[Active vitamin D3 analog].

Vitamin D is a fat-soluble vitamin and exerts effects on skeletal and extraskeletal health in children and adults of all ages. Vitamin D insufficiency is related to low muscle strength, increasing body sway, falls in the elderly. Supplementation with vitamin D reduces risk of osteoporotic fracture, and improves muscle strength and postural balance to prevent the elderly from fall. The preferred vitamin D analog for daily supplementation is cholecalciferol (vitamin D3). The active form of vitamin D3 is 1,25-dihydroxy-vitamin D3. Alfacalcidol, calcitriol and eldecalcitol are used to treat osteoporosis in Japan. Randomized placebo-controlled, double-blinded clinical trial for osteoporotic subjects showed that eldecalcitol is more efficacious to increase bone mineral density and prevent vertebral and wrist fractures in osteoporotic patients with vitamin D sufficiency than alfacalcidol.

Hypothalamic orexin prevents non-alcoholic steatohepatitis and hepatocellular carcinoma in obesity.

Non-alcoholic steatohepatitis (NASH) occasionally occurs under obesity; however, factors modulating the natural history of fatty liver disease remain unknown. Since hypothalamic orexin that regulates physical activity and autonomic balance prevents obesity, we investigate its role in NASH development. Male orexin-deficient mice fed a high-fat diet (HFD) show severe obesity and progression of NASH with fibrosis in the liver. Hepatic fibrosis also develops in ovariectomized orexin-deficient females fed an HFD but not ovariectomized wild-type controls. Moreover, long-term HFD feeding causes hepatocellular carcinoma (HCC) in orexin-deficient mice. Intracerebroventricular injection of orexin A or pharmacogenetic activation of orexin neurons acutely activates hepatic mTOR-sXbp1 pathway to prevent endoplasmic reticulum (ER) stress, a NASH-causing factor. Daily supplementation of orexin A attenuates hepatic ER stress and inflammation in orexin-deficient mice fed an HFD, and autonomic ganglionic blocker suppresses the orexin actions. These results suggest that hypothalamic orexin is an essential factor for preventing NASH and associated HCC under obesity.

The Contribution of Deleterious Rare Alleles in ENPP1 and Osteomalacia Causative Genes to Atypical Femoral Fracture.

CONTEXT: Atypical femoral fractures (AFFs) are very rare atraumatic or mild trauma fractures in the subtrochanteric region or femoral shaft. Some unique genetic variants in Asian populations might confer susceptibility to AFF, since the incidence of AFFs is higher in Asian populations. OBJECTIVE: Because rare variants have been found to be causative in some diseases and the roles of osteomalacia causative genes have not been reported, we investigated rare variants in genes causing abnormal mineralization. METHODS: Exome sequencing was performed to detect variants in gene coding and boundary regions, and the frequencies of deleterious rare alleles were compared between Japanese patients with AFF (n = 42) and controls of the 4.7KJPN panel of Tohoku Medical Megabank by whole genome sequencing (n = 4773). RESULTS: The frequency of the deleterious rare allele of ENPP1 was significantly increased in AFF (P = .0012, corrected P [Pc] = .0155, OR 4.73, 95% CI 2.15-10.40). In multigene panel analysis, the frequencies of deleterious rare alleles of candidate genes were increased in AFF (P = .0025, OR 2.72, 95% CI 1.49-4.93). Principal component analysis of bone metabolism markers identified a subgroup of patients with AFF with higher frequencies of deleterious rare alleles in ENPP1 (P = 4.69 × 10-5, Pc = .0006, OR 8.47, 95% CI 3.76-19.09) and the candidate genes (P = 1.08 × 10-5, OR 5.21, 95% CI 2.76-9.86). CONCLUSION: AFF is associated with genes including ENPP1 that cause abnormal mineralization, suggesting that osteomalacia is an underlying condition predisposing to AFF and that higher incident rates of AFFs in Asian populations might be explained by the genetic risk factors including ENPP1.

Cybernic treatment with wearable cyborg Hybrid Assistive Limb (HAL) improves ambulatory function in patients with slowly progressive rare neuromuscular diseases: a multicentre, randomised, controlled crossover trial for efficacy and safety (NCY-3001).

