Immune and stress factors in the pathophysiology of the mdx mouse model of Duchenne Muscular Dystrophy

Duchenne Muscular Dystrophy (DMD) is a fatal multi-system neuromuscular disease caused by loss of dystrophin. The loss of dystrophin from membranes of contractile muscle cells and the dysregulation of the DAPC, induces chronic inflammation due to tissue necrosis and eventual replacement with collagen which weakens muscular force and strength. Dystrophin deficiency may cause under-diagnosed features of DMD include mood disorders such as depression and anxiety and dysfunction of the gastrointestinal tract. The first study in the thesis examined mood in the dystrophin-deficient mdx mouse model of DMD and examined the effects of the tri-cyclic antidepressant, amitriptyline on behaviours. Amitriptyline had anti-depressant and anxiolytic effects in the mdx mice possibly through effects on stress factors such as corticotrophin-releasing factor (CRF). This antidepressant also reduced skeletal muscle inflammation and caused a reduction in circulating interleukin (IL)-6 levels. In the second and third studies, we specifically blocked IL-6 signalling and used Urocortin 2, CRFR2 agonist to investigate their potential as therapeutic targets in mdx mice pathophysiology. Isometric and isotonic contractile properties of the diaphragm, were compared in mdx mice treated with anti IL-6 receptor antibodies (anti IL-6R) and/or Urocortin 2. Deficits in force production, work and power detected in mdx mice were improved with treatment. In study three I investigated contractile properties in gastrointestinal smooth muscle. As compared to wild type mice, mdx mice had slower faecal transit times, shorter colons with thickened muscle layers and increased contractile activity in response to recombinant IL-6. Blocking IL-6 signalling resulted in an increase in colon length, normalised faecal output times and a reduction in IL-6-evoked contractile activity. The findings from these studies indicate that for both diaphragm and gastrointestinal function in a dystrophin-deficient model, targeting of IL-6 and CRFR2 signalling has beneficial therapeutic effects.

A binding site for the cyclic adenosine 3',5'-monophosphate-response element-binding protein as a regulatory element in the grp78 promoter.

The 78-kDa glucose-regulated protein (GRP78) is ubiquitously expressed in many cell types. Its promoter contains multiple protein-binding sites and functional elements. In this study we examined a high affinity protein-binding site spanning bp -198 to -180 of the rat grp78 promoter, using nuclear extracts from both B-lymphoid and HeLa cells. This region contains a sequence TGACGTGA which, with the exception of one base, is identical to the cAMP-response element (CRE). Site-directed mutagenesis reveals that this sequence functions as a major basal level regulatory element in hamster fibroblast cells and is also necessary to maintain high promoter activity under stress-induced conditions. By gel mobility shift analysis, we detect two specific protein complexes. The major specific complex I, while immunologically distinct from the 42-kDa CRE-binding protein (CREB), binds most strongly to the grp site, but also exhibits affinity for the CRE consensus sequence. As such, complex I may consist of other members of the CREB/activating transcription factor protein family. The minor specific complex II consists of CREB or a protein antigenically related to it. A nonspecific complex III consists of the Ku autoantigen, an abundant 70- to 80-kDa protein complex in HeLa nuclear extracts. By cotransfection experiments, we demonstrate that in F9 teratocarcinoma cells, the grp78 promoter can be transactivated by the phosphorylated CREB or when the CREB-transfected cells are treated with the calcium ionophore A23187. The differential regulation of the grp78 gene by cAMP in specific cell types and tissues is discussed.

Consequences of heat stress on reproduction of cattle

Heat stress represents one of the major environmental factors that adversely affect the reproductive performance of cattle. In this paper the behavioral adjustments, physical mechanisms and physiological responses to heat loss are described; bos indicus adaptive advantages with respect to bos Taurus, pathophysiology of heat stress and heat stress effects in animal reproduction, both the male and the female.

