Desarrollo y caracterización de un material celular de alta porosidad con base cementicia activada mediante agentes inorgánicos

Esta tesis ha estudiado los morteros celulares, centrándose en la experimentación con pastas de cemento aireadas (PCA) con polvo de aluminio como agente expansor. El objetivo es el desarrollo de un material cementicio con una baja conductividad térmica que sirva como aislamiento térmico. La naturaleza inorgánica del material lo hace incombustible, en contraste con las espumas poliméricas existentes en el mercado, cuya aplicación en cámaras ventiladas ha sido prohibida por normativas de construcción tanto a nivel nacional como internacional. Las posibles aplicaciones son con proyección neumática o en paneles prefabricados. Se han ensayado dos series de pastas de cemento con polvo de aluminio: - Serie WPC/CAC/CH. Mezcla de referencia con cemento blanco (WPC), cemento de aluminato cálcico (CAC) y cal aérea (CH) en proporción 5:1:4. - Serie OPC/CH. Mezcla de referencia con cemento portland con cenizas volantes (OPC) y cal aérea (CH) en proporción OPC/CH de 4:1 A las mezclas de referencia se le han añadido adiciones de metacaolín (MK) (10 y 20%) o sepiolita (SP) (1 y 2%) para observar el efecto que producen tanto en el mortero fresco como en el mortero endurecido. Se ha estudiado la reología de las pastas en estado fresco, analizando el proceso de expansión de las pastas, registrando los valores de tensión de fluencia, aire ocluido y temperatura durante la expansión. Con los valores obtenidos se ha discutido la influencia de las adiciones utilizadas en la cinética de corrosión del polvo de aluminio que genera la expansión, concluyendo que las adiciones puzolánicas (CV y MK) y la SP reducen mucho el periodo de inducción, lo que provoca poros más grandes y mayor cantidad de aire ocluido. Asimismo se ha analizado la relación entre la tensión de fluencia y el contenido de aire ocluido, deduciendo que a mayor tensión de fluencia en el momento de iniciarse la expansión, menor tamaño de poros y contenido de aire ocluido. Finalmente, se han obtenido las densidades y capacidades de retención de agua de los morteros frescos. Para caracterizar la red porosa de las pastas aireadas endurecidas, se obtuvieron tanto las densidades reales, netas, aparentes y relativas como las porosidades abiertas, cerradas y totales con ensayos hídricos. Finalmente se obtuvieron imágenes de los poros con tomografía axial computerizada para obtener las porosimetrías de las muestras. La caracterización de la red porosa ha servido para terminar de analizar lo observado en la evolución de la expansión del mortero fresco. Se ha analizado la influencia de la red porosa en la conductividad térmica, obtenida con caja caliente, comparándola con la existente en la literatura existente tanto de morteros celulares como de espumas poliméricas. Se concluye que los valores de conductividad térmica conseguida están en el mínimo posible para un material celular de base cementicia. La microestructura se ha estudiado con microscopía electrónica de barrido, difracción de rayos X y ensayos térmicos TG/ATD, observando que los productos de hidratación encontrados coinciden con los que se producen en morteros sin airear. Las imágenes SEM y los resultados de ultrasonidos han servido para analizar la presencia de microfisuras de retracción en las pastas aireadas, observando que en las muestras con adiciones de MK y SP, se reduce la presencia de microfisuras. ABSTRACT This thesis has studied cellular mortars, focusing in testing aerated cement pastes with aluminum powder as expansive agent. The purpose is the development of a cementitious material with low thermal conductivity that can be used as thermal isolation. Inorganic nature of this material makes it non-combustible, in contrast with polymeric foams in market, whose application in ventilated double skin façade systems has been banned by building standards, both domestically and internationally. Possible uses for this material are pneumatically sprayed applications and precast panels. Two series of batches with aluminum powder have been tested: - WPC/CAC/CH series. Reference paste with white portland cement (WPC), calcium aluminate cement (CAC) and lime (CH) with 5:1:4 ratio. - OPC/CH series. Reference paste with portland cement with fly ash (OPC) and lime (CH) with 4:1 ratio. Metakaolin (MK) (10 and 20%) or sepiolite (SP) (1 and 2%) additions were used in reference pastes to characterize the effect in fresh and hardened mortar. Rheology in fresh pastes was studied, expansion process of pastes was analyzed, recording yield stress, entrained air and temperature values during expansion. Recorded values were used to discuss influence of additions on reaction kinetics of aluminum powder corrosion, that produces expansion.. Conclusion is that pozzolanic additions (FA and MK) and SP greatly reduce induction period, producing bigger pores and more entrained air. Relation between yield stress and entrained air has been also analyzed, observing that the bigger yield stress at beginning of expansion, the smaller pores size and the lower entrained air values. Finally density and water retention of fresh mortars were obtained. Pore network in hardened aerated cement pastes was characterized by imbibition methods providing true, bulk and relative density, and providing also open, closed and total porosity. Finally, pore system imaging were obtained with computerized axial tomography to study porosimetry of specimens. Pore network characterization was useful to complete facts analysis observed in expansion of fresh mortars. Influence of pore network in thermal conductivity, checked in hot box, was analyzed comparing with those existing values in cellular mortar and polymeric foams researches. It was concluded that thermal conductivity values achieved are close to minimum possible in a cementitious cellular material. Microstructure was studied with Scanning Electron Microscopy, X-Ray Diffractometry and TG-DTA analysis, observing that hydration phases found, are those produced in non aerated mortar. SEM imaging and ultrasound results were useful to analyze shrinkage microcracks in aerated cement pastes, concluding that microcrack presence in specimens with MK and SP additions were reduced.

