The Type 2 Diabetes Factor Methylglyoxal Mediates Axon Initial Segment Shortening and Alters Neuronal Function at the Cellular and Network Levels.

Recent evidence suggests that alteration of axon initial segment (AIS) geometry (i.e., length or location along the axon) contributes to CNS dysfunction in neurological diseases. For example, AIS length is shorter in the prefrontal cortex of type 2 diabetic mice with cognitive impairment. To determine the key type 2 diabetes-related factor that produces AIS shortening we modified levels of insulin, glucose, or the reactive glucose metabolite methylglyoxal in cultures of dissociated cortices from male and female mice and quantified AIS geometry using immunofluorescent imaging of the AIS proteins AnkyrinG and ßIV spectrin. Neither insulin nor glucose modification altered AIS length. Exposure to 100 but not 1 or 10 µm methylglyoxal for 24 h resulted in accumulation of the methylglyoxal-derived advanced glycation end-product hydroimidazolone and produced reversible AIS shortening without cell death. Methylglyoxal-evoked AIS shortening occurred in both excitatory and putative inhibitory neuron populations and in the presence of tetrodotoxin (TTX). In single-cell recordings resting membrane potential was depolarized at 0.5-3 h and returned to normal at 24 h. In multielectrode array (MEA) recordings methylglyoxal produced an immediate ∼300% increase in spiking and bursting rates that returned to normal within 2 min, followed by a ∼20% reduction of network activity at 0.5-3 h and restoration of activity to baseline levels at 24 h. AIS length was unchanged at 0.5-3 h despite the presence of depolarization and network activity reduction. Nevertheless, these results suggest that methylglyoxal could be a key mediator of AIS shortening and disruptor of neuronal function during type 2 diabetes.

Methylglyoxal Disrupts Paranodal Axoglial Junctions via Calpain Activation.

Nodes of Ranvier and associated paranodal and juxtaparanodal domains along myelinated axons are essential for normal function of the peripheral and central nervous systems. Disruption of these domains as well as increases in the reactive carbonyl species methylglyoxal are implicated as a pathophysiology common to a wide variety of neurological diseases. Here, using an ex vivo nerve exposure model, we show that increasing methylglyoxal produces paranodal disruption, evidenced by disorganized immunostaining of axoglial cell-adhesion proteins, in both sciatic and optic nerves from wild-type mice. Consistent with previous studies showing that increase of methylglyoxal can alter intracellular calcium homeostasis, we found upregulated activity of the calcium-activated protease calpain in sciatic nerves after methylglyoxal exposure. Methylglyoxal exposure altered clusters of proteins that are known as calpain substrates: ezrin in Schwann cell microvilli at the perinodal area and zonula occludens 1 in Schwann cell autotypic junctions at paranodes. Finally, treatment with the calpain inhibitor calpeptin ameliorated methylglyoxal-evoked ezrin loss and paranodal disruption in both sciatic and optic nerves. Our findings strongly suggest that elevated methylglyoxal levels and subsequent calpain activation contribute to the disruption of specialized axoglial domains along myelinated nerve fibers in neurological diseases.

Validation of 3D surface reconstruction of vertebrae and spinal column using 3D ultrasound data--a pilot study.

Adolescent idiopathic scoliosis (AIS) is a three-dimensional deformity of spine associated with vertebra rotation. The Cobb angle and axial vertebral rotation are important parameters to assess the severity of scoliosis. However, the vertebral rotation is seldom measured from radiographs due to time consuming. Different techniques have been developed to extract 3D spinal information. Among many techniques, ultrasound imaging is a promising method. This pilot study reported an image processing method to reconstruct the posterior surface of vertebrae from 3D ultrasound data. Three cadaver vertebrae, a Sawbones spine phantom, and a spine from a child with AIS were used to validate the development. The in-vitro result showed the surface of the reconstructed image was visually similar to the original objects. The dimension measurement error was <5 mm and the Pearson correlation was >0.99. The results also showed a high accuracy in vertebral rotation with errors of 0.8 ± 0.3°, 2.8 ± 0.3° and 3.6 ± 0.5° for the rotation values of 0°, 15° and 30°, respectively. Meanwhile, the difference in the Cobb angle between the phantom and the image was 4° and the vertebral rotation at the apex was 2°. The Cobb angle measured from the in-vivo ultrasound image was 4° different from the radiograph.

Intra- and Interobserver Reliability of the Cobb Angle-Vertebral Rotation Angle-Spinous Process Angle for Adolescent Idiopathic Scoliosis.

STUDY DESIGN: A reliability analysis of Cobb angle, vertebral rotation (VR), and spinous process angle (SPA) measurements in adolescent idiopathic scoliosis. OBJECTIVE: To determine the intra- and interobserver reliability of semi-automated digital radiograph measurements. SUMMARY OF BACKGROUND DATA: Cobb angle measurements on posteroanterior radiographs are commonly used to determine the severity of scoliosis. Vertebral rotation helps assess scoliosis 3-dimensionally and has a role in predicting curve progression. Recent studies have shown that the spinous process angle is a useful parameter in assessing scoliosis when using ultrasound imaging. Because the reliability of SPA measurements on radiographs has yet to be determined, it is important to compare the reliability of these 3 parameters (Cobb angle, VR, and SPA) using a computer assisted semi-automated method. METHODS: Sixty posteroanterior radiographs of patients with adolescent idiopathic scoliosis were obtained and measured twice by 3 observers who were blinded to their previous measurements, using an in-house developed program. Measurements were obtained using a semi-automated method to minimize variability resulting from observer reliability. The intra- and interobserver reliabilities were analyzed using intra-class correlation coefficients (ICCs) as well as Bland-Altman's bias and limits of agreement. RESULTS: Over 350 (intra) and 90 (inter) sets of curves with an average Cobb angle of 26° ± 9° (range, 10° to 44°) were compared for each parameter. Intra-observer reliabilities for each parameter were excellent (ICC[2,1], .82; 1.00), with mean absolute differences under 3° between most measurements. Interobserver reliability (ICC[2,1], .72; .95) was mostly good to excellent, with mean absolute differences ranging from 2.0° to 5.6°. CONCLUSIONS: Both the intra- and interobserver assessment of the Cobb, VR, and SPA from the semi-automated measurements had clinically acceptable reliability ranges and may be considered for clinical implementation. Additional studies will be conducted to determine the accuracy and sensitivity to change of these scoliosis severity measurements.