BACKGROUND: Rare neuromuscular diseases such as spinal muscular atrophy, spinal bulbar muscular atrophy, muscular dystrophy, Charcot-Marie-Tooth disease, distal myopathy, sporadic inclusion body myositis, congenital myopathy, and amyotrophic lateral sclerosis lead to incurable amyotrophy and consequent loss of ambulation. Thus far, no therapeutic approaches have been successful in recovering the ambulatory ability. Thus, the aim of this trial was to evaluate the efficacy and safety of cybernic treatment with a wearable cyborg Hybrid Assistive Limb (HAL, Lower Limb Type) in improving the ambulatory function in those patients. RESULTS: We conducted an open-label, randomised, controlled crossover trial to test HAL at nine hospitals between March 6, 2013 and August 8, 2014. Eligible patients were older than 18 years and had a diagnosis of neuromuscular disease as specified above. They were unable to walk for 10 m independently and had neither respiratory failure nor rapid deterioration in gait. The primary endpoint was the distance passed during a two-minute walk test (2MWT). The secondary endpoints were walking speed, cadence, and step length during the 10-m walk test (10MWT), muscle strength by manual muscle testing (MMT), and a series of functional measures. Adverse events and failures/problems/errors with HAL were also evaluated. Thirty patients were randomly assigned to groups A or B, with each group of 15 receiving both treatments in a crossover design. The efficacy of a 40-min walking program performed nine times was compared between HAL plus a hoist and a hoist only. The final analysis included 13 and 11 patients in groups A and B, respectively. Cybernic treatment with HAL resulted in a 10.066% significantly improved distance in 2MWT (95% confidence interval, 0.667-19.464; p = 0.0369) compared with the hoist only treatment. Among the secondary endpoints, the total scores of MMT and cadence at 10MWT were the only ones that showed significant improvement. The only adverse effects were slight to mild myalgia, back pain, and contact skin troubles, which were easily remedied. CONCLUSIONS: HAL is a new treatment device for walking exercise, proven to be more effective than the conventional method in patients with incurable neuromuscular diseases. TRIAL REGISTRATION: JMACTR, JMA-IIA00156.

Efficacy, tolerability, and safety of risedronate in Japanese patients with Paget's disease of bone.

This study evaluated the clinical efficacy of treatment with oral risedronate (17.5 mg once daily) for 8 weeks in 11 Japanese patients with Paget's disease of bone (PDB). Risedronate suppressed the excessive bone turnover associated with PDB and improved several biochemical markers, including serum alkaline phosphatase (ALP), serum bone-specific ALP (BALP), urinary deoxypyridinoline (DPD), and urinary cross-linked N-telopeptide of type 1 collagen (NTX). These markers began to decrease within about 2 weeks after the initiation of treatment in most patients, and the response persisted for up to 40 weeks after the cessation of treatment. Risedronate reduced pain by week 24 in most patients. According to quantitative bone scintigraphy, the lesion with the highest radioisotope (RI) uptake showed a decrease of uptake from 12.7 +/- 6.8 to 6.0 +/- 2.3 (mean +/- SD) in week 24, although each lesion of patients with polyostotic disease had a different scintigraphic response. Overall, risedronate at a dose of 17.5 mg once daily was well tolerated by patients with PDB, even though the dosage was seven times higher than that approved for the treatment of osteoporosis in Japan (2.5 mg once daily). In conclusion, treatment with high-dose risedronate for 8 weeks resulted in clinically significant and sustained improvement of biochemical markers of bone turnover for 48 weeks in patients with PDB, and this improvement was associated with a decrease of RI uptake by Paget's bone lesions and with reduced pain.

Continuous local infusion of fibroblast growth factor-2 enhances consolidation of the bone segment lengthened by distraction osteogenesis in rabbit experiment.

