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.

Pannexin1 hemichannels are critical for HIV infection of human primary CD4+ T lymphocytes.

HIV is a major public health issue, and infection of CD4(+) T lymphocytes is one of its key features. Whereas several cellular proteins have been identified that facilitate viral infection and replication, the role of hemichannels in these processes has not been fully characterized. We now show that the HIV isolates, R5 and X4, induced a transient-early (5-30 min) and a later, persistent (48-120 h) opening of Panx1 hemichannels, which was dependent on the binding of HIV to CD4 and CCR5/CXCR4 receptors. Blocking Panx1 hemichannels by reducing their opening or protein expression inhibited HIV replication in CD4(+) T lymphocytes. Thus, our findings demonstrate that Panx1 hemichannels play an essential role in HIV infection.

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.

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

El traumatismo encéfalo craneano (TEC) es un problema de salud de distribución mundial, y es especialmente prevalente en la población adulta joven. Es característica la presencia de uno o más focos de daño, que luego progresan hacia áreas inicialmente no lesionadas, mediante cascadas de respuesta inflamatoria, excitotoxicidad, condiciones de falla energética, y la participación de la glía amplificando la respuesta tisular al daño inicial. Esta progresión es, en teoría, susceptible de una intervención terapéutica. Sin embargo, hasta ahora todos los estudios con fármacos neuroprotectores han fracasado, no existiendo un tratamiento especí-fico efectivo. Los resultados negativos se explican en parte por el empleo de una estrategia centrada solo en las neuronas, sin considerar otras células participantes, u otros mecanismos patogénicos. Para cambiar este panorama, es necesario re-enfocar el problema a través de una mejor comprensión de los mecanismos que determinan la progresión del daño. En esta revisión discutiremos los principales mecanismos biológicos involucrados en la progresión del daño tisular post-trauma. Se aborda la fisiopatología general de los tipos de traumatismos, mecanismos celulares del daño secundario incluyendo inflamación, apoptosis, tumefacción celular, excitoxicidad, y participación de la glía en la propagación del daño. Se destaca el papel de laglía en cada uno de los mecanismos celulares mencionados. Se incluyen algunas aproximaciones terapéuticas relacionadas con los mecanismos descritos. Se finaliza con un diagrama general que resume los principales aspectos discutido (AU)

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 (AU)

[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.

Gap junction hemichannels in astrocytes of the CNS.

Connexins are protein subunits that oligomerize into hexamers called connexons, gap junction hemichannels or just hemichannels. Because some gap junction channels are permeable to negatively and/or positively charged molecules up to approximately 1kDa in size, it was thought that hemichannels should not open to the extracellular space. A growing amount of evidence indicates that opening of hemichannels does occur under both physiological and pathological conditions in astrocytes and other cell types. Electrophysiological studies indicate that hemichannels have a low open probability under physiological conditions but may have a much higher open probability under certain pathological conditions. Some of the physiological behaviours of astrocytes that have been attributed to gap junctions may, in fact, be mediated by hemichannels. Hemichannels constituted of Cx43, the main connexin expressed by astrocytes, are permeable to small physiologically significant molecules, such as ATP, NAD+ and glutamate, and may mediate paracrine as well as autocrine signalling. Hemichannels tend to be closed by negative membrane potentials, high concentrations of extracellular Ca2+ and intracellular H+ ions, gap junction blockers and protein phosphorylation. Hemichannels tend to be opened by positive membrane potentials and low extracellular Ca2+, and possibly by as yet unidentified cytoplasmic signalling molecules. Exacerbated hemichannel opening occurs in metabolically inhibited cells, including cortical astrocytes, which contributes to the loss of chemical gradients across the plasma membrane and speeds cell death.

Expression of connexin43 in mouse Leydig, Sertoli, and germinal cells at different stages of postnatal development.

