Fórmulas para el cálculo del factor de impacto de estructuras semienterradas en líneas de ferrocarril de alta velocidad = Dynamic factors for underpasses in high speed train lines

Los pasos inferiores son muy numerosos en las líneas de ferrocarril. Su comportamiento dinámico ha recibido mucha menos atención que el de otras estructuras como los puentes, pero su elevado número hace que su estudio sea económicamente relevante con vista a optimizar su forma, manteniendo la seguridad. El proyecto de puentes según el Eurocódigo incluye comprobaciones de estados límite de tensiones bajo carga dinámica. En el caso de pasos inferiores, las comprobaciones pueden resultar tan costosas como aquellas de puentes, pese a que su coste es mucho menor. Por tanto, se impone la búsqueda de unas reglas de cálculo simplificado que pongan en consonancia el coste de la estructura con el esfuerzo necesario para su proyecto. Este artículo propone un conjunto de reglas basadas en un estudio paramétrico = Underpasses are common in modern railway lines. Wildlife corridors and drainage conduits often fall into this category of partially buried structures. Their dynamic behavior has received far less attention than that of other structures such as bridges, but their large number makes their study an interesting challenge from the viewpoint of safety and cost savings. The bridge design rules in accordance with the Eurocode involve checks on stresses according to dynamic loading. In the case of underpasses, those checks may be as much as those for bridges. Therefore, simplified design rules may align the design effort with their cost. Such a set of rules may provide estimations of response parameters based on the key parameters influencing the result. This paper contains a proposal based on a parametric study.

A model for setting services on auxiliary bus lines under congestion

In this paper, a mathematical programming model and a heuristically derived solution is described to assist with the efficient planning of services for a set of auxiliary bus lines (a bus-bridging system) during disruptions of metro and rapid transit lines. The model can be considered static and takes into account the average flows of passengers over a given period of time (i.e., the peak morning traffic hour) Auxiliary bus services must accommodate very high demand levels, and the model presented is able to take into account the operation of a bus-bridging system under congested conditions. A general analysis of the congestion in public transportation lines is presented, and the results are applied to the design of a bus-bridging system. A nonlinear integer mathematical programming model and a suitable approximation of this model are then formulated. This approximated model can be solved by a heuristic procedure that has been shown to be computationally viable. The output of the model is as follows: (a) the number of bus units to assign to each of the candidate lines of the bus-bridging system; (b) the routes to be followed by users passengers of each of the origin–destination pairs; (c) the operational conditions of the components of the bus-bridging system, including the passenger load of each of the line segments, the degree of saturation of the bus stops relative to their bus input flows, the bus service times at bus stops and the passenger waiting times at bus stops. The model is able to take into account bounds with regard to the maximum number of passengers waiting at bus stops and the space available at bus stops for the queueing of bus units. This paper demonstrates the applicability of the model with two realistic test cases: a railway corridor in Madrid and a metro line in Barcelona Planificación de los servicios de lineas auxiliares de autobuses durante las incidencias de las redes de metro y cercanías. El modelo estudia el problema bajo condiciones de alta demanda y condiciones de congestión. El modelo no lineal resultante es resuelto mediante heurísticas que demuestran su utilidad. Se demuestran los resultados en dos corredores de las ciudades de Barcelona y Madrid.

Detector de banda ultraancha en tecnología microstrip hasta 40 GHz

Detectors designing is a key aspect for the development of the new millimeter wave systems. In this paper two detectors in microstrip technology are presented. They use zero bias Schottky diodes to detect signals from low frequency to 40 GHz. High sensibility, flat frequency response and ultrabroadband are the main features of these designs. They are also cheap and easy to mount because they have been built using microstrip technology. This paper explains most technological questions which must be taken into account to design such detectors.

