Electrical Stimulation-Based Twitch Exercise Suppresses Progression of Immobilization-Induced Muscle Fibrosis via Downregulation of PGC-1?/VEGF Pathway.

This study aimed to determine whether electrical stimulation-based twitch exercise is effective in inhibiting the progression of immobilization-induced muscle fibrosis. 19 Wistar rats were randomly divided into a control group (n=6), an immobilization group (n=6; with immobilization only), and a Belt group (n=7; with immobilization and twitch exercise through the belt electrode device, beginning 2 weeks after immobilization). The bilateral soleus muscles were harvested after the experimental period. The right soleus muscles were used for histological analysis, and the left soleus muscles were used for biochemical and molecular biological analysis. As a result, in the picrosirius red images, the perimysium and endomysium were thicker in both the immobilization and Belt groups compared to the control group. However, the perimysium and endomysium thickening were suppressed in the Belt group. The hydroxyproline content and alpha-SMA, TGF-beta1, and HIF-1alpha mRNA expressions were significantly higher in the immobilization and belt groups than in the control group. These expressions were significantly lower in the Belt group than in the immobilization group. The capillary-to-myofiber ratio and the mRNA expressions of VEGF and PGC-1alpha were significantly lower in the immobilization and belt groups than in the control group, these were significantly higher in the Belt group than in the immobilization group. From these results, Electrical stimulation-based twitch exercise using the belt electrode device may prevent the progression of immobilization-induced muscle fibrosis caused by downregulating PGC-1alpha/VEGF pathway, we surmised that this intervention strategy might be effective against the progression of muscle contracture. Keywords: Immobilization, Skeletal muscle, Fibrosis, Electrical stimulation-based twitch exercise, PGC-1alpha/VEGF pathway.

Utilising TMS-EEG to Assess the Response to Cerebellar-Brain Inhibition.

Cerebellar-brain inhibition (CBI) is a transcranial magnetic stimulation (TMS) paradigm indexing excitability of cerebellar projections to motor cortex (M1). Stimulation involved with CBI is often considered to be uncomfortable, and alternative ways to index connectivity between cerebellum and the cortex would be valuable. We therefore sought to assess the utility of electroencephalography in conjunction with TMS (combined TMS-EEG) to record the response to CBI. A total of 33 volunteers (25.7 ± 4.9 years, 20 females) participated across three experiments. These investigated EEG responses to CBI induced with a figure-of-eight (F8; experiment 1) or double cone (DC; experiment 2) conditioning coil over cerebellum, in addition to multisensory sham stimulation (experiment 3). Both F8 and DC coils suppressed early TMS-evoked EEG potentials (TEPs) produced by TMS to M1 (P < 0.05). Furthermore, the TEP produced by CBI stimulation was related to the motor inhibitory response to CBI recorded in a hand muscle (P < 0.05), but only when using the DC coil. Multisensory sham stimulation failed to modify the M1 TEP. Cerebellar conditioning produced changes in the M1 TEP that were not apparent following sham stimulation, and that were related to the motor inhibitory effects of CBI. Our findings therefore suggest that it is possible to index the response to CBI using TMS-EEG. In addition, while both F8 and DC coils appear to recruit cerebellar projections, the nature of these may be different.

Magnetization control by angular momentum transfer from surface acoustic wave to ferromagnetic spin moments.

Interconversion between electron spin and other forms of angular momentum is useful for spin-based information processing. Well-studied examples of this are the conversion of photon angular momentum and rotation into ferromagnetic moment. Recently, several theoretical studies have suggested that the circular vibration of atoms work as phonon angular momentum; however, conversion between phonon angular momentum and spin-moment has yet to be demonstrated. Here, we demonstrate that the phonon angular momentum of surface acoustic wave can control the magnetization of a ferromagnetic Ni film by means of the phononic-to-electronic conversion of angular momentum in a Ni/LiNbO3 hybrid device. The result clearly shows that the phonon angular momentum is useful for increasing the functionality of spintronic devices.

Intra-articular human deciduous dental pulp stem cell administration vs. pharmacological therapy in experimental osteoarthritis rat model.

