An in vitro traumatic injury model to examine the response of neurons to a hydrodynamically-induced deformation

A novel in vitro system was developed to examine the effects of traumatic mechanical loading on individual cells. The cell shearing injury device (CSID) is a parallel disk viscometer that applies fluid shear stress with variable onset rate. The CSID was used in conjunction with microscopy and biochemical techniques to obtain a quantitative expression of the deformation and functional response of neurons to injury. Analytical and numerical approximations of the shear stress at the bottom disk were compared to determine the contribution of secondary flows. A significant portion of the shear stress was directed in the r-direction during start-up, and therefore the full Navier-Stokes equation was necessary to accurately describe the transient shear stress. When shear stress was applied at a high rate (800 dyne cm-2 sec-1) to cultured neurons, a range of cell membrane strains (0.01 to 0.53) was obtained, suggesting inhomogeneity in cellular response. Functionally, cytosolic calcium and extracellular lactate dehydrogenase levels increased in response to high strain rate (> 1 sec-1) loading, compared with quasistatic (< 1 sec-1) loading. In addition, a subpopulation of the culture subjected to rapid deformation subsequently died. These strain rates are relevant to those shown to occur in traumatic injury, and, as such, the CSID is an appropriate model for studying the biomechanics and pathophysiology of neuronal injury.     

Strain Rate Sensitivity of Epoxy Resin in Tensile and Shear Loading

The mechanical response of E-862 and PR-520 resins is investigated in tensile and shear loadings. At both types of loading the resins are tested at strain rates of about 5×10?5, 2, and 450–700?s?1. In addition, dynamic shear modulus tests are carried out at various frequencies and temperatures, and tensile stress relaxation tests are conducted at room temperature. The results show that the toughened PR-520 resin can carry higher stresses than the untoughened E-862 resin. Strain rate has a significant effect on the response of both resins. In shear, both resins show a ductile response with maximum stress that is increasing with strain rate. In tension, a ductile response is observed at low strain rate ( ～ 5×10?5?s?1), and brittle response is observed at the medium and high strain rates (2 and 700?s?1). The hydrostatic component of the stress in the tensile tests causes premature failure in the E-862 resin. Localized deformation develops in the PR-520 resin when loaded in shear. An internal state variable constitutive model is proposed for modeling the response of the resins. The model includes a state variable that accounts for the effect of the hydrostatic component of the stress on the deformation.     

Approximation of Nonlinear Unloading Effects in the Strain Rate Dependent Deformation Analysis of Polymer Matrix Materials Utilizing a State Variable Approach

An experimental and analytical program is carried out to explore key behaviors in the loading and unloading behavior of polymers. Specifically, the effects of strain rate and hydrostatic stresses on the nonlinear portions of the deformation response are examined. Tension, compression, and shear load only and load/unload tests are conducted on a representative polymer across a range of strain rates, and key features of the experimental results are identified. To conduct a preliminary exploration of how the key features of the deformation response could be simulated analytically, a previously developed set of constitutive equations, which were developed to analyze the strain rate dependent, nonlinear deformation of polymers including the effects of hydrostatic stresses, were modified in order to approximate key features of the nonlinear unloading behavior observed in the polymer. The constitutive relations are based on state variable constitutive equations originally developed for metals. The nonlinear unloading observed in the experiments is approximated by reducing the unloading modulus of the material as the effective inelastic strain is increased. The effects of the hydrostatic stress state on the unloading modulus are also simulated analytically. To examine the revised formulation, the loading and load/unload responses of the representative polymer in tension, compression, and shear are examined at several strain rates. Results computed using the developed constitutive equations were found to correlate reasonably well with the experimental data.     

Reactive oxygen species in reoxygenation injury of rat brain capillary endothelial cells

OBJECTIVE: To clarify the mechanism of anoxia/reoxygenation (A/R) injury of rat brain capillary endothelial cells (BCEC). METHODS: BCEC isolated from Sprague-Dawley rats by enzymatic treatment and centrifugation were subjected to anoxia (95% N2, 5% CO2) for 20 minutes and then to reoxygenation (95% air, 5% CO2) for 3 hours. Enzyme inhibitors, including oxypurinol, indomethacin, and N(G)-nitro-L-arginine methyl ester, or specific free-radical scavengers, such as superoxide dismutase, catalase, and the ferric iron chelator deferoxamine, were added before A/R injury. The BCEC were incubated in a range of Ca2+ concentrations from 1 to 0.01 mmol/L during A/R injury. Cytotoxicity was assayed by release of intracellular lactate dehydrogenase (LDH). RESULTS: With A/R injury, LDH release from the control group (no protective agents) significantly increased (44.8 +/- 3.3%), compared with a small increase in a normoxic group. BCEC treated with oxypurinol, indomethacin, or N(G)-nitro-L-arginine methyl ester showed suppression of LDH release. LDH release was almost totally suppressed by superoxide dismutase and partially by catalase or deferoxamine. The LDH release was partly dependent on calcium concentration. CONCLUSION: BCEC subjected to A/R become potent generators of free radicals, especially superoxide anion. Free radical production depends on both xanthine oxidase and cyclooxygenase pathways. Peroxynitrite and extracellular Ca2+ both contribute importantly to reoxygenation injury of BCEC.     

