretion systems (T3SS) to deliver effector proteins directly into the host cell cytoplasm. Substrate proteins of these systems are mostly soluble, however, a number of hydrophobic T3SS substrates are known to insert into host cell membranes, e. g. to form pores or receptors. Here, we discuss how bacteria ensure secretion of these membrane proteins through T3SS and prevent erroneous insertion into the bacterial inner membrane.
Kannibalismus und Brudermord: Warum Bakterien Artgenossen töten(S. 260)
Diana Wolf, Philipp Popp, Thorsten Mascher
Antibiotics are classically viewed as ‘biological weapons’ that allow bacteria to thrive and survive in competitive environments. Intuitively, they therefore target cells of different, competing species. A number of recent reports shed light on a novel facet of antibiotic action by discovering antimicrobial compounds that target the producing species itself. The relevance of these processes, termed cannibalism or fratricide, is discussed in an evolutionary and ecological context.
Stoffwechsel 2.0: Retrosynthese des mikrobiellen Metabolismus(S. 264)
Tobias J. Erb
VAAM-Forschungspreis 2017 Synthetic biology is a fast growing field with big promise for biotechnological applications. In our laboratory, we aim at realizing synthetic metabolism in a truly de novo approach via metabolic retrosynthesis. This includes the discovery and engineering of novel enzymes to sustain our designer pathways, the in vitro reconstitution and optimization of the synthetic routes, as well as the transplantation of these artificial pathways into living organisms.
Vom Einsatz polymerbasierter Lipidnanodiscs sowie in situ-Methoden(S. 267)
Oliver P. Ernst, Jana Bröcker
The structure determination of membrane proteins by X-ray crystallography is plagued by two major bottlenecks: the use of compromising detergents for protein extraction and purification as well as the necessity to harvest small crystals. With the aid of polymer-bounded lipid nanodiscs, membrane proteins can be crystallized without ever removing the protein from a lipid bilayer. New in situ strategies, where data are collected from unperturbed crystals, avoid crystal harvesting.
Etablierung filamentöser Mischkulturen für Bioprozesse(S. 270)
Miriam A. Rosenbaum, Ivan Schlembach, Lars Regestein
Defined filamenteous co-cultures have a high potential to translate the extraordinary economy of natural multi-species biomass conversion to biotechnological processes. This article highlights the methodological challenges for the development and engineering of consolidated bioprocesses for the direct conversion of cellulose to target biochemicals with filamenteous mixed cultures.
Acetatmessung mit MIR-Transmissionsspektroskopie bei der E. coli-Kultur(S. 273)
Johannes Hofmann, Ralf Masuch, Andreas Wolf, Anne Steinkämper, Richard Biener
A new approach of MIR transmission spectroscopy for online monitoring of substrates and metabolites during a cultivation process is described. For the quantitative analysis a calibration-free predictive algorithm has been developed. This method allows the simultaneous determination of substrates such as glucose, ammonia, phosphate, sulphate and proline and of by-products such as acetate, lactate, succinate and pyruvate.
HCS-Mikroskopie – ein Schlüssel zu intrazellulären Schmerzmechanismen(S. 277)
Tim Hucho, Jörg Isensee
Pain sensitivity is strongly modulated by intracellular signalling. The analysis of pain signalling is hampered by small numbers of neurons, high cellular heterogeneity, and near absence of transfectability. HCS microscopy allowed us to proof that nociceptive neurons express specific signalling components, to analyze signalling activity of endogenous signalling components with single cell resolution, and thereby to suggest blockers of Nav1.7 as novel therapeutic approach to boost the efficacy of opioids.
Markerfreie Karyotypisierung von Chromosomen durch spektrales Imaging(S. 282)
Rudolf W. Kessler, Edwin Ostertag, Karsten Rebner
Karyotyping is currently based on staining techniques. These methods lack reproducibility, are time-consuming and require complex expert knowledge. This paper introduces a new marker-free, fully automated and reliable screening procedure for chromosomes. The method is based on multimodal, spatially resolved spectroscopy combined with multivariate data analysis and can easily be integrated into a standard light microscope.