Experimental tibial lengthening was achieved in 61 rabbits to examine the effect of continuous local infusion of recombinant human fibroblast growth factor-2 (rhFGF-2) on bone healing of the lengthened segment. The tibial diaphysis was separated by osteotomy and was subjected to slow progressive distraction (rate: 0.35 mm/12 h) using a monolateral external fixator. There were a lag phase for 1 week, a distraction phase for 2 weeks, and a consolidation phase for 5 weeks in this experiment. At various stages of distraction, rhFGF-2 was infused continuously for 2 weeks into the lengthened segment (rate: 14.28 microg/60 microl/day) using an osmotic pump implanted under the skin. Bone healing was significantly accelerated when rhFGF-2 was infused in the beginning of consolidation phase, but not in the distraction phase or in the lag phase. Infusion of normal saline (N/S) using the same osmotic pump had no effect. Dual-energy X-ray absorptiometry (DXA) and peripheral quantitative computerized tomography (pQCT) studies demonstrated that rhFGF-2-treated tibia had increased bone mineral density (BMD), bone mineral content (BMC) and cortical bone thickness (CBT) when compared with N/S-treated tibia. Three-point bending test demonstrated that rhFGF-2-treated bone had significantly stronger mechanical properties than N/S-treated bone. Finally, distribution of the infused materials was checked by using Indian ink or radio-opaque. The dyes distributed widely but exclusively in the lengthened segment. Based on these results, we conclude that direct delivery of rhFGF-2 into the lengthened segment can shorten the consolidation phase of limb lengthening and the method is applicable to the clinical treatment.

Differences of therapeutic effects on regional bone mineral density and markers of bone mineral metabolism between alendronate and alfacalcidol in Japanese osteoporotic women.

We studied the differences of therapeutic effects on regional bone mineral density (BMD) and markers of bone mineral metabolism between alendronate and alfacalcidol in Japanese osteoporotic women. Ninety-two Japanese women suffering from primary osteoporosis without osteoporotic fractures, aged 55 to 81 years, were divided into two groups: women treated orally with alendronate for one-year (5mg/day) (alendronate group, n=35) and women treated orally with alfacalcidol for one year (0.5 microg/day) (alfacalcidol group, n=57). The mean BMD of the 2nd to 4th lumbar vertebrae (L2-4BMD) and regional BMD were measured using dual energy X-ray absorptiometry. In the alendronate group, the percentage changes of L2-4BMD, lumbar spine BMD, thoracic spine BMD, pelvis BMD in the alendronate group were 106.3+/-4.6%, 104.2+/-6.6%, 107.1+/-10.4%, 107.1+/-10.5%, respectively. The percentage changes of L2-4BMD and regional BMD except for head BMD in the alendronate group were significantly greater than those in the alfacalcidol group. In the alfacalcidol group, L2-4BMD, thoracic spine BMD and lumbar spine BMD were maintained at respective pretreatment levels, whereas other regional BMD were decreased. Both serum bone-specific alkaline phosphatase and urinary type I collagen cross-linked N-telopeptide of the alendronate group were decreased, whereas these markers of bone mineral metabolism of alfacalcidol group were increased compared with the respective pre-treatment levels. The results suggest that one-year treatment with alendronate increased L2-4BMD, lumbar spine BMD, thoracic spine BMD and pelvis BMD, and that markers of both bone formation and bone resorption were decreased following one-year treatment with alendronate.

Unilateral chronic insufficiency of anterior cruciate ligament decreases bone mineral content and lean mass of the injured lower extremity.

We studied the effects of unilateral chronic anterior cruciate ligament (ACL) injury on bone size, bone mineral content (BMC), bone mineral density (BMD), soft tissue composition and muscle strength of the injured lower extremity in Japanese 21 men and 12 women aged 15 to 39 years. Bone area, BMD, BMC, lean mass and fat mass of lower extremity were measured using dual energy X-ray absorptiometry. The isometric and isokinetic muscle strength was assessed by an isokinetic machine.BMC, lean mass, circumference of the thigh and circumference of the lower leg of the injured lower extremity were significantly smaller than those of the intact lower extremity (p=0.0002, p<0.0001, p<0.0001, p=0.0131). In contrast, fat mass and %Fat of the injured lower extremity was significantly greater than that of the intact lower extremity (p=0.0301, p<0.0001). Bone area and BMD did not produce significant difference. These findings suggest that chronic insufficiency of ACL decreases BMC and lean mass of the injured lower extremity.