OBJECTIVE: The aim of the present study was to compare the molecular and morphological effects of diacerein and glucosamine-chondroitin drug treatment and intra-articular injection therapy of human deciduous dental pulp stem cells (hDPSCs) in a rat knee model of induced osteoarthritis (OA). MATERIALS AND METHODS: Thirty-six adult male rats were randomly separated into six groups: Control group (without induction of OA), osteoarthritis group 60 (induction of OA, saline gavage started on day 14 and performed for 60 days, followed by euthanasia), osteoarthritis group (induction of OA and euthanasia after 14 days), diacerein group, glucosamine-chondroitin group, and mesenchymal stem cell group. The drug-treated groups were gavaged with 50 mg/kg of diacerein and 400/500 mg/kg of glucosamine-chondroitin starting on dat 14 for 60 days. The cell therapy-treated group received an intra-articular single dose of 8 × 105 hDPSCs on day 14, and euthanasia was performed after 60 days. Lateral femoral condyles were collected and prepared for immunohistochemistry and light microscopy procedures. RESULTS: The morphological features and immunoexpression of SOX-5, IHH, MMP-8, MMP-13, and Type II collagen were statistically analysed. Our data suggest that hDPSC therapy contributes more actively and effectively in the structural reorganization of lateral femoral condyles. In contrast, the glucosamine-chondroitin sulphate treatment was more effective in inflammatory control, while diacerein showed better results associated with the maintenance of the primordial cartilage. CONCLUSIONS: The positive therapeutic effect of daily administered conventional drugs can be confirmed in a rat model of OA. However, one single dose of locally administered hDPSCs provides significant improvement in tissue regeneration in an OA model.

New proposal to revise the classification for squamous cell carcinoma of the external auditory canal and middle ear.

BACKGROUND: The prognosis of patients with advanced squamous cell carcinoma of the external auditory canal and middle ear has been improved by advances in skull base surgery and multidrug chemoradiotherapy during the last two decades. METHODS: Ninety-five patients with squamous cell carcinoma of the external auditory canal and middle ear who were treated between 1998 and 2017 were enrolled. The number of patients with tumour stages T1, T2, T3 and T4 was 15, 22, 24 and 34, respectively. Oncological outcomes and prognostic factors were retrospectively investigated. RESULTS: Among patients with T4 disease, invasion of the brain (p = 0.024), carotid artery (p = 0.049) and/or jugular vein (p = 0.040) were significant predictors of poor prognosis. The five-year overall survival rate of patients with at least one of these factors (T4b) was significantly lower than that of patients without these factors (T4a) (25.5 vs 65.5 per cent, p = 0.049). CONCLUSION: It is proposed that stage T4 be subclassified into T4a and T4b according to the prognostic factors.

The effects of wheel-running using the upper limbs following immobilization after inducing arthritis in the knees of rats.

This study investigated the effects of wheel-running using the upper limbs following immobilization after inducing arthritis in the knees of rats. Forty male Wistar rats (aged 8 weeks) divided into four groups randomly: arthritis (AR), immobilization after arthritis (Im), wheel-running exercise with the upper limbs following immobilization after arthritis induction (Im+Ex) and sham arthritis induction (Con). The knee joints of the Im and Im+Ex groups were immobilized with a cast for 4 weeks. In the Im+Ex group, wheel-running exercise was administered for 60 min/day (5 times/week). The swelling and the pressure pain threshold (PPT) of the knee joint were evaluated for observing the condition of inflammatory symptoms in affected area, and the paw withdraw response (PWR) was evaluated for observing the condition of secondary hyperalgesia in distant area. Especially, in order to evaluate histological inflammation in the knee joint, the number of macrophage (CD68-positive cells) in the synovium was examined. The expression of calcitonin gene-related peptide (CGRP) in the spinal dorsal horn (L2-3 and L4-5) was examined to evaluate central sensitization. The Im+Ex group showed a significantly better recovery than the Im group in the swelling, PPTs, and PWRs. Additionally, CGRP expression of the spinal dorsal horn (L2-3 and L4-5) in the Im+Ex group was significantly decreased compared with the Im group. According to the results, upper limb exercise can decrease pain in the affected area, reduce hyperalgesia in distant areas, and suppress the central sensitization in the spinal dorsal horn by triggering exercise-induced hypoalgesia (EIH).