Heightened maternal inflammation is linked to placental oxidative and nitrosative stress associated with fetal growth restriction in the rat

Deficient trophoblast invasion and spiral artery remodeling are associated with pregnancy complications such as pre-eclampsia (PE) and fetal growth restriction (FGR). Using a model in which pregnant Wistar rats are given daily, low-dose, injections of bacterial lipopolysaccharide (LPS; 10 – 40 µg/kg) on gestational days (GD) 13.5 – 16.5, our group has shown that abnormal maternal inflammation is causally linked to shallow trophoblast invasion, deficient spiral artery remodeling, and altered utero-placental hemodynamics leading to FGR/PE; these alterations were shown to be mediated by TNF-a. The present research evaluated certain consequences of decreased placental perfusion; this was accomplished by examining placental alterations indicative of decreased placental perfusion. Additionally, the role of glyceryl trinitrate (GTN) was determined as a potential therapeutic to prevent the consequences of decreased placental perfusion. Results indicated that dams experiencing heightened maternal inflammation showed significantly greater expression of hypoxia-inducible factor-1a (HIF-1a) and nitrotyrosine, both of which are markers of decreased perfusion and oxidative/nitrosative stress. Contrary to expectations, inflammation did not appear to affect nitric oxide (NO) bioavailability, as revealed by a lack of change in placental or plasma levels of cyclic guanosine monophosphate (cGMP). However, continuous transdermal administration of GTN (25 µg/hr) on GD 12.5 – 16.5 prevented the accumulation of HIF-1a and nitrotyrosine in placentas from LPS-treated rats. These results support the concept that maternal inflammation contributes to placental hypoxia and oxidative/nitrosative stress. Additionally, they indicate that GTN has potential applications in the treatment and/or prevention of pregnancy complications associated with abnormal maternal inflammation.

Constitutive model for localized Lüders-like stress-induced martensitic transformation and super-elastic behaviors of laser-welded NiTi wires

The application of the shape memory alloy NiTi in micro-electro-mechanical-systems (MEMSs) is extensive nowadays. In MEMS, complex while precise motion control is always vital. This makes the degradation of the functional properties of NiTi during cycling loading such as the appearance of residual strain become a serious problem to study, in particular for laser micro-welded NiTi in real applications. Although many experimental efforts have been put to study the mechanical properties of laser welded NiTi, surprisingly, up to the best of our understanding, there has not been attempts to quantitatively model the laser-welded NiTi under mechanical cycling in spite of the accurate prediction required in applications and the large number of constitutive models to quantify the thermo-mechanical behavior of shape memory alloys. As the first attempt to fill the gap, we employ a recent constitutive model, which describes the localized SIMT in NiTi under cyclic deformation; with suitable modifications to model the mechanical behavior of the laser welded NiTi under cyclic tension. The simulation of the model on a range of tensile cyclic deformation is consistent with the results of a series of experiments. From this, we conclude that the plastic deformation localized in the welded regions (WZ and HAZs) of the NiTi weldment can explain most of the extra amount of residual strain appearing in welded NiTi compared to the bare one. Meanwhile, contrary to common belief, we find that the ability of the weldment to memorize its transformation history, sometimes known as ‘return point memory’, still remains unchanged basically though the effective working limit of this ability reduces to within 6% deformation.

1-D constitutive model for evolution of stress-induced R-phase and localized Lüders-like stress-induced martensitic transformation of super-elastic NiTi wires

