Critical Role of Two-Dimensional Island-Mediated Growth on the Formation of Semiconductor Heterointerfaces

We experimentally demonstrate a sigmoidal variation of the composition profile across semiconductor heterointerfaces. The wide range of material systems (III-arsenides, III-antimonides, III-V quaternary compounds, III-nitrides) exhibiting such a profile suggests a universal behavior. We show that sigmoidal profiles emerge from a simple model of cooperative growth mediated by twodimensional island formation, wherein cooperative effects are described by a specific functional dependence of the sticking coefficient on the surface coverage. Experimental results confirm that, except in the very early stages, island growth prevails over nucleation as the mechanism governing the interface development and ultimately determines the sigmoidal shape of the chemical profile in these two-dimensional grown layers. In agreement with our experimental findings, the model also predicts a minimum value of the interfacial width, with the minimum attainable value depending on the chemical identity of the species.

Evaluación de la contribución y el impacto de las tecnologías del habla en la detección automática del Síndrome de la Apnea Obstructiva del Sueño = Contributions and impact assessment of speech technologies on the automatic detection of severe Obstructive Sleep Apnea syndrome

La presente Tesis analiza las posibilidades que ofrecen en la actualidad las tecnologías del habla para la detección de patologías clínicas asociadas a la vía aérea superior. El estudio del habla que tradicionalmente cubre tanto la producción como el proceso de transformación del mensaje y las señales involucradas, desde el emisor hasta alcanzar al receptor, ofrece una vía de estudio alternativa para estas patologías. El hecho de que la señal emitida no solo contiene este mensaje, sino también información acerca del locutor, ha motivado el desarrollo de sistemas orientados a la identificación y verificación de la identidad de los locutores. Estos trabajos han recibido recientemente un nuevo impulso, orientándose tanto hacia la caracterización de rasgos que son comunes a varios locutores, como a las diferencias existentes entre grabaciones de un mismo locutor. Los primeros resultan especialmente relevantes para esta Tesis dado que estos rasgos podrían evidenciar la presencia de características relacionadas con una cierta condición común a varios locutores, independiente de su identidad. Tal es el caso que se enfrenta en esta Tesis, donde los rasgos identificados se relacionarían con una de la patología particular y directamente vinculada con el sistema de físico de conformación del habla. El caso del Síndrome de Apneas Hipopneas durante el Sueno (SAHS) resulta paradigmático. Se trata de una patología con una elevada prevalencia mundo, que aumenta con la edad. Los pacientes de esta patología experimentan episodios de cese involuntario de la respiración durante el sueño, que se prolongan durante varios segundos y que se reproducen a lo largo de la noche impidiendo el correcto descanso. En el caso de la apnea obstructiva, estos episodios se deben a la imposibilidad de mantener un camino abierto a través de la vía aérea, de forma que el flujo de aire se ve interrumpido. En la actualidad, el diagnostico de estos pacientes se realiza a través de un estudio polisomnográfico, que se centra en el análisis de los episodios de apnea durante el sueño, requiriendo que el paciente permanezca en el hospital durante una noche. La complejidad y el elevado coste de estos procedimientos, unidos a las crecientes listas de espera, han evidenciado la necesidad de contar con técnicas rápidas de detección, que si bien podrían no obtener tasas tan elevadas, permitirían reorganizar las listas de espera en función del grado de severidad de la patología en cada paciente. Entre otros, los sistemas de diagnostico por imagen, así como la caracterización antropométrica de los pacientes, han evidenciado la existencia de patrones anatómicos que tendrían influencia directa sobre el habla. Los trabajos dedicados al estudio del SAHS en lo relativo a como esta afecta al habla han sido escasos y algunos de ellos incluso contradictorios. Sin embargo, desde finales de la década de 1980 se conoce la existencia de patrones específicos relativos a la articulación, la fonación y la resonancia. Sin embargo, su descripción resultaba difícilmente aprovechable a través de un sistema de reconocimiento automático, pero apuntaba la existencia de un nexo entre voz y SAHS. En los últimos anos las técnicas de procesado automático han permitido el desarrollo de sistemas automáticos que ya son capaces de identificar diferencias significativas en el habla de los pacientes del SAHS, y que los distinguen de los locutores sanos. Por contra, poco se conoce acerca de la conexión entre estos nuevos resultados, los sé que habían obtenido en el pasado y la patogénesis del SAHS. Esta Tesis continua la labor desarrollada en este ámbito considerando específicamente: el estudio de la forma en que el SAHS afecta el habla de los pacientes, la mejora en las tasas de clasificación automática y la combinación de la información obtenida con los predictores utilizados por los especialistas clínicos en sus evaluaciones preliminares. Las dos primeras tareas plantean problemas simbióticos, pero diferentes. Mientras el estudio de la conexión entre el SAHS y el habla requiere de modelos acotados que puedan ser interpretados con facilidad, los sistemas de reconocimiento se sirven de un elevado número de dimensiones para la caracterización y posterior identificación de patrones. Así, la primera tarea debe permitirnos avanzar en la segunda, al igual que la incorporación de los predictores utilizados por los especialistas clínicos. La Tesis aborda el estudio tanto del habla continua como del