Antioxidants, reactive oxygen and nitrogen species, gene induction and mitochondrial function

Redox-sensitive cell signalling Thiol groups and the regulation of gene expression Redox-sensitive signal transduction pathways Protein kinases Protein phosphatases Lipids and phospholipases Antioxidant (electrophile) response element Intracellular calcium signalling Transcription factors NF-?B AP-1 p53 Cellular responses to oxidative stress Cellular responses to change in redox state Proliferation Cell death Immune cell function Reactive oxygen and nitrogen species – good or bad? Reactive oxygen species and cell death Reactive oxygen species and inflammation Are specific reactive oxygen species and antioxidants involved in modulating cellular responses? Specific effects of dietary antioxidants in cell regulation Carotenoids Vitamin E Flavonoids Inducers of phase II enzymes Disease states affected Oxidants, antioxidants and mitochondria Introduction Mitochondrial generation of reactive oxygen and nitrogen species Mitochondria and apoptosis Mitochondria and antioxidant defences Key role of mitochondrial GSH in the defence against oxidative damage Mitochondrial oxidative damage Direct oxidative damage to the mitochondrial electron transport chain Nitric oxide and damage to mitochondria Effects of nutrients on mitochondria Caloric restriction and antioxidants Lipids Antioxidants Techniques and approaches Mitochondrial techniques cDNA microarray approaches Proteomics approaches Transgenic mice as tools in antioxidant research Gene knockout and over expression Transgenic reporter mice Conclusions Future research needs

Receptor for activated C Kinase 1 protein binds to and activates the human estrogen receptor α

The classical concept of estrogen receptor (ER) activation is that steroid passes the cell membrane, binds to its specific protein receptor in the cell's cytoplasm and the steroid-receptor complex travels to the nucleus where it activates responsive genes. This basic idea has been challenged by results of experiments demonstrating insulin-like growth factor 1 (IGF-1) activation of the ER in the complete absence of estrogen suggesting at least one other mechanism of ER activation not involving steroid. One explanation is that activation of the cell surface IGF-1 receptor leads to synthesis of an intracellular protein(s) able to bind to and stimulate the ER. Based on results using the two-hybrid system, coimmunoprecipitation and transfection-luciferase assays, we herein show that one of these proteins could well be receptor for activated C kinase 1 (RACK-1). Using the human ER type α (ER-α) as bait, a cloned complementary deoxyribonucleic acid (cDNA) library from IGF-1 treated human breast cancer MCF-7 cells was screened for ER-α - protein interactions. Many positive clones were obtained which contained the RACK-1 cDNA sequence. Coimmunoprecipitation of in-vitro translation products of the ER-α and RACK-1 confirmed the interaction between the two proteins. Transfection studies using the estrogen response element spliced to a luciferase reporter gene revealed that constitutive RACK-1 expression was able to powerfully stimulate ER-α activity under estrogen-free conditions. This effect could be enhanced by 17β-estradiol (E2) and blocked by tamoxifen, an E2 antagonist. These results show that RACK-1 is able to activate the ER-α in the absence of E2, although together with the latter, enhanced effects occur. Since RACK-1 gene expression is stimulated by IGF-1, it is distinctly possible that RACK-1 is the mediator of the stimulatory effects of IGF-1 on ER-α. © 2014 JMS.

Early transplantation of mesenchymal stem cells after spinal cord injury relieves pain hypersensitivity through suppression of pain-related signaling cascades and reduced inflammatory cell recruitment