The TRANSPLANTA collection of Arabidopsis lines: a resource for functional analysis of transcription factors based on their conditional over-expression

Transcription factors (TFs) are key regulators of gene expression in all organisms. In eukaryotes, TFs are often represented by functionally redundant members of large gene families. Overexpression might prove a means to unveil the biological functions of redundant TFs; however, constitutive overexpression of TFs frequently causes severe developmental defects, preventing their functional characterization. Conditional overexpression strategies help to overcome this problem. Here, we report on the TRANSPLANTA collection of Arabidopsis lines, each expressing one of 949 TFs under the control of a β–estradiol-inducible promoter. Thus far, 1636 independent homozygous lines, representing an average of 2.6 lines for every TF, have been produced for the inducible expression of 634 TFs. Along with a GUS-GFP reporter, randomly selected TRANSPLANTA lines were tested and confirmed for conditional transgene expression upon β–estradiol treatment. As a proof of concept for the exploitation of this resource, β–estradiol-induced proliferation of root hairs, dark-induced senescence, anthocyanin accumulation and dwarfism were observed in lines conditionally expressing full-length cDNAs encoding RHD6, WRKY22, MYB123/TT2 and MYB26, respectively, in agreement with previously reported phenotypes conferred by these TFs. Further screening performed with other TRANSPLANTA lines allowed the identification of TFs involved in different plant biological processes, illustrating that the collection is a powerful resource for the functional characterization of TFs. For instance, ANAC058 and a TINY/AP2 TF were identified as modulators of ABA-mediated germination potential, and RAP2.10/DEAR4 was identified as a regulator of cell death in the hypocotyl–root transition zone. Seeds of TRANSPLANTA lines have been deposited at the Nottingham Arabidopsis Stock Centre for further distribution.

Diseño de una red de división / combinación de potencia de N-puertos sobre una octava en banda X mediante divisores Wilkinson multietapa y asimétricos implementados en tecnología Microstrip

Se muestra el proceso de diseño y medida de una red de división/combinación de potencia de gran ancho de banda (5 a 10 GHz) basado en divisores de potencia Wilkinson asimétricos y multi-sección. Se comparan las ventajas e inconvenientes de las distintas formas de conectar los divisores de potencia, proponiendo un diseño general que depende de la tecnología de fabricación que se disponga. Se detalla el proceso de diseño de los divisores de potencia del circuito, así como las estrategias a seguir para plasmar los parámetros del circuito en un modelo de simulación,mostrando los problemas que surgen y las soluciones que se tomaron. Finalmente se muestran las medidas el circuito fabricado, comparando su respuesta con la de la simulación.

Improving the pass-band return loss in liquid crystal dual-mode bandpass filters by microstrip patch reshaping

In this paper, the design and experimental characterization of a tunable microstrip bandpass filter based on liquid crystal technology are presented. A reshaped microstrip dual-mode filter structure has been used in order to improve the device performance. Specifically, the aim is to increase the pass-band return loss of the filter by narrowing the filter bandwidth. Simulations confirm the improvement of using this new structure, achieving a pass-band return loss increase of 1.5 dB at least. Because of the anisotropic properties of LC molecules, a filter central frequency shift from 4.688 GHz to 5.045 GHz, which means a relative tuning range of 7.3%, is measured when an external AC voltage from 0 Vrms to 15 Vrms is applied to the device.

Analysis and design of reconfigurable multimode multiport microstrip antennas for mimo systems and diversity schemes

