Monitoring adaptation of skin tissue oxygenation during cycling ergometer exercise by frequency-domain diffuse optical spectroscopy.

In addition to supplying oxygen molecule O2 for metabolic functions during the adaptation to exercise, blood also plays a critical role in heat dissipation for core temperature stabilization. This study investigates the status of hemodynamic oxygenation in the forearm's skin tissue of three participants during a complete ergometer exercise from the resting to exercising, and to recovering conditions using a three-wavelength frequency-domain diffuse reflectance spectroscopy (FD DRS) alongside the monitoring of heartbeat rate and skin temperature. The FD DRS system was synchronized with radiofrequency (RF)-modulated input photon sources and the respective output to extract time-course absorption and scattering coefficients of the skin tissue, which, through the fitting of lambert's law of absorbance, can be used to determine the concentration of oxygenated/deoxygenated hemoglobin molecules, and consequentially, the oxygen saturation of skin tissue and total hemoglobin (THb) concentration. Expressly, a sudden jump in heartbeat rate at the beginning of the exercise, a temporal lag of the rising edge of skin temperature behind that of the THb concentration in the procession of step-wise incremental working intensity, and the uprising of THb in the exhaustion zone in responses to the physiological adaptation to exercise were identified. Finally, conclusive remarks were drawn that the FD DRS system is useful in extracting the hemodynamic properties of forearm skin which is often being neglected in previous exercise physiology studies by DRS-related techniques. The detailed variation of hemodynamic and optical scattering parameters of forearm skin elucidated in the studies can be applied for the analysis of athletes' physiological status, and may be a potential reference for the design of future wearable devices.

Bioinformatics and experimental findings reveal the therapeutic actions and targets of pachymic acid against cystitis glandularis.

Pachymic acid (PA), a bioactive ingredient isolated from Poria cocos Wolf, is reported with potential benefits of anti-inflammatory, anti-oxidative actions. It is reasoned that PA may play the potential benefits against cystitis glandularis (CG), an inflammation of the bladder tissue. In this study, we aimed to apply the network pharmacology and molecular docking analyses to reveal concrete anti-CG targets and mechanisms of PA, and then the bioinformatic findings were verified by using clinical and animal samples. The methodological data from network pharmacology approach showed that 303 and 243 reporting targets of CG and PA, and other 31 shared targets of CG and PA were identified. Subsequently, all top targets of PA against CG were screened out, including cyclooxygenase-2, epidermal growth factor receptor, tumor antigen p53 (TP53), tumor necrosis factor-alpha (TNF), interleukin-1 (IL-1) beta, proto-oncogene c-jun. Molecular docking data demonstrated that PA exerted potent bonding capacities with TNF, TP53 proteins in CG. In human study, the findings suggested that overactivated TNF-α expression and suppressed TP53 activation were detected in CG samples. In animal study, PA-treated mice showed reduced intravesical IL-1, IL-6 levels, and lactate dehydrogenase content, downregulated TNF-α and upregulated TP53 proteins in bladder samples. Taken together, our bioinformatics and experimental findings identify the key anti-CG biotargets and mechanisms of PA. More markedly, these pivotal pharmacological targets of PA against CG have been screened out and verified by using computational and experimental analyses.

Targeted and sustained Sox9 expression in mouse hypertrophic chondrocytes causes severe and spontaneous osteoarthritis by perturbing cartilage homeostasis.

Sox9 is the master transcription factor essential for cartilage development and homeostasis. To investigate the specific role of Sox9 during chondrocyte hypertrophy, we generated a novel Col10a1-Sox9 transgenic mouse model, in which Sox9 is specifically expressed in hypertrophic chondrocytes driven by a well-characterized 10-kb Col10a1 promoter. These mice were viable and fertile, and appeared normal at birth. However, they developed dwarfism by ten weeks of age. The histological analysis of the growth plates from these transgenic mice demonstrated an abnormal growth plate architecture and a significantly reduced amount of trabecular bone and mineral content in the primary spongiosa. Real-time qPCR analysis revealed the reduced expression of Col10a1, and increased expressions of adipogenic differentiation markers in primary hypertrophic chondrocytes isolated from transgenic mice. Concomitantly, the transgenic mouse chondrocyte cultures had increased lipid droplet accumulation. Unexpectedly, we also observed an increased incidence of spontaneous osteoarthritis (OA) development in the transgenic mice by X-ray analysis, micro-computed tomography scanning, and histological examination of knee joints. The manifestation of OA in Col10a1-Sox9 transgenic mice began by six-months of age, and worsened by eleven-months of age. In conclusion, we provide strong evidence that the proper spatiotemporal expression of Sox9 is necessary for normal adult hypertrophic cartilage homeostasis, and that the aberrant expression of Sox9 might lead to spontaneous OA development.