Bone marrow-derived mesenchymal stem cells (BMSC) modulate inflammatory/immune responses and promote motor functional recovery after spinal cord injury (SCI). However, the effects of BMSC transplantation on central neuropathic pain and neuronal hyperexcitability after SCI remain elusive. This is of importance because BMSC-based therapies have been proposed for clinical treatment. We investigated the effects of BMSC transplantation on pain hypersensitivity in green fluorescent protein (GFP)-positive bone marrow-chimeric mice subjected to a contusion SCI, and the mechanisms of such effects. BMSC transplantation at day 3 post-SCI improved motor function and relieved SCI-induced hypersensitivities to mechanical and thermal stimulation. The pain improvements were mediated by suppression of protein kinase C-γ and phosphocyclic AMP response element binding protein expression in dorsal horn neurons. BMSC transplants significantly reduced levels of p-p38 mitogen-activated protein kinase and extracellular signal-regulated kinase (p-ERK1/2) in both hematogenous macrophages and resident microglia and significantly reduced the infiltration of CD11b and GFP double-positive hematogenous macrophages without decreasing the CD11b-positive and GFP-negative activated spinal-microglia population. BMSC transplants prevented hematogenous macrophages recruitment by restoration of the blood-spinal cord barrier (BSCB), which was associated with decreased levels of (a) inflammatory cytokines (tumor necrosis factor-α, interleukin-6); (b) mediators of early secondary vascular pathogenesis (matrix metallopeptidase 9); (c) macrophage recruiting factors (CCL2, CCL5, and CXCL10), but increased levels of a microglial stimulating factor (granulocyte-macrophage colony-stimulating factor). These findings support the use of BMSC transplants for SCI treatment. Furthermore, they suggest that BMSC reduce neuropathic pain through a variety of related mechanisms that include neuronal sparing and restoration of the disturbed BSCB, mediated through modulation of the activity of spinal-resident microglia and the activity and recruitment of hematogenous macrophages.

Short-chain fatty acid acetate stimulates adipogenesis and mitochondrial biogenesis via GPR43 in brown adipocytes

Short-chain fatty acids play crucial roles in a range of physiological functions. However, the effects of short-chain fatty acids on brown adipose tissue have not been fully investigated. We examined the role of acetate, a short-chain fatty acid formed by fermentation in the gut, in the regulation of brown adipocyte metabolism. Our results show that acetate up-regulates adipocyte protein 2, peroxisomal proliferator-activated receptor-γ coactivator-1α, and uncoupling protein-1 expression and affects the morphological changes of brown adipocytes during adipogenesis. Moreover, an increase in mitochondrial biogenesis was observed after acetate treatment. Acetate also elicited the activation of ERK and cAMP response element-binding protein, and these responses were sensitive to G(i/o)-type G protein inactivator, Gβγ-subunit inhibitor, phospholipase C inhibitor, and MAPK kinase inhibitor, indicating a role for the G(i/o)βγ/phospholipase C/protein kinase C/MAPK kinase signaling pathway in these responses. These effects of acetate were mimicked by treatment with 4-chloro-α-(1-methylethyl)-N-2-thiazolylbenzeneacetamide, a synthetic G protein-coupled receptor 43 (GPR43) agonist and were impaired in GPR43 knockdown cells. Taken together, our results indicate that acetate may have important physiological roles in brown adipocytes through the activation of GPR43.

Updating finite element models using measured vibration data

The modelling of mechanical structures using finite element analysis has become an indispensable stage in the design of new components and products. Once the theoretical design has been optimised a prototype may be constructed and tested. What can the engineer do if the measured and theoretically predicted vibration characteristics of the structure are significantly different? This thesis considers the problems of changing the parameters of the finite element model to improve the correlation between a physical structure and its mathematical model. Two new methods are introduced to perform the systematic parameter updating. The first uses the measured modal model to derive the parameter values with the minimum variance. The user must provide estimates for the variance of the theoretical parameter values and the measured data. Previous authors using similar methods have assumed that the estimated parameters and measured modal properties are statistically independent. This will generally be the case during the first iteration but will not be the case subsequently. The second method updates the parameters directly from the frequency response functions. The order of the finite element model of the structure is reduced as a function of the unknown parameters. A method related to a weighted equation error algorithm is used to update the parameters. After each iteration the weighting changes so that on convergence the output error is minimised. The suggested methods are extensively tested using simulated data. An H frame is then used to demonstrate the algorithms on a physical structure.

Measuring aggressiveness and anger, but not aggression? A response to the CAAS critique

Objectives: The Competitive Aggressiveness and Anger Scale (CAAS) was developed to measure antecedents of aggression in sport. The critique attacks the CAAS on three points: (1) the definition of aggression in sport adopted, (2) the‘‘one size fits all’’ element in the thinking behind the scale’s development, (3) the nature of the CAAS Anger and Aggressiveness items. The objectives of this response is to address misunderstandings in the critique. Methods: We identified a number of false assumptions that undermine the validity of the critique and attempt to clarify our position with respect to the criticisms made. Results: (1) The CAAS is being criticised for a definition that it did not use. (2) We accepted that the CAAS may not be suitable for everyone in our limitations section and fully accept the limitations of any scale. We have since undertaken a large research project to establish whether the scale is valid across and within specific sports. (3) The fundamental misunderstanding inherent throughout the critique is that the CAAS was designed as a measure of aggression, rather than anger and aggressiveness, rendering the critique of its items redundant. Conclusions: The critique misrepresents the authors of the CAAS and fails to present a coherent argument against its use. We hope to clarify our position here. The evidence to date suggests that the CAAS is a valid measure of anger and aggressiveness in many sports and that these concepts reliably differentiate players who admit unsanctioned aggression from those who do not.