Characterization of a Mo/Au thermometer for ATHENA

The first dark characterization of a thermometer fabricated with our Mo/Au bilayers to be used as a transition edge sensor is presented. High-quality, stress-free Mo layers, whose thickness is used to tune the critical temperature (TC ) down to 100 mK, are deposited by sputtering at room temperature (RT ) on Si3N4 bulk and membranes, and protected from degradation with a 15-nm sputtered Au layer. An extra layer of high-quality Au is deposited by ex situ e-beam to ensure low residual resistance. The thermometer is patterned on a membrane using standard photolithographic techniques and wet etching processes, and is contacted through Mo paths, displaying a sharp superconducting transition (α ≈ 600). Results show a good coupling between Mo and Au layers and excellent TC reproducibility, allowing to accurately correlate dM o and TC . Since dAu is bigger than ξM for all analyzed samples, bilayer residual resistance can be modified without affecting TC . Finally, first current to voltage measurements at different temperatures are measured and analyzed, obtaining the corresponding characterization parameters.

Ultra high temperature latent heat energy storage and thermophotovoltaic energy conversion

A conceptual energy storage system design that utilizes ultra high temperature phase change materials is presented. In this system, the energy is stored in the form of latent heat and converted to electricity upon demand by TPV (thermophotovoltaic) cells. Silicon is considered in this study as PCM (phase change material) due to its extremely high latent heat (1800 J/g or 500 Wh/kg), melting point (1410 C), thermal conductivity (~25 W/mK), low cost (less than $2/kg or $4/kWh) and abundance on earth. The proposed system enables an enormous thermal energy storage density of ~1 MWh/m3, which is 10e20 times higher than that of lead-acid batteries, 2e6 times than that of Li-ion batteries and 5e10 times than that of the current state of the art LHTES systems utilized in CSP (concentrated solar power) applications. The discharge efficiency of the system is ultimately determined by the TPV converter, which theoretically can exceed 50%. However, realistic discharge efficiencies utilizing single junction TPV cells are in the range of 20e45%, depending on the semiconductor bandgap and quality, and the photon recycling efficiency. This concept has the potential to achieve output electric energy densities in the range of 200-450 kWhe/m3, which is comparable to the best performing state of the art Lithium-ion batteries.

Comportamiento térmico y mecánico del yeso con adiciones de aerogel de sílice granulado