habla sostenida, con el fin de aprovechar las sinergias y diferencias existentes entre ambas. En el análisis del habla continua se tomo como punto de partida un esquema que ya fue evaluado con anterioridad, y sobre el cual se ha tratado la evaluación y optimización de la representación del habla, así como la caracterización de los patrones específicos asociados al SAHS. Ello ha evidenciado la conexión entre el SAHS y los elementos fundamentales de la señal de voz: los formantes. Los resultados obtenidos demuestran que el éxito de estos sistemas se debe, fundamentalmente, a la capacidad de estas representaciones para describir dichas componentes, obviando las dimensiones ruidosas o con poca capacidad discriminativa. El esquema resultante ofrece una tasa de error por debajo del 18%, sirviéndose de clasificadores notablemente menos complejos que los descritos en el estado del arte y de una única grabación de voz de corta duración. En relación a la conexión entre el SAHS y los patrones observados, fue necesario considerar las diferencias inter- e intra-grupo, centrándonos en la articulación característica del locutor, sustituyendo los complejos modelos de clasificación por el estudio de los promedios espectrales. El resultado apunta con claridad hacia ciertas regiones del eje de frecuencias, sugiriendo la existencia de un estrechamiento sistemático en la sección del tracto en la región de la orofaringe, ya prevista en la patogénesis de este síndrome. En cuanto al habla sostenida, se han reproducido los estudios realizados sobre el habla continua en grabaciones de la vocal /a/ sostenida. Los resultados son cualitativamente análogos a los anteriores, si bien en este caso las tasas de clasificación resultan ser más bajas. Con el objetivo de identificar el sentido de este resultado se reprodujo el estudio de los promedios espectrales y de la variabilidad inter e intra-grupo. Ambos estudios mostraron importantes diferencias con los anteriores que podrían explicar estos resultados. Sin embargo, el habla sostenida ofrece otras oportunidades al establecer un entorno controlado para el estudio de la fonación, que también había sido identificada como una fuente de información para la detección del SAHS. De su estudio se pudo observar que, en el conjunto de datos disponibles, no existen variaciones que pudieran asociarse fácilmente con la fonación. Únicamente aquellas dimensiones que describen la distribución de energía a lo largo del eje de frecuencia evidenciaron diferencias significativas, apuntando, una vez más, en la dirección de las resonancias espectrales. Analizados los resultados anteriores, la Tesis afronta la fusión de ambas fuentes de información en un único sistema de clasificación. Con ello es posible mejorar las tasas de clasificación, bajo la hipótesis de que la información presente en el habla continua y el habla sostenida es fundamentalmente distinta. Esta tarea se realizo a través de un sencillo esquema de fusión que obtuvo un 88.6% de aciertos en clasificación (tasa de error del 11.4%), lo que representa una mejora significativa respecto al estado del arte. Finalmente, la combinación de este clasificador con los predictores utilizados por los especialistas clínicos ofreció una tasa del 91.3% (tasa de error de 8.7%), que se encuentra dentro del margen ofrecido por esquemas más costosos e intrusivos, y que a diferencia del propuesto, no pueden ser utilizados en la evaluación previa de los pacientes. Con todo, la Tesis ofrece una visión clara sobre la relación entre el SAHS y el habla, evidenciando el grado de madurez alcanzado por la tecnología del habla en la caracterización y detección del SAHS, poniendo de manifiesto que su uso para la evaluación de los pacientes ya sería posible, y dejando la puerta abierta a futuras investigaciones que continúen el trabajo aquí iniciado. ABSTRACT This Thesis explores the potential of speech technologies for the detection of clinical disorders connected to the upper airway. The study of speech traditionally covers both the production process and post processing of the signals involved, from the speaker up to the listener, offering an alternative path to study these pathologies. The fact that utterances embed not just the encoded message but also information about the speaker, has motivated the development of automatic systems oriented to the identification and verificaton the speaker’s identity. These have recently been boosted and reoriented either towards the characterization of traits that are common to several speakers, or to the differences between records of the same speaker collected under different conditions. The first are particularly relevant to this Thesis as these patterns could reveal the presence of features that are related to a common condition shared among different speakers, regardless of their identity. Such is the case faced in this Thesis, where the traits identified would relate to a particular pathology, directly connected to the speech production system. The Obstructive Sleep Apnea syndrome (OSA) is a paradigmatic case for analysis. It is a disorder with high prevalence among adults and affecting a larger number of them as they grow older. Patients suffering from this disorder experience episodes of involuntary cessation of breath during sleep that may last a few seconds and reproduce throughout the night, preventing proper rest. In the case of obstructive apnea, these episodes are related to the collapse of the pharynx, which interrupts the air flow. Currently, OSA diagnosis is done through a polysomnographic study, which focuses on the analysis of apnea episodes during sleep, requiring the patient to stay at the hospital for the whole night. The complexity and high cost of the procedures involved, combined with the waiting lists, have evidenced the need for screening techniques, which perhaps would not achieve