Switching of ferroelectric liquid crystal doped with cetyltrimethylammonium bromide-assisted CdS nanostructures

Large scale high yield cadmium sulfide (CdS) nanowires with uniform diameter were synthesized using a rapid and simple solvo-chemical and hydrothermal route assisted by the surfactant cetyltrimethylammonium bromide (CTAB). Unique CdS nanowires of different morphologies could be selectively produced by only varying the concentration of CTAB in the reaction system with cadmium acetate, sulfur powder and ethylenediamine. We obtained CdS nanowires with diameters of 64–65 nm and lengths of up to several micrometers. A comparative study of the optical properties of ferroelectric liquid crystal (FLC) Felix-017/100 doped with 1% of CdS nanowires was performed. Response times of the order of from 160 to 180 μs, rotational viscosities of the order of from 5000 to 3000 mN s m−2 and polarizations of the order of from 10 to 70 nC cm−2 were measured. We also observed an anti-ferroelectric to ferroelectric transition for CdS doped FLC instead of the ferroelectric to paraelectric transition for pure FLC.

Analysis of gain-switching in two-section tapered lasers

We analyze the gain-switching dynamics of two-section tapered lasers by means of a simplified three-rate-equation model. The goal is to improve the understanding of the underlying physics and to optimize the device geometry to achieve high power short duration optical pulses.

Wide bandwidth envelope trackers with reduced switching frequency for RF power amplifier