Plastic and viscous deformation of metals

Deformation characteristics of tensile specimens of several alloys, including electrolytic copper, α-brass, and 304 stainless steel, have been studied by application of stress and measurement of change of length in a soft tensile machine. By means of experiments in which the stress rate is reduced suddenly from a positive value to zero and the strain rate measured, both during loading and during creep, it is found that permanent deformation consists of two components, a plastic component for which the strain rate is a function of stress and stress rate, and a viscous component which is functionally dependent on stress and temperature. Plastic deformation is relatively more evident at increasing stress rate but declines in importance through the series copper, a-brass, and stainless steel. As a consequence, for a fixed strain rate during loading, the initial creep rate is low in copper and little creep occurs; in stainless steel, however, the initial creep rate is nearly equal to the loading strain rate and creep is pronounced. The theory is not fully developed but is based on a competition between thermal and mechanical release of dislocation segments from obstacles or sources. Release produces a strain increment which may be small or large depending on the relative values of stress and structural resistance. Plastic deformation occurs when the applied stress is close to the mechanical threshold, mechanical release is relatively easy, and the strain consists, at a given strain rate, of a few large strain increments per unit time. For viscous flow the relative stress is low, thermal release easy, and the strain rate is composed of many small strain increments in each unit of time.     

Room temperature creep of high strength steels

The room-temperature creep behaviour of three high strength steels has been investigated. Several parameters such as creep stress, loading rate, stress history and heat treatment has been altered and their influence on the low temperature creep has been reported. The primary creep in all three alloys agreed well with the logarithmic creep law and the creep mechanism has been identified as pure dislocation creep. Higher stresses and high loading rates led to increased creep strains and strain rates. Reloading after a period of creep resulted in significantly decreased creep strains and no recovery of the time dependent deformation could be detected. The yield strength of the materials per se had no influence on the room temperature creep whereas the same material with decreased 0.2% offset strength showed significantly reduced time dependent deformation. The possible interaction between primary creep and stress corrosion cracking has been discussed.     

Advancing age alters intracellular calcium buffering in rat adrenergic nerves

There is a marked increase with advancing age of stimulation-evoked neurotransmitter release from vascular adrenergic nerves in the rat, an effect correlated with increased levels of plasma norepinephrine. This increase in norepinephrine release could not be accounted for by an alteration in neuronal and extraneuronal uptake of norepinephrine or a decline in feedback inhibition of release by prejunctional alpha2-adrenergic receptors. Measurement of intracellular calcium in fura-2-labeled superior cervical ganglion cells revealed elevated K+-evoked calcium transients in old compared to young neurons. Blockade of mitochondrial calcium uptake with dinitrophenol resulted in increased calcium transients in old neurons only. Furthermore, following blockade of mitochondrial calcium uptake the rate of return of calcium to resting levels was reduced to a greater degree in old cells as compared to young cells. The effects of dinitrophenol in old cells were attenuated when extracellular calcium was reduced. These findings suggest that older cells are more dependent on mitochondrial calcium buffering, perhaps due to changes in ATP dependent calcium uptake. Increased calcium transients as a result of altered intracellular calcium buffering offer a reasonable explanation for our previous observation of increased stimulation evoked norepinephrine release.     

Shear stress-induced release of PGE2 and PGI2 by vascular smooth muscle cells

This study addresses the direct effect of fluid flow shear stress on production of the vascular mediators, PGE2 and PGI2 by vascular smooth muscle cells (SMC). Results indicate that shear stress increases PGE2 and PGI2 release in SMC. The production patterns, however, differ between PGE2 and PGI2. For PGE2, the rate of production is moderate for the first three hours after the onset of shear stress, then dramatically increases between the fourth and fifth hours, returning to basal levels in the sixth hour. On the other hand, the rate for PGI2 production is maximal right after the onset of shear and remains elevated for the first three hours. The rate then plateaus and remains at a moderate level during the next three hours. The results also indicate that SMC production of PGI2 is more sensitive to shear stress than PGE2 production since a level of 0.5 dynes/cm2 produces a maximal PGI2 release whereas 1 dyne/cm2 produces only 1/4 the response seen at 20 dynes/cm2 for PGE2. The physiological implications of fluid shear stress regulation of SMC are discussed.     

