Methods of Tree Appraisal: A Review of Their Features and Application Possibilities

Se revisan los métodas de valoración de arbolado

Artificial pancreas using a personalized rule-based controller achieves overnight normoglycemia in patients with type 1 diabetes

Objective: This study assessed the efficacy of a closed-loop (CL) system consisting of a predictive rule-based algorithm (pRBA) on achieving nocturnal and postprandial normoglycemia in patients with type 1 diabetes mellitus (T1DM). The algorithm is personalized for each patient’s data using two different strategies to control nocturnal and postprandial periods. Research Design and Methods: We performed a randomized crossover clinical study in which 10 T1DM patients treated with continuous subcutaneous insulin infusion (CSII) spent two nonconsecutive nights in the research facility: one with their usual CSII pattern (open-loop [OL]) and one controlled by the pRBA (CL). The CL period lasted from 10 p.m. to 10 a.m., including overnight control, and control of breakfast. Venous samples for blood glucose (BG) measurement were collected every 20 min. Results: Time spent in normoglycemia (BG, 3.9–8.0 mmol/L) during the nocturnal period (12 a.m.–8 a.m.), expressed as median (interquartile range), increased from 66.6% (8.3–75%) with OL to 95.8% (73–100%) using the CL algorithm (P<0.05). Median time in hypoglycemia (BG, <3.9 mmol/L) was reduced from 4.2% (0–21%) in the OL night to 0.0% (0.0–0.0%) in the CL night (P<0.05). Nine hypoglycemic events (<3.9 mmol/L) were recorded with OL compared with one using CL. The postprandial glycemic excursion was not lower when the CL system was used in comparison with conventional preprandial bolus: time in target (3.9–10.0 mmol/L) 58.3% (29.1–87.5%) versus 50.0% (50–100%). Conclusions: A highly precise personalized pRBA obtains nocturnal normoglycemia, without significant hypoglycemia, in T1DM patients. There appears to be no clear benefit of CL over prandial bolus on the postprandial glycemia

Dominance intensity methods for ranking of alternatives in mcdm with imprecise information

Los decisores cada vez se enfrentan a problemas más complejos en los que tomar una decisión implica tener que considerar simultáneamente muchos criterios que normalmente son conflictivos entre sí. En la mayoría de los problemas de decisión es necesario considerar criterios económicos, sociales y medioambientales. La Teoría de la Decisión proporciona el marco adecuado para poder ayudar a los decisores a resolver estos problemas de decisión complejos, al permitir considerar conjuntamente la incertidumbre existente sobre las consecuencias de cada alternativa en los diferentes atributos y la imprecisión sobre las preferencias de los decisores. En esta tesis doctoral nos centramos en la imprecisión de las preferencias de los decisores cuando éstas pueden ser representadas mediante una función de utilidad multiatributo aditiva. Por lo tanto, consideramos imprecisión tanto en los pesos como en las funciones de utilidad componentes de cada atributo. Se ha considerado el caso en que la imprecisión puede ser representada por intervalos de valores o bien mediante información ordinal, en lugar de proporcionar valores concretos. En este sentido, hemos propuesto métodos que permiten ordenar las diferentes alternativas basados en los conceptos de intensidad de dominación o intensidad de preferencia, los cuales intentan medir la fuerza con la que cada alternativa es preferida al resto. Para todos los métodos propuestos se ha analizado su comportamiento y se ha comparado con los más relevantes existentes en la literatura científica que pueden ser aplicados para resolver este tipo de problemas. Para ello, se ha realizado un estudio de simulación en el que se han usado dos medidas de eficiencia (hit ratio y coeficiente de correlación de Kendall) para comparar los diferentes métodos. ABSTRACT Decision makers increasingly face complex decision-making problems where they have to simultaneously consider many often conflicting criteria. In most decision-making problems it is necessary to consider economic, social and environmental criteria. Decision making theory provides an adequate framework for helping decision makers to make complex decisions where they can jointly consider the uncertainty about the performance of each alternative for each attribute, and the imprecision of the decision maker's preferences. In this PhD thesis we focus on the imprecision of the decision maker's preferences represented by an additive multiattribute utility function. Therefore, we consider the imprecision of weights, as well as of component utility functions for each attribute. We consider the case in which the imprecision is represented by ranges of values or by ordinal information rather than precise values. In this respect, we propose methods for ranking alternatives based on notions of dominance intensity, also known as preference intensity, which attempt to measure how much more preferred each alternative is to the others. The performance of the propose methods has been analyzed and compared against the leading existing methods that are applicable to this type of problem. For this purpose, we conducted a simulation study using two efficiency measures (hit ratio and Kendall correlation coefficient) to compare the different methods.