Las comunicaciones inalámbricas han transformado profundamente la forma en la que la gente se comunica en el día a día y es, sin lugar a dudas, una de las tecnologías de nuestro tiempo que más rápidamente evoluciona. Este rápido crecimiento implica retos enormes en la tecnología subyacente, debido y entre otros motivos, a la gran demanda de capacidad de los nuevos servicios inalámbricos. Los sistemas Multiple Input Multiple Output (MIMO) han despertado mucho interés como medio de mejorar el rendimiento global del sistema, satisfaciendo de este modo y en cierta medida los nuevo requisitos exigidos. De hecho, el papel relevante de esta tecnología en los actuales esfuerzos de estandarización internacionales pone de manifiesto esta utilidad. Los sistemas MIMO sacan provecho de los grados de libertad espaciales, disponibles a través del entorno multitrayecto, para mejorar el rendimiento de la comunicación con una destacable eficiencia espectral. Con el fin de alcanzar esta mejora en el rendimiento, la diversidad espacial y por diagrama han sido empleadas tradicionalmente para reducir la correlación entre los elementos radiantes, ya que una correlación baja es condición necesaria, si bien no suficiente, para dicha mejora. Tomando como referencia, o punto de partida, las técnicas empleadas para obtener diversidad por diagrama, esta tesis doctoral surge de la búsqueda de la obtención de diversidad por diagrama y/o multiplexación espacial a través del comportamiento multimodal de la antena microstrip, proponiendo para ello un modelo cuasi analítico original para el análisis y diseño de antenas microstrip multipuerto, multimodo y reconfigurables. Este novedoso enfoque en este campo, en vez de recurrir a simulaciones de onda completa por medio de herramientas comerciales tal y como se emplea en las publicaciones existentes, reduce significativamente el esfuerzo global de análisis y diseño, en este último caso por medio de guías de diseño generales. Con el fin de lograr el objetivo planteado y después de una revisión de los principales conceptos de los sistemas MIMO que se emplearán más adelante, se fija la atención en encontrar, implementar y verificar la corrección y exactitud de un modelo analítico que sirva de base sobre la cual añadir las mejoras necesarias para obtener las características buscadas del modelo cuasi analítico propuesto. Posteriormente y partiendo del modelo analítico base seleccionado, se exploran en profundidad y en diferentes entornos multitrayecto, las posibilidades en cuanto a rendimiento se refiere de diversidad por diagrama y multiplexación espacial, proporcionadas por el comportamiento multimodal de las antenas parche microstrip sin cargar. Puesto que cada modo de la cavidad tiene su propia frecuencia de resonancia, es necesario encontrar formas de desplazar la frecuencia de resonancia de cada modo empleado para ubicarlas en la misma banda de frecuencia, manteniendo cada modo al mismo tiempo tan independiente como sea posible. Este objetivo puede lograrse cargando adecuadamente la cavidad con cargas reactivas, o alterando la geometría del parche radiante. Por consiguiente, la atención en este punto se fija en el diseño, implementación y verificación de un modelo cuasi analítico para el análisis de antenas parche microstrip multipuerto, multimodo y cargadas que permita llevar a cabo la tarea indicada, el cuál es una de las contribuciones principales de esta tesis doctoral. Finalmente y basándose en el conocimiento adquirido a través del modelo cuasi analítico, se proporcionan y aplican guías generales para el diseño de antenas microstrip multipuerto, multimodo y reconfigurables para sistemas MIMO, con el fin de mejorar su diversidad por diagrama y/o su capacidad por medio del comportamiento multimodal de las antenas parche microstrip. Se debe destacar que el trabajo presentado en esta tesis doctoral ha dado lugar a una publicación en una revista técnica internacional de un alto factor de impacto. De igual manera, el trabajo también ha sido presentado en algunas de las más importantes conferencias internacionales en el ámbito de las antenas ABSTRACT Wireless communications have deeply transformed the way people communicate on daily basis and it is undoubtedly one of the most rapidly evolving technologies of our time. This fast growing behaviour involves huge challenges on the bearing technology, due to and among others reasons, the high demanding capacity of new wireless services. MIMO systems have given rise to considerable interest as a means to enhance the overall system performance, thus satisfying somehow the new demanding requirements. Indeed, the significant role of this technology on current international standardization efforts, highlights this usefulness. MIMO systems make profit from the spatial degrees of freedom available through the multipath scenario to improve the communication performance with a remarkable spectral efficiency. In order to achieve this performance improvement, spatial and pattern diversity have been traditionally used to decrease the correlation between antenna elements, as low correlation is a necessary but not sufficient condition. Taking as a reference, or starting point, the techniques used to achieve pattern diversity, this Philosophiae Doctor (Ph.D.) arises from the pursuit of obtaining pattern diversity and/or spatial multiplexing capabilities through the multimode microstrip behaviour, thus proposing a novel quasi analytical model for the analysis and design of reconfigurable multimode multiport microstrip antennas. This innovative approach on this field, instead of resorting to full-wave simulations through commercial tools as done in the available publications, significantly reduces the overall analysis and design effort, in this last case through comprehensive design guidelines. In order to achieve this goal and after a review of the main concepts of MIMO systems which will be followed used, the spotlight is fixed on finding, implementing and verifying the correctness and accuracy of a base quasi analytical model over which add the necessary enhancements to obtain the sought features of the quasi analytical model proposed. Afterwards and starting from the base quasi analytical model selected, the pattern diversity and spatial multiplexing performance capabilities provided by the multimode behaviour of unloaded microstrip patch antennas under different multipath environments are fully explored. As each cavity mode has its own resonant frequency, it is required to find ways to displace the resonant frequency of each used mode to place them at the same frequency band while keeping each mode as independent as possible. This objective can be accomplished with an appropriate loading of the cavity with reactive loads, or through the alteration of the geometry of the radiation patch. Thus, the focus is set at this point on the design, implementation and verification of a quasi analytical model for the analysis of loaded multimode multiport microstrip patch antennas to carry out the aforementioned task, which is one of the main contributions of this Ph.D. Finally and based on the knowledge acquired through the quasi analytical model, comprehensive guidelines to design reconfigurable multimode MIMO microstrip antennas to improve the spatial multiplexing and/or diversity system performance by means of the multimode microstrip patch antenna behaviour are given and applied. It shall be highlighted that the work presented in this Ph.D. has given rise to a publication in an international technical journal of high impact factor. Moreover, the work has also been presented at some of the most important international conferences in antenna area.