Loss of jab1 in osteochondral progenitor cells severely impairs embryonic limb development in mice.

The transcriptional cofactor Jab1 controls cell proliferation, apoptosis, and differentiation in diverse developmental processes by regulating the activity of various transcription factors. To determine the role of Jab1 during early limb development, we developed a novel Jab1(flox/flox) ; Prx1-Cre conditional Knockout (cKO) mutant mouse model in which Jab1 was deleted in the osteochondral progenitor cells of the limb buds. Jab1 cKO mutant mice displayed drastically shortened limbs at birth. The short-limb defect became apparent in Jab1 cKO mutants at E15.5 and increasingly worsened thereafter. By E18.5, Jab1 cKO mutant mice exhibited significantly shorter limbs with: very few hypertrophic chondrocytes, disorganized chondrocyte columns, much smaller primary ossification centers, and significantly increased apoptosis. Real-time RT-PCR analysis showed decreased expression of Sox9, Col2a1, Ihh, and Col10a1 in Jab1 cKO mutant long bones, indicating impaired chondrogenesis. Furthermore, in a micromass culture model of early limb mesenchyme cells, alcian blue staining showed a significant decrease in chondrogenesis in Jab1 cKO limb bud cells. The expression of Sox9 and its downstream targets Col2a1 and Aggrecan, as well as BMP signaling downstream targets, Noggin, Id1, and Ihh, were significantly decreased in Jab1 cKO micromass cultures. Moreover, over-expression of SOX9 in Jab1 cKO micromass cultures partially restored Col2a1and Aggrecan expression. Jab1-deficient micromass cultures also exhibited decreased BMP signaling response and reduced BMP-specific reporter activity ex vivo. In summary, our study demonstrates that Jab1 is an essential regulator of early embryonic limb development in vivo, likely in part by co-activating Sox9 and BMP signaling.

The transcriptional co-regulator Jab1 is crucial for chondrocyte differentiation in vivo.

The evolutionarily conserved transcriptional cofactor Jab1 plays critical roles in cell differentiation, proliferation, and apoptosis by modulating the activity of diverse factors and regulating the output of various signaling pathways. Although Jab1 can interact with the bone morphogenetic protein (BMP) downstream effector Smad5 to repress BMP signaling in vitro, the role of Jab1 in BMP-mediated skeletogenesis in vivo is still poorly understood. As a key regulator of skeletogenesis, BMP signaling regulates the critical Ihh-Pthrp feedback loop to promote chondrocyte hypertrophy. In this study, we utilized the loxP/Cre system to delineate the specific role of Jab1 in cartilage formation. Strikingly, Jab1 chondrocyte-specific knockout Jab1(flox/flox); Col2a1-Cre (cKO) mutants exhibited neonatal lethal chondrodysplasia with severe dwarfism. In the mutant embryos, all the skeletal elements developed via endochondral ossification were extremely small with severely disorganized chondrocyte columns. Jab1 cKO chondrocytes exhibited increased apoptosis, G2 phase cell cycle arrest, and increased expression of hypertrophic chondrocyte markers Col10a1 and Runx2. Jab1 can also inhibit the transcriptional activity of Runx2, a key regulator of chondrocyte hypertrophy. Notably, our study reveals that Jab1 is likely a novel inhibitor of BMP signaling in chondrocytes in vivo. In Jab1 cKO chondrocytes, there was heightened expression of BMP signaling components including Gdf10/Bmp3b and of BMP targets during chondrocyte hypertrophy such as Ihh. Furthermore, Jab1 cKO chondrocytes exhibited an enhanced response to exogenous BMP treatment. Together, our study demonstrates that Jab1 represses chondrocyte hypertrophy in vivo, likely in part by downregulating BMP signaling and Runx2 activity.

Ectopic expression of SOX9 in osteoblasts alters bone mechanical properties.