NiTi alloys have been widely used in the applications for micro-electro-mechanical-systems (MEMS), which often involve some precise and complex motion control. However, when using the NiTi alloys in MEMS application, the main problem to be considered is the degradation of functional property during cycling loading. This also stresses the importance of accurate prediction of the functional behavior of NiTi alloys. In the last two decades, a large number of constitutive models have been proposed to achieve the task. A portion of them focused on the deformation behavior of NiTi alloys under cyclic loading, which is a practical and non-negligible situation. Despite of the scale of modeling studies of the field in NiTi alloys, two experimental observations under uniaxial tension loading have not received proper attentions. First, a deviation from linearity well before the stress-induced martensitic transformation (SIMT) has not been modeled. Recent experiments confirmed that it is caused by the formation of stress-induced R phase. Second, the influence of the well-known localized Lüders-like SIMT on the macroscopic behavior of NiTi alloys, in particular the residual strain during cyclic loading, has not been addressed. In response, we develop a 1-D phenomenological constitutive model for NiTi alloys with two novel features: the formation of stress-induced R phase and the explicit modeling of the localized Lüders-like SIMT. The derived constitutive relations are simple and at the same time sufficient to describe the behavior of NiTi alloys. The accumulation of residual strain caused by R phase under different loading schemes is accurately described by the proposed model. Also, the residual strain caused by irreversible SIMT at different maximum loading strain under cyclic tension loading in individual samples can be explained by and fitted into a single equation in the proposed model. These results show that the proposed model successfully captures the behavior of R phase and the essence of localized SIMT.

Evaluation of Cyclic Deformation Behavior of Laser-welded Shape Memory NiTi Alloys at Different Working Temperatures

Post-weld heat-treatment (PWHT) has been established as one of the cost-effective ways to improve the functional properties, namely shape memory and super-elastic effects (SME and SE), of laser-welded NiTi alloys. However, the functional performance of the laser-welded joint at different working temperatures has not been explored yet. The purpose of this study is to investigate the effect of different working temperatures on the functional properties of the laser-welded NiTi alloys before and after PWHT by applying cyclic deformation tests. Two laser-welded samples: as-welded and heat-treated sample (after PWHT at 350 oC or 623 K) were tested in this work at room temperature, 50 oC (or 323 K) and 75 oC (or 348 K) respectively. The samples were cyclically loaded and unloaded for 10 cycles up to 4 % strain. The critical stress to induce the martensitic transformation and the residual strain after the cyclic tests were recorded. The results indicate that the heat-treated sample exhibited better functional properties than the as-welded sample at room temperature and 50 oC (or 323 K). However, both the as-welded and heat-treated samples failed in the cyclic tests at 75 oC (or 348 K). These findings are important to determine the feasible working temperature range for the laser-welded NiTi components to exhibit desirable functional properties in engineering applications involving cyclic loading.

Cyclic nanoindentation and nano-impact fatigue characterisation of functionally graded TiN/TiNi shape memory film

This paper investigates the mechanism of nanoscale fatigue of functionally graded TiN/TiNi films using nano-impact and multiple-loading-cycle nanoindentation tests. The functionally graded films were deposited on silicon substrate, in which TiNi films maintain shape memory and pseudo elastic behavior, while a modified TiN surface layer provides tribological and anti-corrosion properties. Nanomechanical tests were performed to comprehend the localized film performance and failure modes of the functionally graded film using NanoTestTM equipped with Berkovich and conical indenter between 100 μN to 500 mN loads. The loading mechanism and load history are critical to define film failure modes (i.e. backward depth deviation) including the shape memory effect of the functionally graded layer. The results are sensitive to the applied load, loading type (e.g. semi-static, dynamic) and probe geometry. Based on indentation force-depth profiles, depth-time data and post-test surface observations of films, it is concluded that the shape of the nanoindenter is critical in inducing the localized indentation stress and film failure, including shape recovery at the lower load range. Elastic-plastic finite element (FE) simulation during nanoindentation loading indicated that the location of subsurface maximum stress near the interface influences the backward depth deviation type of film failure. A standalone, molecular dynamics simulation was performed with the help of a long range potential energy function to simulate the tensile test of TiN nanowire with two different aspect ratios to investigate the theory of its failure mechanism.