outstanding performance rates but would allow clinicians to reorganize these lists ranking patients according to the severity of their condition. Among others, imaging diagnosis and anthropometric characterization of patients have evidenced the existence of anatomical patterns related to OSA that have direct influence on speech. Contributions devoted to the study of how this disorder affects scpeech are scarce and somehow contradictory. However, since the late 1980s the existence of specific patterns related to articulation, phonation and resonance is known. By that time these descriptions were virtually useless when coming to the development of an automatic system, but pointed out the existence of a link between speech and OSA. In recent years automatic processing techniques have evolved and are now able to identify significant differences in the speech of OSAS patients when compared to records from healthy subjects. Nevertheless, little is known about the connection between these new results with those published in the past and the pathogenesis of the OSA syndrome. This Thesis is aimed to progress beyond the previous research done in this area by addressing: the study of how OSA affects patients’ speech, the enhancement of automatic OSA classification based on speech analysis, and its integration with the information embedded in the predictors generally used by clinicians in preliminary patients’ examination. The first two tasks, though may appear symbiotic at first, are quite different. While studying the connection between speech and OSA requires simple narrow models that can be easily interpreted, classification requires larger models including a large number dimensions for the characterization and posterior identification of the observed patterns. Anyhow, it is clear that any progress made in the first task should allow us to improve our performance on the second one, and that the incorporation of the predictors used by clinicians shall contribute in this same direction. The Thesis considers both continuous and sustained speech analysis, to exploit the synergies and differences between them. On continuous speech analysis, a conventional speech processing scheme, designed and evaluated before this Thesis, was taken as a baseline. Over this initial system several alternative representations of the speech information were proposed, optimized and tested to select those more suitable for the characterization of OSA-specific patterns. Evidences were found on the existence of a connection between OSA and the fundamental constituents of the speech: the formants. Experimental results proved that the success of the proposed solution is well explained by the ability of speech representations to describe these specific OSA-related components, ignoring the noisy ones as well those presenting low discrimination capabilities. The resulting scheme obtained a 18% error rate, on a classification scheme significantly less complex than those described in the literature and operating on a single speech record. Regarding the connection between OSA and the observed patterns, it was necessary to consider inter-and intra-group differences for this analysis, and to focus on the articulation, replacing the complex classification models by the long-term average spectra. Results clearly point to certain regions on the frequency axis, suggesting the existence of a systematic narrowing in the vocal tract section at the oropharynx. This was already described in the pathogenesis of this syndrome. Regarding sustained speech, similar experiments as those conducted on continuous speech were reproduced on sustained phonations of vowel / a /. Results were qualitatively similar to the previous ones, though in this case perfomance rates were found to be noticeably lower. Trying to derive further knowledge from this result, experiments on the long-term average spectra and intraand inter-group variability ratios were also reproduced on sustained speech records. Results on both experiments showed significant differences from the previous ones obtained from continuous speech which could explain the differences observed on peformance. However, sustained speech also provided the opportunity to study phonation within the controlled framework it provides. This was also identified in the literature as a source of information for the detection of OSA. In this study it was found that, for the available dataset, no sistematic differences related to phonation could be found between the two groups of speakers. Only those dimensions which relate energy distribution along the frequency axis provided significant differences, pointing once again towards the direction of resonant components. Once classification schemes on both continuous and sustained speech were developed, the Thesis addressed their combination into a single classification system. Under the assumption that the information in continuous and sustained speech is fundamentally different, it should be possible to successfully merge the two of them. This was tested through a simple fusion scheme which obtained a 88.6% correct classification (11.4% error rate), which represents a significant improvement over the state of the art. Finally, the combination of this classifier with the variables used by clinicians obtained a 91.3% accuracy (8.7% error rate). This is within the range of alternative, but costly and intrusive schemes, which unlike the one proposed can not be used in the preliminary assessment of patients’ condition. In the end, this Thesis has shed new light on the underlying connection between OSA and speech, and evidenced the degree of maturity reached by speech technology on OSA characterization and detection, leaving the door open for future research which shall continue in the multiple directions that have been pointed out and left as future work.