Patents and Economic Activity: The Need to Develop Prediction Methods for Patent Applications

Since the decade of the 1980’s the literature on economic development began paying attention to the cases of countries which were industrialized after the first industrial revolution. One of the most relevant aspects analyzed has been the role of technology as a factor which promotes or delays the process of catching up with technology leaders. As result of this interest, new and more adequate indicators were identified to provide a coherent explanation for technological activities and their relationship with economic efficiency. Although the earliest studies focused on analyzing the activities of research and development (R&D), recently the focus of analysis has shifted to another type of variables, more oriented towards the processes of innovation and the gathering of knowledge and capabilities, in which patents provide relevant information.

Health monitoring of web plastifying dampers subjected to cyclic loading through vibration tests

This article investigates experimentally the application of health monitoring techniques to assess the damage on a particular kind of hysteretic (metallic) damper called web plastifying dampers, which are subjected to cyclic loading. In general terms, hysteretic dampers are increasingly used as passive control systems in advanced earthquake-resistant structures. Nonparametric statistical processing of the signals obtained from simple vibration tests of the web plastifying damper is used here to propose an area index damage. This area index damage is compared with an alternative energy-based index of damage proposed in past research that is based on the decomposition of the load?displacement curve experienced by the damper. Index of damage has been proven to accurately predict the level of damage and the proximity to failure of web plastifying damper, but obtaining the load?displacement curve for its direct calculation requires the use of costly instrumentation. For this reason, the aim of this study is to estimate index of damage indirectly from simple vibration tests, calling for much simpler and cheaper instrumentation, through an auxiliary index called area index damage. Web plastifying damper is a particular type of hysteretic damper that uses the out-of-plane plastic deformation of the web of I-section steel segments as a source of energy dissipation. Four I-section steel segments with similar geometry were subjected to the same pattern of cyclic loading, and the damage was evaluated with the index of damage and area index damage indexes at several stages of the loading process. A good correlation was found between area index damage and index of damage. Based on this correlation, simple formulae are proposed to estimate index of damage from the area index damage.

Time-multiscale methods for the simulation of slow transport problems in atomistic systems

En esta tesis presentamos una teoría adaptada a la simulación de fenómenos lentos de transporte en sistemas atomísticos. En primer lugar, desarrollamos el marco teórico para modelizar colectividades estadísticas de equilibrio. A continuación, lo adaptamos para construir modelos de colectividades estadísticas fuera de equilibrio. Esta teoría reposa sobre los principios de la mecánica estadística, en particular el principio de máxima entropía de Jaynes, utilizado tanto para sistemas en equilibrio como fuera de equilibrio, y la teoría de las aproximaciones del campo medio. Expresamos matemáticamente el problema como un principio variacional en el que maximizamos una entropía libre, en lugar de una energía libre. La formulación propuesta permite definir equivalentes atomísticos de variables macroscópicas como la temperatura y la fracción molar. De esta forma podemos considerar campos macroscópicos no uniformes. Completamos el marco teórico con reglas de cuadratura de Monte Carlo, gracias a las cuales obtenemos modelos computables. A continuación, desarrollamos el conjunto completo de ecuaciones que gobiernan procesos de transporte. Deducimos la desigualdad de disipación entrópica a partir de fuerzas y flujos termodinámicos discretos. Esta desigualdad nos permite identificar la estructura que deben cumplir los potenciales cinéticos discretos. Dichos potenciales acoplan las tasas de variación en el tiempo de las variables microscópicas con las fuerzas correspondientes. Estos potenciales cinéticos deben ser completados con una relación fenomenológica, del tipo definido por la teoría de Onsanger. Por último, aportamos validaciones numéricas. Con ellas ilustramos la capacidad de la teoría presentada para simular propiedades de equilibrio y segregación superficial en aleaciones metálicas. Primero, simulamos propiedades termodinámicas de equilibrio en el sistema atomístico. A continuación evaluamos la habilidad del modelo para reproducir procesos de transporte en sistemas complejos que duran tiempos largos con respecto a los tiempos característicos a escala atómica. ABSTRACT In this work, we formulate a theory to address simulations of slow time transport effects in atomic systems. We first develop this theoretical framework in the context of equilibrium of atomic ensembles, based on statistical mechanics. We then adapt it to model ensembles away from equilibrium. The theory stands on Jaynes' maximum entropy principle, valid for the treatment of both, systems in equilibrium and away from equilibrium and on meanfield approximation theory. It is expressed in the entropy formulation as a variational principle. We interpret atomistic equivalents of macroscopic variables such as the temperature and the molar fractions, wich are not required to be uniform, but can vary from particle to particle. We complement this theory with Monte Carlo summation rules for further approximation. In addition, we provide a framework for studying transport processes with the full set of equations driving the evolution of the system. We first derive a dissipation inequality for the entropic production involving discrete thermodynamic forces and fluxes. This discrete dissipation inequality identifies the adequate structure for discrete kinetic potentials which couple the microscopic field rates to the corresponding driving forces. Those kinetic potentials must finally be expressed as a phenomenological rule of the Onsanger Type. We present several validation cases, illustrating equilibrium properties and surface segregation of metallic alloys. We first assess the ability of a simple meanfield model to reproduce thermodynamic equilibrium properties in systems with atomic resolution. Then, we evaluate the ability of the model to reproduce a long-term transport process in complex systems.