La presente tesis doctoral aborda el estudio de un nuevo material mineral, compuesto principalmente por una matriz de yeso (proveniente de un conglomerante industrial basado en sulfato de calcio multifase) y partículas de aerogel de sílice hidrófugo mesoporoso, compatibilizadas mediante un surfactante polimérico, debido a su alto carácter hidrófugo. La investigación se centra en conocer los factores que influyen en las propiedades mecánicas y conductividad térmica del material compuesto generado. Este estudio pretende contribuir al conocimiento sobre el desarrollo de nuevos morteros de elevado aislamiento térmico que puedan ser utilizados en la rehabilitación energética de edificios de viviendas existentes, debido a que estos representan gran parte del consumo energético del parque de viviendas de España, aunque también a nivel internacional. De los materiales utilizados para desarrollar los morteros estudiados, el yeso, además de ser un material muy abundante, especialmente en España, requiere una menor cantidad de energía para la fabricación de un conglomerante (debido a una menor temperatura de fabricación), en comparación con el cemento o la cal, por lo que presenta una menor huella de carbono que estos últimos. Por otro lado, el aerogel de sílice hidrófugo mesoporoso es, de acuerdo con la documentación disponible, el material que posee actualmente la mayor capacidad de aislamiento térmico en el mercado. El desarrollo de nuevos morteros minerales con una capacidad de aislamiento térmico mayor que los materiales aislantes utilizados tradicionalmente, tiene una aplicación relevante en los casos de rehabilitación energética de edificios históricos y patrimoniales, en los que se requiere la aplicación del aislamiento por el interior de la fachada, ya que este tipo de soluciones tienen el inconveniente de reducir el espacio habitable de las áreas involucradas, especialmente en zonas climáticas en las que el aislamiento térmico puede suponer un espesor considerable, por lo que es ideal utilizar materiales de altas prestaciones de aislamiento térmico capaces de aportar el mismo nivel de aislamiento (o incluso mayor), pero en un espesor considerablemente menor. La investigación se desarrolla en tres etapas: bibliográfica, experimental y de simulación. La primera etapa, parte del estudio de la bibliografía existente, relacionada con materiales aislantes, incluyendo soluciones basadas, tanto en morteros aislantes, como en paneles de aislamiento térmico. La segunda, de carácter experimental, se centra en estudiar la influencia de la microestrucrura y macroestructura, del nuevo material mineral, en las propiedades físicas elementales, mecánicas y conductividad térmica del compuesto. La tercera etapa, mediante una simulación del consumo energético, consiste en cuantificar teóricamente el potencial ahorro energético que puede aportar este material en un caso de rehabilitación energética en particular. La investigación experimental se centró principalmente en conocer los factores principales que influyen en las propiedades mecánicas y conductividad térmica de los materiales compuestos minerales desarrollados en esta tesis. Para ello, se llevó a cabo una caracterización de los materiales de estudio, así como el desarrollo de distintas muestras de ensayo, de tal forma que se pudo estudiar, tanto la hidratación del yeso en los compuestos, como su posterior microestructura y macroestructura, aspectos fundamentales para el entendimiento de las propiedades mecánicas y conductividad térmica del compuesto aislante. De este modo, se pudieron conocer y cuantificar, los factores que influyen en las propiedades estudiadas, aportando una base de conocimiento y entendimiento de este tipo de compuestos minerales con aerogel de sílice hidrófugo, no existiendo estudios publicados hasta el momento de finalización de esta tesis, con la aproximación al material propuesta en este estudio, ni con yeso (basado en sulfato de calcio multifase), ni con otro tipo de conglomerantes. Particularmente, se determinó la influencia que tiene la incorporación de partículas de aerogel de sílice hidrófugo, en grandes proporciones en volumen, en un compuesto mineral basado en distintas fases de sulfato de calcio. No obstante, para llevar a cabo las mezclas, fue necesario utilizar un surfactante para compatibilizar este tipo de partículas, con el conglomerante basado en agua. El uso de este tipo de aditivos tiene una influencia, no solo en el aerogel, sino en las propiedades del compuesto en general, dependiendo de su concentración, por lo que se establecieron dos porcentajes de adición: la primera, determinada a partir de la cantidad mínima necesaria para compatibilizar las mezclas (0,1% del agua de amasado), y la segunda, como límite superior, la concentración utilizada habitualmente a nivel industrial para estabilizar burbujas de aire en hormigones espumados (5%). El surfactante utilizado mostró la capacidad de modificar la superficie del aerogel, cambiando el comportamiento de las partículas frente al agua, permitiendo una invasión parcial de su estructura porosa, por parte del agua de amasado. Este comportamiento supone un aumento muy importante en la relación agua/yeso, afectando el hábito cristalino e influenciando negativamente las propiedades mecánicas de la matriz de yeso, presentando un efecto aún notable a mayor concentración de surfactante (5%). En cuanto a las propiedades finales alcanzadas, fue posible lograr un compuesto mineral ultraligero (200 kg/m3), con alrededor de un 60% de aerogel en volumen y de alta capacidad aislante (0,028 W/m•K), presentando una conductividad térmica notablemente menor que los morteros aislantes del mercado, e incluso también menor que la de los aislantes tradicionales basado en las lanas minerales o EPS; no obstante, con la limitante de presentar bajas propiedades mecánicas, condicionando su posible aplicación futura. Entre los factores principales relacionados con las propiedades mecánicas, se encontró que estas dependen exponencialmente del volumen de yeso en el compuesto; no obstante, factores de segundo orden, como el grado de hidratación, o una mejor distribución del conglomerante entre las partículas de aerogel, debido al aumento de la superficie específica del polvo mineral, pueden aumentar las propiedades mecánicas entre el doble y el triple, dependiendo del volumen de aerogel en cuestión. Además, se encontró que el aerogel, en conjunto con el surfactante, es capaz de introducir una gran cantidad de aire (0,70 m3 por cada m3 de aerogel), que unido al agua evaporada (no consumida por el conglomerante durante la hidratación), el volumen de aire total alcanza, generalmente, un 40%, independientemente de la cantidad de aerogel en la mezcla. De este modo, el aire introducido en la matriz desplaza las proporciones en volumen del aerogel y del yeso, disminuyendo, tanto las propiedades mecánicas, como la capacidad aislante de compuesto mineral. Por otro lado, la conductividad térmica mostró tener una dependencia directa de la contribución de las tres fases principales en el compuesto: yeso, aerogel y aire ocluido. De este modo, se pudo desarrollar un modelo matemático, adaptado de uno existente, capaz de calcular, con bastante precisión, la relación de los tres componentes mencionados, en la conductividad térmica de los compuestos, para el rango de volúmenes y materiales utilizados en esta tesis. Finalmente, la simulación del consumo energético realizada a una vivienda típica de España, de los años 1900 a 1959 (basada en muros de ladrillo macizo), para las zonas climáticas estudiadas (A, D y E), permitió observar el potencial ahorro energético que puede aportar este material, dependiendo de su espesor, como aislamiento interior de los muros de fachada. Particularmente, para la zona A, se determinó