Atypical Lipomatous Tumor/Well-Differentiated Liposarcoma Developed in a Patient with Progressive Muscular Dystrophy: A Case Report and Review of the Literature.

BACKGROUND: Atypical lipomatous tumor/well-differentiated liposarcoma (ALT/WDLS) is an intermediate or locally aggressive form of adipocytic soft tissue sarcoma. Muscular dystrophy (MD) is characterized by progressive muscle atrophy and its replacement by adipose and fibrous tissue. Recently, some authors have reported that MD genes are related to neoplastic formation, but there have been no detailed clinical reports of ALT associated with MD. CASE PRESENTATION: A 73-year-old woman with a diagnosis of limb-girdle MD visited our department for recurrence of a huge tumor in her left thigh. She had undergone resection of a lipoma at the same site more than 20 years earlier. Imaging studies revealed a lipomatous tumor in her left thigh. We performed marginal resection including the adjacent muscles. Histological diagnosis was atypical lipomatous tumor. The postoperative course was uneventful, with no recurrence at 36 months of follow-up. CONCLUSION: We encountered a huge atypical tumor in a patient with MD. This is the first detailed report to describe an association between ALT and MD. We hypothesize that degenerative changes occurring in adipose tissue during muscle atrophy can cause lipomatous neoplasms and moreover that the mutation of MD-related genes may lead to the proliferation of tumor cells or to malignancy.

Bisphosphonate modulates morphological and mechanical properties in distraction osteogenesis through inhibition of bone resorption.

Despite the general clinical acceptance of distraction osteogenesis and much attention to bone formation in this method, little is recognized about activated bone resorption in the regenerated bone. The purpose of this study was to demonstrate the simultaneously activated bone resorption with activated bone formation and to investigate the role and efficacy of bisphosphonate in distraction osteogenesis. Left tibiae of 54 immature rabbits were lengthened for 3 weeks at a rate of 0.7 mm/day after a 1-week lag. Regenerated bone was quantitatively investigated by radiographic bone density, bone histomorphometry, and three-point bending testing. Animals received either vehicle or nitrogen-containing bisphosphonate (N-BP), YM529/ONO5920 at doses of 0.4 mg/kg/w or 0.004 mg/kg/w for 6 weeks. Regenerated bone of the vehicle group showed a radiologically characteristic zone structure containing the osteopenic zones adjacent to the sclerotic zones. The regenerated bone of the 0.4-mg/kg/w group showed no osteopenic zones during the course and eventually became homogeneously radiodense. The bone volume corresponding to the osteopenic zone of this group was 5.6-fold greater compared with that of the vehicle group. The lengthened bone strength of this group was 3.3-fold greater in ultimate force than that of the vehicle group and equivalent to the contralateral tibia. The 0.004-mg/kg/w group had no substantial differences compared with the vehicle group, despite radiological enhancement of the mineralized front as well as somewhat delayed bone resorption. These results demonstrate that not only bone formation but also bone resorption is highly activated in the regenerated bone, implying high bone turnover. Sufficient N-BP caused a notable modulation in morphological properties of the regenerated bone through inhibition of highly activated bone resorption and eventually increased mechanical properties.

[Therapeutic effects of risedronate on Paget's disease of bone].

Paget's disease of bone (PDB) is a chronic disorder characterized by focal abnormalities of bone turnover. The characteristic feature of PDB is excessive osteoclastic bone resorption coupled to increased and disorganized bone formation. Although PDB is rare in Japan, it is quite common in most European countries, except for Scandinavia. The recommended regimens for bisphosphonates use in various parts of the world. Calcitonin and etidronate are approved by the Ministry of Health, Labour and Welfare in Japan to treat PDB. Risedronate is currently under development for use in the treatment of PDB in Japan.

[Paget's disease of bone].