Connexin 43 (Cx43) is the most abundant and ubiquitously distributed gap junction protein in testicular cells. Lack of Cx43 expression results in male infertility. We investigated whether Cx43 is expressed and regulated in Leydig, Sertoli and germinal cells at different stages of postnatal development. Cx43 was detected using three different antibodies shown by immunoblotting to be highly specific. At different postnatal ages Cx43 localization was compared in serial or double labeled testicular cryosections with immunocytochemical distribution of steroidogenic enzyme, 3 betahydroxysteroid-dehydrogenase (3betaHSD), Mullerian inhibitory hormone (MIH), and germinal nuclear cell antigen (GNCA1), which are specific markers of interstitial Leydig, Sertoli and germinal cells, respectively. In the interstitium, round cell clumps (RCC) with lipid droplets positive for 3betaHSD and Cx43 were frequently found at intertubular areas at birth and Cx43 was mainly localized at cell membrane appositions. From day 3, the number and size of 3betaHSD-positive RCC started to decrease, and reached a minimum at 7-14 dpp; Cx43 expressed by them is progressively downregulated. From day 21 an increase in the size and number of RCC positive for Cx43 and 3betaHSD was found that continued at 24, 26 and 28 days and reached a maximum at 35 and 60 dpp. Biphasic expression of interstitial Cx43 and 3betaHSD was also found to be positively and temporally correlated with fluctuations in intratesticular testosterone content at all ages studied. In the seminiferous cord (SC), Cx43 was expressed at birth between adjacent Sertoli cells (MIH positive) localized at the periphery, as well as in their cytoplasm projections that surround centrally localized gonocytes. From days 3 to 7, Cx43 labeling increased in Sertoli cells mainly at their apical border. At day 14, Cx43 distribution in Sertoli cells changed from apical to basal in parallel to migration of germinal (GNCA1-positive) cells from the periphery to the center of the SC. At all these ages, Cx43 was also localized at cell borders between Sertoli and germinal cells. In conclusion, this study demonstrates that Cx43 in Leydig cells is regulated during postnatal development in an age and functional dependent manner. In the tubule, it is demonstrated that Cx43 is modulated in Sertoli cells during the neonatal and prepubertal period. We also provide evidence for the first time that Cx43-gap junctions communicate between Sertoli and germinal cells before and during the first wave of spermatogenesis.

Microglia at brain stab wounds express connexin 43 and in vitro form functional gap junctions after treatment with interferon-gamma and tumor necrosis factor-alpha.

Gap junctional communication between microglia was investigated at rat brain stab wounds and in primary cultures of rat and mouse cells. Under resting conditions, rat microglia (FITC-isolectin-B4-reactive cells) were sparsely distributed in the neocortex, and most (95%) were not immunoreactive for Cx43, a gap junction protein subunit. At brain stab wounds, microglia progressively accumulated over several days and formed aggregates that frequently showed Cx43 immunoreactivity at interfaces between cells. In primary culture, microglia showed low levels of Cx43 determined by Western blotting, diffuse intracellular Cx43 immunoreactivity, and a low incidence of dye coupling. Treatment with the immunostimulant bacterial lipopolysaccharide (LPS) or the cytokines interferon-gamma (INF-gamma) or tumor necrosis factor-alpha (TNF-alpha) one at a time did not increase the incidence of dye coupling. However, microglia treated with INF-gamma plus LPS showed a dramatic increase in dye coupling that was prevented by coapplication of an anti-TNF-alpha antibody, suggesting the release and autocrine action of TNF-alpha. Treatment with INF-gamma plus TNF-alpha also greatly increased the incidence of dye coupling and the Cx43 levels with translocation of Cx43 to cell-cell contacts. The cytokine-induced dye coupling was reversibly inhibited by 18 alpha-glycyrrhetinic acid, a gap junction blocker. Cultured mouse microglia also expressed Cx43 and developed dye coupling upon treatment with cytokines, but microglia from homozygous Cx43-deficient mice did not develop significant dye coupling after treatment with either INF-gamma plus LPS or INF-gamma plus TNF-alpha. This report demonstrates that microglia can communicate with each other through gap junctions that are induced by inflammatory cytokines, a process that may be important in the elaboration of the inflammatory response.

Influence of operating conditions and vessel size on oxygen transfer during cellulase production.

The production of low-cost cellulase enzyme is a key step in the development of an enzymatic-based process for conversion of lignocellulosic biomass to ethanol. Although abundant information is available on cellulase production, little of this work has examined oxygen transfer. We investigated oxygen transfer during the growth of Trichoderma reesei, a cellulase-producing microorganism, on soluble and insoluble substrates in vessel sizes from 7 to 9000 L. Oxygen uptake rates and volumetric mass transfer coefficients (kLa) were determined using mass spectroscopy to measure off gas composition. Experimentally measured kLa values were found to compare favorably with a kLa correlation available in the literature for a non-Newtonian fermentation broth during the period of heavy cell growth.

Short- and long-term regulation of rat carotid body gap junctions by cAMP. Identification of connexin43, a gap junction subunit.

Intact and cultured carotid bodies (CBs) of the rat were used in this study. Applications of membrane-permeant db-cAMP to cultured carotid bodies increased electric coupling between most glomus cells (increasing junctional conductance) probably by opening preformed intercellular channels. This a short-term effect of the nucleotide, increasing gating between glomus cells. When cultures and intact carotid bodies were treated with membrane-permeant 8Br-cAMP for 3 h or more (to increase cytosolic cAMP), there was enhanced gap junction formation and better dye spread between carotid body cells. Connexin43 (CX43) was identified by immunocytochemical methods as forming part of the intercellular channels between carotid body cells, and the expression of Cx43 increased by cAMP. This is a long-term effect, inducing the formation of gap junctions. Thus, cAMP had short and long-term effects on the intercellular junctions of the carotid body. Long-term formation of gap junctions may be important in modulating carotid body functions during stimulation by chronic hypoxia.

Gap junctions in cells of the immune system: structure, regulation and possible functional roles.

