Antimicrobial activity of the volatile compound 3,5-dichloro-4-methoxybenzaldehyde, produced by the mushroom Porostereum spadiceum, against plant-pathogenic bacteria and fungi.

AIMS: In this study, volatile compounds released from mycelia of some aromatic mushrooms were investigated for their inhibitory activity against plant-pathogenic bacteria and fungi. METHODS AND RESULTS: A screening revealed that volatile compounds from mycelia of Porostereum spadiceum remarkably inhibited the colony formation of plant-pathogenic bacteria, including Clavibacter michiganensis subsp. michiganensis and Ralstonia solanacearum while also inhibiting the conidial germination of plant-pathogenic fungi including Alternaria brassicicola and Colletotrichum orbiculare. The volatile compounds were isolated from the culture filtrate of P. spadiceum, and 3,4-dichloro-4-methoxybenzaldehyde (DCMB) was identified as a major compound. DCMB significantly inhibited bacterial colonization at 10 µg ml-1 and fungal conidial germination at 0·1-1 µg ml-1 as a vapour. CONCLUSIONS: This is the first report on the production of the volatile compound DCMB by P. spadiceum and on the antimicrobial activity of DCMB against plant-pathogenic bacteria and fungi at low concentrations. It may be possible to use the compound as an agent for protecting crops from bacterial and fungal diseases during cultivation and storage. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides an understanding of antimicrobial activity of the mushroom volatile compound that may be useful as a novel biological control agent for protecting various plant diseases.

Effects of voluntary running exercise on skeletal muscle properties in nonobese rats with type 2 diabetes.

The skeletal muscles of animals and humans with type 2 diabetes have decreased oxidative capacity. Aerobic exercise can improve muscle oxidative capacity, but no data are available on the amount of exercise required. We investigated the effects of voluntary running exercise and running distance on the skeletal muscle properties of nonobese rats with type 2 diabetes. Six-week-old male diabetic Goto-Kakizaki rats were divided into nonexercised (GK) and exercised (GK-Ex) groups. The rats in the GK-Ex group were permitted voluntary running exercise on wheels for 6 weeks. Age-matched male Wistar rats (WR) were used as nondiabetic controls. Fasting blood glucose and HbA1c levels were higher in the GK and GK-Ex groups than in the WR group and lower in the GK-Ex group than in the GK group. Succinate dehydrogenase (SDH) activity and peroxisome proliferator-activated receptor gamma coactivator-1alpha (Pgc-1alpha) mRNA levels in the soleus and plantaris muscles were higher in the WR and GK-Ex groups than in the GK group. HbA1c and total cholesterol levels were negatively correlated with running distance and SDH activity and Pgc-1alpha mRNA levels in the soleus muscle were positively correlated with running distance. The onset and progression of diabetes in nonobese diabetic rats were effectively inhibited by running longer distances.

Effects of collagen hydrolysate on the tibialis anterior muscle and femur in senescence-accelerated mouse prone 6.

OBJECTIVES: Effects of collagen hydrolysate (CHD) on the oxidative capacity of the tibialis anterior muscle and the cortical and trabecular density of the femur were investigated in senescence-accelerated mouse prone 6 (SAMP6). METHODS: Sixteen-week-old male SAMP6 mice were divided into control (CON) and CHD groups. The CON group was given normal water, while the CHD group was given water containing CHD. Fibre cross-sectional areas (CSAs), fibre succinate dehydrogenase (SDH) staining intensity, and SDH activity of the tibialis anterior muscle were determined at 42 and 60 weeks of age. The cortical and trabecular density of the femur and serum osteocalcin levels were also determined. RESULTS: The fibre SDH staining intensity and muscle SDH activity were higher in the CHD group at 60 weeks of age than in the age-matched CON group. The cortical and trabecular density and serum osteocalcin levels were greater in the CHD group at 60 weeks of age than in the age-matched CON group. CONCLUSION: CHD inhibited th age-induced decrease in muscle oxidative capacity and bone density of SAMP6 mice. There is a possibility that CHD is effective for inhibition of age-induced degeneration in the musculoskeletal system.