Survey on student motivation and approval of educational environment at Universidad Politécnica de Madrid within the European Higher Education area implementation

A recent study elaborated by Vicerrectorado de Ordenación Académica y Planificación Estratégica of Technical University of Madrid (UPM) defines the satisfaction of the university student body as "the response that the University offers to the expectations and demands of service of the students, considered in a general way ". Besides an indicator of academic and institutional insertion of the student, the assessment of student engagement allows us to adapt the academic offer and the extension services of the University to the real needs of the students. The process of convergence towards the European Higher Education Area (EHEA) raises the need to form in competitions, that is to say, of developing in our students capacities and knowledge beyond the purely theoretical-practical thing. Therefore, the perception and experience of the educational process and environment by the students is an important issue to be addressed to accomplish their expectations and achieve a curriculum accordingly to EHEA expectations. The present study aims to explore the student motivation and approval of the educational environment at the UPM. To this end a total of 97 students enrolled in the undergraduate program of Civil Engineering, Computer Engineering and Agronomic Engineering at UPM were surveyed. The survey consisted of 40 questions divided in three blocks. The first one of 20 questions of personal character in that they were gathering, besides the sex and the age, the degree of fulfilment, implication and dedication with the institution and the academic tasks. In the second block we identify 10 questions related to the perception of the student on the teaching quality, and finally a block of 10 questions regarding the Bologna Process. The students personal motivation was moderately high, with a score of 3.6 (all scores are provided on a 5-point scale), being the most valuable items obtaining a university degree (4,3) and the friendship between students (4,2). Any significant difference was shown between sexes (P=0.23) since the averages for this block of questions were of 3.7±0.3 and 3.5±0.4 for women and men respectively. The students are moderately satisfied with their graduate studies with an average score of 3,2, being the questions that reflect a minor satisfaction the research profile of the teachers (2,8) and the organization of the Schools (2,9). The best valued questions are related to the usefulness and quality of the degrees, with 3,5 and 3,4 respectively, and to the interest of the courses within the degree (3,4). For sexes, the results of this block of questions are similar (3.1±0.3 and 3.2±0.3 for men and women respectively=0.79). Also, there were no differences (P=0.39) between the students who arrange work and studies or do not work (3.1±0.2 and 3.2±0.3 respectively). In conclusion, students at UPM present an acceptable degree of motivation and satisfaction with regard to the studies and services that offer their respective Schools. Both characteristics receive the same value both for men and for women and so much for students who arrange work and studies as for those who devote themselves only to studying. In a significant way, students who are more engaged and are in-class attendants present the major degree of satisfaction.Overall, there is a great lack of information regarding the Bologna Process. In fact to the majority, they would like to know more on what it is, what it means and what changes will involve its implementation.

