SNAP-25, but not SNAP-23, is essential for photoreceptor development, survival, and function in mice.

SNARE-mediated vesicular transport is thought to play roles in photoreceptor glutamate exocytosis and photopigment delivery. However, the functions of Synaptosomal-associated protein (SNAP) isoforms in photoreceptors are unknown. Here, we revisit the expression of SNAP-23 and SNAP-25 and generate photoreceptor-specific knockout mice to investigate their roles. Although we find that SNAP-23 shows weak mRNA expression in photoreceptors, SNAP-23 removal does not affect retinal morphology or vision. SNAP-25 mRNA is developmentally regulated and undergoes mRNA trafficking to photoreceptor inner segments at postnatal day 9 (P9). SNAP-25 knockout photoreceptors develop normally until P9 but degenerate by P14 resulting in severe retinal thinning. Photoreceptor loss in SNAP-25 knockout mice is associated with abolished electroretinograms and vision loss. We find mistrafficked photopigments, enlarged synaptic vesicles, and abnormal synaptic ribbons which potentially underlie photoreceptor degeneration. Our results conclude that SNAP-25, but not SNAP-23, mediates photopigment delivery and synaptic functioning required for photoreceptor development, survival, and function.

Progenitor division and cell autonomous neurosecretion are required for rod photoreceptor sublaminar positioning.

Migration is essential for the laminar stratification and connectivity of neurons in the central nervous system. In the retina, photoreceptors (PRs) migrate to positions according to birthdate, with early-born cells localizing to the basal-most side of the outer nuclear layer. It was proposed that apical progenitor mitoses physically drive these basal translocations non-cell autonomously, but direct evidence is lacking, and whether other mechanisms participate is unknown. Here, combining loss- or gain-of-function assays to manipulate cell cycle regulators (Sonic hedgehog, Cdkn1a/p21) with an in vivo lentiviral labelling strategy, we demonstrate that progenitor division is one of two forces driving basal translocation of rod soma. Indeed, replacing Shh activity rescues abnormal rod translocation in retinal explants. Unexpectedly, we show that rod differentiation also promotes rod soma translocation. While outer segment function or formation is dispensable, Crx and SNARE-dependent synaptic function are essential. Thus, both non-cell and cell autonomous mechanisms underpin PR soma sublaminar positioning in the mammalian retina.

The Gluteus Medius Experiences Significant Excursion With Hip Flexion.

Purpose: To evaluate the effect of hip flexion and rotation on excursion of the gluteus medius (Gmed) and minimus (Gmin) myotendinous unit. Methods: Seven hips from 4 cadaveric specimens (males, 68.5 ± 18.3 years old) were dissected to isolate the Gmed posterior and lateral insertions and the Gmin proximal and distal insertions. Sutures were placed from tendon insertions through origins created in the iliac fossa to simulate the myotendinous unit. A load of 10N was applied. Myotendinous excursion was measured at 10° hip extension and 0°, 45°, and 90° of hip flexion in neutral rotation, and from maximum internal and external rotation in 90° flexion. The amount of abduction and rotation was also measured at each flexion position with 20N applied to each tendon. Repeated-measures analysis of variance with Tukey post hoc was used for statistics. Results: Gmed-lateral excursion ranged from 2.4 ± 0.4 mm in 10° of hip extension to 23.0 ± 1.5 mm in 90° of flexion (P < .001), and Gmed-posterior excursion ranged from 0.92 ± 0.5 mm in 10° of extension to 38.1 ± 1.1 mm in 90° of flexion (P < .001). Gmin excursion shortened with hip flexion from 4.2 ± 0.3 mm in 10° of extension to -0.2 ± 1.5 mm in 90° of flexion (Gmin-prox: P = .525, Gmin-distal: P < .001). At 90° flexion from maximum internal to maximum external rotation, Gmin-distal and proximal demonstrated a 92.6% and 51.3% increase in excursion, respectively (P < .001). Gmed-lateral and Gmed-posterior demonstrated 49.4% and 23.1% increase in excursion with external rotation, respectively (P < .001). Conclusions: The Gmed myotendinous unit undergoes significant excursion with hip flexion, whereas both Gmed and Gmin had significant excursion with femoral external rotation in 90° flexion. Clinical Relevance: It is important to understand whether active or passive hip flexion or rotation in the early postoperative period causes excessive strain to an abductor tendon repair. We found that consideration should be given to limit flexion after Gmed repair and external rotation after Gmed or Gmin repairs.

Hydrogel assisted photoreceptor delivery inhibits material transfer.