El requerimiento de proveer alta frecuencia de datos en los modernos sistema de comunicación inalámbricos resulta en complejas señales moduladas de radio-frequencia (RF) con un gran ancho de banda y alto ratio pico-promedio (PAPR). Para garantizar la linealidad del comportamiento, los amplificadores lineales de potencia comunes funcionan típicamente entre 4 y 10 dB de back-o_ desde la máxima potencia de salida, ocasionando una baja eficiencia del sistema. La eliminación y restauración de la evolvente (EER) y el seguimiento de la evolvente (ET) son dos prometedoras técnicas para resolver el problema de la eficiencia. Tanto en EER como en ET, es complicado diseñar un amplificador de potencia que sea eficiente para señales de RF de alto ancho de banda y alto PAPR. Una propuesta común para los amplificadores de potencia es incluir un convertidor de potencia de muy alta eficiencia operando a frecuencias más altas que el ancho de banda de la señal RF. En este caso, la potencia perdida del convertidor ocasionado por la alta frecuencia desaconseja su práctica cuando el ancho de banda es muy alto. La solución a este problema es el enfoque de esta disertación que presenta dos arquitecturas de amplificador evolvente: convertidor híbrido-serie con una técnica de evolvente lenta y un convertidor multinivel basado en un convertidor reductor multifase con control de tiempo mínimo. En la primera arquitectura, una topología híbrida está compuesta de una convertidor reductor conmutado y un regulador lineal en serie que trabajan juntos para ajustar la tensión de salida para seguir a la evolvente con precisión. Un algoritmo de generación de una evolvente lenta crea una forma de onda con una pendiente limitada que es menor que la pendiente máxima de la evolvente original. La salida del convertidor reductor sigue esa forma de onda en vez de la evolvente original usando una menor frecuencia de conmutación, porque la forma de onda no sólo tiene una pendiente reducida sino también un menor ancho de banda. De esta forma, el regulador lineal se usa para filtrar la forma de onda tiene una pérdida de potencia adicional. Dependiendo de cuánto se puede reducir la pendiente de la evolvente para producir la forma de onda, existe un trade-off entre la pérdida de potencia del convertidor reductor relacionada con la frecuencia de conmutación y el regulador lineal. El punto óptimo referido a la menor pérdida de potencia total del amplificador de evolvente es capaz de identificarse con la ayuda de modelo preciso de pérdidas que es una combinación de modelos comportamentales y analíticos de pérdidas. Además, se analiza el efecto en la respuesta del filtro de salida del convertidor reductor. Un filtro de dampeo paralelo extra es necesario para eliminar la oscilación resonante del filtro de salida porque el convertidor reductor opera en lazo abierto. La segunda arquitectura es un amplificador de evolvente de seguimiento de tensión multinivel. Al contrario que los convertidores que usan multi-fuentes, un convertidor reductor multifase se emplea para generar la tensión multinivel. En régimen permanente, el convertidor reductor opera en puntos del ciclo de trabajo con cancelación completa del rizado. El número de niveles de tensión es igual al número de fases de acuerdo a las características del entrelazamiento del convertidor reductor. En la transición, un control de tiempo mínimo (MTC) para convertidores multifase es novedosamente propuesto y desarrollado para cambiar la tensión de salida del convertidor reductor entre diferentes niveles. A diferencia de controles convencionales de tiempo mínimo para convertidores multifase con inductancia equivalente, el propuesto MTC considera el rizado de corriente por cada fase basado en un desfase fijo que resulta en diferentes esquemas de control entre las fases. La ventaja de este control es que todas las corrientes vuelven a su fase en régimen permanente después de la transición para que la siguiente transición pueda empezar muy pronto, lo que es muy favorable para la aplicación de seguimiento de tensión multinivel. Además, el control es independiente de la carga y no es afectado por corrientes de fase desbalanceadas. Al igual que en la primera arquitectura, hay una etapa lineal con la misma función, conectada en serie con el convertidor reductor multifase. Dado que tanto el régimen permanente como el estado de transición del convertidor no están fuertemente relacionados con la frecuencia de conmutación, la frecuencia de conmutación puede ser reducida para el alto ancho de banda de la evolvente, la cual es la principal consideración de esta arquitectura. La optimización de la segunda arquitectura para más alto anchos de banda de la evolvente es presentada incluyendo el diseño del filtro de salida, la frecuencia de conmutación y el número de fases. El área de diseño del filtro está restringido por la transición rápida y el mínimo pulso del hardware. La rápida transición necesita un filtro pequeño pero la limitación del pulso mínimo del hardware lleva el diseño en el sentido contrario. La frecuencia de conmutación del convertidor afecta principalmente a la limitación del mínimo pulso y a las pérdidas de potencia. Con una menor frecuencia de conmutación, el ancho de pulso en la transición es más pequeño. El número de fases relativo a la aplicación específica puede ser optimizado en términos de la eficiencia global. Otro aspecto de la optimización es mejorar la estrategia de control. La transición permite seguir algunas partes de la evolvente que son más rápidas de lo que el hardware puede soportar al precio de complejidad. El nuevo método de sincronización de la transición incrementa la frecuencia de la transición, permitiendo que la tensión multinivel esté más cerca de la evolvente. Ambas estrategias permiten que el convertidor pueda seguir una evolvente con un ancho de banda más alto que la limitación de la etapa de potencia. El modelo de pérdidas del amplificador de evolvente se ha detallado y validado mediante medidas. El mecanismo de pérdidas de potencia del convertidor reductor tiene que incluir las transiciones en tiempo real, lo cual es diferente del clásico modelos