The radiation biology of boron neutron capture therapy

Cerebral ischemia leads to a massive increase in cytoplasmic calcium activity resulting from an influx of calcium ions into cells and a release of calcium from mitochondria and endoplasmic reticulum (ER). It is widely believed that this increase in cytoplasmic calcium activity plays a major role in ischemic cell injury in neurons. Recently, this concept was modified, taking into account that disturbances occurring during ischemia are potentially reversible: it then was proposed that after reversible ischemia, calcium ions are taken up by mitochondria, leading to disturbances of oxidative phosphorylation, formation of free radicals, and deterioration of mitochondrial functions. The current review focuses on the possible role of disturbances of ER calcium homeostasis in the pathologic process culminating in ischemic cell injury. The ER is a subcellular compartment that fulfills important functions such as the folding and processing of proteins, all of which are strictly calcium dependent. ER calcium activity is therefore relatively high, lying in the lower millimolar range (i.e., close to that of the extracellular space). Depletion of ER calcium stores is a severe form of stress to which cells react with a highly conserved stress response, the most important changes being a suppression of global protein synthesis and activation of stress gene expression. The response of cells to disturbances of ER calcium homeostasis is almost identical to their response to transient ischemia, implying common underlying mechanisms. Many observations from experimental studies indicate that disturbances of ER calcium homeostasis are involved in the pathologic process leading to ischemic cell injury. Evidence also has been presented that depletion of ER calcium stores alone is sufficient to activate the process of programmed cell death. Furthermore, it has been shown that activation of the ER-resident stress response system by a sublethal form of stress affords tolerance to other, potentially lethal insults. Also, disturbances of ER function have been implicated in the development of degenerative disorders such as prion disease and Alzheimer's disease. Thus, disturbances of the functioning of the ER may be a common denominator of neuronal cell injury in a wide variety of acute and chronic pathologic states of the brain. Finally, there is evidence that ER calcium homeostasis plays a key role in maintaining cells in their physiologic state, since depletion of ER calcium stores causes growth arrest and cell death, whereas cells in which the regulatory link between ER calcium homeostasis and protein synthesis has been blocked enter a state of uncontrolled proliferation.     

Loading Rate Effect on Nanohardness of Soda-Lime-Silica Glass

To understand how hardness, the key design parameter for applications of brittle solids such as glass concerning contact deformation, is affected by loading rate variation, nanoindentation with a Berkovich tip was used to measure the nanohardness of a 330-μm-thick soda-lime-silica glass as a function of loading rate (1 to 1000 mN·s⁻¹). The results showed for the very first time that, with variations in the loading rate, there was a 6 to 9 pct increase in the nanohardness of glass up to a threshold loading rate (TLR), whereafter it did not appreciably increase with further increase in loading rate. Further, the nanohardness data showed an indentation size effect (ISE) that obeyed the Meyer’s law. These observations were explained in terms of a strong shear stress component developed just beneath the nanoindenter and the related shear-induced deformation processes at local microstructural scale weak links. The significant or insignificant presence of shear-induced serrations in load depth plots and corresponding scanning electron microscopic evidence of a strong or mild presence of shear deformation bands in and around the nanoindentation cavity supported such a rationalization. Finally, a qualitative picture was developed for different deformation processes induced at various loading rates in glass.     

Arachidonic acid protects against hypoxic injury in rat proximal tubules

Free fatty acids (FFA) and lysophospholipids accumulate during hypoxia (H) in rat proximal tubular epithelial cells partly as a result of increased phospholipase A2 (PLA2) activity. The role of FFA in mediating hypoxic injury and modulating PLA2 activity is not clear. In the present study, the effect of several FFA including arachidonic acid (AA, 20:4) on hypoxia-induced injury and PLA2 activity was assessed in freshly isolated rat proximal tubules. Hypoxia (H) was induced in the presence of either an unsaturated free fatty acid (uFFA) [AA or linoleic acid (LA, 18:2)] or a saturated FFA (sFFA) [palmitic (PA, 16:0) or myristic acid (MA, 14:0)]. Cell membrane injury was assessed by measuring lactate dehydrogenase release (LDH). AA markedly reduced LDH release during hypoxia in a dose dependent manner, while sFFA had no protective effect. LA showed similar protection to that observed with AA. AA did not affect buffer calcium concentration, buffer pH, intracellular pH or intracellular calcium concentration. Neither inhibiting the cyclooxygenase pathway with indomethacin, nor the lipoxygenase pathway with nordihydroguaiaretic acid (NDGA) had any effect on the AA observed cytoprotection. In vitro PLA2 activity in the control tubular extracts was compared to that following addition of AA or PA. PLA2 activity decreased significantly with AA but not with PA in a dose dependent manner. These data suggest that: (1) AA protects against hypoxic injury in rat proximal tubules. (2) This cytoprotection is not specific for AA and other uFFA have a similar effect. (3) AA significantly inhibits PLA2 activity, (4) AA induced cytoprotection may be related to a negative feedback inhibition of PLA2 activity.     