Estudio dinámico de cimentaciones en obras portuarias

Los diques de abrigo verticales son estructuras monolíticas que sirven para reflejar el oleaje creando, de este modo, una zona abrigada en el lado tierra. Son estructuras de contención de gravedad, es decir, su peso es el elemento resistente fundamental. Las solicitaciones sobre estas estructuras son de tipo dinámico (oleaje) y consisten, fundamentalmente, en un empuje frontal y una subpresión en la base que varían en el tiempo. Habitualmente, las acciones sobre los diques de abrigo se establecen mediante fórmulas empíricas, que se describen en la presente tesis si bien, para obras de especial importancia, suelen medirse en modelos reducidos de laboratorio. Cuando el cimiento en que han de apoyarse estas estructuras no presenta una resistencia al corte suficiente, el apoyo se realiza en banquetas que redistribuyen la carga y que están formadas por materiales granulares. En la práctica habitual para conocer la estabilidad de estas estructuras frente a un temporal, tras establecer las acciones de cálculo (empuje frontal y subpresión) se efectúa un cálculo pseudoestático en el que se consideran condiciones drenadas o no drenadas del cimiento en función de su permeabilidad. Se conoce que en los suelos saturados, bajo cargas cíclicas, tiende a producirse una elevación de las presiones intersticiales y una reducción de tensiones efectivas así como una degradación del módulo de deformación tangencial en función del número de ciclos de carga, pudiéndose producir el fenómeno conocido como licuefacción (arenas) o movilidad cíclica (suelos más finos). El objeto de la tesis es explorar la posibilidad, con la tecnología actual, de analizar la estabilidad de los diques de abrigo verticales en cuyo cimiento existen suelos blandos, proponiendo un procedimiento para evaluar la estabilidad dinámica en este tipo de obras. Para ello se han revisado los procedimientos actualmente utilizados para definir las acciones de cálculo, los principales modelos de comportamiento dinámicos de suelos saturados disponibles y los procedimientos de cálculo. Una vez investigado el estado del arte sobre este tema, se propone un procedimiento de cálculo en el que, utilizando el programa comercial FLAC, se establecen las acciones cíclicas sobre un dique de abrigo vertical tipo con distintas condiciones de apoyo, aplicando, para el cimiento, un modelo de comportamiento tipo hiperbólico con generación de presiones intersticiales cuyos parámetros pueden obtenerse de ensayos de campo y laboratorio. Por último, una vez descrito el procedimiento, se aplica a un caso real en el que se produjo un fallo en la cimentación que desembocó en el hundimiento de parte de un dique vertical situado en el puerto de Barcelona, presentándose los resultados obtenidos del análisis efectuado y comparándolos con los obtenidos utilizando los métodos de cálculo habituales. Vertical breakwaters are monolithic structures built to reflect sea waves, thereby providing a sheltered area on the land side. They are gravity retaining structures, that is, their own weight is their basic resisting mechanism. Loads acting on these structures are dynamic (waves) and consist essentially in a frontal thrust and an uplift pressure on the base, which both vary over time. Usually, actions in breakwater design are established by empirical formulas, which are described in this thesis. For works of particular importance, such forces are measured in small-scale laboratory tests. When there are no soils with enough shear strength under the planned vertical breakwaters, they usually rest on granular berms which redistribute the load. Nowadays, after establishing the acting forces on the breakwater (front push and uplift pressure), a pseudostatic calculation (with drained or undrained conditions depending on the foundation permeability) is normally done to analyze the stability of these structures against storm waves. It is known that pore pressures tend to rise in saturated soils under cyclic loading and, consequently, there is a reduction of effective stress. A degradation of the shear modulus also occurs depending on the number of load cycles. All of these effects can bring about the phenomenon known as liquefaction in sands or cyclic mobility in fine-grained soils. The aim of the thesis is to explore the possibility that current technology provides to analyze the stability of vertical breakwaters founded on soft soils, and to suggest a method to evaluate the dynamic stability in this type of works. To this end, a review has been made of procedures currently used to define the actions in calculations, the main models of dynamic behaviour of saturated soils available and of calculation procedures. Once the state of the art on this subject has been reviewed, a method of calculation is proposed that uses the commercial program FLAC and is applied to a typical vertical breakwater on a range of different foundation conditions. For the foundation soil, a hyperbolic constitutive model with pore pressure generation has been employed, whose parameters can be obtained from field and laboratory tests. Finally, the described procedure is applied to an actual case where a foundation failure occurred that led to the sinking of several caissons in a vertical breakwater located in the port of Barcelona. The results obtained with the proposed method are compared with those obtained using conventional methods.