Intra-articular human deciduous dental pulp stem cell administration vs. pharmacological therapy in experimental osteoarthritis rat model

OBJECTIVE: The aim of the present study was to compare the molecular and morphological effects of diacerein and glucosamine-chondroitin drug treatment and intra-articular injection herapy of human deciduous dental pulp stem cells (hDPSCs) in a rat knee model of induced osteoarthritis OA. MATERIALS AND METHODS: Thirty-six adult male rats were randomly separated into six groups: Control group without induction of OA, osteoarthritis group 60 induction of OA, saline gavage started on day 14 and performed for 60 days, followed by euthanasia, osteoarthritis group induction of OA and euthanasia after 14 days, diacerein group, glucosamine-chondroitin group, and mesenchymal stem cell group. The drug-treated groups were gavaged with 50 mg/kg of diacerein and 400/500 mg/kg of glucosamine-chondroitin starting on dat 14 for 60 days. The cell therapy-treated group received an intra-articular single dose of 8 × 105 hDPSCs on day 14, and euthanasia was performed after 60 days. Lateral femoral condyles were collected and prepared for immunohistochemistry and light microscopy procedures. RESULTS: The morphological features and immunoexpression of SOX-5, IHH, MMP-8, MMP-13, and Type II collagen were statistically analysed. Our data suggest that hDPSC therapy contributes more actively and effectively in the structural reorganization of lateral femoral condyles. In contrast, the glucosamine-chondroitin sulphate treatment was more effective in inflammatory control, while diacerein showed better results associated with the maintenance of the primordial cartilage. CONCLUSIONS: The positive therapeutic effect of daily administered conventional drugs can be confirmed in a rat model of OA. However, one single dose of locally administered hDPSCs provides significant improvement in tissue regeneration in an OA model.

Development of Duodenal Ulcers due to the Discontinuation of Proton Pump Inhibitors After the Induction of Sofosbuvir Plus Ledipasvir Therapy: A Report of Two Cases.

Sofosbuvir plus ledipasvir (SOF-LDV) combination therapy is a promising therapy for post-transplant hepatitis C virus (HCV) reinfection. It is known that gastric pH elevation induces lower absorption of ledipasvir; therefore, the use of proton pump inhibitors (PPIs) should be considered regarding dose reduction after SOF-LDV therapy induction. Here, we report two patients who developed duodenal ulcers due to the discontinuation of PPIs after the induction of SOF-LDV therapy for post-transplant HCV reinfection. The first patient was a 71-year-old man who had undergone living donor liver transplantation due to HCV-related liver cirrhosis. Lansoprazole, 30 mg daily, was discontinued upon SOF-LDV therapy induction. Seven days after SOF-LDV therapy induction, gastrointestinal endoscopy revealed the presence of a duodenal ulcer. The second patient was a 54-year-old man who had undergone living donor liver transplantation due to HCV-related end-stage liver disease. Similar to the first patient, rabeprazole sodium was discontinued upon the induction of SOF-LDV therapy. Eighteen days after SOF-LDV therapy induction, gastrointestinal endoscopy revealed the presence of a duodenal ulcer. In both cases, these duodenal ulcers improved after the resumption of the administration of PPIs, and a sustained virologic response at 12 weeks was achieved by SOF-LDV therapy with PPI use. Thus, PPI use should be continued consistently during SOF-LDV therapy for post-transplant HCV reinfection.

Dynamic encapsulation of corannulene molecules into a single-walled carbon nanotube.

The morphology of corannulene molecules encapsulated in a single-walled carbon nanotube (SWCNT) is addressed using atomistic simulations. Firstly, dynamic simulation (DS) of encapsulation of corannulene molecules into a SWCNT is performed using a molecular dynamics (MD) method. It is revealed that corannulene molecules encapsulated in a SWCNT tend to form concave-concave (CC) dimers, and these dimers make stacks tilting against the SWCNT axis or take an arrangement such that their convex surfaces face the inner wall of the SWCNT. This tendency arises from the fact that the van der Waals interactions between the convex surfaces of the corannulene molecules and the inner wall of the SWCNT dominate in their dynamic encapsulation into the SWCNT, and CC dimers are favored based on the energetics. Next, conjugate gradient (CG) energy minimizations starting from two kinds of initial arrangement of corannulene molecules in a SWCNT, concave-convex (CV) and CC/convex-convex (VV) arrangements, are performed. The CG energy minimizations confirm the result of DS that CC dimers are the structural motif of corannulene molecules in a SWCNT. From the final configurations of both the simulations, the tilt angles and intermolecular distances of the stacked molecules are calculated. With increasing the SWCNT diameter, the tilt angles decrease while the intermolecular distances remain almost constant. The tilt angles of the stacked corannulene molecules are approximately expressed by a semi-analytical formula which is derived on the basis of a geometrical constraint condition.