de pérdidas de un convertidor reductor síncrono. Este modelo estima la eficiencia del sistema y juega un papel muy importante en el proceso de optimización. Finalmente, la segunda arquitectura del amplificador de evolvente se integra con el amplificador de clase F. La medida del sistema EER prueba el ahorro de energía con el amplificador de evolvente propuesto sin perjudicar la linealidad del sistema. ABSTRACT The requirement of delivering high data rates in modern wireless communication systems results in complex modulated RF signals with wide bandwidth and high peak-to-average ratio (PAPR). In order to guarantee the linearity performance, the conventional linear power amplifiers typically work at 4 to 10 dB back-off from the maximum output power, leading to low system efficiency. The envelope elimination and restoration (EER) and envelope tracking (ET) are two promising techniques to overcome the efficiency problem. In both EER and ET, it is challenging to design efficient envelope amplifier for wide bandwidth and high PAPR RF signals. An usual approach for envelope amplifier includes a high-efficiency switching power converter operating at a frequency higher than the RF signal's bandwidth. In this case, the power loss of converter caused by high switching operation becomes unbearable for system efficiency when signal bandwidth is very wide. The solution of this problem is the focus of this dissertation that presents two architectures of envelope amplifier: a hybrid series converter with slow-envelope technique and a multilevel converter based on a multiphase buck converter with the minimum time control. In the first architecture, a hybrid topology is composed of a switched buck converter and a linear regulator in series that work together to adjust the output voltage to track the envelope with accuracy. A slow envelope generation algorithm yields a waveform with limited slew rate that is lower than the maximum slew rate of the original envelope. The buck converter's output follows this waveform instead of the original envelope using lower switching frequency, because the waveform has not only reduced slew rate but also reduced bandwidth. In this way, the linear regulator used to filter the waveform has additional power loss. Depending on how much reduction of the slew rate of envelope in order to obtain that waveform, there is a trade-off between the power loss of buck converter related to the switching frequency and the power loss of linear regulator. The optimal point referring to the lowest total power loss of this envelope amplifier is identified with the help of a precise power loss model that is a combination of behavioral and analytic loss model. In addition, the output filter's effect on the response is analyzed. An extra parallel damping filter is needed to eliminate the resonant oscillation of output filter L and C, because the buck converter operates in open loop. The second architecture is a multilevel voltage tracking envelope amplifier. Unlike the converters using multi-sources, a multiphase buck converter is employed to generate the multilevel voltage. In the steady state, the buck converter operates at complete ripple cancellation points of duty cycle. The number of the voltage levels is equal to the number of phases according the characteristics of interleaved buck converter. In the transition, a minimum time control (MTC) for multiphase converter is originally proposed and developed for changing the output voltage of buck converter between different levels. As opposed to conventional minimum time control for multiphase converter with equivalent inductance, the proposed MTC considers the current ripple of each phase based on the fixed phase shift resulting in different control schemes among the phases. The advantage of this control is that all the phase current return to the steady state after the transition so that the next transition can be triggered very soon, which is very favorable for the application of multilevel voltage tracking. Besides, the control is independent on the load condition and not affected by the unbalance of phase current. Like the first architecture, there is also a linear stage with the same function, connected in series with the multiphase buck converter. Since both steady state and transition state of the converter are not strongly related to the switching frequency, it can be reduced for wide bandwidth envelope which is the main consideration of this architecture. The optimization of the second architecture for wider bandwidth envelope is presented including the output filter design, switching frequency and the number of phases. The filter design area is restrained by fast transition and the minimum pulse of hardware. The fast transition needs small filter but the minimum pulse of hardware limitation pushes the filter in opposite way. The converter switching frequency mainly affects the minimum pulse limitation and the power loss. With lower switching frequency, the pulse width in the transition is smaller. The number of phases related to specific application can be optimized in terms of overall efficiency. Another aspect of optimization is improving control strategy. Transition shift allows tracking some parts of envelope that are faster than the hardware can support at the price of complexity. The new transition synchronization method increases the frequency of transition, allowing the multilevel voltage to be closer to the envelope. Both control strategies push the converter to track wider bandwidth envelope than the limitation of power stage. The power loss model of envelope amplifier is detailed and validated by measurements. The power loss mechanism of buck converter has to include the transitions in real time operation, which is different from classical power loss model of synchronous buck converter. This model estimates the system efficiency and play a very important role in optimization process. Finally, the second envelope amplifier architecture is integrated with a Class F amplifier. EER system measurement proves the power saving with the proposed envelope amplifier without disrupting the linearity performance.