Osteoporosis is a common skeletal disease characterized by low bone mass and microarchitectural deterioration of bone tissue, with a consequent increase in bone fragility and susceptibility to fracture. We previously demonstrated that Col1a1-SOX9 transgenic (TG) mice, in which SOX9 specifically expresses in osteoblasts driven by a 2.3-kb Col1a1 promoter, display osteopenia during the early postnatal stage. In this study, to further analyze the osteopenia phenotype and especially the effect of the osteoblast-specific expression of SOX9 on bone mechanical properties, we performed bone geometry and mechanical property analysis of long bones from Col1a1-SOX9 TG mice and wild-type littermates (WT) at different time points. Interestingly, after body weight adjustment, TG mice had similar whole-bone strength as WT mice but significantly thinner cortical bone, lower elastic modulus, and higher moment of inertia. Thus, osteoblast-specific SOX9 expression results in altered bone structure and material properties. Furthermore, the expression levels of Pcna, Col1a1, osteocalcin, and the Opg/Rankl ratio in TG mice were significantly lower until 4 months of age compared with WT mice, suggesting that TG mice have dysregulated bone homeostasis. Finally, bone marrow stromal cells (MSCs) isolated from TG mice display enhanced adipocyte differentiation and decreased osteoblast differentiation in vitro, suggesting that osteoblast-specific expression of SOX9 can lead to altered mesenchymal stem cell differentiation potentials. In conclusion, our study implies that SOX9 activity has to be tightly regulated in the adult skeleton to ensure optimal bone quality.

Rescue of impaired fracture healing in COX-2-/- mice via activation of prostaglandin E2 receptor subtype 4.

Although the essential role of cyclooxygenase (COX)-2 in fracture healing is known, the targeted genes and molecular pathways remain unclear. Using prostaglandin E2 receptor (EP)2 and EP4 agonists, we examined the effects of EP receptor activation in compensation for the lack of COX-2 during fracture healing. In a fracture-healing model, COX-2(-/-) mice showed delayed initiation and impaired endochondral bone repair, accompanied by a severe angiogenesis deficiency. The EP4 agonist markedly improved the impaired healing in COX-2(-/-) mice, as evidenced by restoration of bony callus formation on day 14, a near complete reversal of bone formation, and an approximately 70% improvement of angiogenesis in the COX-2(-/-) callus. In comparison, the EP2 agonist only marginally enhanced bone formation in COX-2(-/-) mice. To determine the differential roles of EP2 and EP4 receptors on COX-2-mediated fracture repair, the effects of selective EP agonists on chondrogenesis were examined in E11.5 long-term limb bud micromass cultures. Only the EP4 agonist significantly increased cartilage nodule formation similar to that observed during prostaglandin E2 treatment. The prostaglandin E2/EP4 agonist also stimulated MMP-9 expression in bone marrow stromal cell cultures. The EP4 agonist further restored the reduction of MMP-9 expression in the COX-2(-/-) fracture callus. Taken together, our studies demonstrate that EP2 and EP4 have differential functions during endochondral bone repair. Activation of EP4, but not EP2 rescued impaired bone fracture healing in COX-2(-/-) mice.

Neural network based pattern matching and spike detection tools and services--in the CARMEN neuroinformatics project.

In the study of information flow in the nervous system, component processes can be investigated using a range of electrophysiological and imaging techniques. Although data is difficult and expensive to produce, it is rarely shared and collaboratively exploited. The Code Analysis, Repository and Modelling for e-Neuroscience (CARMEN) project addresses this challenge through the provision of a virtual neuroscience laboratory: an infrastructure for sharing data, tools and services. Central to the CARMEN concept are federated CARMEN nodes, which provide: data and metadata storage, new, thirdparty and legacy services, and tools. In this paper, we describe the CARMEN project as well as the node infrastructure and an associated thick client tool for pattern visualisation and searching, the Signal Data Explorer (SDE). We also discuss new spike detection methods, which are central to the services provided by CARMEN. The SDE is a client application which can be used to explore data in the CARMEN repository, providing data visualization, signal processing and a pattern matching capability. It performs extremely fast pattern matching and can be used to search for complex conditions composed of many different patterns across the large datasets that are typical in neuroinformatics. Searches can also be constrained by specifying text based metadata filters. Spike detection services which use wavelet and morphology techniques are discussed, and have been shown to outperform traditional thresholding and template based systems. A number of different spike detection and sorting techniques will be deployed as services within the CARMEN infrastructure, to allow users to benchmark their performance against a wide range of reference datasets.

Polyethylene wear against alumina and zirconia heads in cemented total hip arthroplasty.

We compared the polyethylene wear of acetabular sockets articulated with 22.225-mm alumina heads with the polyethylene wear of those articulated with 22.225-mm zirconia heads in cemented total hip arthroplasty during a mean follow-up period of 5.4 years. Using a computer-aided technique, we measured polyethylene wear radiologically in 46 hips with alumina heads and 58 hips with zirconia heads. The preoperative diagnosis in all cases was osteoarthritis. The mean linear wear rate and mean volumetric wear rate of polyethylene sockets against zirconia heads were 0.133 mm/y and 39.8 mm(3)/y, respectively, significantly greater (P < .01) than the wear rates against alumina heads (0.078 mm/y and 24.2 mm(3)/y, respectively). Age at operation, patient body weight as well as height, thickness of polyethylene, and socket abduction angle did not influence the wear rates. We speculate that the excessive polyethylene wear was caused by phase transformation of zirconia, leading to an increase of surface roughness.