Computational and Experimental Modelling of Mooring Lines Dynamics for Offshore Floating Wind Turbines

El cálculo de cargas de aerogeneradores flotantes requiere herramientas de simulación en el dominio del tiempo que consideren todos los fenómenos que afectan al sistema, como la aerodinámica, la dinámica estructural, la hidrodinámica, las estrategias de control y la dinámica de las líneas de fondeo. Todos estos efectos están acoplados entre sí y se influyen mutuamente. Las herramientas integradas se utilizan para calcular las cargas extremas y de fatiga que son empleadas para dimensionar estructuralmente los diferentes componentes del aerogenerador. Por esta razón, un cálculo preciso de las cargas influye de manera importante en la optimización de los componentes y en el coste final del aerogenerador flotante. En particular, el sistema de fondeo tiene gran impacto en la dinámica global del sistema. Muchos códigos integrados para la simulación de aerogeneradores flotantes utilizan modelos simplificados que no consideran los efectos dinámicos de las líneas de fondeo. Una simulación precisa de las líneas de fondeo dentro de los modelos integrados puede resultar fundamental para obtener resultados fiables de la dinámica del sistema y de los niveles de cargas en los diferentes componentes. Sin embargo, el impacto que incluir la dinámica de los fondeos tiene en la simulación integrada y en las cargas todavía no ha sido cuantificada rigurosamente. El objetivo principal de esta investigación es el desarrollo de un modelo dinámico para la simulación de líneas de fondeo con precisión, validarlo con medidas en un tanque de ensayos e integrarlo en un código de simulación para aerogeneradores flotantes. Finalmente, esta herramienta, experimentalmente validada, es utilizada para cuantificar el impacto que un modelos dinámicos de líneas de fondeo tienen en la computación de las cargas de fatiga y extremas de aerogeneradores flotantes en comparación con un modelo cuasi-estático. Esta es una información muy útil para los futuros diseñadores a la hora de decidir qué modelo de líneas de fondeo es el adecuado, dependiendo del tipo de plataforma y de los resultados esperados. El código dinámico de líneas de fondeo desarrollado en esta investigación se basa en el método de los Elementos Finitos, utilizando en concreto un modelo ”Lumped Mass” para aumentar su eficiencia de computación. Los experimentos realizados para la validación del código se realizaron en el tanque del École Céntrale de Nantes (ECN), en Francia, y consistieron en sumergir una cadena con uno de sus extremos anclados en el fondo del tanque y excitar el extremo suspendido con movimientos armónicos de diferentes periodos. El código demostró su capacidad para predecir la tensión y los movimientos en diferentes posiciones a lo largo de la longitud de la línea con gran precisión. Los resultados indicaron la importancia de capturar la dinámica de las líneas de fondeo para la predicción de la tensión especialmente en movimientos de alta frecuencia. Finalmente, el código se utilizó en una exhaustiva evaluación del efecto que la dinámica de las líneas de fondeo tiene sobre las cargas extremas y de fatiga de diferentes conceptos de aerogeneradores flotantes. Las cargas se calcularon para tres tipologías de aerogenerador flotante (semisumergible, ”spar-buoy” y ”tension leg platform”) y se compararon con las cargas obtenidas utilizando un modelo cuasi-estático de líneas de fondeo. Se lanzaron y postprocesaron más de 20.000 casos de carga definidos por la norma IEC 61400-3 siguiendo todos los requerimientos que una entidad certificadora requeriría a un diseñador industrial de aerogeneradores flotantes. Los resultados mostraron que el impacto de la dinámica de las líneas de fondeo, tanto en las cargas de fatiga como en las extremas, se incrementa conforme se consideran elementos situados más cerca de la plataforma: las cargas en la pala y en el eje sólo son ligeramente modificadas por la dinámica de las líneas, las cargas en la base de la torre pueden cambiar significativamente dependiendo del tipo de plataforma y, finalmente, la tensión en las líneas de fondeo depende fuertemente de la dinámica de las líneas, tanto en fatiga como en extremas, en todos los conceptos de plataforma que se han evaluado. ABSTRACT The load calculation of floating offshore wind turbine requires time-domain simulation tools taking into account all the phenomena that affect the system such as aerodynamics, structural dynamics, hydrodynamics, control actions and the mooring lines dynamics. These effects present couplings and are mutually influenced. The results provided by integrated simulation