Insulin-Degrading Enzyme Efficiently Degrades polyQ Peptides but not Expanded polyQ Huntingtin Fragments.

Background: Huntington's disease is an inheritable autosomal dominant disorder caused by an expanded CAG trinucleotide repeat within the Huntingtin gene, leading to a polyglutamine (polyQ) expansion in the mutant protein. Objective: A potential therapeutic approach for delaying or preventing the onset of the disease involves enhancing the degradation of the aggregation-prone polyQ-expanded N-terminal mutant huntingtin (mHTT) exon1 fragment. A few proteases and peptidases have been identified that are able to cleave polyQ fragments with low efficiency. This study aims to identify a potent polyQ-degrading endopeptidase. Methods: Here we used quenched polyQ peptides to identify a polyQ-degrading endopeptidase. Next we investigated its role on HTT turnover, using purified polyQ-expanded HTT fragments and striatal cells expressing mHTT exon1 peptides. Results: We identified insulin-degrading enzyme (IDE) as a novel endopeptidase for degrading polyQ peptides. IDE was, however, ineffective in reducing purified polyQ-expanded HTT fragments. Similarly, in striatal cells expressing mHTT exon1 peptides, IDE did not enhance mHTT turnover. Conclusions: This study shows that despite IDE's efficiency in degrading polyQ peptides, it does not contribute to the direct degradation of polyQ-expanded mHTT fragments.

Medial talar resection: how much remains stable?

PURPOSE: Pathologies of the medial talus (e.g., fractures, tarsal coalitions) can lead to symptomatic problems such as pain and nonunion. Bony resection may be a good solution for both. It is unclear how much of the medial talus can be taken before the subtalar joint becomes unstable. The aim of this study was to evaluate the effect a limited resection of the medial talar facet and the anteromedial portion of the posterior talar facet has on subtalar stability. METHODS: Eight fresh-frozen human cadaveric lower limbs were mounted in a frame for simulated weight-bearing. Computed tomography scans were obtained under 700 N single-legged stance loading, with the foot in neutral, 15° inversion, and 15° eversion positions. A sequential resection of 10, 20, and 30% of the medial facet and the anteromedial portion of the posterior talar facet to the calcaneus, based on the intact talus width, was performed. Measurements of subtalar vertical angulation, talar subluxation, coronal posterior facet angle and talocalcaneal (Kite) angle in the anteroposterior and lateral view were performed. RESULTS: Gross clinical instability was not observed in any of the specimens. No significant differences were detected in the measurements between the resected and intact states (P ≥ 0.10) as well as among the resected states (P ≥ 0.11). CONCLUSION: In a biomechanical setting, resecting up to 30% of the medial facet and anteromedial portion of the posterior facet based on the intact talus width-does not result in any measurable instability of the subtalar joint in presence of intact ligamentous structures. LEVEL OF EVIDENCE: V.

3D geometry of femoral reaming for bone graft harvesting.

The reamer-irrigator-aspirator (RIA) technique allows to collect large bone graft amounts without the drawbacks of iliac crest harvesting. Nevertheless, clinical cases with occurrence of femur fractures have been reported. Therefore, this study aimed to systematically investigate the three-dimensional geometry of the reamed bone as a function of the reaming diameter and its influence on the associated potential fracture pattern. Forty-five intact fresh-frozen human cadaveric femora underwent computed tomography (CT). They were randomized to three groups (n = 15) for reaming at a diameter of either 1.5 mm (Group 1), 2.5 mm (Group 2) or 4.0 mm (Group 3) larger than their isthmus using RIA. Reaming was followed by a second CT scan, biomechanical testing until failure and a third CT scan. All CT scans of each femur were aligned via rigid registration, and fracture lines were visualized. Subsequently, a decrease in wall thickness, cross-sectional area, and harvested bone volume have been evaluated. The total volume of the bone graft was significantly higher for Group 3 (7.8 ± 2.9 ml) compared to Group 1 (2.9 ± 1.1 ml) and Group 2 (3.0 ± 1.1 ml). The maximal relative decrease of the wall thickness was located medially (72.7%) in the third (61.4%), fourth (18.2%) and second (9.1%) eighth for all groups. As the diameter of the reaming increased, an overlap of the fracture line with the maximal relative decrease in wall thickness and a maximal average relative decrease of the cross-sectional area became more frequent. This suggests that a reaming-associated fracture is most likely to occur in this region.

Influence of the Reamer-Irrigator-Aspirator diameter on femoral bone strength and amount of harvested bone graft - a biomechanical cadaveric study.

BACKGROUND: Treatment of large bone defects is still related to unsolved problems in orthopaedic trauma surgery. Minimally invasive intramedullary reaming with the use of the Reamer-Irrigator-Aspirator (RIA) device allows autograft harvesting of large bone graft amounts from the medullary canal of the femur. The aim of this study was to investigate the influence of RIA diameter on femoral bone strength and amount of harvested bone graft in a human cadaveric model. METHODS: Forty-five pairs human cadaveric femora were randomized to 3 paired groups with 15 pairs each. One femur of each pair was reamed with RIA at a diameter of either 1.5 mm (group 1), 2.5 mm (group 2) or 4.0 mm (group 3) larger than its isthmus, whereas its contralateral femur was left intact without reaming. The amount of harvested bone graft was determined for each specimen and all femora were destructively tested in internal rotation under 750 N axial compression to calculate their torsional stiffness and torque at failure. RESULTS: Significant reduction in torsional stiffness was detected after reaming in group 3 (p = 0.03) in contrast to groups 1 and 2 where no such significant reduction was observed (p ≥ 0.34). Torque at failure was significantly reduced after reaming in all 3 groups (p ≤ 0.04). Collected bone graft amount in group 3 was significantly bigger compared to groups 1 and 2 (p ≤ 0.04). CONCLUSIONS: Reaming with RIA diameter of 4.0 mm larger than the isthmus of the femur seems to influence considerably its torsional stiffness, however, it allows harvesting of a significantly bigger bone graft amount.

Cannabinoid receptor-interacting protein Crip1a modulates CB1 receptor signaling in mouse hippocampus.

The cannabinoid type 1 receptor (Cnr1, CB1R) mediates a plethora of physiological functions in the central nervous system as a presynaptic modulator of neurotransmitter release. The recently identified cannabinoid receptor-interacting protein 1a (Cnrip1a, CRIP1a) binds to the C-terminal domain of CB1R, a region known to be important for receptor desensitization and internalization. Evidence that CRIP1a and CB1R interact in vivo has been reported, but the neuroanatomical distribution of CRIP1a is unknown. Moreover, while alterations of hippocampal CRIP1a levels following limbic seizures indicate a role in controlling excessive neuronal activity, the physiological function of CRIP1a in vivo has not been investigated. In this study, we analyzed the spatial distribution of CRIP1a in the hippocampus and examined CRIP1a as a potential modulator of CB1R signaling. We found that Cnrip1a mRNA is co-expressed with Cnr1 mRNA in pyramidal neurons and interneurons of the hippocampal formation. CRIP1a protein profiles were largely segregated from CB1R profiles in mossy cell terminals but not in hippocampal CA1 region. CB1R activation induced relocalization to close proximity with CRIP1a. Adeno-associated virus-mediated overexpression of CRIP1a specifically in the hippocampus revealed that CRIP1a modulates CB1R activity by enhancing cannabinoid-induced G protein activation. CRIP1a overexpression extended the depression of excitatory currents by cannabinoids in pyramidal neurons of the hippocampus and diminished the severity of chemically induced acute epileptiform seizures. Collectively, our data indicate that CRIP1a enhances hippocampal CB1R signaling in vivo.