Tsuyoshi Miyazaki

Antibodies against Muscle-Specific Kinase Impair Both Presynaptic and Postsynaptic Functions in a Murine Model of Myasthenia Gravis

Antibodies against acetylcholine receptors (AChRs) cause pathogenicity in myasthenia gravis (MG) patients through complement pathway-mediated destruction of postsynaptic membranes at neuromuscular junctions (NMJs). However, antibodies against muscle-specific kinase (MuSK), which constitute a major subclass of antibodies found in MG patients, do not activate the complement pathway. To investigate the pathophysiology of MuSK-MG and establish an experimental autoimmune MG (EAMG) model, we injected MuSK protein into mice deficient in complement component five (C5). MuSK-injected mice simultaneously developed severe muscle weakness, accompanied by an electromyographic pattern such as is typically observed in MG patients. In addition, we observed morphological and functional defects in the NMJs of EAMG mice, demonstrating that complement activation is not necessary for the onset of MuSK-MG. Furthermore, MuSK-injected mice exhibited acetylcholinesterase (AChE) inhibitor-evoked cholinergic hypersensitivity, as is observed in MuSK-MG patients, and a decrease in both AChE and the AChE-anchoring protein collagen Q at postsynaptic membranes. These findings suggest that MuSK is indispensable for the maintenance of NMJ structure and function, and that disruption of MuSK activity by autoantibodies causes MG. This mouse model of EAMG could be used to develop appropriate medications for the treatment of MuSK-MG in humans.

Effects of Alendronate on Bone Metabolism in Glucocorticoid-Induced Osteoporosis Measured by 18F-Fluoride PET: A Prospective Study

Osteoporosis represents a significant side effect of glucocorticoid therapy, and alendronate has been reported to prevent this glucocorticoid-induced osteoporosis. Functional imaging with 18F-fluoride PET allows quantitative analysis of bone metabolism in specific skeletal regions. However, only a few studies have quantitatively determined bone turnover and metabolism in glucocorticoid-induced osteoporosis by radiologic imaging techniques including PET. The aim of this study was to examine changes in regional bone remodeling and turnover as measured by 18F-fluoride PET, the relationship between these measured changes and conventional bone metabolism parameters, and the effect of alendronate treatment. Methods: The study group consisted of 24 postmenopausal women (mean age, 59.7 y) who had various diseases, excluding rheumatoid arthritis, and had been treated with 10 mg or more of oral glucocorticoids (prednisolone equivalent) per day for more than 6 mo. Treatment with 5 mg of alendronate per day began at the time of study entry and continued for 12 mo. 18F-fluoride PET was performed at baseline, 3 mo, and 12 mo to determine localized bone turnover, and the results were compared with other bone metabolism parameters. Results: Lumbar spine standardized uptake values (SUVs) were significantly lower (P < 0.05) in the osteoporotic group (T-score ≤ −2.5) than in the group that was healthy or osteopenic (T-score > −2.5). Patients treated with alendronate for 12 mo exhibited significant decreases in serum bone-specific alkaline phosphate (P < 0.05), urinary N-telopeptide for type I collagen (P < 0.01), lumbar spine SUV (P < 0.01), and femoral neck SUV (P < 0.01) in association with a gradual increase in bone mineral density (BMD) of the lumbar spine relative to the baseline value (P < 0.05). Although there was a significant correlation between BMD and SUV in the lumbar spine at baseline (P < 0.05), there was no correlation between the 2 variables at 12 mo of treatment with alendronate. Conclusion: Alendronate treatment resulted in significant decreases in bone metabolism and turnover in the lumbar spine. It also led to an increase in BMD of the lumbar spine in patients with glucocorticoid-induced osteoporosis. Our findings suggest that antiresorptive therapy has a direct bone-metabolism effect on skeletal kinetics in glucocorticoid-induced osteoporosis at the clinically important site of the lumbar spine.

Joint proprioception before and after total knee arthroplasty.

To investigate the effects of total knee arthroplasty on joint proprioception, the absolute angular error of the knee in 38 consecutive patients before and after total knee arthroplasty for osteoarthritis and in 23 age-matched control subjects were examined. Varus and valgus laxity of the knee and muscle strengths of the thigh were measured using appropriate instruments. There were no significant differences in absolute angular error before and after total knee arthroplasty, independent of retaining or substituting the posterior cruciate ligament. The absolute angular error of the knee with a normal appearing anterior cruciate ligament was larger than that with a missing anterior cruciate ligament before total knee arthroplasty and decreased significantly after surgery. The absolute angular error correlated with the varus and valgus laxity of the knee, but did not correlate with the strength of thigh muscles after total knee arthroplasty. These results suggest that deficiency of the anterior cruciate ligament may not adversely affect proprioception in severe knee osteoarthritis. In addition, proper ligament balance may partly contribute to better proprioception after total knee arthroplasty.