tools are used to compute the fatigue and ultimate loads needed for the structural design of the different components of the wind turbine. For this reason, their accuracy has an important influence on the optimization of the components and the final cost of the floating wind turbine. In particular, the mooring system greatly affects the global dynamics of the floater. Many integrated codes for the simulation of floating wind turbines use simplified approaches that do not consider the mooring line dynamics. An accurate simulation of the mooring system within the integrated codes can be fundamental to obtain reliable results of the system dynamics and the loads. The impact of taking into account the mooring line dynamics in the integrated simulation still has not been thoroughly quantified. The main objective of this research consists on the development of an accurate dynamic model for the simulation of mooring lines, validate it against wave tank tests and then integrate it in a simulation code for floating wind turbines. This experimentally validated tool is finally used to quantify the impact that dynamic mooring models have on the computation of fatigue and ultimate loads of floating wind turbines in comparison with quasi-static tools. This information will be very useful for future designers to decide which mooring model is adequate depending on the platform type and the expected results. The dynamic mooring lines code developed in this research is based in the Finite Element Method and is oriented to the achievement of a computationally efficient code, selecting a Lumped Mass approach. The experimental tests performed for the validation of the code were carried out at the `Ecole Centrale de Nantes (ECN) wave tank in France, consisting of a chain submerged into a water basin, anchored at the bottom of the basin, where the suspension point of the chain was excited with harmonic motions of different periods. The code showed its ability to predict the tension and the motions at several positions along the length of the line with high accuracy. The results demonstrated the importance of capturing the evolution of the mooring dynamics for the prediction of the line tension, especially for the high frequency motions. Finally, the code was used for an extensive assessment of the effect of mooring dynamics on the computation of fatigue and ultimate loads for different floating wind turbines. The loads were computed for three platforms topologies (semisubmersible, spar-buoy and tension leg platform) and compared with the loads provided using a quasi-static mooring model. More than 20,000 load cases were launched and postprocessed following the IEC 61400-3 guideline and fulfilling the conditions that a certification entity would require to an offshore wind turbine designer. The results showed that the impact of mooring dynamics in both fatigue and ultimate loads increases as elements located closer to the platform are evaluated; the blade and the shaft loads are only slightly modified by the mooring dynamics in all the platform designs, the tower base loads can be significantly affected depending on the platform concept and the mooring lines tension strongly depends on the lines dynamics both in fatigue and extreme loads in all the platform concepts evaluated.

Down-regulation of transmembrane carbonic anhydrases in renal cell carcinoma cell lines by wild-type von Hippel-Lindau transgenes

To discover genes involved in von Hippel-Lindau (VHL)-mediated carcinogenesis, we used renal cell carcinoma cell lines stably transfected with wild-type VHL-expressing transgenes. Large-scale RNA differential display technology applied to these cell lines identified several differentially expressed genes, including an alpha carbonic anhydrase gene, termed CA12. The deduced protein sequence was classified as a one-pass transmembrane CA possessing an apparently intact catalytic domain in the extracellular CA module. Reintroduced wild-type VHL strongly inhibited the overexpression of the CA12 gene in the parental renal cell carcinoma cell lines. Similar results were obtained with CA9, encoding another transmembrane CA with an intact catalytic domain. Although both domains of the VHL protein contribute to regulation of CA12 expression, the elongin binding domain alone could effectively regulate CA9 expression. We mapped CA12 and CA9 loci to chromosome bands 15q22 and 17q21.2 respectively, regions prone to amplification in some human cancers. Additional experiments are needed to define the role of CA IX and CA XII enzymes in the regulation of pH in the extracellular microenvironment and its potential impact on cancer cell growth.