Cell therapy holds tremendous promise for vision restoration; yet donor cell survival and integration continue to limit efficacy of these strategies. Transplanted photoreceptors, which mediate light sensitivity in the retina, transfer cytoplasmic components to host photoreceptors instead of integrating into the tissue. Donor cell material transfer could, therefore, function as a protein augmentation strategy to restore photoreceptor function. Biomaterials, such as hyaluronan-based hydrogels, can support donor cell survival but have not been evaluated for effects on material transfer. With increased survival, we hypothesized that we would achieve greater material transfer; however, the opposite occurred. Photoreceptors delivered to the subretinal space in mice in a hyaluronan and methylcellulose (HAMC) hydrogel showed reduced material transfer. We examined mitochondria transfer in vitro and cytosolic protein transfer in vivo and demonstrate that HAMC significantly reduced transfer in both contexts, which we ascribe to reduced cell-cell contact. Nanotube-like donor cell protrusions were significantly reduced in the hydrogel-transplanted photoreceptors compared to the saline control group, which suggests that HAMC limits the contact required to the host retina for transfer. Thus, HAMC can be used to manipulate the behaviour of transplanted donor cells in cell therapy strategies.

Characterization of the Ability of Low-Cost GNSS Receiver to Detect Spoofing Using Clock Bias.

The aim of this paper was to propose a method to characterize the ability of a GNSS user to detect a spoofing attack from the behavior of the clock bias. Spoofing interference is not a new issue, especially in military GNSS, although it is a new challenge for civil GNSS, since it is currently implemented and used in many everyday applications. For this reason, it is still a topical issue, especially for receivers that only have access to high-level data (PVT,CN0). To address this important issue, after conducting a study of the receiver clock polarization calculation process, this led to the development of a very basic Matlab model that emulates a spoofing attack at the computational level. Using this model, we were able to observe that the clock bias is affected by the attack. However, the amplitude of this disturbance depends on two factors: the distance between the spoofer and the target and the synchronization between the clock that generates the spoofing signal and the reference clock of the constellation. To validate this observation, more or less synchronized spoofing attacks were carried out on a fixed commercial GNSS receiver with the use of GNSS signal simulators and also with a moving target. We propose then a method to characterize the capacity of detecting a spoofing attack with the clock bias behavior. We present the application of this method for two commercial receivers of the same manufacturer from different generations.

Should We Repair the Pronator Quadratus in a Distal Radius Fracture with an Ulnar Styloid Base Fracture? A Biomechanical Study.

Background Merits of repairing the pronator quadratus (PQ) to restore distal radioulnar joint (DRUJ) stability after a volar approach remain controversial. Distal radius fractures are often associated with ulnar styloid fractures (USF). When involving the fovea, this USF can lead to a DRUJ instability. The PQ repair may be key in reducing this DRUJ instability. Methods This study aims to assess the biomechanical role of PQ repair in a cadaveric model of USF. In 17 forearm specimens, a USF including the fovea was executed. Positional changes of the DRUJ in forearm rotation and dorso-palmar (DP) translation were measured with variable loads (0, 2.5, and 5N) applied to the PQ origin. Results Forearm rotation and DP-translation decreased significantly with PQ loading of 5N, changing on average by 5 degrees and 0.6 mm, respectively. Conclusion We found a significant decrease in forearm rotation and DP-translation comparing a fully loaded PQ to an unloaded PQ in our cadaveric model.

Comparison of Three Different Internal Brace Augmentation Techniques for Scapholunate Dissociation: A Cadaveric Biomechanical Study.

Internal bracing (IB) is an augmentation method using high-strength nonabsorbable tape. However, there is no detailed information about the direction, location, or number of IBs required for scapholunate interosseous ligament (SLIL) injury repair. Thus, this study compared the biomechanical characteristics of short-transverse IB, long-oblique IB, and the combination of short-transverse and long-oblique (Combo) IB for SLIL injury in a biomechanical cadaveric model. We prepared nine fresh-frozen full upper extremity cadaveric specimens for this study. The scapholunate distance, scapholunate angle, and radioscaphoid angle were measured using the MicroScribe digitizing system with the SLIL intact, after scapholunate dissociation and the three different reconstructions. Three-dimensional digital records were obtained in six wrist positions in each experimental condition. Short-transverse IB had a similar effect compared with long-oblique IB in addressing the widening of the scapholunate distance. However, both were less effective than Combo IB. For scaphoid flexion deformity, short-transverse IB had minimal effect, while long-oblique IB had a similar effect compared to Combo IB. Combo IB was the most effective for improving distraction intensity and rotational strength. This study provides important information about the biomechanical characteristics of three different IB methods for SLIL injury and may be useful to clinicians in treating scapholunate dissociation.