Lexical phrases in telecommunication blogs: a genre analysis

The study presented in this paper aims to provide a description of telecommunication blogs as a genre. Lexical phrases are analysed in order to reach conclusions regarding the nature of the language in these texts and the extent to which the results obtained are comparable with other written or conversational discourse types. Although the departing hypothesis is that the articles in blogs are basically transactional and their main objective is to transfer information, the conclusions point to interaction as a distinctive characteristic of this type of discourse and also to a more careful organization in the comments to the blogs entries than originally expected. RESUMEN. Este trabajo presenta una descripción de los blogs de Telecomunicación como género. A través del análisis de frases léxicas se llega a conclusiones sobre algunas de las características que definen la lengua que se usa en este tipo de textos y se comparan los resultados obtenidos con otros previos sobre el discurso escrito y el conversacional. Aunque la hipótesis de partida es que los artículos de los blogs son básicamente transaccionales, donde el principal objetivo es transmitir información, los resultados llevan a conclusiones sobre la importancia de la interacción en este tipo de discurso y también apuntan a una mayor organización de la esperable en las entradas de los comentarios al artículo principal del blog.

Application of Agent Technology for Fault Diagnosis of Telecommunication Networks

La presente tesis doctoral contribuye al problema del diagnóstico autonómico de fallos en redes de telecomunicación. En las redes de telecomunicación actuales, las operadoras realizan tareas de diagnóstico de forma manual. Dichas operaciones deben ser llevadas a cabo por ingenieros altamente cualificados que cada vez tienen más dificultades a la hora de gestionar debidamente el crecimiento exponencial de la red tanto en tamaño, complejidad y heterogeneidad. Además, el advenimiento del Internet del Futuro hace que la demanda de sistemas que simplifiquen y automaticen la gestión de las redes de telecomunicación se haya incrementado en los últimos años. Para extraer el conocimiento necesario para desarrollar las soluciones propuestas y facilitar su adopción por los operadores de red, se propone una metodología de pruebas de aceptación para sistemas multi-agente enfocada en simplificar la comunicación entre los diferentes grupos de trabajo involucrados en todo proyecto de desarrollo software: clientes y desarrolladores. Para contribuir a la solución del problema del diagnóstico autonómico de fallos, se propone una arquitectura de agente capaz de diagnosticar fallos en redes de telecomunicación de manera autónoma. Dicha arquitectura extiende el modelo de agente Belief-Desire- Intention (BDI) con diferentes modelos de diagnóstico que gestionan las diferentes sub-tareas del proceso. La arquitectura propuesta combina diferentes técnicas de razonamiento para alcanzar su propósito gracias a un modelo estructural de la red, que usa razonamiento basado en ontologías, y un modelo causal de fallos, que usa razonamiento Bayesiano para gestionar debidamente la incertidumbre del proceso de diagnóstico. Para asegurar la adecuación de la arquitectura propuesta en situaciones de gran complejidad y heterogeneidad, se propone un marco de argumentación que permite diagnosticar a agentes que estén ejecutando en dominios federados. Para la aplicación de este marco en un sistema multi-agente, se propone un protocolo de coordinación en el que los agentes dialogan hasta alcanzar una conclusión para un caso de diagnóstico concreto. Como trabajos futuros, se consideran la extensión de la arquitectura para abordar otros problemas de gestión como el auto-descubrimiento o la auto-optimización, el uso de técnicas de reputación dentro del marco de argumentación para mejorar la extensibilidad del sistema de diagnóstico en entornos federados y la aplicación de las arquitecturas propuestas en las arquitecturas de red emergentes, como SDN, que ofrecen mayor capacidad de interacción con la red. ABSTRACT This PhD thesis contributes to the problem of autonomic fault diagnosis of telecommunication networks. Nowadays, in telecommunication networks, operators perform manual diagnosis tasks. Those operations must be carried out by high skilled network engineers which have increasing difficulties to properly manage the growing of those networks, both in size, complexity and heterogeneity. Moreover, the advent of the Future Internet makes the demand of solutions which simplifies and automates the telecommunication network management has been increased in recent years. To collect the domain knowledge required to developed the proposed solutions and to simplify its adoption by the operators, an agile testing methodology is defined for multiagent systems. This methodology is focused on the communication gap between the different work groups involved in any software development project, stakeholders and developers. To contribute to overcoming the problem of autonomic fault diagnosis, an agent architecture for fault diagnosis of telecommunication networks is defined. That architecture extends the Belief-Desire-Intention (BDI) agent model with different diagnostic models which handle the different subtasks of the process. The proposed architecture combines different reasoning techniques to achieve its objective using a structural model of the network, which uses ontology-based reasoning, and a causal model, which uses Bayesian reasoning to properly handle the uncertainty of the diagnosis process. To ensure the suitability of the proposed architecture in complex and heterogeneous environments, an argumentation framework is defined. This framework allows agents to perform fault diagnosis in federated domains. To apply this framework in a multi-agent system, a coordination protocol is defined. This protocol is used by agents to dialogue until a reliable conclusion for a specific diagnosis case is reached. Future work comprises the further extension of the agent architecture to approach other managements problems, such as self-discovery or self-optimisation; the application of reputation techniques in the argumentation framework to improve the extensibility of the diagnostic system in federated domains; and the application of the proposed agent architecture in emergent networking architectures, such as SDN, which offers new capabilities of control for the network.