Paget's disease of bone (PDB) is a chronic disorder characterized by focal abnormalities of bone turnover. The symptoms of PDB include bone pain, bone deformity, osteoarthritis, and an increase in risk of pathological fracture and neurological problems. PDB is the second most common metabolic bone disease in European countries, although PDB is uncommon in Japan. PDB shows geographical and ethnic clustering. Based on this, two hypotheses of etiology of PDB have been proposed. One hypothesis is that PDB results from a slow virus infection of osteoclasts with paramyxovirus. The other is that PDB is a genetic disease. The development of bisphosphonate has brought major changes to the treatment of PDB. In Japan, etidronate and calcitonin are approved by the Ministry of Health, Labour and Welfare for treating PDB. Risedronate is under development for treating PDB in Japan.

Effects of constitution, atraumatic vertebral fracture and aging on bone mineral density and soft tissue composition in women.

Constitution, atraumatic vertebral fracture and aging affect bone mineral density (BMD) and soft tissue composition. The high body weight of obese women involves a high mechanical load being exerted on weight bearing bones compared with thin women, which probably contributes to their higher BMD and the lower incidence of fractures in obese women compared with thin women. Atraumatic vertebral fracture (AVF) is a typical osteoporotic fracture and its favorite site of AVF is the vertebral bodies of the thoracolumbar region. The BMD of weight bearing bones is lower in patients with AVF than in patients without AVF, whereas there is no significant difference in soft tissue composition between the two. The regional and total BMD decrease with advancing age. The magnitude of the decrease in lumbar and thoracic BMD is high compared with other regional BMD, and total fat mass and total lean mass decline with age to their respective minimal level. The high rate of decrease in lumbar and thoracic BMD appears to be due to the high content of trabecular bone compared with other regional bones.

Effects of unilateral sciatic neurectomy on growing rat femur as assessed by peripheral quantitative computed tomography, Fourier transform infrared spectroscopy and bending test.

We studied the effects of unilateral sciatic neurectomy (USN) on the development of the femoral shaft in the 30 growing Wistar-derived rats aged 5 weeks. Rats were allocated to three groups. One of these was immediately used for measurements, and the remaining 2 groups underwent USN of internal control. Specimens obtained from each group were divided into 2 subgroups: left femurs of each group served as the control subgroup (CONT) and right femurs from each group as the USN-operated subgroup (USN-OP). The bone mineral density (BMD), bone mineral content (BMC), bone area, periosteal circumference and endosteal circumference were measured by peripheral quantitative computed tomography (pQCT) and the mineral/matrix ratio was evaluated by Fourier transform infrared spectroscopy (FTIR). A three-point bending test was performed to analyze the biomechanical effects of sciatic neurectomy. USN-OP showed a significant decrease in cortical BMC, bone area, and periosteal circumference compared with CONT. The mineral/matrix ratio of cortical bone did not differ significantly between USN-OP and CONT. Strength and stiffness were significantly decreased in USN-OP compared with CONT. The results showed that USN inhibited periosteal bone formation, but has no significant effects on the mineral/matrix ratio of cortical bone in femurs.

Intergenerational comparison of total and regional bone mineral density and soft tissue composition in Japanese women without vertebral fractures.

We measured total and regional bone mineral density (BMD) and soft tissue mass in 115 healthy Japanese women without vertebral fractures. The subjects, aged 20 to 75 years, were divided into four age groups: 20's to 40's group (n=33), 50's group (n=26), 60's group (n=26), and 70's group (n=30). BMD was measured by dual energy X-ray absorptiometry (DXA). The evaluated regions were the head, arms, legs, ribs, thoracic vertebrae, lumbar vertebrae and pelvis. The total and regional BMDs were the highest in the 20's to 40's group, and they decreased with age, and reached their respective lowest values in the 70's group. The decrease in BMDs of the spine and pelvis was the most prominent of all regional BMDs. Total and regional lean mass and fat mass were the highest in the 20's to 40's group, and they decreased to their respective lowest values in the 70's group. The results showed that the decrease in BMD of the spine and pelvis was the most prominent of all regional BMDs associated with a decrease in the total and regional lean mass and total and regional fat mass.

Biophysic evaluation of bone quality-application of Fourier transform infrared spectroscopy and phosphorus-31 solid-state nuclear magnetic resonance spectroscopy.