Synthesis of nano-TaO(x) oxygen reduction reaction catalysts on multi-walled carbon nanotubes connected via a decomposition of oxy-tantalum phthalocyanine.

Nano-TaOx particles were supported on multi-walled carbon nanotubes via the thermal decomposition of oxy-tantalum phthalocyanine. The phthalocyanine-derived carbon connected TaOx particles with the nanotube-support to provide a conductive path. The oxygen reduction reaction activity, which solely originated from TaOx, was above 0.9 V with larger currents than conventional TaOx particles in acidic media.

Observations of crack propagation along a Zr-doped alumina grain boundary.

Structural ceramics are typically used in polycrystalline form. It is well known that polycrystalline ceramics often show the intergranular fracture. To improve their mechanical properties, transition metals can be used as dopants into a bulk material, which tend to segregate into the grain boundaries[1]. However, the effect of dopant segregation on grain boundary fracture is still uncertain. In order to investigate the fracture behavior of a dopant-segregated grain boundary, we observed the crack propagation of a Zr-doped alumina grain boundary by in situ nanoindentation in a transmission electron microscope (TEM), and characterized the fracture surface by scanning TEM (STEM).An alumina bicrystal with a Zr-doped Σ13 grain boundary was fabricated by diffusion bonding at 1500(o)C for 10 hours in air, where a face of one crystal was coated by Zr metal in advance to the bonding (Fig. 1a). A TEM sample was prepared from the bicrystal by mechanical grinding and Ar ion milling. For in situ indentation, the sample had a free edge perpendicular to the grain boundary (Fig. 1b). The indentation experiment was performed by using a double-tilt indentation holder (Nanofactory) and JEM-2010 (200kV, JEOL). The fracture surface was further observed by high angle annular dark field (HAADF) STEM (ARM-200F, 200kV, JEOL).jmicro;63/suppl_1/i20-a/DFU064F1F1DFU064F1Fig. 1.(a) Schematic illustrations of bicrystal fabrication by diffusion bonding and (b) Bright field TEM image showing the geometric arrangement of the in situ nanoindentation experiment In the in situ TEM nanoindentation experiment, at first a crack was introduced in bulk close to the grain boundary and propagated with the amount of indentation. After the crack reached the grain boundary, it preferentially propagated along the grain boundary. To identify the crack pass at the atomic level, the STEM analysis was performed. We found that three-atomic-layer Zr was formed in the unbroken region of the grain boundary, whereas one to three Zr layers remained on the fracture surface. This indicates that the crack propagated within the segregation region of Zr in the grain boundary. In the presentation, we will discuss the crack propagation behavior and the atomic structure of the fracture surfaces in detail.

Pharmacokinetics and bone resorption evaluation of a novel Cathepsin K inhibitor (VEL-0230) in healthy adult horses.

Plasma pharmacokinetic (PK) and bone resorption biomarker [carboxy-terminal cross-linking telopeptide of type I collagen (CTX-1)] analyses were performed following single and multiple oral dose protocols of a Cathepsin K inhibitor (VEL-0230) in horses. Outcomes included plasma and urine drug and CTX-1 concentrations. In the dose range study, 2, 4, and 8 mg/kg body weight (b.w.) doses were administered in a Latin square design to three mares and evaluated for 1 week. Based on the PK characteristics of VEL-0230, 4 mg/kg b.w. was selected for the dose interval study in which 3.25 days (d) and 7 days dose intervals were evaluated over three administrations using four exercising horses in a Latin square design. The 3.25 days and 7 days dose intervals provided a rapid inhibition of bone resorption based on plasma CTX-1. CTX-1 inhibition prior to next dose administration was not different from baseline in the 3.25 days and 7 days protocols, and for the first 3 days but the sustained CTX-1 inhibition in the 7 days protocol along with the cost and logistic benefits for weekly administration made the 7 days protocol preferable. Weekly administration of VEL-0230 may provide effective inhibition of bone resorption in young exercising horses that returns to baseline within 7 days after drug withdrawal even after multiple doses.

Inflammatory effects of autologous, genetically modified autologous, allogeneic, and xenogeneic mesenchymal stem cells after intra-articular injection in horses.