Evaluation of the PV cell operation temperature in the process of fast switching to open-circuit mode

A procedure for measuring the overheating temperature (ΔT ) of a p-n junction area in the structure of photovoltaic (PV) cells converting laser or solar radiations relative to the ambient temperature has been proposed for the conditions of connecting to an electric load. The basis of the procedure is the measurement of the open-circuit voltage (VO C ) during the initial time period after the fast disconnection of the external resistive load. The simultaneous temperature control on an external heated part of a PV module gives the means for determining the value of VO C at ambient temperature. Comparing it with that measured after switching OFF the load makes the calculation of ΔT possible. Calibration data on the VO C = f(T ) dependences for single-junction AlGaAs/GaAs and triple-junction InGaP/GaAs/Ge PV cells are presented. The temperature dynamics in the PV cells has been determined under flash illumination and during fast commutation of the load. Temperature measurements were taken in two cases: converting continuous laser power by single-junction cells and converting solar power by triple-junction cells operating in the concentrator modules.

Conditional switching of vascular endothelial growth factor (VEGF) expression in tumors: Induction of endothelial cell shedding and regression of hemangioblastoma-like vessels by VEGF withdrawal

We have recently shown that VEGF functions as a survival factor for newly formed vessels during developmental neovascularization, but is not required for maintenance of mature vessels. Reasoning that expanding tumors contain a significant fraction of newly formed and remodeling vessels, we examined whether abrupt withdrawal of VEGF will result in regression of preformed tumor vessels. Using a tetracycline-regulated VEGF expression system in xenografted C6 glioma cells, we showed that shutting off VEGF production leads to detachment of endothelial cells from the walls of preformed vessels and their subsequent death by apoptosis. Vascular collapse then leads to hemorrhages and extensive tumor necrosis. These results suggest that enforced withdrawal of vascular survival factors can be applied to target preformed tumor vasculature in established tumors. The system was also used to examine phenotypes resulting from over-expression of VEGF. When expression of the transfected VEGF cDNA was continuously “on,” tumors became hyper-vascularized with abnormally large vessels, presumably arising from excessive fusions. Tumors were significantly less necrotic, suggesting that necrosis in these tumors is the result of insufficient angiogenesis.

Calcium-dependent switching of the specificity of phosphoinositide binding to synaptotagmin