Effect of blood gas derangement on QTc dispersion in severe chronic obstructive pulmonary disease: evidence of an electropathy?

Mature myocardium utilizes calcium released by the sarcoplasmic reticulum (SR) for cell contraction. Transient exposure of mature myocytes to caffeine is known to directly trigger Ca2+ release from the SR. In contrast, neonatal rabbit heart cells rely on transsarcolemmal Ca2+ influx for tension generation. SR function is decreased in immature heart and appears to play a minimal role as a calcium source. Accordingly, we hypothesized that neonatal rabbit myocytes would not respond to a caffeine pulse. Isolated neonatal and adult myocytes were paced to load the SR with calcium and then exposed to a 1-s pulse of 10 mM caffeine. As previously described, adult myocytes exhibited a brisk contraction in response to caffeine. Unexpectedly, neonatal myocytes also exhibited a similar, brisk response. These caffeine-induced contractions were not dependent on extracellular Ca2+ but were dependent upon the loading of SR Ca2+ stores. When SR Ca2+ stores were depleted by exposure to caffeine, mature myocytes exhibited only small, slow contractions in response to electrical field stimulation. Replenishing the SR Ca2+ stores resulted in normal, brisk contractions. In contrast, electrically stimulated contractions in immature myocytes were largely unaffected by caffeine-induced SR depletion. Thus, although neonatal myocytes are capable of loading and releasing calcium from the SR, such SR calcium release is not normally required for contraction in the developing heart. The minor role of SR Ca2+ release in immature rabbit heart may not result from immaturity of the SR, but rather from an inadequate mechanism to trigger SR calcium release.     

Fructus corni attenuates oxidative stress in macrophages and endothelial cells

The antioxidant effect of a Chinese medicinal herb, Fructus corni extract (FCE), was investigated using models of oxidative stress in macrophages and vascular endothelial cells. Murine macrophages (J774) were incubated with FCE at 37 degrees C and 5% CO2 for 1 hr. Oxidative burst was triggered by zymosan and measured with a fluorescent probe. FCE exhibited a concentration- dependent suppression of oxidative burst. Confluent monolayers of bovine pulmonary artery endothelial cells (PAEC) were preincubated with FCE for 20 hrs, washed, and then exposed to an organic oxidant t-butyl hydroperoxide (tBHP) for 2 hrs. Cell viability was assessed by methylthiazol tetrazolium (MTT) assay, and cell injury by the release of intracellular lactate dehydrogenase (LDH). Lipid peroxidation products of PAEC were determined by measuring thiobarbituric acid-reactive substances (TBARS). Exposure of PAEC to tBHP resulted in decreased cell viability, increased LDH release, and elevated TBARS. Preincubation of PAEC with FCE significantly reversed these changes. Our results demonstrated that FCE can protect vascular endothelial cells from oxidant injury. The data thus suggest that Fructus corni may be useful for the prevention and/or treatment of disorders associated with oxidative damage.     

渗透剪切作用下黄土的力学特征

黄土高原地区黄土在灌溉作用下,逐渐达到饱和状态,饱和中,陡坡类黄土坡体自重增加引起下滑力增加. 该过程持续进行后,坡体内部同时发生渗流和剪切过程,导致坡体的变形不断增大,直至破坏后形成滑坡. 本文选取黑方台4.29滑坡为研究对象,在现场调查的基础上,利用滑坡后壁原状黄土试样,基于三轴试验设置10组共60个原状样对饱和黄土的渗透剪切行为进行模拟. 试验中设置了0.5、0.1和0.05 mm·min-1三个不同的加载速率对黄土试样进行剪切,为比较分析,对0.1 mm·min-1剪切速率试样设置了0、1、2和5 m几个不同水头进行了试验. 试验结果表明:饱和黄土在渗流与剪切耦合作用下,表现出应变硬化特征,渗透作用明显降低了黄土的强度,尤其是黄土黏聚力降低,其降幅达5.24%~63.35%. 对已有强度指标拟合后获得黄土在渗透剪切工况下的强度修正公式.      