Maßgeschneiderte Methoden und Technologien für high content imaging(S. 286)
Daniel F. Gilbert
High content imaging has become an indispensable and probably the most frequently applied research tool in modern cell biology. To make use of its versatile applicability, the available instrumental infrastructure has to meet specific requirements which are defined by the employed experimental approach and increase along with the complexity of the assay technique used for assessing a specific biological question. Exemplary assay techniques and technology of various complexity are portrayed in this article.
Anwendungen & Produkte
Vereinzelung von prokaryotischen und eukaryotischen Zellen(S. 298)
André Gross, Stefan Zimmermann, Julian Riba, Benjamin Steimle
Single-cell analysis emerged as a promising tool to analyze the heterogeneity of complex cell populations by characterizing each individual cell. Conventional isolation workflows or instruments are often not able to isolate both eukaryotic and prokaryotic cells or require complex and costintensive devices. The single-cell printer (SCP) – an instrument for automated single-cell isolation and deposition – allows precise cell deposition for subsequent molecular analyses on the single-cell level, and efficient clonal cell line development.
Miniaturisierte menschliche Organe auf dem Chip(S. 301)
Tobias Hasenberg, Eva-Maria Dehne, Roland Lauster, Uwe Marx
New regulations and novel substance classes made the evaluation and authorisation process of drugs, cosmetics and chemicals not only more complicated but laborious. A microphysiological system was developed as a new foundation for substance discovery and risk assessment. Its microfluidic setup allows the combination of any 3D organ model and their interaction.
Mikroskopie und Analyse der Wundheilungsassays(S. 303)
Ulf Rädler, Joel Mailliet, Brad Larson, Leonie Rieger
The scratch wound assay is a powerful method to analyze cell migration which plays a central role in the wound healing process. In this application, we tracked the ability of HT-1080 fibrosarcoma cells to move into a created gap and heal the wound. The obtained results show that the combination of an easy experimental setup, time-lapse imaging and precise data analysis facilitate the study of collective cell migration, crucial for the development of novel therapeutics.
Bioplastik aus Licht und Luft – das Konzept einer synthetischen Ko-Kultur(S. 338)
Hannes Löwe, Andreas Kremling, Katharina PflügerGrau
Some of the most challenging tasks of our times are to combat global warming, to avoid further contamination of the environment with plastics, and to provide enough food for an increasing world population. A promising contribution to these overwhelming problems can be made by modern biotechnology. Here, we present the concept of a modular co-culture in which the feedstock is produced from CO2 and light by a cyanobacterium and converted directly to bioplastic (PHA) by a heterotrophic strain.
Biogene Treibstoffe aus Biomassepyrolyse(S. 341)
Nikolaus Schwaiger, Klara Treusch, Matthäus Siebenhofer
The bioCRACK process can convert as much as 18 percent of biomass into fuel intermediates for diesel production. This process was operated in pilot scale over two years. Beside fuel intermediates, pyrolysis oil is formed in the bioCRACK process. In the second technology development step pyrolysis oil was upgraded by continuous hydrodeoxygenation in lab scale. Results show a carbon yield from pyrolysis oil of 50 percent. The boiling range of upgraded pyrolysis oil is between diesel and gasoline.
Produktion pflanzlicher Polyphenole mit Corynebacterium glutamicum(S. 344)
Jan Marienhagen, Nicolai Kallscheuer
Many plant polyphenols demonstrate important pharmacological activities, but extraction from natural plant sources is usually limited by low abundance. After identification and elimination of the responsible degradation pathways, production of a broad range of different stilbenes and flavonoids could be achieved in Corynebacterium glutamicum. Furthermore, a synthetic pathway could be established in C. glutamicum, which allows for plant polyphenol synthesis from cheap 4-hydroxybenzoic acids.