OBJECTIVES: To compare the clinical and inflammatory joint responses to intra-articular injection of bone marrow-derived mesenchymal stem cells (MSC) including autologous, genetically modified autologous, allogeneic, or xenogeneic cells in horses. METHODS: Six five-year-old Thoroughbred mares had one fetlock joint injected with Gey's balanced salt solution as the vehicle control. Each fetlock joint of each horse was subsequently injected with 15 million MSC from the described MSC groups, and were assessed for 28 days for clinical and inflammatory parameters representing synovitis, joint swelling, and pain. RESULTS: There were not any significant differences between autologous and genetically modified autologous MSC for synovial fluid total nucleated cell count, total protein, interleukin (IL)-6, IL-10, fetlock circumference, oedema score, pain-free range-of-motion, and soluble gene products that were detected for at least two days. Allogeneic and xenogeneic MSC produced a greater increase in peak of inflammation at 24 hours than either autologous MSC group. CLINICAL SIGNIFICANCE: Genetically engineered MSC can act as vehicles to deliver gene products to the joint; further investigation into the therapeutic potential of this cell therapy is warranted. Intra-articular MSC injection resulted in a moderate acute inflammatory joint response that was greater for allogeneic and xenogeneic MSC than autologous MSC. Clinical management of this response may minimize this effect.

Protective effects of astaxanthin on capillary regression in atrophied soleus muscle of rats.

AIM: The capillary regression in skeletal muscles associated with a chronic decrease in activity is related to a dysfunction of endocapillary cells induced by over-expression of oxidative stress. We hypothesized that treatment with astaxanthin, an antioxidant, would attenuate the oxidative stress induced by decreased skeletal muscle use, and that this attenuation would prevent the associated capillary regression. The purpose of the present study was to investigate the antioxidant and preventive effects of astaxanthin on capillary regression in the soleus muscle during hindlimb unloading. METHODS: Twenty-four adult male Wistar rats were assigned randomly either to a control, control plus astaxanthin treatment, hindlimb unloaded or hindlimb unloaded plus astaxanthin treatment group for 7 days. RESULTS: Hindlimb unloading resulted in a decrease in mean soleus absolute weight, capillary number, volume and luminal diameter. The accumulation of reactive oxygen species and the over-expression of superoxide dismutase (SOD-1), a decrease in the levels of vascular endothelial growth factor (VEGF) and its receptors, an inhibition of the angiopoietin pathway and an increase of thrombospondin-1 (TSP-1), as an anti-angiogenic factor were showed. Administration of astaxanthin attenuated the changes in SOD-1 and VEGF, up-regulated the angiogenic factors and reduced the capillary regression in the soleus of hindlimb unloaded rats. In addition, the VEGF-to-TSP1 ratio was higher in the astaxanthin treated groups than in the control and HU groups. CONCLUSION: These results suggest that astaxanthin may be an effective treatment to counter the detrimental effects of a chronic decrease in skeletal muscle use on the capillary network and associated angiogenic pathways.

Comparative efficacy of dermal fibroblast-mediated and direct adenoviral bone morphogenetic protein-2 gene therapy for bone regeneration in an equine rib model.

Cell-mediated and direct adenoviral (Ad) vector gene therapies can induce bone regeneration, including dermal fibroblasts (DFbs). We compared two effective therapies, DFb-mediated and direct Ad vector delivery of bone morphogenetic protein-2 (BMP2), for relative efficacy in bone regeneration. Equine rib drill defects were treated by percutaneous injection of either DFb-BMP2 or an Ad-BMP2 vector. At week 6, both DFb-BMP2- and Ad-BMP2-treated rib defects had greater bone filling volume and mineral density, with DFb-BMP2 inducing greater bone volume and maturity in the cortical bone aspect of the defect than Ad-BMP2. The transplantation of DFb alone induced modest bone formation. Increased mineral density and bone turnover were evident in the cortical and cancellous bone directly adjacent to the healing drill defects treated with either DFb-BMP2 or Ad-BMP2. Using our cell/vector dosage and model, BMP2, whether delivered by the DFb vector or direct Ad vector, induced greater and robust bone regeneration. DFb-mediated BMP2 therapy promoted greater cortical bone regeneration than did direct gene delivery, possibly because of an increased cellularity of the bone healing site. BMP2 delivery, regardless of gene delivery method, increased the mineral density of the neighboring bone, which may be beneficial clinically in repairing or weak bone.