Coping strategies and risk of cardiovascular disease incidence and mortality: the Japan Public Health Center-based prospective Study.

AIMS: Coping strategies may be significantly associated with health outcomes. This is the first study to investigate the association between baseline coping strategies and cardiovascular disease (CVD) incidence and mortality in a general population cohort. METHODS AND RESULTS: The Japan Public Health Center-based prospective Study asked questions on coping in its third follow-up survey (2000-04). Analyses on CVD incidence and mortality included 57 017 subjects aged 50-79 without a history of CVD and who provided complete answers on approach- and avoidance-oriented coping behaviours and strategies. Cox regression models, adjusted for confounders, were used to determine hazard ratios (HRs) according to coping style. Mean follow-up time was 7.9 years for incidence and 8.0 years for mortality.The premorbid use of an approach-oriented coping strategy was inversely associated with incidence of stroke (HR = 0.85; 95% CI, 0.73-1.00) and CVD mortality (HR = 0.74; 95% CI, 0.55-0.99). Stroke subtype analyses revealed an inverse association between the approach-oriented coping strategy and incidence of ischaemic stroke (HR = 0.79; 95% CI, 0.64-0.98) and a positive association between the combined coping strategy and incidence of intra-parenchymal haemorrhage (HR = 2.03; 95% CI, 1.01-4.10). Utilizing an avoidance coping strategy was associated with increased mortality from ischaemic heart disease (IHD) only in hypertensive individuals (HR = 3.46; 95% CI, 1.07-11.18). The coping behaviours fantasizing and positive reappraisal were associated with increased risk of CVD incidence (HR = 1.24; 95% CI, 1.03-1.50) and reduced risk of IHD mortality (HR = 0.63; 95% CI, 0.40-0.99), respectively. CONCLUSION: An approach-oriented coping strategy, i.e. proactively dealing with sources of stress, may be associated with significantly reduced stroke incidence and CVD mortality in a Japanese population-based cohort.

Surgical procedure of extracting teeth for obtaining dental pulp for regenerative medicine in swine.

Dental pulp is a potential source of cells that can be used in cell replacement therapy for various nerve disorders, including stroke, spinal cord injury, and peripheral nerve defect. However, the validation of an animal model closely related to humans is needed in translational research. The miniature pig is a suitable experimental model in maxillofacial surgery, because its anatomical structure and size are similar to those of humans. However, the swine tooth is extremely long. The routine closed extraction procedure for harvesting dental pulp tissue causes root fracture. This report describes the details of a surgical procedure for tooth extraction. Four healthy 7-8-month-old male NIBS miniature pigs were used. Two mandibular deciduous right incisors (Di1 and Di2) were extracted in order to obtain dental pulp tissue. Gingival envelope incision with vertical-releasing incision was performed, and a full-thickness mucoperiosteal flap was made. The buccal alveolar bone was exposed and removed by osteotomy. Di1 and Di2 were extracted. Dental pulp tissue was obtained from these extracted teeth by splitting hard tissue. In this procedure, 9.8 ± 2.5 × 10(5) cells were obtained from the mandibular Di1 and Di2 (n = 4).

Development of an analytical method for the determination of sterigmatocystin in grains using LCMS after immunoaffinity column purification.