Intracellular and Extracellular ATP Coordinately Regulate the Inverse Correlation between Osteoclast Survival and Bone Resorption

Background: Mature osteoclasts with a spontaneous tendency toward apoptosis resorb bone efficiently during their short lifespan. Results: Released ATP from intracellular stores has a negative impact on the bone resorption activity of osteoclasts by altering their cytoskeletal structures. Conclusion: ATP depletion leads to osteoclastic bone resorption. Significance: This study provides a new direction for investigating the mechanisms involved in physiological and pathological bone resorption. Osteoclasts, highly differentiated bone-resorbing cells of hematopoietic origin, have two conflicting tendencies: a lower capacity to survive and a higher capacity to execute energy-consuming activities such as bone resorption. Here, we report that when compared with their precursors, mature mitochondria-rich osteoclasts have lower levels of intracellular ATP, which is associated with receptor activator of nuclear factor κ-B ligand (RANKL)-induced Bcl-xL down-regulation. Severe ATP depletion, caused by disrupting mitochondrial transcription factor A (Tfam) gene, leads to increased bone-resorbing activity despite accelerated apoptosis. Although AMP-activated protein kinase (AMPK) activation by ATP depletion is not involved in the regulation of osteoclast function, the release of ATP from intracellular stores negatively regulates bone-resorbing activity through an autocrine/paracrine feedback loop by altering cytoskeletal structures. Furthermore, osteoclasts derived from aged mice exhibit reduced mitochondrial DNA (mtDNA) and intracellular ATP levels with increased bone-resorbing activity, implicating the possible involvement of age-related mitochondrial dysfunction in osteoporosis. Thus, our study provides evidence for a mechanism underlying the control of cellular functions by reciprocal changes in intracellular and extracellular ATP, which regulate the negative correlation between osteoclast survival and bone resorption.

Vertebroplasty-augmented short-segment posterior fixation of osteoporotic vertebral collapse with neurological deficit in the thoracolumbar spine: comparisons with posterior surgery without vertebroplasty and anterior surgery

OBJECTnThe surgical approach and treatment of thoracolumbar osteoporotic vertebral collapse with neurological deficit have not been documented in detail. Anterior surgery provides good decompression and solid fusion, but the surgery-related risk is relatively higher than that associated with the posterior approach. In posterior surgery, the major problem after posterior correction and instrumentation is failure to support the anterior spinal column, leading to loss of correction of kyphosis. The aim of this study was to evaluate the efficacy of reinforcing short-segment posterior fixation with vertebroplasty and to compare the outcome with those of posterior surgery without vertebroplasty and anterior surgery, retrospectively.nnnMETHODSnThe authors studied 83 patients who underwent surgical treatment for a single thoracolumbar osteoporotic vertebral collapse with neurological deficit. Twenty-eight patients treated by posterior surgery combined with vertebroplasty (Group A), 25 patients treated by posterior surgery without vertebroplasty (Group B), and 30 patients treated by anterior surgery (Group C) were followed up for a mean postoperative period of 4.4 years. Neurological outcome, visual analog scale pain score, and radiographic results were compared in the 3 groups.nnnRESULTSnPostoperative (4-6 weeks) and follow-up neurological outcome and visual analog scale scores were not significantly different among the 3 groups. Postoperative kyphotic angle was significantly reduced in Group B compared with Group C (p = 0.007), whereas the kyphotic angle was not significantly different among the 3 groups at follow-up. The mean ± SD loss of correction at follow-up was 4.6° ± 4.5°, 8.6° ± 6.2°, and 4.5° ± 5.9° in Groups A, B, and C, respectively. The correction loss at follow-up in Group B was significantly higher compared with Groups A and C (p = 0.0171 and p = 0.0180, respectively).nnnCONCLUSIONSnThe results suggest that additional reinforcement with vertebroplasty reduces the kyphotic loss and instrumentation failure, compared with patients without the reinforcement of vertebroplasty. Vertebroplasty-augmented short-segment fixation seems to offer immediate spinal stability in patients with thoracolumbar osteoporotic vertebral collapse; the effect seems equivalent to that of anterior reconstruction.

Pathomechanisms of Sciatica in Lumbar Disc Herniation: Effect of Periradicular Adhesive Tissue on Electrophysiological Values by an Intraoperative Straight Leg Raising Test

Study Design. This study is aimed to investigate the changes of nerve root functions during the straight leg raising (SLR) test in vivo. Objective. To investigate the relationship between nerve root movement and the electrophysiological values during an intraoperative SLR test. Summary of Background Data. The SLR test is one of the most significant signs for making a clinical diagnosis of lumbar disc herniation. A recent study showed that intraradicular blood flow apparently decreased during the SLR test in patients with disc herniation. Methods. The study included 32 patients who underwent microdiscectomy. During the surgery, the nerve root motion affected by the hernia was observed during the SLR test. The patients legs were allowed to hang down to the angle at which sciatica had occurred and the change of nerve root action potentials was measured. After removal of the hernia, a similar procedure was repeated. The periradicular specimens collected during surgery were examined by light and electron microscope. Results. In all patients intraoperative microscopy revealed that the hernia was adherent to the dura mater of the nerve roots. During the SLR test, the limitation of nerve root movement occurred by periradicular adhesive tissue and amplitude of action potential showed a sharp decrease at the angle that produced sciatica. After removal of the hernia, all the patients showed smooth gliding of the nerve roots during the test, and there was no marked decrease of amplitude. Our data suggest that temporary ischemic changes in the nerve root cause transient conduction disturbances. Pathologic examination showed that the periradicular tissue consisted of the granulation with vascularization and many inflammatory cell infiltrations. Conclusion. The presence of periradicular fibrosis will compound the nerve root pain by fixing the nerve in one position, thereby increasing the susceptibility of the nerve root to tension or compression.