β-Catenin mutations in cell lines established from human colorectal cancers

β-catenin has functions as both an adhesion and a signaling molecule. Disruption of these functions through mutations of the β-catenin gene (CTNNB1) may be important in the development of colorectal tumors. We examined the entire coding sequence of β-catenin by reverse transcriptase–PCR (RT-PCR) and direct sequencing of 23 human colorectal cancer cell lines from 21 patients. In two cell lines, there was apparent instability of the β-catenin mRNA. Five different mutations (26%) were found in the remaining 21cell lines (from 19 patients). A three-base deletion (codon 45) was identified in the cell line HCT 116, whereas cell lines SW 48, HCA 46, CACO 2, and Colo 201 each contained single-base missense mutations (codons 33, 183, 245, and 287, respectively). All 23 cell lines had full-length β-catenin protein that was detectable by Western blotting and that coprecipitated with E-cadherin. In three of the cell lines with CTNNB1 mutations, complexes of β-catenin with α-catenin and APC were detectable. In SW48 and HCA 46, however, we did not detect complexes of β-catenin protein with α-catenin and APC, respectively. These results show that selection of CTNNB1 mutations occurs in up to 26% of colorectal cancers from which cell lines are derived. In these cases, mutation selection is probably for altered β-catenin function, which may significantly alter intracellular signaling and intercellular adhesion and may serve as a complement to APC mutations in the early stages of tumorigenesis.

Increased sensitivity to apoptotic stimuli in c-abl-deficient progenitor B-cell lines

The protooncogene c-abl encodes a nonreceptor tyrosine kinase whose cellular function is unknown. To study the possible involvement of c-Abl in proliferation, differentiation, and cell cycle regulation of early B cells, long-term lymphoid bone marrow cultures were established from c-abl-deficient mice and their wild-type littermates. Interleukin 7-dependent progenitor B-cell clones and lines expressing B220 and CD43 could be generated from both mutant and wild-type mice. The mutant and wild-type lines displayed no difference in their proliferative capacity as measured by thymidine incorporation in response to various concentrations of interleukin 7. Similarly, c-abl deficiency did not interfere with the ability of mutant clones to differentiate into surface IgM-positive cells in vitro. Analysis of cultures after growth factor deprivation, however, revealed a strikingly accelerated rate of cell death in c-abl mutant cells, due to apoptosis as confirmed by terminal deoxynucleotidyltransferase-mediated UTP nick end labeling analysis. Furthermore, a greater susceptibility to apoptotic cell death in c-abl mutant cells was also observed after glucocorticoid treatment. These results suggest that mutant c-Abl renders the B-cell progenitors more sensitive to apoptosis, and may account for some of the phenotypes observed in c-abl-deficient animals.

NADPH-oxidase and a hydrogen peroxide-sensitive K+ channel may function as an oxygen sensor complex in airway chemoreceptors and small cell lung carcinoma cell lines

Pulmonary neuroepithelial bodies (NEB) are widely distributed throughout the airway mucosa of human and animal lungs. Based on the observation that NEB cells have a candidate oxygen sensor enzyme complex (NADPH oxidase) and an oxygen-sensitive K+ current, it has been suggested that NEB may function as airway chemoreceptors. Here we report that mRNAs for both the hydrogen peroxide sensitive voltage gated potassium channel subunit (KH2O2) KV3.3a and membrane components of NADPH oxidase (gp91phox and p22phox) are coexpressed in the NEB cells of fetal rabbit and neonatal human lungs. Using a microfluorometry and dihydrorhodamine 123 as a probe to assess H2O2 generation, NEB cells exhibited oxidase activity under basal conditions. The oxidase in NEB cells was significantly stimulated by exposure to phorbol esther (0.1 μM) and inhibited by diphenyliodonium (5 μM). Studies using whole-cell voltage clamp showed that the K+ current of cultured fetal rabbit NEB cells exhibited inactivating properties similar to KV3.3a transcripts expressed in Xenopus oocyte model. Exposure of NEB cells to hydrogen peroxide (H2O2, the dismuted by-product of the oxidase) under normoxia resulted in an increase of the outward K+ current indicating that H2O2 could be the transmitter modulating the O2-sensitive K+ channel. Expressed mRNAs or orresponding protein products for the NADPH oxidase membrane cytochrome b as well as mRNA encoding KV3.3a were identified in small cell lung carcinoma cell lines. The studies presented here provide strong evidence for an oxidase-O2 sensitive potassium channel molecular complex operating as an O2 sensor in NEB cells, which function as chemoreceptors in airways and in NEB related tumors. Such a complex may represent an evolutionary conserved biochemical link for a membrane bound O2-signaling mechanism proposed for other cells and life forms.