Design, development and evaluation of a portable recording system to capture dynamic presentations using the teacher's tablet PC

Computers and multimedia equipment have improved a lot in the last years. They have reduced their cost and size while at the same time increased their capabilities. These improvements allowed us to design and implement a portable recording system that also integrates the teacher´s tablet PC to capture what he/she writes on the slides and all that happens in it. This paper explains this system in detail and the validation of the recordings that we did after using it to record all the lectures the “Communications Software” course in our university. The results show that pupils used the recordings for different purposes and consider them useful for a variety of things, especially after missing a lecture.

Identidade profissional: como se veem os professores na escola atual

Dissertação apresentada para a obtenção do grau de mestre em Ciências da Educação - área de Supervisão e Orientação Pegagógica

O PERFIL IDEOLÓGICO DO DOCENTE DA REDE PÚBLICA ESTADUAL: UM ESTUDO DE CASO

Tendo como referencial o sociólogo Luiz Pereira, e levando em consideração as relevantes críticas à visão weberiana sobre a burocracia feitas principalmente por Maurício Tragtenberg, esta dissertação almejou, através de uma pesquisa empírica, mostrar que a construção da identidade profissional do professor sofre influência direta de sua visão ideológica. Partindo de uma análise crítica da visão weberiana de burocracia, o presente trabalho busca traçar o perfil ideológico do professor da rede pública. Para tanto, este trabalho baseia-se em uma pesquisa abrangendo entrevistas, análise de documentos e observações do cotidiano de uma escola pública estadual escolhida como fonte de pesquisa. Pretendeu-se abordar a questão da filiação ideológica dos professores da rede pública, analisando em dois níveis: o da filiação ideológica a uma classe social (a classe média) e o da filiação ideológica a uma categoria social ligada ao Estado (a burocracia estatal). Levou-se, portanto, em consideração a hipótese de uma filiação ideológica dos professores a ambos os grupos, já que, à parte a alta burocracia (filiada ideologicamente às classes dominantes), a maioria do corpo burocrático (média e baixa burocracia) tende a adotar a perspectiva ideológica da classe média.(AU)

O PERFIL IDEOLÓGICO DO DOCENTE DA REDE PÚBLICA ESTADUAL: UM ESTUDO DE CASO

Tendo como referencial o sociólogo Luiz Pereira, e levando em consideração as relevantes críticas à visão weberiana sobre a burocracia feitas principalmente por Maurício Tragtenberg, esta dissertação almejou, através de uma pesquisa empírica, mostrar que a construção da identidade profissional do professor sofre influência direta de sua visão ideológica. Partindo de uma análise crítica da visão weberiana de burocracia, o presente trabalho busca traçar o perfil ideológico do professor da rede pública. Para tanto, este trabalho baseia-se em uma pesquisa abrangendo entrevistas, análise de documentos e observações do cotidiano de uma escola pública estadual escolhida como fonte de pesquisa. Pretendeu-se abordar a questão da filiação ideológica dos professores da rede pública, analisando em dois níveis: o da filiação ideológica a uma classe social (a classe média) e o da filiação ideológica a uma categoria social ligada ao Estado (a burocracia estatal). Levou-se, portanto, em consideração a hipótese de uma filiação ideológica dos professores a ambos os grupos, já que, à parte a alta burocracia (filiada ideologicamente às classes dominantes), a maioria do corpo burocrático (média e baixa burocracia) tende a adotar a perspectiva ideológica da classe média.(AU)