Effects of exposure to hyperbaric oxygen on oxidative stress in rats with type II collagen-induced arthritis.

Arthritis was induced in 9-week-old female Dark Agouti rats by injecting type II collagen. Serum levels of the derivatives of reactive oxygen metabolites (dROMs), which are oxidative stress markers, and C-reactive protein (CRP) in arthritic rats that were exposed to a pressure of 1.25 atmospheres absolute and an oxygen concentration of 36% for 3 weeks (arthritis + HBO group) were compared to those of control rats (control group) and arthritic rats that were not exposed to hyperbaric oxygen (arthritis group). The body weights of the arthritis and arthritis + HBO groups were lower than that of the control group, whereas no difference in the body weight was observed between the arthritis and arthritis + HBO groups. The serum levels of dROMs and CRP in the arthritis group were higher than those in the control and arthritis + HBO groups. No difference in the serum level of CRP was observed between the control and arthritis + HBO groups. These results indicate that the conditions of hyperbaric oxygen exposure used in this study are effective for reducing the levels of reactive oxygen species, which are overproduced during arthritis.

Mechanism of the anti-obesity effects induced by a novel melanin-concentrating hormone 1-receptor antagonist in mice.

BACKGROUND AND PURPOSE: Melanin-concentrating hormone (MCH) is an orexigenic neuropeptide expressed in the lateral hypothalamus that is involved in feeding and body weight regulation. Intracerebroventricular infusion of a peptidic MCH1 receptor antagonist ameliorated obesity in murine models. Recently, small molecule MCH1 receptor antagonists have been developed and characterized for the treatment of obesity. However, little is known of the mechanism of the anti-obesity effects of MCH1 receptor antagonists. EXPERIMENTAL APPROACH: To examine the mechanisms of action of the anti-obesity effect of MCH1 receptor antagonists more precisely, we conducted a pair-feeding study in mice with diet-induced obesity (DIO), chronically treated with an orally active and highly selective MCH1 receptor antagonist and examined changes in mRNA expression levels in liver, brown and white adipose tissues. We also assessed the acute effects of the MCH1 receptor antagonist in energy expenditure under thermoneutral conditions. KEY RESULTS: Treatment with the MCH1 receptor antagonist at 30 mg.kg(-1) for 1 month moderately suppressed feeding and significantly reduced body weight by 24%. In contrast, pair-feeding resulted in a smaller weight reduction of 10%. Treatment with the MCH1 receptor antagonist resulted in a higher body temperature compared with the pair-fed group. TaqMan and calorimetry data suggested that the MCH1 receptor antagonist also stimulated thermogenesis. CONCLUSIONS AND IMPLICATIONS: Our results indicate that an MCH1 receptor antagonist caused anti-obesity effects im mice by acting on both energy intake and energy expenditure.

Protective effects of exercise preconditioning on hindlimb unloading-induced atrophy of rat soleus muscle.

AIM: A chronic decrease in the activation and loading levels of skeletal muscles as occurs with hindlimb unloading (HU) results in a number of detrimental changes. Several proteolytic pathways are involved with an increase in myofibrillar protein degradation associated with HU. Exercise can be used to counter this increase in proteolytic activity and, thus, may be able to protect against some of the detrimental changes associated with chronic decreased use. The purpose of the present study was to determine the potential of a single bout of preconditioning endurance exercise in attenuating the effects of 2 weeks of HU on the mass, phenotype and force-related properties of the soleus muscle in adult rats. METHODS: Male Wistar rats were subjected to HU for 2 weeks. One half of the rats performed a single bout of treadmill exercise for 25 min immediately prior to the 2 weeks of HU. RESULTS: Soleus mass, maximum tetanic tension, myofibrillar protein content, fatigue resistance and percentage of type I (slow) myosin heavy chain were decreased in HU rats. In addition, markers for the cathepsin, calpain, caspase and ATP-ubiquitin-proteasome proteolytic pathways were increased. The preconditioning endurance exercise bout attenuated all of the detrimental changes associated with HU, and increased HSP72 mRNA expression and protein levels. CONCLUSION: These findings indicate that exercise preconditioning may be an effective countermeasure to the detrimental effects of chronic decreases in activation and loading levels on skeletal muscles and that an elevation in HSP72 may be one of the mechanisms associated with these responses.

