Mechanical characterization of Co/Cu multilayered nanowires.

The mechanical deformation properties of (110) Co/Cu multilayered nanowires were studied by Molecular Dynamics under uniaxial tensile and compressive stresses. The potential of the immiscible CoCu system was modeled by a second-moment tight-binding approximation. Stress-strain curves at different conditions were obtained and the elastic modulus and yield stress were analyzed. Both magnitudes are approximately independent of the strain rate, except at high values. They decrease linearly with increasing temperature. Below a volume-to-surface-area ratio, their values drastically increase and diverge from the bulk values. If the thickness of the Cu sublayers increases, the Young's modulus and yield stress decrease, although in a different way. The elastic modulus decreases linearly and the yield stress falls steeply whenever Cu is present in the nanowire, since the lattice distortion takes place firstly and fundamentally in Cu sublayers. The change in the axial stress at the interface is little significant on average and rather localized. Unlike, the transverse stress has a non-uniform distribution along the Cu sublayer, especially at the yield point. The Young's modulus and yield stress are larger in tension than in compression. Under tensile stress, nanowires slip via partial dislocation nucleation and propagation. Unlike, compressive deformation of nanowires takes place via both partial and full dislocations.

[Biological mechanisms involved in the spread of traumatic brain damage]. / Mecanismos biológicos involucrados en la propagación del daño en el traumatismo encéfalo craneano.

Traumatic brain injury (TBI) is a worldwide health problem that is especially prevalent in young adults. It is characterized by one or more primary injury foci, with secondary spread to initially not compromised areas via cascades of inflammatory response, excitotoxicity, energy failure conditions, and amplification of the original tissue injury by glia. In theory, such progression of injury should be amenable to management. However, all neuroprotective drug trials have failed, and specific treatments remain lacking. These negative results can be explained by a neuron centered approach, excluding the participation of other cell types and pathogenic mechanisms. To change this situation, it is necessary to secure a better understanding of the biological mechanisms determining damage progression or spread. We discuss the biological mechanisms involved in the progression of post-trauma tissue damage, including the general physiopathology of TBI and cellular mechanisms of secondary damage such as inflammation, apoptosis, cell tumefaction, excitotoxicity, and the role of glia in damage propagation. We highlight the role of glia in each cellular mechanism discussed. Therapeutic approaches related to the described mechanisms have been included. The discussion is completed with a working model showing the convergence of the main topics.

Dependence on temperature and energy of the heteroepitaxy of small metallic nanoclusters.

Deposition at different energies and temperatures of small metallic nanoclusters on metallic substrates is studied by molecular-dynamics simulations. Small-, Co/Cu(001), and large-misfit, Cu/Au(001) and Au/Cu(001), systems are considered. The rise in temperature improves the epitaxial order, although its effect is smaller in large-misfit systems. Thus, by increasing this parameter, non-epitaxial clusters can turn their structure into epitaxial in the case of Co/Cu(001), into aligned in Cu/Au(001), and into layered in Au/Cu(001). Therefore, the characteristics of the alignment are determined by the properties of the material. In addition, the influence of the initial structure is more marked in Co and Cu clusters, since they can reproduce locally other phases. Epitaxy can also be improved if the deposition energy is increased, although the deposited cluster loses its original shape progressively. Its effect is different depending mainly on the degree of misfit. An increase in energy (of up to 0.75 eV/atom) produces similar effects, but more noticeable, as a rise in temperature.

Influence of the cluster orientation on the epitaxy: deposition of Co nanoclusters on Cu(001) surfaces.

Deposition at low energy of 147-atom icosahedral Co nanoclusters on Cu(001) substrates is studied by molecular-dynamics simulations. Atomic interactions were mimicked by a many-body potential based on the tight-binding second-moment approximation. Clusters were rotated by using the two first Euler angles, in the so-called "x-convention," and subsequently, they were deposited on the substrate. The dependence of the degree of epitaxy on these angles has been obtained. Epitaxy is also related to the initial number of (001)-oriented atoms, especially for extreme values of this latter quantity. A better epitaxial matching is connected with a larger spreading index. The explanation of the epitaxial behavior of the supported clusters resides mainly in the dynamical interaction between grains during approximately the first 40 ps. Whenever the newly-formed (001)-oriented grain competes against a large number of grains after the collision, a very low epitaxial matching is obtained.

Proteoglycans and glycosaminoglycans induce gap junction synthesis and function in primary liver cultures.