un espesor óptimo de 1 cm, mientras que para la zona D y E, 3,5 y 3,9 cm respectivamente. En este sentido, el nuevo material estudiado es capaz de disminuir, entre un 35% y un 80%, el espesor de la capa aislante, en comparación con paneles de lana de roca o los morteros minerales de mayor capacidad aislante del mercado español respectivamente. ABSTRACT The present doctoral thesis studies a new mineral-based composite material, composed by a gypsum matrix (based on an industrial multiphase gypsum binder) and mesoporous hydrophobic silica aerogel particles, compatibilized with a polymeric surfactant due to the high hydrophobic character of the insulating particles. This study pretends to contribute to the development of new composite insulating materials that could be used in energy renovation of existing dwellings, in order to reduce their high energy consumption, as they represent a great part of the total energy consumed in Spain, but also internationally. Between the materials used to develop de studied insulating mortars, gypsum, besides being an abundant material, especially in Spain, requires less energy for the manufacture of a mineral binder (due to lower manufacturing temperatures), compared to lime or cement, thus presenting lower carbon footprint. In other hand, the hydrophobic mesoporous silica aerogel, is, according to the existing references, the material with the highest know insulating capacity in the market. The development of new mineral mortars with higher thermal insulation capacity than traditional insulating materials, presents a relevant application in energy retrofitting of historic and cultural heritage buildings, in which implies that the insulating material should be installed as an internal layer, rather than as an external insulating system. This type of solution involves a reduced internal useful area, especially in climatic zones where the demand for thermal insulation is higher, and so the insulating layer thickness, being idealistic to use materials with very high insulating properties, in order to reach same insulating level (or higher), but in lower thickness than the provided by traditional insulating materials. This research is developed in three main stages: bibliographic, experimental and simulation. The first stage starts by studying the existing references regarding thermally insulating materials, including existing insulating mortars and insulating panels. The second stage, mainly experimental, is centered in the study of the the influence of the microstructure and macrostructure in the physical and mechanical properties, and also in the thermal conductivity of the new mineral-based material. The thirds stage, through energy simulation, consists in theoretically quantifying the energy savings potential that can provide this type of insulating material, in a particular energy retrofitting case study. The experimental research is mainly focused in the study of the factors that influence the mechanical properties and the thermal conductivity of the thermal insulating mineral composites developed in this thesis. For this, the characterization of the studied materials has been performed, as well as the development of several experimental samples, in order to study the hydration of the mineral binder within the composites, but also the final microstructure and macrostructure, fundamental aspects for the understanding of the composite’s mechanical and insulating properties. Thus, is was possible to determine and quantify the factors that influence the studied material properties, providing a knowledge base and understanding of mineral composites that comprises mesoporous hydrophobic silica aerogel particles, being the first study up to date regarding the specific approach of the present study, regarding not just multiphase calcium sulfate plaster, but also other mineral binders. Particularly, the influence of the incorporation of hydrophobic silica aerogel particles, in high volume ratios into a mineral compound, based on different phases of calcium sulfate has been determined. However, to perform mixing, it is necessary to use a surfactant in order to compatibilize these particles with the water-based mineral binder. The use of such additives has an influence, not only in the aerogel, but the overall properties of the compound, so two different surfactant concentration has been studied: the first, the minimum amount of surfactant (used in this thesis) in order to develop the slurries (0.1% concentration of the mixing water), and the second, as the upper limit, the concentration usually used industrially to stabilize air bubbles in foamed concrete (5%). One of the side effects of using such additive, was the modification of the aerogel particles, by changing their behavior in respect to water, generating a partial invasion of the aerogel’s porous structure, by the mixing water. This behavior produces a very important increase in water/binder ratios, affecting the crystal habit and negatively influencing the mechanical properties of the gypsum matrix. This effect further increased when a higher concentration of surfactant (5%) is used. Regarding final materials properties, it was possible to achieve an ultra-lightweight mineral composite (200 kg/m3), with around 60% by volume of aerogel, presenting a very high insulating capacity (0.028 W/m•K), a noticeable lower thermal conductivity compared to the insulating mortars and traditional thermal insulating panels on the market, such as mineral wool or EPS; however, the limiting factor for future’s material application in buildings, is related to the very low mechanical properties achieved. Among the main factors related to the mechanical properties, it has been found an exponential correlation to the volume of gypsum in the composite. However, second-order factors such as the degree of hydration, or a better distribution of the binder between the aerogel particles, due to the increased surface area of the mineral powder, can increase the mechanical properties between two to three times, depending aerogel volume involved. In addition, it was found that the aerogel, together with the surfactant, is able to entrain a large amount of air volume (around 0.70 m3 per m3 of aerogel), which together with the evaporated water (not consumed by the binder during hydration), can reach generally around 40% of entrained air within the gypsum matrix, regardless of the amount of aerogel in the mixture. Thus, the entrained air into the matrix displaces the volume proportions of the aerogel and gypsum, reducing both mechanical and insulating properties of the mineral composite. On the other hand, it has been observed a direct contribution of three main phases into the thermal conductivity of the composite: gypsum, aerogel and entrained air. Thus, it was possible to develop a mathematical model (adapted from an existing one), capable of calculating quite accurate the thermal conductivity of such mineral composites, from the ratio these three components and for the range of volumes and materials used in this thesis. Finally, the energy simulation performed to a typical Spanish dwelling, from the years 1900 to 1959 (mainly constructed with massive clay bricks), within three climatic zones of Spain (A, D and E), showed the energy savings potential that can provide this type of insulating material, depending on the thickness of the applied layer. Particularly, for the climatic A zone, it has been found an optimal layer thickness of 1 cm, while for zone D and E, 3.5 and 3.9 cm respectively. In this manner, the new studied materials is capable of decreasing the thickness of the insulating layer by 35% and 80%, compared with rock wool panels or mineral mortars with the highest insulating performance of the Spanish market respectively.