Blockade of MCH1 receptor signalling ameliorates obesity and related hepatic steatosis in ovariectomized mice.

BACKGROUND AND PURPOSE: Melanin-concentrating hormone (MCH) is a cyclic orexigenic neuropeptide predominantly expressed in the lateral hypothalamus. We investigated the roles of MCH1 receptor signalling in ovariectomy (OVX)-induced obesity in female C57BL/6J mice, an animal model of postmenopausal obesity. EXPERIMENTAL APPROACH: The effects of blocking signalling via the MCH1 receptor on OVX-induced obesity was investigated by using Mch1r deficient (KO) mice and chronic treatment with a selective MCH1 receptor antagonist. KEY RESULTS: OVX induced body weight gain and increases in the weight of visceral fat and of liver; these effects were attenuated following OVX in Mch1r KO mice. OVX-induced triglyceride (TG) accumulation and elevated expression of lipogenic genes were significantly ameliorated in the liver of Mch1r KO mice. In agreement with these results, chronic i.c.v. infusion of a selective MCH1 receptor antagonist significantly reduced body weight gain, visceral fat and liver weights in OVX mice, and hepatic TG contents and lipogenic gene expression levels were normalized. CONCLUSION AND IMPLICATIONS: Our results indicate that MCH1 receptor signalling is involved in the development of fatty liver, as well as obesity, in OVX mice, and suggest a therapeutic potential for MCH1 receptor antagonists in the treatment of obesity and fatty liver.

Hyperbaric exposure with high oxygen concentration improves altered fiber types in the plantaris muscle of diabetic Goto-Kakizaki rats.

Hyperbaric exposure with high oxygen concentration inhibits a growth-related increase in the glucose and insulin of diabetic rats. In this study, 5-week-old diabetic Goto-Kakizaki rats were exposed to a hyperbaric environment (1.25 atmospheric pressure) with a high oxygen concentration (36%) for 6 h daily. Fiber type distributions and oxidative enzyme activities in the fast-twitch plantaris muscle of Goto-Kakizaki rats were examined after hyperbaric exposure for 4 weeks. The percentages of high-oxidative type I and type IIA fibers increased and that of low-oxidative type IIB fibers decreased after hyperbaric exposure. Furthermore, the fiber oxidative enzyme activity increased after hyperbaric exposure, regardless of fiber type. It is concluded that altered patterns of fiber types in the plantaris muscle of diabetic rats shift toward normal, which is observed in nondiabetic rats, following hyperbaric exposure with high oxygen concentration.

Head torsion technique for detailed observation of larynx and hypopharynx.

We introduce here an easy but effective method for detailed observation of the larynx and hypopharynx. During the endoscopic observation, the patient's head is turned to one side. Anatomical structures on the same side of the endolarynx, such as the laryngeal ventricle and inferior surface of the vocal fold, are easily observed. In addition, observation of the opposite side of the hypopharynx also becomes easier. Such head turning is also useful in patients with an oblique larynx, in whom the epiglottis obstructs insertion of the endoscope. This is a simple but very effective technique for laryngeal and hypopharyngeal observation.

Effects of hyperbaric exposure with high oxygen concentration on the physical activity of developing rats.

The effects of hyperbaric exposure with high oxygen concentration on the physical activity of developing male rats were investigated. Five-week-old male rats were exposed to an atmospheric pressure of 1.25 with an oxygen concentration of 36.0% for 12 h (7.00-19.00 h) and exercised voluntarily for 12 h (19.00-7.00 h) daily for 8 weeks. The voluntary running activities were compared with those in age-matched rats without hyperbaric exposure. In addition, the properties of the soleus and plantaris muscle fibers and their spinal motoneurons were examined. The voluntary running activities of rats with or without hyperbaric exposure increased during development. However, the mean voluntary running activities were higher in rats with hyperbaric exposure (7,104 m/day) than in those without hyperbaric exposure (4,932 m/day). The oxidative capacities of the soleus and plantaris muscle fibers and their spinal motoneurons increased following hyperbaric exposure. It is suggested that adaptations of neuromuscular units to hyperbaric exposure with high oxygen concentration enhance the metabolism, and thus, the function of neuromuscular units is promoted.