Natural killer cell lines kill autologous β2-microglobulin-deficient melanoma cells: Implications for cancer immunotherapy

Cancer vaccines used to generate specific cytotoxic T lymphocytes are not effective against tumor cells that have lost or suppressed expression of their class I major histocompatibility complex proteins. This loss is common in some cancers and particularly in metastatic lesions. We show that β2-microglobulin-deficient class I-negative melanoma variants derived from patients undergoing specific T cell therapy are lysed by heterologous as well as autologous natural killer (NK) lines and clones, but not by specific T cells. Moreover, the minor NK cell fraction but not the major T cell fraction derived from heterologous lymphokine activated killer cells kills those tumor cell lines. ICAM-1 expression by the different class I protein deficient tumors was correlated with their sensitivity to lysis by NK cells. Adoptive autologous NK therapy may be an important supplement to consider in the design of new cancer immunotherapies.

The intrinsic radioresistance of glioblastoma-derived cell lines is associated with a failure of p53 to induce p21BAX expression

Radiation is the primary modality of therapy for all commonly occurring malignant brain tumors, including medulloblastoma and glioblastoma. These two brain tumors, however, have a distinctly different response to radiation therapy. Medulloblastoma is very sensitive to radiation therapy, whereas glioblastoma is highly resistant, and the long-term survival of medulloblastoma patients exceeds 50%, while there are few long-term survivors among glioblastoma patients. p53-mediated apoptosis is thought to be an important mechanism mediating the cytotoxic response of tumors to radiotherapy. In this study, we compared the response to radiation of five cell lines that have wild-type p53: three derived from glioblastoma and two derived from medulloblastoma. We found that the medulloblastoma-derived cell lines underwent extensive radiation-induced apoptotic cell death, while those from glioblastomas did not exhibit significant radiation-induced apoptosis. p53-mediated induction of p21BAX is thought to be a key component of the pathway mediating apoptosis after the exposure of cells to cytotoxins, and the expression of mRNA encoding p21BAX was correlated with these cell lines undergoing radiation-induced apoptosis. The failure of p53 to induce p21BAX expression in glioblastoma-derived cell lines is likely to be of biologic significance, since inhibition of p21BAX induction in medulloblastoma resulted in a loss of radiation-induced apoptosis, while forced expression of p21BAX in glioblastoma was sufficient to induce apoptosis. The failure of p53 to induce p21BAX in glioblastoma-derived cell lines suggests a distinct mechanism of radioresistance and may represent a critical factor in determining therapeutic responsiveness to radiation in glioblastomas.

TAK, an HIV Tat-associated kinase, is a member of the cyclin-dependent family of protein kinases and is induced by activation of peripheral blood lymphocytes and differentiation of promonocytic cell lines

We have previously identified a cellular protein kinase activity termed TAK that specifically associates with the HIV types 1 and 2 Tat proteins. TAK hyperphosphorylates the carboxyl-terminal domain of the large subunit of RNA polymerase II in vitro in a manner believed to activate transcription [Herrmann, C. H. & Rice, A. P. (1995) J. Virol. 69, 1612–1620]. We show here that the catalytic subunit of TAK is a known human kinase previously named PITALRE, which is a member of the cyclin-dependent family of proteins. We also show that TAK activity is elevated upon activation of peripheral blood mononuclear cells and peripheral blood lymphocytes and upon differentiation of U1 and U937 promonocytic cell lines to macrophages. Therefore, in HIV-infected individuals TAK may be induced in T cells following activation and in macrophages following differentiation, thus contributing to high levels of viral transcription and the escape from latency of transcriptionally silent proviruses.