Intercellular communication via gap junctions, as measured by dye and electrical coupling, disappears within 12 h in primary rat hepatocytes cultured in serum-supplemented media or within 24 h in cells in a serum-free, hormonally defined medium (HDM) designed for hepatocytes. Glucagon and linoleic acid/BSA were the primary factors in the HDM responsible for the extended life span of the electrical coupling. After 24 h of culture, no hormone or growth factor tested could restore the expression of gap junctions. After 4-5 d of culture, the incidence of coupling was undetectable in a serum-supplemented medium and was only 4-5% in HDM alone. However, treatment with glycosaminoglycans or proteoglycans of 24-h cultures, having no detectable gap junction protein, resulted in synthesis of gap junction protein and of reexpression of electrical and dye coupling within 48 h. Most glycosaminoglycans were inactive (heparan sulfates, chondroitin-6 sulfates) or only weakly active (dermatan sulfates, chondroitin 4-sulfates, hyaluronates), the weakly active group increasing the incidence of coupling to 10-30% with the addition of 50-100 micrograms/ml of the factor. Treatment of the cells with 50-100 micrograms/ml of heparins derived from lung or intestine resulted in cells with intermediate levels of coupling (30-50%). By contrast, 10-20 micrograms/ml of chondroitin sulfate proteoglycan, dermatan sulfate proteoglycan, or liver-derived heparin resulted in dye coupling in 80-100% of the cells, with numerous cells showing dye spread from a single injected cell. Sulfated polysaccharides of glucose (dextran sulfates) or of galactose (carrageenans) were inactive or only weakly active except for lambda-carrageenan, which induced up to 70% coupling (albeit no multiple coupling in the cultures). The abundance of mRNA (Northern blots) encoding gap junction protein and the amounts of the 27-kD gap junction polypeptide (Western blots) correlated with the degree of electrical and dye coupling indicating that the active glycosaminoglycans and proteoglycans are inducing synthesis and expression of gap junctions. Thus, proteoglycans and glycosaminoglycans, especially those found in abundance in the extracellular matrix of liver cells, are important in the regulation of expression of gap junctions and, thereby, in the regulation of intercellular communication in the liver. The relative potencies of heparins from different tissue sources at inducing gap junction expression are suggestive of functional tissue specificity for these glycosaminoglycans.

Carbon tetrachloride at hepatotoxic levels blocks reversibly gap junctions between rat hepatocytes.

Electrical coupling and dye coupling between pairs of rat hepatocytes were reversibly reduced by brief exposure to halogenated methanes (CBrCl3, CCl4, and CHCl3). The potency of different halomethanes in uncoupling hepatocytes was comparable to their hepatotoxicity in vivo, and the rank order was the same as that of their tendency to form free radicals. The effect of carbon tetrachloride (CCl4) on hepatocytes was substantially reduced by prior treatment with SKF 525A, an inhibitor of cytochrome P-450, and by exposure to the reducing reagent beta-mercaptoethanol. Halomethane uncoupling occurred with or without extracellular calcium and did not change intracellular concentrations of calcium and hydrogen ions or the phosphorylation state of the main gap-junctional protein. Thus the uncoupling appears to depend on cytochrome P-450 oxidative metabolism in which free radicals are generated and may result from oxidation of the gap-junctional protein or of a regulatory molecule that leads to closure of gap-junctional channels. Decreases in junctional conductance may be a rapid cellular response to injury that protects healthy cells by uncoupling them from unhealthy ones.

SURVEILLANCE OF ENVIRONMENTAL DOSE RATE WITH THE COVARIANCE MATRIX.

The objective of this study is the operational verification of an environmental dose rate monitoring network composed by several sensors. The verification of the study has been tested on the CIEMAT's Radiological Network, establishing a records quality assurance of the detectors using the covariance matrix and the eigenvalues. The technique has revealed an underlying records malfunction, which have not been appreciated by applying conventional surveillance. In this sense, the malfunction has economic and security consequences, which can be minimized with an alternative methodology, which guarantees the radiological protection of a local area according to the Regulatory Agency.

USING THE HERMITE POLYNOMIALS IN RADIOLOGICAL MONITORING NETWORKS.

The most interesting events in Radiological Monitoring Network correspond to higher values of H*(10). The higher doses cause skewness in the probability density function (PDF) of the records, which there are not Gaussian anymore. Within this work the probability of having a dose >2 standard deviations is proposed as surveillance of higher doses. Such probability is estimated by using the Hermite polynomials for reconstructing the PDF. The result is that the probability is ~6 ± 1%, much >2.5% corresponding to Gaussian PDFs, which may be of interest in the design of alarm level for higher doses.