Long-term follow-up study of bioactive bone cement in canine total hip arthroplasty.

We report a long-term study of bioactive bone cement (BABC) in a canine total hip arthroplasty (THA) at follow-up of 8 years. Previous studies have shown excellent biomechanical and histological results at follow-up of 6, 12, and 24 months. In the present study, THA was performed in a beagle dog using BABC consisting of an apatite- and wollastonite-containing glass ceramic (AW-GC) powder and SiO2 powder as the filler and a bisphenol-a-glycidyl methacrylate (Bis-GMA) based resin as the organic matrix. Histological examination showed direct bonding between BABC and bone without any intervening soft tissue layer at the BABC-bone interface. A reactive layer, through which BABC bonded to the bone, was observed thicker at 8 years than it was at 24 weeks. No adverse effects of BABC were observed. BABC maintained the high bioactivity and direct bonding to bone in a canine THA at follow-up of 8 years.

PGE2 signal through EP2 promotes the growth of articular chondrocytes.

UNLABELLED: EP2 was identified as the major PGE2 receptor expressed in articular cartilage. An EP2 agonist increased intracellular cAMP in articular chondrocytes, stimulating DNA synthesis in both monolayer and 3D cultures. Hence, the EP2 agonist may be a potent therapeutic agent for degenerative cartilage diseases. INTRODUCTION: Prostaglandin E2 (PGE2) exhibits pleiotropic effects in various types of tissue through four types of receptors, EP1-4. We examined the expression of EPs and effects of agonists for each EP on articular chondrocytes. MATERIALS AND METHODS: The expression of each EP in articular chondrocytes was examined by immunohistochemistry and RT-PCR. A chondrocyte cell line, MMA2, was established from articular cartilage of p53(-/-) mice and used to analyze the effects of agonists for each EP. A search for molecules downstream of the PGE2 signal through the EP2 agonist was made by cDNA microarray analysis. The growth-promoting effect of the EP2 agonist on chondrocytes surrounded by cartilage matrix was examined in an organ culture of rat femora. RESULTS AND CONCLUSION: EP2 was identified as the major EP expressed in articular cartilage. Treatment of MMA2 cells with specific agonists for each EP showed that only the EP2 agonist significantly increased intracellular cAMP levels in a dose-dependent manner. Gene expression profiling of MMA2 revealed a set of genes upregulated by the EP2 agonist, including several growth-promoting and apoptosis-protecting genes such as the cyclin D1, fibronectin, integrin alpha5, AP2alpha, and 14-3-3gamma genes. The upregulation of these genes by the EP2 agonist was confirmed in human articular chondrocytes by quantitative mRNA analysis. On treatment with the EP2 agonist, human articular chondrocytes showed an increase in the incorporation of 5-bromo-2-deoxyuracil (BrdU), and the organ culture of rat femora showed an increase of proliferating cell nuclear antigen (PCNA) staining in articular chondrocytes surrounded by cartilage matrix, suggesting growth-promoting effects of the PGE2 signal through EP2 in articular cartilage. These results suggested that the PGE2 signal through EP2 enhances the growth of articular chondrocytes, and the EP2 agonist is a candidate for a new therapeutic compound for the treatment of degenerative cartilage diseases.

Histological and mechanical investigation of the bone-bonding ability of anodically oxidized titanium in rabbits.

The purpose of this study was to histologically and mechanically investigate the in vivo bone-bonding ability of anodically oxidized titanium (AO Ti) with an anatase crystal layer on its surface. AO Ti plates, anodically oxidized at 155 V in 1 M H2SO4, were implanted into the proximal metaphyses of mature rabbit tibiae for 4, 8, 16, and 24 weeks and investigated by light microscopy, scanning electron microscopy and detaching test. High bone-bonding ability, comparable to our previous study data of the bioactive titanium produced by sodium-free alkali and heat treatment, was observed at the early stages of implantation. However, no substantial increase was demonstrated. AO Ti plates bonded to bone directly, with no intervening soft tissue layer, and no breakage of the AO Ti layer was observed. The AO Ti layer was porous through to the titanium substrate, while the porosity was low. Apatite-like deposition into the pores of the AO layer was observed only in the superficial zone. The lack of improvement of bone-bonding ability in the later stages of implantation may be attributed to the low porosity and to the superficial ingrowths of apatite-like deposits into the pores of the AO Ti layer.