Long-term potentiation and dual-component quantal signaling in the dentate gyrus

Long-term potentiation (LTP) of excitatory transmission is an important candidate cellular mechanism for the storage of memories in the mammalian brain. The subcellular phenomena that underlie the persistent increase in synaptic strength, however, are incompletely understood. A potentially powerful method to detect a presynaptic increase in glutamate release is to examine the effect of LTP induction on the rate at which the use-dependent blocker MK-801 attenuates successive N-methyl-d-aspartic acid (NMDA) receptor-mediated synaptic signals. This method, however, has given apparently contradictory results when applied in hippocampal CA1. The inconsistency could be explained if NMDA receptors were opened by glutamate not only released from local presynaptic terminals, but also diffusing from synapses on neighboring cells where LTP was not induced. Here we examine the effect of pairing-induced LTP on the MK-801 blocking rate in two afferent inputs to dentate granule cells. LTP in the medial perforant path is associated with a significant increase in the MK-801 blocking rate, implying a presynaptic increase in glutamate release probability. An enhanced MK-801 blocking rate is not seen, however, in the lateral perforant path. This result still could be compatible with a presynaptic contribution to LTP in the lateral perforant path if intersynaptic cross-talk occurred. In support of this hypothesis, we show that NMDA receptors consistently sense more quanta of glutamate than do α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors. In the medial perforant path, in contrast, there is no significant difference in the number of quanta mediated by the two receptors. These results support a presynaptic contribution to LTP and imply that differences in intersynaptic cross-talk can complicate the interpretation of experiments designed to detect changes in transmitter release.