ADVANCED SURVEILLANCE OF ENVIROMENTAL RADIATION IN AUTOMATIC NETWORKS.

The objective of this study is the verification of the operation of a radiation monitoring network conformed by several sensors. The malfunction of a surveillance network has security and economic consequences, which derive from its maintenance and could be avoided with an early detection. The proposed method is based on a kind of multivariate distance, and the verification for the methodology has been tested at CIEMAT's local radiological early warning network.

The Influence of Radon (Gas and Progeny) and Weather Conditions on Ambient Dose Equivalent Rate.

The purpose of this study is to identify the influence of radon (gas and progeny) on the ambient dose equivalent rate measured at the reference station ESMERALDA, where continuous measurements of the ambient dose equivalent rate (every 10 min) combined with activity concentration measurements of radon gas and radon progeny as well as meteorological parameters have been collected. This study has been performed using a correlation study based on a principal components analysis and the Spearman's rank correlation coefficient.

Identification of gap junctions in the oviduct and regulation of connexins during development and by sexual hormones.

Gap junctions in epithelial cells of the oviduct were identified by immunohistochemistry and Western blotting. Immunohistochemical studies showed that rat, hamster, mouse, and human oviducts contained connexin26 and connexin43 but not connexin32. The content of both connexins in the oviduct depended on the cell type, state of maturation and hormone status. During ontogeny, the epithelial and smooth muscle cells of immature rat oviducts (< 30 days) contained a low amount of connexin43, and connexin26 was undetectable. In mature oviducts (> 30 days), however, connexin26 was detected only in the isthmus and in localized regions of the ampullar epithelial layer. Moreover, at this age, the amount of connexin43 was high in both cell strata throughout the entire organ. During the estrous cycle, levels of connexin43 in the isthmic but not in the ampullar segment were higher in the proestrous and estrous day than at diestrous days. In addition, estrogen treatment produced a significant increase in total and phosphorylated isoforms of connexin43 levels in oviducts of pregnant rats. The estrogen effect was prevented by the simultaneous administration of progesterone which by itself did not affect the levels of connexin43. The high content of connexins found between oviductal cells as well as their responsiveness to hormone regulation, suggest that gap junctions might be involved in coordinating oviductal cell functions such as smooth muscle contraction and epithelial ciliary beat.

Connexin43 in MDCK cells: regulation by a tumor-promoting phorbol ester and Ca2+.

Prior to confluence, cultures of Madin Darby canine kidney (MDCK) cells expressed gap junctional communication, as assessed by fluorescent dye transfer, as well as relatively high levels of an anti-connexin43 immunoreactive component referred to as connexin43 (Cx43). After confluence, dye coupling and levels of Cx43 were dramatically reduced. Immunofluorescence analysis of the distribution of Cx43 in subconfluent cultures showed punctate labeling on the plasma membrane at regions of cell apposition and a more diffuse labeling in perinuclear regions. Western blots of total cell homogenates showed that the dephosphorylated form of Cx43 was more abundant than the phosphorylated forms. Phosphorylation of Cx43 was not significantly affected by 8-Bromo-cAMP or 8-Bromo-cGMP. However, 12-O-tetradecanoylphorbol-13-acetate (TPA) inhibited dye coupling and induced an increase in the amount of phosphorylated forms of Cx43 at the expense of the dephosphorylated form. This effect occurred as rapidly as 5 min after TPA treatment without apparent changes in distribution of Cx43 or cell morphology. These results suggest that second messenger pathways involving protein kinase C, but not cAMP- or cGMP-dependent protein kinase, led to changes in electrophoretic mobility of Cx43, revealed by Western blot, consistent with an alteration in the state of phosphorylation of the gap junction protein. Treatments with staurosporine, a protein kinase inhibitor, or okadaic acid, a protein phosphatase inhibitor, either alone or in combination with TPA, indicated that the abundance of the dephosphorylated form of Cx43 in MDCK cells was due to low kinase activity. It was also found that lowering the concentration of extracellular Ca2+, which reduced cell contact, did not affect the abundance, the state of phosphorylation, or the TPA-induced phosphorylation of Cx43. These results suggest that neither extracellular Ca2+ nor cell contact is required for basal or TPA-induced phosphorylation of Cx43.

On the mechanisms of cell uncoupling induced by a tumor promoter phorbol ester in clone 9 cells, a rat liver epithelial cell line.