3,4-Diaminopyridine improves neuromuscular transmission in a MuSK antibody-induced mouse model of myasthenia gravis

This study investigated the effect of 3,4-diaminopyridine (3,4-DAP), a potent potentiator of transmitter release, on neuromuscular transmission in vivo in a mouse model of myasthenia gravis (MG) caused by antibodies against muscle-specific kinase (MuSK; MuSK-MG) and ex vivo in diaphragm muscle from these mice. 3,4-DAP significantly improved neuromuscular transmission, predominantly by increasing acetylcholine (ACh) release, supporting presynaptic potentiation as an effective treatment strategy for MuSK-MG patients who have defective transmitter release. In MuSK-MG, we suggest that only low-dose acetylcholinesterase (AChE) inhibitors be used to avoid side effects, and we propose that 3,4-DAP may be effective as a symptomatic therapy.

A review of denosumab for the treatment of osteoporosis.

Osteoporosis is an age-related systemic skeletal disease characterized by low bone mass and microarchitectural deterioration of bone tissue, with a consequent increase in bone fragility. Bone remodeling involves two types of cells: osteoblasts and osteoclasts. Receptor activator of nuclear factor-κB ligand (RANKL) is a key regulator of the formation and function of bone-resorbing osteoclasts, and its cell surface receptor, receptor activator of nuclear factor-κB (RANK), is expressed by both osteoclast precursors and mature osteoclasts. Denosumab is a fully human monoclonal anti-RANKL antibody that inhibits the binding of RANKL to RANK, thereby decreasing osteoclastogenesis and bone-resorbing activity of mature osteoclasts. Although there are many medications available for the treatment of osteoporosis, inhibition of RANKL by denosumab has been shown to significantly affect bone metabolism. Denosumab appears to be a promising, highly effective, and safe parenteral therapy with good adherence for osteoporosis. Moreover, denosumab may be cost-effective therapy compared with existing alternatives. Therefore, in this review, we focus on studies of denosumab and the risks and benefits identified for this type of treatment for osteoporosis.

Muscle weakness and neuromuscular junctions in aging and disease

A critical issue in todays super‐aging society is the need to reduce the burden of family care while continuing to make our medical institutions supportive. A rapidly emerging, major health concern is the debilitating effect of muscle weakness and atrophy from aging, termed sarcopenia; however, the molecular basis of this condition is not well understood. Our research aim is to elucidate the molecular mechanisms of age‐related muscle atrophy and to devise new measures for preventing and treating this disability. A promising treatment for muscle atrophy is the promotion of muscle regeneration by recruiting stem cells into the targeted region. The first requirement is to understand how the motor system, which consists of muscles and motoneurons, is maintained to accomplish that goal. Recent studies in the field of neuroscience have focused on neuromuscular junctions (NMJ), which play important roles in the maintenance of both motor nerves and muscle fibers. Signaling between muscles and motoneurons at NMJ supports interactions within the motor system. To understand the mechanisms involved, we focus our research on the pathogenic processes underlying neuromuscular diseases. The well‐known autoimmune disease, myasthenia gravis (MG), serves as a model not only for tracking the pathogenesis and treatment outcomes of all autoimmune diseases, but also for understanding synaptic functions in maintaining the motor system. Here, we describe recent insights into the molecular mechanisms required for the maintenance of NMJ and the related causes of muscle atrophy. Geriatr Gerontol Int 2010; 10 (Suppl. 1): S137–S147.

Effects of graded mechanical compression of rabbit sciatic nerve on nerve blood flow and electrophysiological properties

Entrapment neuropathy is a frequent clinical problem that can be caused by, among other factors, mechanical compression; however, exactly how a compressive force affects the peripheral nerves remains poorly understood. In this study, using a rabbit model of sciatic nerve injury (n=12), we evaluated the time-course of changes in intraneural blood flow, compound nerve action potentials, and functioning of the blood-nerve barrier during graded mechanical compression. Nerve injury was applied using a compressor equipped with a custom-made pressure transducer. Cessation of intraneural blood flow was noted at a mean compressive force of 0.457+/-0.022 N (+/-SEM), and the compound action potential became zero at 0.486+/-0.031 N. Marked extravasation of Evans blue albumin was noted after 20 min of intraneural ischemia. The functional changes induced by compression are likely due to intraneural edema, which could subsequently result in impairment of nerve function. These changes may be critical factors in the development of symptoms associated with nerve compression.