The mycotoxin sterigmatocystin (STC) is produced mainly by some Aspergillus and Penicillium fungi; it naturally contaminates cereals, peanuts, and products derived from these crops, and is both mutagenic and carcinogenic. As an intermediate of aflatoxin (AF) biosynthesis, its structure is similar to that of AF. Although immunoaffinity columns (IACs) are a popular approach to sample clean-up, no IAC is commercially available for STC, but a commercially available IAC for AF shows cross reactivity to STC. We here developed a new method for analyzing STC in grains using such an IAC and liquid chromatography mass spectrometry (LCMS), and validated this method using six different grains. The STC limit of detection (signal-to-noise ratio, S/N = 3) was 2.5 pg (1.0 µg/kg in the product), and the calibration curve was linear in the range of 7.5-375 pg (3.0-150 µg/kg in the product). The within-day recovery of STC from samples spiked with STC at 5.0 and 50 µg/kg was 83.2-102.5% and the RSDr (relative standard deviation of repeatability) of these samples was 1.9-6.5%; the RSDr of STC-pretreated grain samples was 3.1-14.0%. Average recovery of STC from samples spiked with STC in the range of 5.0-100 µg/kg STC was 83.2-102.5%, with an RSDr of 0.24-6.5%; the RSDr of STC-pretreated grain samples was 2.4-14.0%. In an intermediate precision study, the average STC recovery from STC-spiked samples by three analysts was 95.2-107.5%, with RSDRi (intermediate precision) of 4.0-7.1%; the RSDRi of the STC-pretreated samples was 4.8-10.4%. Thus, the proposed method was effective for STC analysis in grains, and holds potential for a novel application of a commercial IAC, intended for AFs, in STC analysis.

Particle therapy using carbon ions or protons as a definitive therapy for patients with primary sacral chordoma.

OBJECTIVE: This study retrospectively evaluated the efficacy and toxicity of particle therapy using carbon ions or protons for primary sacral chordomas. METHODS: We evaluated 23 patients with primary sacral chordoma treated with carbon ion therapy (CIT) or proton therapy (PT) between July 2005 and June 2011 at the Hyogo Ion Beam Medical Center, Hyogo, Japan. The median patient age was 72 years. 14 patients were treated with 70.4 Gy equivalents (GyE) in 16 fractions and 9 were treated with 70.4 GyE in 32 fractions. CIT was used for 16 patients, and PT was used for 7 patients. RESULTS: The median follow-up period was 38 months. At 3 years, local control (LC), overall survival (OS) and progression-free survival (PFS) for all patients were 94%, 83% and 68%, respectively. The log-rank test revealed that male sex was significantly related to better PFS (p=0.029). No other factors, including dose fractionation and ion type, were significant for LC, OS or PFS. In nine patients, ≥ Grade 3 acute dermatitis was observed, and ≥ Grade 3 late toxicities were observed in nine patients. The 32-fraction protocol reduced severe toxicities in both the acute and late phases compared with the 16-fraction protocol. CONCLUSION: Particle therapy for patients with sacral chordoma showed favourable LC and OS. Severe toxicities were successfully reduced by modifying the dose fractionation and treatment planning in the later treatment era. Thus, this therapeutic modality should be considered useful and safe. ADVANCES IN KNOWLEDGE: This is the first study including both CIT and PT for sacral chordomas.

Inactivation of koi-herpesvirus in water using bacteria isolated from carp intestines and carp habitats.

Since its first outbreak in Japan in 2003, koi-herpesvirus (KHV) remains a challenge to the carp Cyprinus carpio L. breeding industry. In this study, inactivation of KHV in water from carp habitats (carp habitat water) was investigated with the aim of developing a model for rapidly inactivating the pathogen in aquaculture effluent. Experiments with live fish showed that, in carp habitat water, KHV lost its infectivity within 3 days. Indications were that inactivation of KHV was caused by the antagonistic activity of bacteria (anti-KHV bacteria) in the water from carp habitats. Carp habitat water and the intestinal contents of carp were therefore screened for anti-KHV bacteria. Of 581 bacterial isolates, 23 showed anti-KHV activity. An effluent treatment model for the disinfection of KHV in aquaculture effluent water using anti-KHV bacteria was developed and evaluated. The model showed a decrease in cumulative mortality and in the number of KHV genome copies in kidney tissue of fish injected with treated effluent compared with a positive control. It is thought that anti-KHV bacteria isolated from the intestinal contents of carp and from carp habitat water can be used to control KHV outbreaks.

Transplantation of a tooth involved in dentigerous cyst.

