A negative regulatory mechanism involving 14-3-3zeta limits signaling downstream of ROCK to regulate tissue stiffness in epidermal homeostasis

ROCK signaling causes epidermal hyper-proliferation by increasing ECM production, elevating dermal stiffness, and enhancing Fak-mediated mechano-transduction signaling. Elevated dermal stiffness in turn causes ROCK activation, establishing mechano-reciprocity, a positive feedback loop that can promote tumors. We have identified a negative feedback mechanism that limits excessive ROCK signaling during wound healing and is lost in squamous cell carcinomas (SCCs). Signal flux through ROCK was selectively tuned down by increased levels of 14-3-3ζ, which interacted with Mypt1, a ROCK signaling antagonist. In 14-3-3ζ(-/-) mice, unrestrained ROCK signaling at wound margins elevated ECM production and reduced ECM remodeling, increasing dermal stiffness and causing rapid wound healing. Conversely, 14-3-3ζ deficiency enhanced cutaneous SCC size. Significantly, inhibiting 14-3-3ζ with a novel pharmacological agent accelerated wound healing 2-fold. Patient samples of chronic non-healing wounds overexpressed 14-3-3ζ, while cutaneous SCCs had reduced 14-3-3ζ. These results reveal a novel 14-3-3ζ-dependent mechanism that negatively regulates mechano-reciprocity, suggesting new therapeutic opportunities.Jasreen Kular, Kaitlin G. Scheer, Natasha T. Pyne, Amr H. Allam, Anthony N. Pollard, Astrid Magenau, Rebecca L. Wright, Natasha Kolesnikoff, Paul A. Moretti, Lena Wullkopf, Frank C. Stomski, Allison J. Cowin, Joanna M. Woodcock, Michele A. Grimbaldeston, Stuart M. Pitson, Paul Timpson, Hayley S. Ramshaw, Angel F. Lopez, and Michael S. Samue

Analytics in Microfluidic Systems

Viefhues M. Analytics in Microfluidic Systems. In: Advances in Biochemical Engineering/Biotechnology. Berlin ; Heidelberg: Springer ; 2020.Microfluidic analysis proved to be very sufficient in supporting biotechnological studies. This is due to the wide range of new analysis methods that provide further insight into biotechnological questions but also to intrinsic advantages of the systems themselves. To name two of them, only very small sample amounts are needed, and the analytics are very fast. In this overview paper, microfluidic analysis methods are introduced with a special focus on electric analysis methods. The aim of this work is to shed light on the special advantages of miniaturized electrical analysis in microfluidics; the main theoretical aspects of the methods are given together with the potential scientific insight that can be gained by the respective methods