Feasibility and utility of population-level geospatial injury profiling : prospective, national cohort study

The Health Services Research Unit receives funding from the Chief Scientist Office of the Scottish Government Health and Social Care Directorates. The opinions expressed in this article are those of the authors alone. The GEOS study was funded by the North of Scotland Planning Group.

Apoptosis and proliferation of dentate gyrus neurons after single and intermittent limbic seizures

Neuronal apoptosis was observed in the rat dentate gyrus in two experimental models of human limbic epilepsy. Five hours after one hippocampal kindling stimulation, a marked increase of in situ terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling (TUNEL) of fragmented DNA was observed in nuclei located within and on the hilar border of the granule cell layer and in the polymorphic region. Forty kindling stimulations with 5-min interval produced higher numbers of labeled nuclei compared with one stimulation. The increase of TUNEL-positive nuclei was prevented by the protein synthesis inhibitor cycloheximide but not affected by the N-methyl-d-aspartate receptor antagonist MK-801. Kainic acid-induced seizures lead to a pattern of labeling in the hippocampal formation identical to that evoked by kindling. A large proportion of cells displaying TUNEL-positive nuclei was double-labeled by the neuron-specific antigen NeuN, demonstrating the neuronal identity of apoptotic cells. Either 1 or 40 kindling stimulations also gave rise to a marked increase of the number of cells double-labeled with the mitotic marker bromodeoxyuridine and NeuN in the subgranular zone and on the hilar border of the dentate granule cell layer. The present data show that single and intermittent, brief seizures induce both apoptotic death and proliferation of dentate gyrus neurons. We hypothesize that these processes, occurring early during epileptogenesis, are primary events in the development of hippocampal pathology in animals and possibly also in patients suffering from temporal lobe epilepsy.

Temperature, ordering, and equilibrium with time-dependent confining forces

The concepts of temperature and equilibrium are not well defined in systems of particles with time-varying external forces. An example is a radio frequency ion trap, with the ions laser cooled into an ordered solid, characteristic of sub-mK temperatures, whereas the kinetic energies associated with the fast coherent motion in the trap are up to 7 orders of magnitude higher. Simulations with 1,000 ions reach equilibrium between the degrees of freedom when only aperiodic displacements (secular motion) are considered. The coupling of the periodic driven motion associated with the confinement to the nonperiodic random motion of the ions is very small at low temperatures and increases quadratically with temperature.

Lineage commitment in the progeny of murine hematopoietic preprogenitor cells: Influence of thrombopoietin and interleukin 5

Normal mouse marrow cells were stimulated by stem cell factor (SCF) to form dispersed or multicentric blast colonies containing progenitor cells committed to various hematopoietic lineages. Combination of the eosinophil-specific regulator interleukin 5 with SCF increased the frequency of colonies containing eosinophil-committed progenitor cells with multicentric but not dispersed blast colonies. Combination of thrombopoietin with SCF increased the frequency of colonies containing megakaryocyte-committed progenitor cells with both types of blast colony. Neither interleukin 5 nor thrombopoietin significantly altered the number or total cell content of blast colonies or progenitor cell numbers in blast colonies from those stimulated by SCF alone. No correlation was observed between total progenitor cell content and the presence or absence of either eosinophil or megakaryocyte progenitors in either type of blast colony. The data argue against a random process as being responsible for the formation of particular committed progenitor cells or the possibility that lineage-specific regulators merely enhance survival of such committed progenitor cells formed in developing blast colonies.

Identification in mice of two isoforms of the cysteinyl leukotriene 1 receptor that result from alternative splicing

Two classes of human G protein-coupled receptors, cysteinyl leukotriene 1 (CysLT1) and CysLT2 receptors, recently have been characterized and cloned. Because the CysLT1 receptor blockers are effective in treating human bronchial asthma and the mouse is often used to model human diseases, we isolated the mouse CysLT1 receptor from a mouse lung cDNA library and found two isoforms. A short isoform cDNA containing two exons encodes a polypeptide of 339 aa with 87.3% amino acid identity to the human CysLT1 receptor. A long isoform has two additional exons and an in-frame upstream start codon resulting in a 13-aa extension at the N terminus. Northern blot analysis revealed that the mouse CysLT1 receptor mRNA is expressed in lung and skin; and reverse transcription–PCR showed wide expression of the long isoform with the strongest presence in lung and skin. The gene for the mouse CysLT1 receptor was mapped to band XD. Leukotriene (LT) D4 induced intracellular calcium mobilization in Chinese hamster ovary cells stably expressing either isoform of the mouse CysLT1 receptor cDNA. This agonist effect of LTD4 was fully inhibited by the CysLT1 receptor antagonist, MK-571. Microsomal membranes from each transformant showed a single class of binding sites for [3H]LTD4; and the binding was blocked by unlabeled LTs, with the rank order of affinities being LTD4 >> LTE4 = LTC4 >> LTB4. Thus, the dominant mouse isoform with the N-terminal amino acid extension encoded by an additional exon has the same ligand response profile as the spliced form and the human receptor.