CASE REPORT: To salvage an uneruputed tooth within a dentigerous cyst, the retransplantation of the unerupted premolar involved in the dentigerous cyst was performed. In the present case a dentigerous cyst with the impacted maxillary second premolar of a 10-year-old boy was removed, because of the missing eruption after marsupialisation of the premolar 4 months earlier. Covering the crown and the one third of root, the cyst was removed from the tooth, and the premolar was retransplanted into a cavity created between the premolar and first molar in the maxilla. The tooth was fixed with resin and a twisted wire for 1 month. Two years after transplantation, the tooth responded positively to pulp vitality tests but did not move toward the occlusal line, and dental X-ray showed a lamina dura, mineralisation of the pulp cavity, no root resorption and development.

Correlation of circulating dehydroepiandrosterone with activated protein C generation and carotid intima-media thickness in male patients with type 2 diabetes.

AIMS: Dehydroepiandrosterone exerts a protective effect against cardiovascular diseases. However, the relationship of dehydroepiandrosterone with the anticoagulant factor activated protein C, generated by the thrombin-thrombomodulin complex on vascular endothelial cells, remains unknown. This study aimed at studying the relationship between dehydroepiandrosterone and activated protein C generation in patients with Type 2 diabetes. METHODS: Sixty-two male patients with Type 2 diabetes were enrolled in this study. Data obtained from 40 healthy male subjects were used as controls. The plasma levels of dehydroepiandrosterone, the activated protein C-protein C inhibitor complex, high-sensitivity C-reactive protein and monocyte chemoattractant protein-1 were measured by enzyme immunoassays. Carotid intima-media thickness was measured by ultrasonography. RESULTS: The plasma levels of dehydroepiandrosterone (5.15 ± 2.81 vs. 3.76 ± 2.16 ng/ml; P < 0.005) and the activated protein C-protein C inhibitor complex (1.90 ± 1.07 vs. 1.02 ± 0.51 ng/ml; P < 0.001) were significantly lower in patients with diabetes than in normal subjects. Univariate analysis showed a significant correlation of the plasma level of dehydroepiandrosterone with that of the activated protein C-protein C inhibitor complex (r = 0.48, P < 0.001), high-sensitivity C-reactive protein (r = -0.30, P < 0.05) and with the mean intima-media thickness (r = -0.28, P < 0.05) in patients with diabetes. Stepwise multiple regression analysis showed that the plasma level of dehydroepiandrosterone is significantly correlated with the plasma levels of the activated protein C-protein C inhibitor complex (F = 18.06) and high-sensitivity C-reactive protein (F = 4.94). There was no correlation between the plasma levels of dehydroepiandrosterone and monocyte chemoattractant protein-1. CONCLUSIONS: These results suggest that lower circulating levels of dehydroepiandrosterone are associated with decreased activated protein C generation and higher intima-media thickness in patients with Type 2 diabetes.

Effect of tooth bleaching on bond strength of enamel-dentin cavities restored with silorane- and dimethacrylate-based materials.

The aim of this study was to evaluate the influence of tooth bleaching on the push-out bond strength of a composite resin based on dimethacrylates and silorane to cavities that involve both enamel and dentin. A total of 80 bovine incisors were sectioned on the buccal surface to obtain specimens (10 × 10 mm) presenting enamel and dentin (1-mm thick each substrate). The specimens were randomly distributed into eight groups (n=10), according to the bleaching protocol (1--none; 2--10% carbamide peroxide [CP] for 21 days, six hours each day; 3--three applications of 35% hydrogen peroxide [HP] in 15-minute sessions, one session every seven days for three weeks; 4--10% CP for 18 days, six hours each day + three applications of 35% HP in 15-minute sessions, one session every seven days for three weeks) and the restorative system applied (Adper Single Bond 2 + Filtek Supreme; Filtek Silorane adhesive and composite resin). After treatment, cavities were made (1.2-mm diameter on dentin; 1.5-mm diameter on enamel) with a diamond bur. At 24 hours after restoration, a push-out bond strength test was performed at a crosshead speed of 0.5 mm/min. The bleaching treatments did not significantly affect the bond strengths of either restorative system to enamel-dentin. Regardless of the bleaching treatment, the dimethacrylate-based resin system exhibited significantly higher bond strengths to enamel-dentin than did the silorane-based system.