Comparison of cell body size and oxidative enzyme activity in motoneurons between the cervical and lumbar segments in the rat spinal cord after spaceflight and recovery.

The cell body sizes and succinate dehydrogenase (SDH) activities of motoneurons in the dorsolateral region of the ventral horn at the cervical and lumbar segments in the rat spinal cord were determined following 9 days of spaceflight with or without 10 days of recovery on Earth. The motoneurons were divided into three types based on their cell body sizes; small-, medium-, and large-sized motoneurons. In control rats, there was no difference in the cell body size or SDH activity of small- and large-sized motoneurons between the cervical and lumbar segments. The SDH activity of medium-sized motoneurons in control rats was higher in the lumbar segment than in the cervical segment, while the cell body sizes of medium-sized motoneurons were identical. The SDH activity of medium-sized motoneurons in the lumbar segment decreased to a level similar to that in the cervical segment of control rats following spaceflight. In addition, the decreased SDH activity of medium-sized motoneurons persisted for at least 10 days of recovery on Earth. It is concluded that spaceflight selectively affects the SDH activity of medium-sized motoneurons in the lumbar segment of the spinal cord, which presumably innervate skeletal muscles having an antigravity function.

Detection of COL1A1-PDGFB fusion transcripts and platelet-derived growth factor alpha and beta receptors in giant cell fibroblastoma of the postsacrococcygeal region.

We describe a 2-year-old girl with recurrent giant cell fibroblastoma (GCF) of the postsacrococcygeal region. Both the initial and recurrent tumours contained solid and angiectoid areas. The former was composed of loosely arranged wavy spindle cells and giant cells with a well-vascularized myxoid to collagenous stroma. The angiectoid spaces were often lined by multinucleated giant cells. Immunohistochemically, the tumour cells and small vessels in the tumour tissue were positive for platelet-derived growth factor (PDGF) alpha and beta receptors. Molecular analysis revealed fusion of collagen type Ialpha1 exon 26 with PDGF-B chain exon 2 that induced unscheduled production of PDGF-BB. These findings suggest that PDGF and its receptors significantly contribute to the development of GCF in both an autocrine and a paracrine manner.

Effects of running exercise on fibre-type distribution of soleus and plantaris muscles in diabetic Otsuka Long-Evans Tokushima fatty rats.

AIM: Effect of running exercise on fibre-type distributions of the slow soleus and fast plantaris muscles was investigated in male Otsuka Long-Evans Tokushima fatty rats (OLETF) as an animal model of spontaneous type 2 diabetes mellitus. METHODS: Five-week-old OLETF rats were allowed to exercise voluntarily in running wheels for 32 days and the data were compared with those of age-matched non-exercised OLETF and non-diabetic Long-Evans Tokushima Otsuka rats (LETO). RESULTS: In the soleus muscle, a higher percentage of type I fibres was observed in non-exercised OLETF rats compared with LETO rats, and there were no type IIA fibres in non-exercised OLETF rats. In the plantaris muscle, a higher percentage of type IIB fibres and a lower percentage of type I and type IIA fibres were observed in non-exercised OLETF rats compared with LETO rats. In contrast, there were no differences in the fibre-type distribution of soleus and plantaris muscles between exercised OLETF and LETO rats. The body weight and type I fibre percentage of the soleus muscle were related to the running distance in exercised OLETF rats. White adipose tissue weight, HbA(1c) and blood insulin and glucose concentrations were lower in exercised OLETF rats than in non-exercised OLETF rats, irrespective of the running distance. There was a difference in the gene-expression pattern of the soleus muscle among LETO rats, non-exercised OLETF and exercised OLETF rats. CONCLUSION: Running exercise can inhibit diabetes-associated type shifting of fibres, which is more apparent with postnatal growth, in skeletal muscles of diabetic OLETF rats, as a result of mRNA expression change in muscle.