Gap junction channels are sites of cytoplasmic communication between contacting cells. In vertebrates, they consist of protein subunits denoted connexins (Cxs) which are encoded by a gene family. According to their Cx composition, gap junction channels show different gating and permeability properties that define which ions and small molecules permeate them. Differences in Cx primary sequences suggest that channels composed of different Cxs are regulated differentially by intracellular pathways under specific physiological conditions. Functional roles of gap junction channels could be defined by the relative importance of permeant substances, resulting in coordination of electrical and/or metabolic cellular responses. Cells of the native and specific immune systems establish transient homo- and heterocellular contacts at various steps of the immune response. Morphological and functional studies reported during the last three decades have revealed that many intercellular contacts between cells in the immune response present gap junctions or "gap junction-like" structures. Partial characterization of the molecular composition of some of these plasma membrane structures and regulatory mechanisms that control them have been published recently. Studies designed to elucidate their physiological roles suggest that they might permit coordination of cellular events which favor the effective and timely response of the immune system.

Regulation of astrocyte gap junctions by hypoxia-reoxygenation.

Confluent cultures of rat cortical astrocytes were subjected to 12-h hypoxia (<1% O(2)) followed by reoxygenation. Just after hypoxia, the cellular distribution, phosphorylation state and levels of connexin43 (Cx43), as well as the extent of dye coupling were as in control conditions. Nonetheless, 15-30 min after reoxygenation, dye coupling was transiently reduced by approximately 70%. The reduction in dye coupling occurred without changes in the state of phosphorylation or levels of Cx43. Nevertheless, it was correlated with a decrease in Cx43 reactivity found at membrane appositions and the appearance of intracellular Cx43-positive vesicle-like structures of variable size, suggesting internalization of gap junction channels. Reoxygenation-induced cellular uncoupling and redistribution of Cx43 were prevented by melatonin (500 microM), a potent-free radical scavenger, or indomethacin (50 microM), an inhibitor of the cyclooxygenase-dependent arachidonic acid metabolism. In astrocytes cultured under normoxia, the state of phosphorylation of Cx43 was not affected by antimycin A, a blocker of the mitochondrial oxidative metabolism, but phosphorylation was drastically reduced by iodoacetate, a blocker of anaerobic glycolysis. Thus, these results strongly suggest that reoxygenation-induced uncoupling is mediated by arachidonic acid byproducts that induce, at least, disorganization of Cx43 gap junction channels.

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.

Gap junctions in cells of the immune system: structure, regulation and possible functional roles

Gap junction channels are sites of cytoplasmic communication between contacting cells. In vertebrates, they consist of protein subunits denoted connexins (Cxs) which are encoded by a gene family. According to their Cx composition, gap junction channels show different gating and permeability properties that define which ions and small molecules permeate them. Differences in Cx primary sequences suggest that channels composed of different Cxs are regulated differentially by intracellular pathways under specific physiological conditions. Functional roles of gap junction channels could be defined by the relative importance of permeant substances, resulting in coordination of electrical and/or metabolic cellular responses. Cells of the native and specific immune systems establish transient homo- and heterocellular contacts at various steps of the immune response. Morphological and functional studies reported during the last three decades have revealed that many intercellular contacts between cells in the immune response present gap junctions or "gap junction-like" structures. Partial characterization of the molecular composition of some of these plasma membrane structures and regulatory mechanisms that control them have been published recently. Studies designed to elucidate their physiological roles suggest that they might permit coordination of cellular events which favor the effective and timely response of the immune system

Use of antibodies in the analysis of connexin 43 turnover and phosphorylation.

A series of antipeptide antibodies designed to recognize specific sequences of the gap junction protein connexin 43 (Cx43) were developed and characterized immunochemically and immunohistologically. These antibodies bound to gap junctions and, on Western blots, to 43-kDa (often resolved as a doublet) and 41-kDa proteins in samples from heart, leptomeningeal cells, and brain. Relatively little of the 41-kDa protein was detectable in heart homogenates. Cultured rat leptomeningeal cells expressed high levels of the gap junction protein Cx43 and were used to analyze its turnover and phosphorylation. Pulse-chase experiments in leptomeningeal cells with [(35)S]methionine indicated that the 41-kDa form of connexin 43 was the first immunoprecipitable translation product. Radiolabel subsequently appeared in the lower band of the doublet at 43 kDa, followed by a shift into the higher band and turnover of the protein with a t(1/2) of 2.7 h. Pulse-chase labeling with [(32)P]P(i) indicated that phosphorylation of connexin 43 was limited to the 43-kDa protein, with a t(1/2) of 1.7 h. Treatment with alkaline phosphatase shifted the apparent molecular mass of the 43-kDa protein doublet such that it comigrated with the 41-kDa form. Hence, the 43-kDa protein observed on Western blots of both leptomeningeal cells and heart arises by phosphorylation of the 41 kDa precursor. Phosphorylation of serine residues accounts for most, if not all, of Cx43 phosphorylation in this system.