It is known that in Clone 9 (C9) cells, intercellular gap junctional communication (IGJC) is rapidly blocked by the tumor promoter phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA), but it recovers spontaneously a few hours later and becomes refractory to TPA (Yada et al., J. Membr. Biol. 88, 217-232 (1985)). We now report that gap junctions between C9 cells contain at least two junctional proteins, connexin26 (Cx26) and connexin43 (Cx43), and that the TPA-induced changes in IGJC correlate temporally to changes in the state of phosphorylation of Cx43. The latter changes were prevented by inhibition of protein kinase C. Phosphoamino acid analysis and two-dimensional tryptic peptide maps of 32P-labeled Cx43 showed that during the TPA-induced phosphorylation at least two of the phosphorylated forms of Cx43 were differentially phosphorylated in seryl residues as compared to control. TPA induced a drastic reduction in junctional conductance as well as a redistribution of unitary gap junction channel event sizes seen in control cells. These changes were associated with retrieval of Cxs from the plasma membrane. Reappearance of gap junctions formed by Cx43 but not by Cx26 accounted for the spontaneous recovery in IGJC. It is proposed that gap junctions between C9 cells contain two types of channels each formed by Cx43 or Cx26 and that they are differentially affected during the action of TPA.

Blockade of electrophysiological properties of muscle fibres by lycorine.

This study concerns the action of the alkaloid lycorine on electrophysiological properties of single sartorius muscle fibres. Lycorine caused a decrease of the resting membrane potential, a lengthening in the rise and decay times of the action potential and a fall of the muscle accommodation. The depolarizing effect was not affected when Na+ is replaced by choline in the Ringer solution. Lycorine did not inhibit the (Na+ + K+) ATPase activity. The results suggest that action of lycorine on the electrophysiological properties can be attributed to an interference with potassium channels.

Effects of sodium pentobarbitone on the bioelectric properties of isolated toad skin.

The effects of sodium pentobarbitone on the transepithelial potential difference and on the short-circuit current in the isolated toad skin were studied under different experimental conditions. Pentobarbitone was consistently effective when placed in the solution bathing the inside surface of the skin. The response to small concentrations (1.2, 3.6 and 6.0 x 10(-5) M) was a transient dose-related increase in the transepithelial potential difference and short-circuit current, and a decrease in both parameters at larger concentrations (3.6 x 10(-4) M). Total skin resistance decreased at small and increased at large concentrations. Analysis of the response in terms of the equivalent electrical circuit indicated that in small concentrations the effect was due to an increase in the driving force of the sodium (Na+) pump although the rate of entry of Na+ through the outer barrier was reduced. The activity of (Na+-K+)ATPase was increased within the first 7 min of the effect of pentobarbitone, thus confirming a direct action of the drug on Na+ transport. In large concentrations the electromotive force of the Na+ pump was decreased and inactivation of (Na+-K+)ATPase became evident. The effect of pentobarbitone was unchanged in six reserpinized toads, which would seem to rule out a catecholamine-mediated effect of the drug. The reduction of Na+-influx could indirectly increase transport efficiency since the rate of transport work should be reduced.

Arachidonic acid-induced dye uncoupling in rat cortical astrocytes is mediated by arachidonic acid byproducts.

Arachidonic acid (AA) induced a concentration- and time-dependent reduction in gap junction-mediated dye coupling between cultured astrocytes. The effect was greatly diminished by inhibition of cyclooxygenases and lipoxygenases. The action of a low concentration of AA (5 microM) was also prevented by Ca2+-free extracellular solution or a high concentration of melatonin, a potent free radical scavenger, but not by Nomega-nitro-l-arginine, a nitric oxide (NO) synthase inhibitor. Thus, this effect may depend on Ca2+ influx and oxygen free radicals but not on NO generation. Cellular uncoupling induced by a high (100 microM), but not a low, AA concentration was rapidly reversed by washing with albumin containing solution. After reversal from 5 min but not 2.5 min inhibition with a high AA concentration dye coupling between astrocytes became refractory to a low concentration of AA, suggesting desensitization of the response elicited by a low concentration of the fatty acid. Dye uncoupling occurred without changes in levels and state of phosphorylation (immunoblotting and 32P-incorporation) of connexin43, the main astrocyte gap junctional protein. However, maximal cell uncoupling induced by a low (Slow action) but not by a high (Fast action) AA concentration was paralleled by a reduction in connexin43 (immunofluorescence) at cell-to-cell contacts. It is proposed that the AA-induced dye uncoupling is mediated by byproducts that induce rapid channel closure or slow removal of connexin43 gap junctions.