The synaptic vesicle membrane protein synaptotagmin (tagmin) is essential for fast, calcium-dependent, neurotransmitter release and is likely to be the calcium sensor for exocytosis, because of its many calcium-dependent properties. Polyphosphoinositides are needed for exocytosis, but it has not been known why. We now provide a possible connection between these observations with the finding that the C2B domain of tagmin I binds phosphatidylinositol-4,5-bisphosphate (PIns-4,5-P2), its isomer phosphatidylinositol-3,4-bisphosphate and phosphatidylinositol-3,4,5-trisphosphate (PIns-3,4,5-P3). Calcium ions switch the specificity of this binding from PIns-3,4,5-P3 (at calcium concentrations found in resting nerve terminals) to PIns-4,5-P2 (at concentration of calcium required for transmitter release). Inositol polyphosphates, known blockers of neurotransmitter release, inhibit the binding of both PIns-4,5-P2 and PIns-3,4,5-P3 to tagmin. Our findings imply that tagmin may operate as a bimodal calcium sensor, switching bound lipids during exocytosis. This connection to polyphosphoinositides, compounds whose levels are physiologically regulated, could be important for long-term memory and learning.

Phenotypic switching in the human pathogenic fungus Cryptococcus neoformans is associated with changes in virulence and pulmonary inflammatory response in rodents

High-frequency reversible changes in colony morphology were observed in three strains of Cryptococcus neoformans. For one strain (SB4, serotype A), this process produced three colony types: smooth (S), wrinkled (W), and serrated (C). The frequency of switching between colony types varied for the individual colony transitions and was as high as 10−3. Mice infected with colony type W died faster than those infected with other colony types. The rat inflammatory response to infection with colony types S, W, and C was C > S > W and ranged from intense granulomatous inflammation with caseous necrosis for infection with type C to minimal inflammation for infection with type W. Infection with the various colony types was associated with different antibody responses to cryptococcal proteins in rats. Analysis of cellular characteristics revealed differences between the three colony types. High-frequency changes in colony morphology were also observed in two additional strains of C. neoformans. For one strain (24067A, serotype D) the switching occurred between smooth and wrinkled colonies. For the other strain (J32A, serotype A), the switching occurred between mucoid and nonmucoid colonies. The findings indicate that C. neoformans undergoes phenotypic switching and that this process can affect virulence and host inflammatory and immune responses. Phenotypic switching may play a role in the ability of this fungus to escape host defenses and establish chronic infections.

The role of prefrontal cortex and posterior parietal cortex in task switching

Human ability to switch from one cognitive task to another involves both endogenous preparation without an external stimulus and exogenous adjustment in response to the external stimulus. In an event-related functional MRI study, participants performed pairs of two tasks that are either the same (task repetition) or different (task switch) from each other. On half of the trials, foreknowledge about task repetition or task switch was available. On the other half, it was not. Endogenous preparation seems to involve lateral prefrontal cortex (BA 46/45) and posterior parietal cortex (BA 40). During preparation, higher activation increases in inferior lateral prefrontal cortex and superior posterior parietal cortex were associated with foreknowledge than with no foreknowledge. Exogenous adjustment seems to involve superior prefrontal cortex (BA 8) and posterior parietal cortex (BA 39/40) in general. During a task switch with no foreknowledge, activations in these areas were relatively higher than during a task repetition with no foreknowledge. These results suggest that endogenous preparation and exogenous adjustment for a task switch may be independent processes involving different brain areas.

The neural basis of task-switching in working memory: Effects of performance and aging

We studied the performance of young and senior subjects on a well known working memory task, the Operation Span. This is a dual-task in which subjects perform a memory task while simultaneously verifying simple equations. Positron-emission tomography scans were taken during performance. Both young and senior subjects demonstrated a cost in accuracy and latency in the Operation Span compared with performing each component task alone (math verification or memory only). Senior subjects were disproportionately impaired relative to young subjects on the dual-task. When brain activation was examined for senior subjects, we found regions in prefrontal cortex that were active in the dual-task, but not in the component tasks. Similar results were obtained for young subjects who performed relatively poorly on the dual-task; however, for young subjects who performed relatively well in the dual-task, we found no prefrontal regions that were active only in the dual-task. Results are discussed as they relate to the executive component of task switching.

Environmental induction and genetic control of surface antigen switching in the nematode Caenorhabditis elegans.