Implementation of an Associative Flow Rule Including Hydrostatic Stress Effects into the High Strain Rate Deformation Analysis of Polymer Matrix Composites

A previously developed analytical formulation has been modified in order to more accurately account for the effects of hydrostatic stresses on the nonlinear, strain rate dependent deformation of polymer matrix composites. State variable constitutive equations originally developed for metals have been modified in order to model the nonlinear, strain rate dependent deformation of polymeric materials. To account for the effects of hydrostatic stresses, which are significant in polymers, the classical J2 plasticity theory definitions of effective stress and effective inelastic strain, along with the equations used to compute the components of the inelastic strain rate tensor, are appropriately modified. To verify the revised formulation, the shear and tensile deformation of a representative polymer are computed across a wide range of strain rates. Results computed using the developed constitutive equations correlate well with experimental data. The polymer constitutive equations are implemented within a strength of materials based micromechanics method to predict the nonlinear, strain rate dependent deformation of polymer matrix composites. The composite mechanics are verified by analyzing the deformation of a representative polymer matrix composite for several fiber orientation angles across a variety of strain rates. The computed values compare well to experimentally obtained results.     

Dent Resistance and Effect of Indentation Loading Rate on Superelastic TiNi Alloy

The large recoverable deformation associated with reversible stress-induced martensitic transformation for superelastic TiNi alloys has been widely exploited in many applications. However, to employ superelastic TiNi in applications where high impact loading is expected, as in bearings, the effect of loading rate on superelasticity needs to be understood. In the current article, the effect of indentation loading rate on dent resistance and superelasticity of TiNi is studied. Indentation tests are performed, at different loading rates on superelastic TiNi alloy and correlated to tensile stress–strain data. It is found that the reversible deformation drops as loading rate is increased and superelasticity diminishes. Based on data collected and results analysis it is proposed that the loss in superelastic behavior under high indentation loading rate is related to retardation of the stress-induced martensitic transformation. Furthermore, a simple heat model was proposed and showed that the temperature rise during indentation is not significant.     

Fracture Analysis of Shear Deformable Bi-Material Interface

Based on a novel split bi-layer shear deformable beam model capable of capturing the local deformation at the crack tip, the explicit closed-form solutions of bi-material interface fracture are presented in this paper. A recently developed novel shear deformable bi-layer beam theory is briefly reviewed, from which the deformation at the crack tip is explicitly derived. A new expression for the energy release rate is then obtained using the J integral, in which several new terms associated with the transverse shear force are present; this represents an improved solution compared to the one from the classical beam model. By exploiting the two concentrated crack tip forces, the general loadings acting at the crack tip are decomposed into two groups which produce only the mode I and mode II energy release rates, respectively; the total energy release rate is thus decomposed into the mode I and II components in a global sense. The stress intensity factor referred to as local decomposition is also obtained including the transverse shear effect. The difference between the global and local mode decompositions is clarified, and a simple relationship between them is provided. The effect of the existence of a thin layer of adhesive on the stress intensity factor is further studied by an asymptotic analysis. A simple and improved expression for the T stress, the nonsingular term of stress at the crack tip, is also given. The fracture parameters of several commonly used interface fracture specimens are summarized. The present fracture analysis including the transverse shear effect is in better agreement with finite element analyses and shows advantages and improved accuracy over the available classical solutions.     

The GABAA receptor is expressed in human neurons derived from a teratocarcinoma cell line

NT2 cells, a human teratocarcinoma cell line, are shown to be differentiated in neuron-like cells (NT2-N cells) by treatment with retinoic acid. The present study identified the neurotransmitter receptors expressed in NT2-N cells using patch-clamp recording. Voltage-sensitive Na+ currents, which are specific for neurons, were observed in NT2-N cells but not in NT2 cells, suggesting that NT2-N cells actually function as neurons. Glutamate receptor agonists, N-methyl-D-aspartate (NMDA) and kainate, evoked whole-cell currents. In addition, gamma-aminobutyric acid (GABA) evoked currents and the currents were inhibited by the selective GABAA receptor antagonist, bicuculline. In outside-out patches, GABA elicited single channel currents with two classes of the slope conductance (26 and 50 pS). No current, however, was induced by ACh, serotonin, or dopamine NT2-N cells, thus, express at least two types of the major excitatory and inhibitory neurotransmitter receptor in the central nervous system, the glutamate and GAGAA receptors, suggesting that these receptors have a crucial role in neurotransmission from the earlier stage of the brain development.