Lichtstrukturierte Matrizen für die räumlich-zeitliche Proteinanordnung(S. 471)
In vivo, cells can grow in 3D and significantly differ from cells grown in 2D. Imitating an extracellular microenvironment, 3D cell culture platforms using dynamic hydrogels have emerged, which can be altered by external stimuli, such as light, to mimic the chemical and physical properties of the native cellular environment. Photo-sensitive matrices enable the non-invasive, on-demand assembly of proteins and allow them to take influence on cells in real-time with high accuracy.
Um die Ecke gedacht: molekulargenetische Diagnose mit Transkriptomanalyse(S. 475)
Transcriptome analysis reveals functional effects of coding as well as non-coding variants on RNA abundance and sequence and can hence ease the identification of pathogenic variants in patients with genetic disorders.
Struktur und Funktion des mechanochemischen Motorproteins Dynamin(S. 481)
Katja Fäber, Oliver Daumke
The GTPase dynamin is a molecular machine that assembles at the neck of clathrin-coated pits and catalyzes the scission of the vesicle neck in a GTPase-dependent fashion . Recent structural work, in combination with biochemical and cell-based experiments, have led to a molecular model of how dynamin functions.
Eine vielseitige Enzymklasse für die Synthese ringförmiger Siderophore(S. 484)
Thomas Böttcher, Sina Rütschlin
The macrocyclic hydroxamate siderophore avaroferrin of Shewanella algae inhibits ferric iron dependent swarming motility of Vibrio alginolyticus. Investigating the biosynthesis of avaroferrin and related siderophores demonstrates the importance of the substrate pool for product formation and reveals an unprecedented flexibility in the substrate range of the responsible synthetases. Exploiting the promiscuity of these enzymes allowed to generate a broad spectrum of 15 different ringsize engineered siderophores some of which also inhibit Vibrio’s swarming behavior.
Modelle für Stresstoleranz und Herausforderung für die Medizin(S. 488)
Katja Sterflinger, Barbara Blasi, Caroline Poyntner, Donatella Tesei, Hakim Tafer
Exophiala dermatitidis is a polyextremophilic fungus able to live in glaciers and in the human environment. This emerging human pathogen causes skin infections, deep mycosis, and it can get neurotropic. Noncoding RNA, small circular RNA, and fusion transcripts are involved in its stress resistance and pathogenicity. Melanin is one of its most important virulence factors. Results from ex-vivo skin infections suggest that melanin might serve as a Trojan horse for the fungus in order to invade human skin.
Markierung im mikrobiellen Stoffwechsel(S. 491)
Lorenz Adrian, Karolina Malgorzata Nowak, Ernest Marco-Urrea
Stable isotopes are increasingly used to trace microbial activities and metabolic pathways and are widely applied in microbiology, biochemistry, and risk assessment. This development is tightly linked to advance of mass spectrometric instrumentation as the key analysis for stable isotopes. Also the concepts for stable isotope tracing are progressing fast. We briefly review such concepts and indicate some of our own studies.
Konformationsbewegungen von aktiven Membrantransportern(S. 495)
Rebecca Mächtel, Christian Gebhardt, Thorben Cordes
Mechanistic understanding of biological and biochemical processes requires methods to analyze structures and states but also conformational dynamics of biomacromolecules at room temperature under physiologically relevant conditions. Single-molecule Förster resonance energy transfer (smFRET) has evolved to a versatile tool for exactly this, the observation of intra- and intermolecular conformational dynamics and interactions of biomacromolecules. We here outline the basic principles and illustrate the applications of smFRET for understanding molecular mechanisms of active membrane transporters.
Markierungsfreies Hochdurchsatzscreening mit Raman-Spektroskopie(S. 499)
Jürgen Popp, Jan Rüger, Iwan W. Schie, Abdullah Saif Mondol, Anuradha Ramoji, Karina Weber
Raman microspectroscopy displays a label-free and non-destructive modality providing highly specific information on single-cell level. Thus, it bears great potential as a standard tool in biomedical studies. We have developed a high-throughput screening Raman spectroscopy platform combining both automated imaging microscopy and spectra acquisition. The system allows for rapid screening of whole cell populations enabling differential white blood cell count and circulating tumor cell detection.
Neue anwenderfreundliche Konzepte zur Zellkultivierung in Mikrochips(S. 504)
Frank Bunge, Sander van den Driesche, Michael J. Vellekoop
Microfluidic cell culture chips enable 3D-cultures or co-cultures as well as integrated sensors. Though, such devices are not yet utilized in biolabs. The reasons such as incompatibilities or inappropriate materials are addressed in this contribution by presenting two novel concepts with integrated porous membranes for gas and nutrient supply. The combination of clean-room technologies and 3D-printing enable production at reasonable costs as well as adapted interfaces for standard equipment.
Potenzial von Magnetpartikeln in der automatisierten Biodetektion(S. 509)
Christian Reis, Stefan Scheuermann, Caroline von Wulfen
The use of magnetic beads is a common tool for various biotechnological procedures. In laboratory automation, magnetizable particles help to purify cells or biomolecules for subsequent cultivation or analysis. Besides that, many devices and methods evolved using intrinsic properties of the beads to quantify biomolecules. Combination of innovative devices with new methods will allow for high sensitive assays and bear the potential of less process steps for analytical procedures in the future.
Anwendungen & Produkte
Isothermale Amplifikation macht’s möglich: Molekularbiologie für zu Hause(S. 522)
Dirk Kuhlmeier, Vicky Tröger, Cindy Baumann
Since more than 30 years, PCR offers an established solution to amplify genetic sequences fast and easy. To go beyond PCR, isothermal methods offer an alternative to simplify the process. Thermocycling devices are obsolete, because methods like LAMP or RPA work at constant and low temperatures. Here, we present the development of the world’s first disposable point-of-care (POC) and HomeCare test utilizing isothermal nucleic acid amplification technology for the detection of sexual transmitted infections.
Künstliche Intelligenz zählt Nervenzellen in Mäusegehirnen(S. 525)
Philipp Schubert, Marie E. Müller, Adrian A. Wanner
The vast amount of data that today’s imaging technologies generate, renders manual annotation and analysis infeasible. Novel workflows combining human and artificial intelligence process multi-terabyte-sized datasets. One example for such a workflow is the automated detection of millions of neurons in 3D whole mouse brain datasets. Our pipeline achieves high accuracy and reduces the required human input by orders of magnitude, enabling high-throughput analysis.