Fatiga en CFRP: Caracterización y Método Óptico para la Estimación del Daño

El desarrollo de nuevas estructuras aeroespaciales optimizadas, utilizan materiales compuestos, para los componentes críticos y subsistemas, principalmente polímeros reforzados con fibra de carbono (CFRP). Un conocimiento profundo del estado de daño por fatiga de estructuras de CFRP avanzado, es esencial para predecir la vida residual y optimizar los intervalos de inspección estructural, reparaciones y/o sustitución de componentes. Las técnicas actuales se basan principalmente en la medición de cargas estructurales a lo largo de la vida útil de la estructura mediante galgas extensométricas eléctricas. Con esos datos, se estima la vida a fatiga utilizando modelos de acumulación de daño. En la presente tesis, se evalúa la metodología convencional para la estimación de la vida a fatiga de un CFRP aeronáutico. Esta metodología está basada en la regla de acumulación de daño lineal de Palmgren-Miner, y es aplicada para determinar la vida a fatiga de estructuras sometidas a cargas de amplitud variable. Se ha realizado una campaña de ensayos con cargas de amplitud constante para caracterizar un CFRP aeronáutico a fatiga, obteniendo las curvas clásicas S-N, en diferentes relaciones de esfuerzo. Se determinaron los diagramas de vida constante, (CLD), también conocidos como diagramas de Goodman, utilizando redes neuronales artificiales debido a la ausencia de modelos coherentes para materiales compuestos. Se ha caracterizado la degradación de la rigidez debido al daño por fatiga. Se ha ensayado un segundo grupo de probetas con secuencias estandarizadas de cargas de amplitud variable, para obtener la vida a fatiga y la degradación de rigidez en condiciones realistas. Las cargas aplicadas son representativas de misiones de aviones de combate (Falstaff), y de aviones de transporte (Twist). La vida a fatiga de las probetas cicladas con cargas de amplitud variable, se comparó con el índice de daño teórico calculado en base a la regla de acumulación de daño lineal convencional. Los resultados obtenidos muestran predicciones no conservativas. Esta tesis también presenta el estudio y desarrollo, de una nueva técnica de no contacto para evaluar el estado de daño por fatiga de estructuras de CFRP por medio de cambios de los parámetros de rugosidad. La rugosidad superficial se puede medir fácilmente en campo con métodos sin contacto, mediante técnicas ópticas tales como speckle y perfilómetros ópticos. En el presente estudio, se han medido parámetros de rugosidad superficial, y el factor de irregularidad de la superficie, a lo largo de la vida de las probetas cicladas con cargas de amplitud constante y variable, Se ha obtenido una buena tendencia de ajuste al correlacionar la magnitud de la rugosidad y el factor de irregularidad de la superficie con la degradación de la rigidez de las probetas fatigadas. Estos resultados sugieren que los cambios en la rugosidad superficial medida en zonas estratégicas de componentes y estructuras hechas de CFRP, podrían ser indicativas del nivel de daño interno debido a cargas de fatiga. Los resultados también sugieren que el método es independiente del tipo de carga de fatiga que ha causado el daño. Esto último hace que esta técnica de medición sea aplicable como inspección para una amplia gama de estructuras de materiales compuestos, desde tanques presurizados con cargas de amplitud constante, estructuras aeronáuticas como alas y colas de aeronaves cicladas con cargas de amplitud variable, hasta aplicaciones industriales como automoción, entre otros. ABSTRACT New optimized aerospace structures use composite materials, mainly carbon fiber reinforced polymer composite (CFRP), for critical components and subsystems. A strong knowledge of the fatigue state of highly advanced (CFRP) structures is essential to predict the residual life and optimize intervals of structural inspection, repairs, and/or replacements. Current techniques are based mostly on measurement of structural loads throughout the service life by electric strain gauge sensors. These sensors are affected by extreme environmental conditions and by fatigue loads in such a way that the sensors and their systems require exhaustive maintenance throughout system life. In the present thesis, the conventional methodology based on linear damage accumulation rules, applied