Cells of the carotid body express connexin43 which is up-regulated by cAMP.

We identified a gap junction protein subunit, connexin43 (Cx43) by immunofluorescence and immunoblotting, in cultured rat carotid body cells and in whole organs. In 1-week-old cultures, all cells were flat but after 3 h exposure to 8Br-cAMP (1 mM), tyrosine hydroxylase (TH) positive cells (chemoreceptors), but not TH negative cells, adopted a round body with multiple thin arborization processes. The incidence of dye coupling between cultured cells of the same type increased from 26% in controls to 73% after treatment with 8Br-cAMP. In control cultures, Cx43 immunoreactivity showed a diffuse perinuclear distribution and after 8Br-cAMP treatment, it was also found at cell-cell contacts. Both 8Br-cAMP-induced dye coupling and cellular redistribution of Cx43 were blocked by pretreatment with actinomycin D (5 microM), a mRNA transcription blocker. Moreover, 3 h exposure to 8Br-cAMP increased the levels of Cx43 in entire organs. We suggest that conditions that promote a sustained increase in cytosolic cAMP up-regulate coupling between carotid body cells in a transcription-dependent manner. The possible functional significance of these findings is discussed.

Regulation of glycogen content in rat pineal gland by norepinephrine.

In the rat pineal gland the glycogen stores were cytochemically localized in astrocytes and pinealocytes. Moreover, it was found that norepinephrine (NE) induced a time- and concentration-dependent reduction in pineal glycogen content and yielded lactic acid. The NE effect was prevented by blocking alpha1- but not alpha2 or beta-adrenoceptors. Activation of alpha2-adrenoceptors induced a small decrease in glycogen levels that could have pre- and postsynaptic components. Activation of beta-adrenoceptors with 10(-12)-10(-3) M isoproterenol (ISO) induced a bell shape concentration-response curve, presumably due to desensitization, since the response induced by 10(-4) M ISO was greater with shorter period of stimulation. On the other hand, activation of alpha1-adrenoceptors with 10(-12)-10(-3) M phenylephrine (PHN) induced a hyperbolic concentration-response curve with a maximum at concentrations above 10(-8) M. Moreover, treatment with ISO drastically reduced the response induced by PHN concentrations lower but not higher than 10(-6) M, favoring a concentration-dependent response between 10(-6) and 10(-4) M PHN, similar to that induced by equimolar NE concentrations. Thus, the NE-induced reduction in glycogen content of the rat pineal gland is mainly mediated by alpha1-adrenoceptors and modulated by intracellular mechanisms activated by beta-adrenoceptors.

Gap junctions in the chicken pineal gland.

The chicken pineal gland, which contains a heterogeneous cell population, sustains a circadian rhythm of activity. Synchronization of cellular activity of heterogeneous cells might be facilitated by gap junctional intercellular channels which are permeable to ions and second messengers. To test this possibility, we looked for morphologically identifiable gap junctions between the different pineal cells, used antibodies and cDNA probes to screen for the presence of connexins, and tested for functional intercellular coupling. By transmission electron microscopy and immunocytochemistry, gap junctions and connexins were observed between pinealocyte cell bodies, stromal cells, astrocytes, and astrocyte and pinealocyte processes. Two gap junctional proteins, connexin43 and connexin45, were detected by immunocytochemistry, immunoblotting and RNA blot analysis. Functional intercellular coupling was observed in the gland by transfer of low molecular weight dyes. Dye transferred between homologous and heterologous cells. These data suggest that homologous and heterologous gap junctions may provide a mechanism for coordination of the cellular responses of the elements of the biological clock which are induced by lighting cues to produce the circadian rhythm of pineal activity.

Pinealocytes in rats: connexin identification and increase in coupling caused by norepinephrine.

Dye coupling was observed between pinealocytes in acutely dissected pineal glands of adult rats. Pinealocytes maintained in culture were also electrically coupled. Connexins 26 and 43 and their respective mRNAs were present but neither connexin32 nor its mRNA were detected. Pinealocytes expressed only connexin26 whereas connexin43 was confined to astrocytes. In 5-day-old cultures of pinealocytes the incidence of dye coupling and level of immunodetectable connexin26 were low, and both were increased by norepinephrine (NE). The increase in incidence of coupling was maximal at around 6 h after treatment and was prevented by inhibitors of protein or mRNA synthesis. NE-induced metabolic and electrical synchronization mediated by gap junctions may favor melatonin secretion.