Investigation of p75 neurotrophin receptor and the role of its post-translational modifications in tumour cell motility and invasiveness

The p75 neurotrophin receptor (p75NTR) is a member of the tumour necrosis factor superfamily, which relies on the recruitment of cytosolic protein partners - including the TNF receptor associated factor 6 (TRAF6) E3 ubiquitin ligase - to produce cellular responses such as apoptosis, survival, and inhibition of neurite outgrowth. Recently,p75NTR was also shown to undergo γ-secretase-mediated regulated intramembrane proteolysis, and the receptor ICD was found to migrate to the nucleus where it regulates gene transcription. Moreover, γ-secretase-mediated proteolysis was shown to be involved in glioblastoma cell migration and invasion. In this study we report that TRAF6-mediated K63-linked polyubiquitination at multiple or alternative lysine residues influences p75NTR-ICD stability in vitro. In addition, we found that TRAF6-mediated ubiquitination of p75NTR is not influenced by inhibition of dynamin. Moreover, we report beta-transducin repeats-containing protein (β-TrCP) as a novel E3- ligase that ubiquitinates p75NTR, which is independent of serine phosphorylation of the p75NTR destruction motif. In contrast to its influence on other substrates, co-expression of β-TrCP did not reduce p75NTR stability. We created U87-MG glioblastoma cell lines stably expressing wild type, γ-secretaseresistant and constitutively cleaved receptor, as well as the ICD-stabilized mutant K301R. Interestingly, only wild-type p75NTR induces increased glioblastoma cell migration, which could be reversed by application of γ-secretase inhibitor. Microarray and qRT-PCR analysis of mRNA transcripts in these cell lines yielded several promising genes that might be involved in glioblastoma cell migration and invasion, such as cadherin 11 and matrix metalloproteinase 12. Analysis of potential transcription factor binding sites revealed that transcription of these genes might be regulated by well known p75NTR signalling cascades such as NF-κB or JNK signalling, which are independent of γ-secretase-mediated cleavage of the receptor. In contrast, while p75NTR overexpression was confirmed in melanoma cell lines and a patient sample of melanoma metastasis to the brain, inhibition of γ-secretase did not influence melanoma cell migration. Collectively, this study provides several avenues to better understand the physiological importance of posttranslational modifications of p75NTR and the significance of the receptor in glioblastoma cell migration and invasion.

CUBEs: decoupled, compact, and monolithic spatial translational compliant parallel manipulators

This paper deals with the conceptual design of decoupled, compact, and monolithic XYZ compliant parallel manipulators (CPMs): CUBEs. Position spaces of compliant P (P: prismatic) joints are first discussed, which are represented by circles about the translational directions. A design method of monolithic XYZ CPMs is then proposed in terms of both the kinematic substitution method and the position spaces. Three types of monolithic XYZ CPMs are finally designed using the proposed method with the help of three classes of kinematical decoupled 3-DOF (degree of freedom) translational parallel mechanisms (TPMs). These monolithic XYZ CPMs include a 3-PPP XYZ CPM composed of identical parallelogram modules (a previously reported design), a novel 3-PPPR (R: revolute) XYZ CPM composed of identical compliant four-beam modules, and a novel 3-PPPRR XYZ CPM. The latter two monolithic designs also have extended lives. It is shown that the proposed design method can be used to design other decoupled and compact XYZ CPMs by using the concept of position spaces, and the resulting XYZ CPM is the most compact one when the fixed ends of the three actuated compliant P joints thereof overlap.

A constraint and position identification (CPI) approach for the synthesis of decoupled spatial translational compliant parallel manipulators

This paper introduces a screw theory based method termed constraint and position identification (CPI) approach to synthesize decoupled spatial translational compliant parallel manipulators (XYZ CPMs) with consideration of actuation isolation. The proposed approach is based on a systematic arrangement of rigid stages and compliant modules in a three-legged XYZ CPM system using the constraint spaces and the position spaces of the compliant modules. The constraint spaces and the position spaces are firstly derived based on the screw theory instead of using the rigid-body mechanism design experience. Additionally, the constraint spaces are classified into different constraint combinations, with typical position spaces depicted via geometric entities. Furthermore, the systematic synthesis process based on the constraint combinations and the geometric entities is demonstrated via several examples. Finally, several novel decoupled XYZ CPMs with monolithic configurations are created and verified by finite elements analysis. The present CPI approach enables experts and beginners to synthesize a variety of decoupled XYZ CPMs with consideration of actuation isolation by selecting an appropriate constraint and an optimal position for each of the compliant modules according to a specific application.

Bifidobacterium longum 1714 as a translational psychobiotic: modulation of stress, electrophysiology and neurocognition in healthy volunteers

The emerging concept of psychobiotics—live microorganisms with a potential mental health benefit—represents a novel approach for the management of stress-related conditions. The majority of studies have focused on animal models. Recent preclinical studies have identified the B. longum 1714 strain as a putative psychobiotic with an impact on stress-related behaviors, physiology and cognitive performance. Whether such preclinical effects could be translated to healthy human volunteers remains unknown. We tested whether psychobiotic consumption could affect the stress response, cognition and brain activity patterns. In a within-participants design, healthy volunteers (N=22) completed cognitive assessments, resting electroencephalography and were exposed to a socially evaluated cold pressor test at baseline, post-placebo and post-psychobiotic. Increases in cortisol output and subjective anxiety in response to the socially evaluated cold pressor test were attenuated. Furthermore, daily reported stress was reduced by psychobiotic consumption. We also observed subtle improvements in hippocampus-dependent visuospatial memory performance, as well as enhanced frontal midline electroencephalographic mobility following psychobiotic consumption. These subtle but clear benefits are in line with the predicted impact from preclinical screening platforms. Our results indicate that consumption of B. longum 1714 is associated with reduced stress and improved memory. Further studies are warranted to evaluate the benefits of this putative psychobiotic in relevant stress-related conditions and to unravel the mechanisms underlying such effects.