Insulin promotes rapid delivery of N-methyl-d- aspartate receptors to the cell surface by exocytosis

Insulin potentiates N-methyl-d-aspartate receptors (NMDARs) in neurons and Xenopus oocytes expressing recombinant NMDARs. The present study shows that insulin induced (i) an increase in channel number times open probability (nPo) in outside-out patches excised from Xenopus oocytes, with no change in mean open time, unitary conductance, or reversal potential, indicating an increase in n and/or Po; (ii) an increase in charge transfer during block of NMDA-elicited currents by the open channel blocker MK-801, indicating increased number of functional NMDARs in the cell membrane with no change in Po; and (iii) increased NR1 surface expression, as indicated by Western blot analysis of surface proteins. Botulinum neurotoxin A greatly reduced insulin potentiation, indicating that insertion of new receptors occurs via SNARE-dependent exocytosis. Thus, insulin potentiation occurs via delivery of new channels to the plasma membrane. NMDARs assembled from mutant subunits lacking all known sites of tyrosine and serine/threonine phosphorylation in their carboxyl-terminal tails exhibited robust insulin potentiation, suggesting that insulin potentiation does not require direct phosphorylation of NMDAR subunits. Because insulin and insulin receptors are localized to glutamatergic synapses in the hippocampus, insulin-regulated trafficking of NMDARs may play a role in synaptic transmission and plasticity, including long-term potentiation.

The Friend of GATA proteins U-shaped, FOG-1, and FOG-2 function as negative regulators of blood, heart, and eye development in Drosophila

Friend of GATA (FOG) proteins regulate GATA factor-activated gene transcription. During vertebrate hematopoiesis, FOG and GATA proteins cooperate to promote erythrocyte and megakaryocyte differentiation. The Drosophila FOG homologue U-shaped (Ush) is expressed similarly in the blood cell anlage during embryogenesis. During hematopoiesis, the acute myeloid leukemia 1 homologue Lozenge and Glial cells missing are required for the production of crystal cells and plasmatocytes, respectively. However, additional factors have been predicted to control crystal cell proliferation. In this report, we show that Ush is expressed in hemocyte precursors and plasmatocytes throughout embryogenesis and larval development, and the GATA factor Serpent is essential for Ush embryonic expression. Furthermore, loss of ush function results in an overproduction of crystal cells, whereas forced expression of Ush reduces this cell population. Murine FOG-1 and FOG-2 also can repress crystal cell production, but a mutant version of FOG-2 lacking a conserved motif that binds the corepressor C-terminal binding protein fails to affect the cell lineage. The GATA factor Pannier (Pnr) is required for eye and heart development in Drosophila. When Ush, FOG-1, FOG-2, or mutant FOG-2 is coexpressed with Pnr during these developmental processes, severe eye and heart phenotypes result, consistent with a conserved negative regulation of Pnr function. These results indicate that the fly and mouse FOG proteins function similarly in three distinct cellular contexts in Drosophila, but may use different mechanisms to regulate genetic events in blood vs. cardial or eye cell lineages.

Arrested development of embryonic red cell precursors in mouse embryos lacking transcription factor GATA-1.