to determine the fatigue life of structures subjected to variable amplitude loads was evaluated for an aeronautical CFRP. A test program with constant amplitude loads has been performed to obtain the classical S-N curves at different stress ratios. Constant life diagrams, CLDs, where determined by means of Artificial Neural Networks due to the absence of consistent models for composites. The stiffness degradation due to fatigue damage has been characterized for coupons under cyclic tensile loads. A second group of coupons have been tested until failure with a standardized sequence of variable amplitude loads, representative of missions for combat aircraft (Falstaff), and representative of commercial flights (Twist), to obtain the fatigue life and the stiffness degradation under realistic conditions. The fatigue life of the coupons cycled with variable amplitude loads were compared to the theoretical damage index calculated based on the conventional linear damage accumulation rule. The obtained results show non-conservative predictions. This thesis also presents the evaluation of a new non-contact technique to evaluate the fatigue damage state of CFRP structures by means of measuring roughness parameters to evaluate changes in the surface topography. Surface roughness can be measured easily on field with non-contact methods by optical techniques such as speckle and optical perfilometers. In the present study, surface roughness parameters, and the surface irregularity factor, have been measured along the life of the coupons cycled with constant and variable amplitude loads of different magnitude. A good agreement has been obtained when correlating the magnitude of the roughness and the surface irregularity factor with the stiffness degradation. These results suggest that the changes on the surface roughness measured in strategic zones of components and structures made of CFRP, could be indicative of the level of internal damage due to fatigue loads. The results also suggest that the method is independent of the type of fatigue load that have caused the damage. It makes this measurement technique applicable for a wide range of inspections of composite materials structures, from pressurized tanks with constant amplitude loads, to variable amplitude loaded aeronautical structures like wings and empennages, up to automotive and other industrial applications.

Health monitoring of a new hysteretic damper subjected to earthquakes on a shaking table

This paper presents the experimental results obtained by applying frequency-domain structural health monitoring techniques to assess the damage suffered on a special type of damper called Web Plastifying Damper (WPD). The WPD is a hysteretic type energy dissipator recently developed for the passive control of structures subjected to earthquakes. It consists of several I-section steel segments connected in parallel. The energy is dissipated through plastic deformations of the web of the I-sections, which constitute the dissipative parts of the damper. WPDs were subjected to successive histories of dynamically-imposed cyclic deformations of increasing magnitude with the shaking table of the University of Granada. To assess the damage to the web of the I-section steel segments after each history of loading, a new damage index called Area Index of Damage (AID) was obtained from simple vibration tests. The vibration signals were acquired by means of piezoelectric sensors attached on the I-sections, and non-parametric statistical methods were applied to calculate AID in terms of changes in frequency response functions. The damage index AID was correlated with another energy-based damage index-ID- which past research has proven to accurately characterize the level of mechanical damage. The ID is rooted in the decomposition of the load-displacement curve experienced by the damper into the so-called skeleton and Bauschinger parts. ID predicts the level of damage and the proximity to failure of the damper accurately, but it requires costly instrumentation. The experiments reported in this paper demonstrate a good correlation between AID and ID in a realistic seismic loading scenario consisting of dynamically applied arbitrary cyclic loads. Based on this correlation, it is possible to estimate ID indirectly from the AID, which calls for much simpler and less expensive instrumentation.