Shape stabilised phase change materials based on a high melt viscosity HDPE and paraffin waxes

Shape stabilised phase change materials (SSPCMs) based on a high density poly(ethylene)(hv-HDPE) with high (H-PW, Tm = 56–58 °C) and low (L-PW, Tm = 18–23 °C) melting point paraffin waxes were readily prepared using twin-screw extrusion. The thermo-physical properties of these materials were assessed using a combination of techniques and their suitability for latent heat thermal energy storage (LHTES) assessed. The melt processing temperature (160 °C) of the HDPE used was well below the onset of thermal decomposition of H-PW (220 °C), but above that for L-PW (130 °C), although the decomposition process extended over a range of 120 °C and the residence time of L-PW in the extruder was <30 s. The SSPCMs prepared had latent heats up to 89 J/g and the enthalpy values for H-PW in the respective blends decreased with increasing H-PW loading, as a consequence of co-crystallisation of H-PW and hv-HDPE. Static and dynamic mechanical analysis confirmed both waxes have a plasticisation effect on this HDPE. Irrespective of the mode of deformation (tension, flexural, compression) modulus and stress decreased with increased wax loading in the blend, but the H-PW blends were mechanically superior to those with L-PW.

Occurrence of ribbons of cyclic water pentamers in a metallo-organic framework formed by spontaneous fixation of CO2

In the reaction of equimolar amounts of copper(II) acetate with 2,2'-dipyridylamine (DPA) in aqueous tetrahydrofuran, in presence of KOH, aerial CO2 is spontaneously fixed to the carbonate anion yielding [Cu(DPA)(CO3)] . 3H(2)O (1). X-ray crystallography shows the presence of zigzag ribbons of cyclic water pentamers in the channels of a chain-like metallo-organic framework. The water ribbons are stabilised by hydrogen bonds to the metallo-organic backbone. Each (H2O)(5) pentamer is approximately planar.

Cyclic extrusion of a lava dome based on a stick-slip mechanism

Lava dome eruptions are sometimes characterised by large periodic fluctuations in extrusion rate over periods of hours that may be accompanied by Vulcanian explosions and pyroclastic flows. We consider a simple system of nonlinear equations describing a 1D flow of lava extrusion through a deep elastic dyke feeding a shallower cylindrical conduit in order to simulate this short-period cyclicity. Stick-slip conditions depending on a critical shear stress are assumed at the wall boundary of the cylindrical conduit. By analogy with the behaviour of industrial polymers in a plastic extruder, the elastic dyke acts like a barrel and the shallower cylindrical portion of the conduit as a die for the flow of magma acting as a polymer. When we applied the model to the Soufrière Hills Volcano, Montserrat, for which the key parameters have been evaluated from previous studies, cyclic extrusions with periods from 3 to 30 h were readily simulated, matching observations. The model also reproduces the reduced period of cycles observed when a major unloading event occurs due to lava dome collapse.

NEONATAL HANDLING AND THE MATERNAL ODOR PREFERENCE IN RAT PUPS: INVOLVEMENT OF MONOAMINES AND CYCLIC AMP RESPONSE ELEMENT-BINDING PROTEIN PATHWAY IN THE OLFACTORY BULB

Early-life environmental events, such as the handling procedure, can induce long-lasting alterations upon several behavioral and neuroendocrine systems. However, the changes within the pups that could be causally related to the effects in adulthood are still poorly understood. In the present study, we analyzed the effects of neonatal handling on behavioral (maternal odor preference) and biochemical (cyclic AMP response element-binding protein (CREB) phosphorylation, noradrenaline (NA), and serotonin (5-HT) levels in the olfactory bulb (OB)) parameters in 7-day-old male and female rat pups. Repeated handling (RH) abolished preference for the maternal odor in female pups compared with nonhandled (NH) and the single-handled (SH) ones, while in RH males the preference was not different than NH and SH groups. In both male and female pups, RH decreased NA activity in the OB, but 5-HT activity increased only in males. Since preference for the maternal odor involves the synergic action of NA and 5-HT in the OB, the maintenance of the behavior in RH males could be related to the increased 5-HT activity, in spite of reduction in the NA activity in the OB. RH did not alter CREB phosphorylation in the OB of both male and females compared with NH pups. The repeated handling procedure can affect the behavior of rat pups in response to the maternal odor and biochemical parameters related to the olfactory learning mechanism. Sex differences were already detected in 7-day-old pups. Although the responsiveness of the hypothalamic-pituitary-adrenal axis to stressors is reduced in the neonatal period, environmental interventions may impact behavioral and biochemical mechanisms relevant to the animal at that early age. (C) 2009 IBRO. Published by Elsevier Ltd. All rights reserved.