Neuronal convergence in early contrast vision: binocular summation is followed by response nonlinearity and area summation

We assessed summation of contrast across eyes and area at detection threshold ( C t). Stimuli were sine-wave gratings (2.5 c/deg) spatially modulated by cosine- and anticosine-phase raised plaids (0.5 c/deg components oriented at ±45°). When presented dichoptically the signal regions were interdigitated across eyes but produced a smooth continuous grating following their linear binocular sum. The average summation ratio ( C t1/([ C t1+2]) for this stimulus pair was 1.64 (4.3 dB). This was only slightly less than the binocular summation found for the same patch type presented to both eyes, and the area summation found for the two different patch types presented to the same eye. We considered 192 model architectures containing each of the following four elements in all possible orders: (i) linear summation or a MAX operator across eyes, (ii) linear summation or a MAX operator across area, (iii) linear or accelerating contrast transduction, and (iv) additive Gaussian, stochastic noise. Formal equivalences reduced this to 62 different models. The most successful four-element model was: linear summation across eyes followed by nonlinear contrast transduction, linear summation across area, and late noise. Model performance was enhanced when additional nonlinearities were placed before binocular summation and after area summation. The implications for models of probability summation and uncertainty are discussed.

All-fiber passively mode-locked femtosecond laser using a 45º-tilted fiber grating polarization element

We report on the demonstration of an all-fiber femtosecond erbium doped fiber laser passively mode-locked using a 45º tilted fiber grating as an in-fiber polarizer in the laser cavity. The laser generates 600 fs pulses with output pulse energies ~1 nJ. Since the 45° tilted grating has a broad polarization response, the laser output has shown a tunabilty in wavelength from 1548 nm to 1562 nm by simply adjusting the polarization controllers in the cavity.

Fibre laser using a microchannel based loss tuning element for refractive index sensing

We have proposed and demonstrated a fibre laser system using a microchannel as a cavity loss tuning element for surrounding medium refractive index (SRI) sensing. A ~6µm width microchannel was created by femtosecond (fs) laser inscription assisted chemical etching in the cavity fibre, which offers a direct access to the external liquids. When the SRI changes, the microchannel behaves as a loss tuning element, hence modulating the laser cavity loss and output power. The results indicate that the presented laser sensing system has a linear response to the SRI with a sensitivity in the order of 10-5. Using higher pump power and more sensitive photodetector, the SRI sensitivity could be further enhanced.

Three-dimensional finite element analysis of the effects of anisotropy on bone mechanical properties measured by nanoindentation

A three-dimensional finite element analysis (FEA) model with elastic-plastic anisotropy was built to investigate the effects of anisotropy on nanoindentation measurements for cortical bone. The FEA model has demonstrated a capability to capture the cortical bone material response under the indentation process. By comparison with the contact area obtained from monitoring the contact profile in FEA simulations, the Oliver-Pharr method was found to underpredict or overpredict the contact area due to the effects of anisotropy. The amount of error (less than 10% for cortical bone) depended on the indentation orientation. The indentation modulus results obtained from FEA simulations at different surface orientations showed a trend similar to experimental results and were also similar to moduli calculated from a mathematical model. The Oliver-Pharr method has been shown to be useful for providing first-order approximations in the analysis of anisotropic mechanical properties of cortical bone, although the indentation modulus is influenced by anisotropy.

Fibre laser using a microchannel based loss tuning element for refractive index sensing

We have proposed and demonstrated a fibre laser system using a microchannel as a cavity loss tuning element for surrounding medium refractive index (SRI) sensing. A ~6µm width microchannel was created by femtosecond (fs) laser inscription assisted chemical etching in the cavity fibre, which offers a direct access to the external liquids. When the SRI changes, the microchannel behaves as a loss tuning element, hence modulating the laser cavity loss and output power. The results indicate that the presented laser sensing system has a linear response to the SRI with a sensitivity in the order of 10-5. Using higher pump power and more sensitive photodetector, the SRI sensitivity could be further enhanced.