An isoform of the rod photoreceptor cyclic nucleotide-gated channel β subunit expressed in olfactory neurons

Sensory transduction in olfactory neurons involves the activation of a cyclic nucleotide-gated (CNG) channel by cAMP. Previous studies identified a CNG channel α subunit (CNG2) and a β subunit (CNG5), which when heterologously expressed form a channel with properties similar but not identical to those of native olfactory neurons. We have cloned a new type of CNG channel β subunit (CNG4.3) from rat olfactory epithelium. CNG4.3 derives from the same gene as the rod photoreceptor β subunit (CNG4.1) but lacks the long, glutamic acid-rich domain found in the N terminus of CNG4.1. Northern blot and in situ hybridization revealed that CNG4.3 is expressed specifically in olfactory neurons. Expression of CNG4.3 in human embryonic kidney 293 cells did not lead to detectable currents. Coexpression of CNG4.3 with CNG2 induced a current with significantly increased sensitivity for cAMP whereas cGMP affinity was not altered. Additionally, CNG4.3 weakened the outward rectification of the current in the presence of extracellular Ca2+, decreased the relative permeability for Ca2+, and enhanced the sensitivity for l-cis diltiazem. Upon coexpression of CNG2, CNG4.3, and CNG5, a conductance with a cAMP sensitivity greater than that of either the CNG2/CNG4.3 or the CNG2/CNG5 channel and near that of native olfactory channel was observed. Our data suggest that CNG4.3 forms a subunit of the native olfactory CNG channel. The expression of various CNG4 isoforms in retina and olfactory epithelium indicates that the CNG4 subunit may be necessary for normal function of both photoreceptor and olfactory CNG channels.

Production of cyclic peptides and proteins in vivo

Combinatorial libraries of synthetic and natural products are an important source of molecular information for the interrogation of biological targets. Methods for the intracellular production of libraries of small, stable molecules would be a valuable addition to existing library technologies by combining the discovery potential inherent in small molecules with the large library sizes that can be realized by intracellular methods. We have explored the use of split inteins (internal proteins) for the intracellular catalysis of peptide backbone cyclization as a method for generating proteins and small peptides that are stabilized against cellular catabolism. The DnaE split intein from Synechocystis sp. PCC6803 was used to cyclize the Escherichia coli enzyme dihydrofolate reductase and to produce the cyclic, eight-amino acid tyrosinase inhibitor pseudostellarin F in bacteria. Cyclic dihydrofolate reductase displayed improved in vitro thermostability, and pseudostellarin F production was readily apparent in vivo through its inhibition of melanin production catalyzed by recombinant Streptomyces antibioticus tyrosinase. The ability to generate and screen for backbone cyclic products in vivo is an important milestone toward the goal of generating intracellular cyclic peptide and protein libraries.

Ablation of P/Q-type Ca2+ channel currents, altered synaptic transmission, and progressive ataxia in mice lacking the α1A-subunit

The Ca2+ channel α1A-subunit is a voltage-gated, pore-forming membrane protein positioned at the intersection of two important lines of research: one exploring the diversity of Ca2+ channels and their physiological roles, and the other pursuing mechanisms of ataxia, dystonia, epilepsy, and migraine. α1A-Subunits are thought to support both P- and Q-type Ca2+ channel currents, but the most direct test, a null mutant, has not been described, nor is it known which changes in neurotransmission might arise from elimination of the predominant Ca2+ delivery system at excitatory nerve terminals. We generated α1A-deficient mice (α1A−/−) and found that they developed a rapidly progressive neurological deficit with specific characteristics of ataxia and dystonia before dying ≈3–4 weeks after birth. P-type currents in Purkinje neurons and P- and Q-type currents in cerebellar granule cells were eliminated completely whereas other Ca2+ channel types, including those involved in triggering transmitter release, also underwent concomitant changes in density. Synaptic transmission in α1A−/− hippocampal slices persisted despite the lack of P/Q-type channels but showed enhanced reliance on N-type and R-type Ca2+ entry. The α1A−/− mice provide a starting point for unraveling neuropathological mechanisms of human diseases generated by mutations in α1A.