The X chromosome-linked transcription factor GATA-1 is expressed specifically in erythroid, mast, megakaryocyte, and eosinophil lineages, as well as in hematopoietic progenitors. Prior studies revealed that gene-disrupted GATA-1- embryonic stem cells give rise to adult (or definitive) erythroid precursors arrested at the proerythroblast stage in vitro and fail to contribute to adult red blood cells in chimeric mice but did not clarify a role in embryonic (or yolk sac derived) erythroid cells. To examine the consequences of GATA-1 loss on embryonic erythropoiesis in vivo, we inactivated the GATA-1 locus in embryonic stem cells by gene targeting and transmitted the mutated allele through the mouse germ line. Male GATA-1- embryos die between embryonic day 10.5 and 11.5 (E10.5-E11.5) of gestation. At E9.5, GATA-1- embryos exhibit extreme pallor yet contain embryonic erythroid cells arrested at an early proerythroblast-like stage of their development. Embryos stain weakly with benzidine reagent, and yolk sac cells express globin RNAs, indicating globin gene activation in the absence of GATA-1. Female heterozygotes (GATA-1+/-) are born pale due to random inactivation of the X chromosome bearing the normal allele. However, these mice recover during the neonatal period, presumably as a result of in vivo selection for progenitors able to express GATA-1. Our findings conclusively establish the essential role for GATA-1 in erythropoiesis within the context of the intact developing mouse and further demonstrate that the block to cellular maturation is similar in GATA-1- embryonic and definitive erythroid precursors. Moreover, the recovery of GATA-1+/- mice from anemia seen at birth provides evidence indicating a role for GATA-1 at the hematopoietic progenitor cell level.

Diazepam binding inhibitor is a potent cholecystokinin-releasing peptide in the intestine.

Pancreatic proteases in the duodenum inhibit the release of cholecystokinin (CCK) and thus exert feedback control of pancreatic exocrine secretion. Exclusion of proteases from the duodenum either by the diversion of bile-pancreatic juice or by the addition of protease inhibitors stimulates exocrine pancreatic secretion. The mechanism by which pancreatic proteases in the duodenum regulate CCK secretion is unknown. In this study, we isolated a trypsin-sensitive peptide that is secreted intraduodenally, releases CCK, and stimulates pancreatic enzyme secretion in rats. This peptide was found to be identical to the porcine diazepam binding inhibitor by peptide sequencing and mass spectrometry analysis. Intraduodenal infusion of 200 ng of synthetic porcine diazepam binding inhibitor1-86 in rats significantly stimulated pancreatic amylase output. Infusion of the CCK antagonist MK-329 completely blocked the diazepam binding inhibitor-stimulated amylase secretion. Similarly, diazepam binding inhibitor33-52 [corrected] also stimulated CCK release and pancreatic secretion in a dose-dependent manner although it was 100 times less potent than the whole peptide. Using a perfusion system containing isolated mucosal cells from the proximal intestine of rats, porcine diazepam binding inhibitor 10(-12) M) dose dependently stimulated CCK secretion. In separate studies, it was demonstrated that luminal secretion of the diazepam binding inhibitor immunoreactivity (7.5 X 10(11) M) could be detected in rat's intestinal washing following the diversion of bile-pancreatic juice. The secretion of this peptide was inhibited by atropine. In conclusion, we have isolated and characterized a CCK-releasing peptide that has a sequence identical to the porcine diazepam binding inhibitor from pig intestinal mucosa and that stimulates CCK release when administered intraduodenally in rat. This peptide may mediate feedback regulation of pancreatic enzyme secretion.

Excitotoxicity in the lung: N-methyl-D-aspartate-induced, nitric oxide-dependent, pulmonary edema is attenuated by vasoactive intestinal peptide and by inhibitors of poly(ADP-ribose) polymerase.

Excitatory amino acid toxicity, resulting from overactivation of N-methyl-D-aspartate (NMDA) glutamate receptors, is a major mechanism of neuronal cell death in acute and chronic neurological diseases. We have investigated whether excitotoxicity may occur in peripheral organs, causing tissue injury, and report that NMDA receptor activation in perfused, ventilated rat lungs triggered acute injury, marked by increased pressures needed to ventilate and perfuse the lung, and by high-permeability edema. The injury was prevented by competitive NMDA receptor antagonists or by channel-blocker MK-801, and was reduced in the presence of Mg2+. As with NMDA toxicity to central neurons, the lung injury was nitric oxide (NO) dependent: it required L-arginine, was associated with increased production of NO, and was attenuated by either of two NO synthase inhibitors. The neuropeptide vasoactive intestinal peptide and inhibitors of poly(ADP-ribose) polymerase also prevented this injury, but without inhibiting NO synthesis, both acting by inhibiting a toxic action of NO that is critical to tissue injury. The findings indicate that: (i) NMDA receptors exist in the lung (and probably elsewhere outside the central nervous system), (ii) excessive activation of these receptors may provoke acute edematous lung injury as seen in the "adult respiratory distress syndrome," and (iii) this injury can be modulated by blockade of one of three critical steps: NMDA receptor binding, inhibition of NO synthesis, or activation of poly(ADP-ribose) polymerase.