Tactile acuity and left/right judgment performance in patients with knee osteoarthritis: A cross-sectional study.

BACKGROUND: Left/right judgment task (LRJT) performance and tactile acuity are impaired in chronic pain conditions, however, evidence is limited for knee osteoarthritis (OA). OBJECTIVE: To compare LRJT performance and the two-point discrimination threshold (TPDT) of chronic knee OA patients with asymptomatic knee and pain-free controls. DESIGN: Cross-sectional study. METHODS: Fifty knee OA patients and 50 age and gender-matched pain-free controls were assessed using the Recognize® application by displaying knee images and a digital caliper for the TPDT of the medial and lateral knee joint line. RESULTS: TPDTs over the lateral joint line in symptomatic (mean difference [MD]: 13.59 mm; 95% confidence interval [CI]: 8.72, 18.46; d = 1.40) and asymptomatic knee (MD: 10.15 mm; 95% CI: 5.08, 15.22; d = 0.99) were significantly increased compared to pain-free controls. Similarly, TPDTs of the medial joint line were significantly increased in symptomatic (MD: 12.19 mm; 95% CI: 7.59, 16.79; d = 1.31) and asymptomatic knee (MD: 7.64 mm; 95% CI: 3.64, 11.64; d = 1.31) compared to pain-free controls. Patients with knee OA were less accurate (MD: 7.80%; 95% CI: 15.32, -0.27; d = 0.52) recognizing images of their symptomatic knee. No correlation was found between pain severity, pain duration, LRJT performance, and TPDTs. Post-hoc analysis revealed no differences in LRJT and TPDTs between patients with and without clinically relevant symptoms of central sensitization. CONCLUSION: Chronic knee OA is associated with increased TPDT for medial and lateral knee joint lines and decreased recognition accuracy performance which should be considered during treatment process.

A Proteomic Screen of Neuronal Cell-Surface Molecules Reveals IgLONs as Structurally Conserved Interaction Modules at the Synapse.

In the developing brain, cell-surface proteins play crucial roles, but their protein-protein interaction network remains largely unknown. A proteomic screen identified 200 interactions, 89 of which were not previously published. Among these interactions, we find that the IgLONs, a family of five cell-surface neuronal proteins implicated in various human disorders, interact as homo- and heterodimers. We reveal their interaction patterns and report the dimeric crystal structures of Neurotrimin (NTRI), IgLON5, and the neuronal growth regulator 1 (NEGR1)/IgLON5 complex. We show that IgLONs maintain an extended conformation and that their dimerization occurs through the first Ig domain of each monomer and is Ca2+ independent. Cell aggregation shows that NTRI and NEGR1 homo- and heterodimerize in trans. Taken together, we report 89 unpublished cell-surface ligand-receptor pairs and describe structural models of trans interactions of IgLONs, showing that their structures are compatible with a model of interaction across the synaptic cleft.

An ELISA-Based Screening Platform for Ligand-Receptor Discovery.

Cell surface molecules are important for development and function of multicellular organisms. Although several methods are available to identify ligand-receptor pairs, ELISA-based methods are particularly amenable to high-throughput screens. ELISA-based methods have high sensitivity and low false-positive rates for detecting protein-protein interaction (PPI) complexes. Here, we provide a detailed protocol for a 384-well ELISA-based PPI screening protocol for the identification of novel cell surface ligand-receptor interactions, together with considerations for validation of PPIs by biophysical methods. This PPI screen has been developed and tested for discovery of novel ligand-receptor pairs between human synaptic adhesion proteins, believed to play crucial roles in many steps of neurodevelopment, from neuronal maturation, to axon guidance, synapse connectivity, and pruning.

Linking a compound-heterozygous Parkin mutant (Q311R and A371T) to Parkinson's disease by using proteomic and molecular approaches.

Parkin is an E3-protein ubiquitin ligase, which plays an important role as a scavenger in cell metabolism. Since the discovery of the link between Parkin and Parkinson's disease, Parkin was placed in the center of Parkinson's disease research. Previously, we isolated a mutant form of the Parkin protein (Q311R and A371T) from a Parkinson's disease patient. In this study, we aimed at characterizing this mutant Parkin protein by using biochemical and proteomic approaches. We used neuroblastoma cells (SH-SY5Y) as our model and created two inducible cell lines that expressed the wild type and the mutant Parkin proteins. We first investigated the effect of expressing both the wild type and the mutant Parkin proteins on the overall proteome by using 2D-DIGE approach. The experiments yielded the identification of 22 differentially regulated proteins, of which 13 were regulated in the mutant Parkin expressing cells. Classification of the identified proteins based on biological process and molecular function revealed that the majority of the regulated proteins belonged to protein folding and energy metabolism. Ingenuity Pathway Analysis predicted the presence of a link between the regulated proteins of the mutant Parkin expressing cells and Parkinson's disease. We also performed biochemical characterization studies on the wild type and the mutant Parkin proteins to make sense out of the differences observed at the proteome level. Both proteins displayed biological activity, had similar stabilities and localized similarly to the cytoplasm and the nucleus in SH-SY5Y cells. The mutant protein, however, was cut by a protease and subjected to a post-translational modification. The observed differences at the proteome level might be due to the differences in processing of the mutant Parkin protein. Overall, we were able to create a possible link between a pair of Parkin mutations to its pertinent disease by using 2D-DIGE in combination with biochemical and molecular approaches.