Interactive cloning with the SH3 domain of N-src identifies a new brain specific ion channel protein, with homology to Eag and cyclic nucleotide-gated channels

We have isolated a novel cDNA, that appears to represent a new class of ion channels, by using the yeast two-hybrid system and the SH3 domain of the neural form of Src (N-src) as a bait. The encoded polypeptide, BCNG-1, is distantly related to cyclic nucleotide-gated channels and the voltage-gated channels, Eag and H-erg. BCNG-1 is expressed exclusively in the brain, as a glycosylated protein of ≈132 kDa. Immunohistochemical analysis indicates that BCNG-1 is preferentially expressed in specific subsets of neurons in the neocortex, hippocampus, and cerebellum, in particular pyramidal neurons and basket cells. Within individual neurons, the BCNG-1 protein is localized to either the dendrites or the axon terminals depending on the cell type. Southern blot analysis shows that several other BCNG-related sequences are present in the mouse genome, indicating the emergence of an entire subfamily of ion channel coding genes. These findings suggest the existence of a new type of ion channel, which is potentially able to modulate membrane excitability in the brain and could respond to regulation by cyclic nucleotides.

A Novel Ras-interacting Protein Required for Chemotaxis and Cyclic Adenosine Monophosphate Signal Relay in Dictyostelium

We have identified a novel Ras-interacting protein from Dictyostelium, RIP3, whose function is required for both chemotaxis and the synthesis and relay of the cyclic AMP (cAMP) chemoattractant signal. rip3 null cells are unable to aggregate and lack receptor activation of adenylyl cyclase but are able, in response to cAMP, to induce aggregation-stage, postaggregative, and cell-type-specific gene expression in suspension culture. In addition, rip3 null cells are unable to properly polarize in a cAMP gradient and chemotaxis is highly impaired. We demonstrate that cAMP stimulation of guanylyl cyclase, which is required for chemotaxis, is reduced ∼60% in rip3 null cells. This reduced activation of guanylyl cyclase may account, in part, for the defect in chemotaxis. When cells are pulsed with cAMP for 5 h to mimic the endogenous cAMP oscillations that occur in wild-type strains, the cells will form aggregates, most of which, however, arrest at the mound stage. Unlike the response seen in wild-type strains, the rip3 null cell aggregates that form under these experimental conditions are very small, which is probably due to the rip3 null cell chemotaxis defect. Many of the phenotypes of the rip3 null cell, including the inability to activate adenylyl cyclase in response to cAMP and defects in chemotaxis, are very similar to those of strains carrying a disruption of the gene encoding the putative Ras exchange factor AleA. We demonstrate that aleA null cells also exhibit a defect in cAMP-mediated activation of guanylyl cyclase similar to that of rip3 null cells. A double-knockout mutant (rip3/aleA null cells) exhibits a further reduction in receptor activation of guanylyl cyclase, and these cells display almost no cell polarization or movement in cAMP gradients. As RIP3 preferentially interacts with an activated form of the Dictyostelium Ras protein RasG, which itself is important for cell movement, we propose that RIP3 and AleA are components of a Ras-regulated pathway involved in integrating chemotaxis and signal relay pathways that are essential for aggregation.

Novel Nuclear Signaling Pathway Mediates Activation of Fibroblast Growth Factor-2 Gene by Type 1 and Type 2 Angiotensin II Receptors

In bovine adrenal medullary cells synergistically acting type 1 and type 2 angiotensin II (AII) receptors activate the fibroblast growth factor-2 (FGF-2) gene through a unique AII-responsive promoter element. Both the type 1 and type 2 AII receptors and the downstream cyclic adenosine 1′,3′-monophosphate- and protein kinase C-dependent signaling pathways activate the FGF-2 promoter through a novel signal-transducing mechanism. This mechanism, which we have named integrative nuclear FGF receptor-1 signaling, involves the nuclear translocation of FGF receptor-1 and its subsequent transactivation of the AII-responsive element in the FGF-2 promoter.