Phase transformation evolution in NiTi shape memory alloy under cyclic nanoindentation loadings at dissimilar rates

Hysteresis energy decreased significantly as nanocrystalline NiTi shape memory alloy was under triangular cyclic nanoindentation loadings at high rate. Jagged curves evidenced discrete stress relaxations. With a large recovery state of maximum deformation in each cycle, this behavior concluded in several nucleation sites of phase transformation in stressed bulk. Additionally, the higher initial propagation velocity of interface and thermal activation volume, and higher levels of phase transition stress in subsequent cycles explained the monotonic decreasing trend of dissipated energy. In contrast, the dissipated energy showed an opposite increasing trend during triangular cyclic loadings at a low rate and 60âsec holding time after each unloading stage. Due to the isothermal loading rate and the holding time, a major part of the released latent heat was transferred during the cyclic loading resulting in an unchanged phase transition stress. This fact with the reorientation phenomenon explained the monotonic increasing trend of hysteresis energy.

Thermomechanical properties of Ni-Ti shape memory wires containing nanoscale precipitates induced by stress-assisted ageing.

This paper systematically examines the thermomechanical properties and phase transformation behaviour of slightly Ni-rich Ni-Ti biomedical shape memory wires containing homogeneously distributed nanoscale precipitates induced by stress-assisted ageing. In contrast to previous studies, particular attention is paid to the role of precipitates in impeding twin boundary movement (TBM) and its underlying mechanisms. The size and volume fraction of precipitates are altered by changing the ageing time. The martensitic transformation temperatures increase with prolonged ageing time, whereas the R-phase transformation temperature remains relatively unchanged. The stress-strain behaviour in different phase regions during both cooling and heating is comprehensively examined, and the underlying mechanisms for the temperature- and thermal-history-dependent behaviour are elucidated with the help of the established stress-temperature phase diagram. The effect of precipitates on TBM is explored by mechanical testing at 133K. It is revealed that the critical stress for TBM (σcr) increases with increasing ageing time. There is a considerable increase of 104MPa in σcr in the sample aged at 773K for 120min under 70MPa compared with the solution-treated sample, owing to the presence of precipitates. The Orowan strengthening model of twinning dislocations is insufficient to account for this increase in σcr. The back stress generation is the predominant mechanism for the interactions between precipitates and twin boundaries during TBM that give rise to the increase in σcr. Such results provide new insights into the thermomechanical properties of precipitate containing Ni-Ti biomedical shape memory wires, which are instructive for developing high-performance biomedical shape memory alloys.

The effect of low cycle fatigue, ratcheting and mean stress relaxation on stress-strain response and microstructural development in a dual phase steel

This study examines the cyclic plastic deformation behavior and microstructural development of a dual phase steel in both symmetric and asymmetric cycling in strain and stress control modes. The low-cycle fatigue (LCF) and mean stress relaxation (MSR) tests show very similar fatigue lifetimes. However, fatigue lifetimes reduce and prominent accumulation of directional strain was observed in ratcheting. A microstructural analysis has revealed that the type of cyclic test carried out has a noticeable impact on the substructural development, and this has been correlated with differences in accumulated tensile strain. Electron backscatter diffraction investigation has shown larger in-grain misorientation for ratcheting specimen in comparison with LCF and MSR specimens. The orientation of ferrite grains was found to have very little effect on their substructural development, and strain localization commonly occurred in the ferrite at the ferrite/martensite interface.