In this review, we focus on findings obtained with biophysic techniques, Fourier transformed infrared (FTIR) spectroscopy and phosphorus-31 solid-state nuclear magnetic resonance (31P solid-state NMR) spectroscopy, which may allow us to evaluate bone quality and to predict bone strength. FTIR measures the absorption energy that produces an increase in the vibrational or rotational energy of atoms or groups of atoms within the molecule. FTIR spectroscopy allows us to examine the relative amount of minerals and matrix content and the arrangement of apatite and organic matrix. FTIR spectroscopy should become an important tool, because the relative amount of minerals and the arrangement of apatite and organic matrix could be a measure for evaluating bone quality. 31P solid-state NMR spectroscopy is useful for evaluating the quality of bone and predicting bone strength by calculating the spine-lattice relaxation time (T1) of bone. 31P solid-state NMR imaging can be used to measure quantitatively the mass of hydroxyapatite. The T1 relaxation time of both bone and deficient hydroxyapatite was much longer than that of pure hydroxyapatite. T1 relaxation time is one of the promising indices of bone quality.

Mineral to matrix ratio determines biomaterial and biomechanical properties of rat femur--application of Fourier transform infrared spectroscopy.

We studied the changes of biomaterial and biomechanical properties of the rat femur during development. Thirty male Wistar rats were allocated to 6 groups: aged 6 weeks (n=5), 9 weeks (n=5), 12 weeks (n=5), 15 weeks (n=5), 24 weeks (n=5), and 36 weeks (n=5). The mineral to matrix ratio (M/M ratio) of rat femur by Fourier transform infrared spectroscopy was 0.97 ± 0.10 at the age of 6 weeks, and reached the maximum of 1.52 ± 0.17 at the age of 36 weeks. Total bone mineral density (BMD) by peripheral quantitative computed tomography of the femoral shaft aged 6 weeks was 479.1 ± 58.7 mg/cm(3), and reached the maximum of 1022.2 ± 42.3 mg/cm(3) at the age of 36 weeks. The ultimate load to failure of the femur of the rat aged 6 weeks by the three-point bending test was 29.6 ± 6.1 N. At the age of 36 weeks, the ultimate load to failure of the rat femur increased to the maximum of 283.5 ± 14.7 N. The results showed that the M/M ratio increased with development as total BMD and bone strength increased. The results suggest that the M/M ratio is one of the determinants of the biomaterial and biomechanical properties of bone.

Long-term outcome of severe stroke patients: is the ADL status at discharge from a stroke center indicative of the long-term outcome?

The objective of this study was to examine the possibility of predicting the long-term outcome of severe stroke patients from the ADL status at discharge. The design is a retrospective observational study in the stroke center at Tokushima University Hospital. Severe stroke patients at discharge with Barthel Index (BI) scores ≤ 40 (N=51) were divided into two subgroups retrospectively, improved and unimproved. The Functional Independence Measure (FIM) and BI were analyzed at discharge, 3 and 6 months after onset. Correlation analysis between BI and FIM showed good correlation, especially between BI and motor FIM (mFIM). In conclusion, the ADL status at discharge from a stroke center is not definitely indicative of the long-term outcome, although good recovery of severe stroke cases was observed in males and younger patients, and in patients with motor FIM score>25. This result shows that the first 3 months after the attack should be regarded as an important time window for intensive rehabilitation.

Three dimensional motion analyses for rehabilitation version of Awa Odori exercise and the expectancy of physical effects.

'Awa Odori Exercise--Rehabilitation version--was developed in 2006 for the new trial of physical exercise for the aging and the impaired person with lower balance performance in Tokushima prefecture, Japan. Public relations of this exercise had been spreading over Tokushima since then. The characteristics of the exercise were highly familiar with most of people in Tokushima because of popularity in original 'Awa Odori'. This study proposed the efficacies of Awa Odori Exercise as a rehabilitation exercise. This exercise expected the flexible balance reinforcements and the substitution for walking training with prevention of fall, bedridden and participating restriction for the old people, also promoting the health in Tokushima.