Nematodes can alter their surface coat protein compositions at the molts between developmental stages or in response to environmental changes; such surface alterations may enable parasitic nematodes to evade host immune defenses during the course of infection. Surface antigen switching mechanisms are presently unknown. In a genetic study of surface antigen switching, we have used a monoclonal antibody, M37, that recognizes a surface antigen on the first larval stage of the free-living nematode Caenorhabditis elegans. We demonstrate that wild-type C. elegans can be induced to display the M37 antigen on a later larval stage by altering the growth conditions. Mutations that result in nonconditional display of this antigen on all four larval stages fall into two classes. One class defines the new gene srf-6 II. The other mutations are in previously identified dauer-constitutive genes involved in transducing environmental signals that modulate formation of the dauer larva, a developmentally arrested dispersal stage. Although surface antigen switching is affected by some of the genes that control dauer formation, these two process can be blocked separately by specific mutations or induced separately by environmental factors. Based on these results, the mechanisms of nematode surface antigen switching can now be investigated directly.

Peroxo-iron and oxenoid-iron species as alternative oxygenating agents in cytochrome P450-catalyzed reactions: switching by threonine-302 to alanine mutagenesis of cytochrome P450 2B4.

Among biological catalysts, cytochrome P450 is unmatched in its multiplicity of isoforms, inducers, substrates, and types of chemical reactions catalyzed. In the present study, evidence is given that this versatility extends to the nature of the active oxidant. Although mechanistic evidence from several laboratories points to a hypervalent iron-oxenoid species in P450-catalyzed oxygenation reactions, Akhtar and colleagues [Akhtar, M., Calder, M. R., Corina, D. L. & Wright, J. N. (1982) Biochem. J. 201, 569-580] proposed that in steroid deformylation effected by P450 aromatase an iron-peroxo species is involved. We have shown more recently that purified liver microsomal P450 cytochromes, including phenobarbital-induced P450 2B4, catalyze the analogous deformylation of a series of xenobiotic aldehydes with olefin formation. The investigation presented here on the effect of site-directed mutagenesis of threonine-302 to alanine on the activities of recombinant P450 2B4 with N-terminal amino acids 2-27 deleted [2B4 (delta2-27)] makes use of evidence from other laboratories that the corresponding mutation in bacterial P450s interferes with the activation of dioxygen to the oxenoid species by blocking proton delivery to the active site. The rates of NADPH oxidation, hydrogen peroxide production, and product formation from four substrates, including formaldehyde from benzphetamine N-demethylation, acetophenone from 1-phenylethanol oxidation, cyclohexanol from cyclohexane hydroxylation, and cyclohexene from cyclohexane carboxaldehyde deformylation, were determined with P450s 2B4, 2B4 (delta2-27), and 2B4 (delta2-27) T302A. Replacement of the threonine residue in the truncated cytochrome gave a 1.6- to 2.5-fold increase in peroxide formation in the presence of a substrate, but resulted in decreased product formation from benzphetamine (9-fold), cyclohexane (4-fold), and 1-phenylethanol (2-fold). In sharp contrast, the deformylation of cyclohexane carboxaldehyde by the T302A mutant was increased about 10-fold. On the basis of these findings and our previous evidence that aldehyde deformylation is supported by added H202, but not by artificial oxidants, we conclude that the iron-peroxy species is the direct oxygen donor. It remains to be established which of the many other oxidative reactions involving P450 utilize this species and the extent to which peroxo-iron and oxenoid-iron function as alternative oxygenating agents with the numerous isoforms of this versatile catalyst.

Formation of chimeric DNA primer extension products by template switching onto an annealed downstream oligonucleotide.

Oligodeoxynucleotide sequences are described that anneal to a template downstream of a priming site. During polymerase-catalyzed extension of the primer, the extending primer shifts from the original template to a segment of the annealed oligonucleotide that acts as an alternative template. The resulting chimeric extended primer has one segment that is complementary to the template and a second segment that is complementary to the oligonucleotide. The influence of the sequence elements of the oligonucleotide and the reaction conditions on template switching have been explored. The sequence requirements for template switching are compared to those for transposon excision.