SER-PROFESSOR: construção de identidade em processo autoformativo

Este estudo teve por objetivo analisar como o ser-professor se narra e interpretase como sujeito na profissão, tendo em vista compreender de que maneira esse profissional constrói a identidade pessoal/profissional, sendo capaz de afirmar-se ou negar-se como sujeito dentro do sistema-escola? Trata-se de uma pesquisa realizada com educadores de 1º e 2º ano do ciclo I e II do Ensino Fundamental da Rede Pública de São Bernardo do Campo e Mestrandos em Educação. Optou-se por entrevistar seis educadores, de diferentes áreas de ensino, tornando-os autores desta pesquisa. A abordagem metodológica de cunho qualitativo foi utilizadas priorizando a narrativa dos educadores. Para esta metodologia recorreu-se às contribuições de Maria Isabel Cunha; Lüdke; André; Connelly e Clandini que explicitam sobre a importância de trabalhar com essa metodologia na perspectiva de interpretar e compreender a construção da identidade docente, sobretudo, na linha Formação de Educadores. A temática formação docente e construção da identidade foi trazida à luz dos estudos de Pimenta; Brzezinski; Freire; Vianna; Lane; Sung; Nóvoa; Ciampa, entre outros. No âmbito da análise, foram priorizadas algumas categorias ao longo desta temática: razões que levaram à escolha da profissão; percepções que o professor tem de sujeito na profissão; condições de ser e estar na profissão ; sistema e construção da identidade: condições de se sujeitar para se auto-afirmar como sujeito profissional. No trato destas categorias e, ao longo desta pesquisa, foram consideradas as percepções dos professores, como sendo elementos valiosos para identificar da construção de identidade docente. Estas considerações fundam-se no aporte teórico-metodológico que apresentam grandes contribuições referentes à temática. A partir daí, são estabelecidas relações entre Psicologia Social e Sociologia que possibilitam identificar nas narrativas dos educadores uma insatisfação perante o sistema. Alguns professores partem do próprio sentimento ao narrar todo desconforto que as instituições propiciam-lhes; outros enfatizam de modo objetivo seu cotidiano, contornando e explicitando diferenças na condição de ser estar na profissão .

SER-PROFESSOR: construção de identidade em processo autoformativo

Este estudo teve por objetivo analisar como o ser-professor se narra e interpretase como sujeito na profissão, tendo em vista compreender de que maneira esse profissional constrói a identidade pessoal/profissional, sendo capaz de afirmar-se ou negar-se como sujeito dentro do sistema-escola? Trata-se de uma pesquisa realizada com educadores de 1º e 2º ano do ciclo I e II do Ensino Fundamental da Rede Pública de São Bernardo do Campo e Mestrandos em Educação. Optou-se por entrevistar seis educadores, de diferentes áreas de ensino, tornando-os autores desta pesquisa. A abordagem metodológica de cunho qualitativo foi utilizadas priorizando a narrativa dos educadores. Para esta metodologia recorreu-se às contribuições de Maria Isabel Cunha; Lüdke; André; Connelly e Clandini que explicitam sobre a importância de trabalhar com essa metodologia na perspectiva de interpretar e compreender a construção da identidade docente, sobretudo, na linha Formação de Educadores. A temática formação docente e construção da identidade foi trazida à luz dos estudos de Pimenta; Brzezinski; Freire; Vianna; Lane; Sung; Nóvoa; Ciampa, entre outros. No âmbito da análise, foram priorizadas algumas categorias ao longo desta temática: razões que levaram à escolha da profissão; percepções que o professor tem de sujeito na profissão; condições de ser e estar na profissão ; sistema e construção da identidade: condições de se sujeitar para se auto-afirmar como sujeito profissional. No trato destas categorias e, ao longo desta pesquisa, foram consideradas as percepções dos professores, como sendo elementos valiosos para identificar da construção de identidade docente. Estas considerações fundam-se no aporte teórico-metodológico que apresentam grandes contribuições referentes à temática. A partir daí, são estabelecidas relações entre Psicologia Social e Sociologia que possibilitam identificar nas narrativas dos educadores uma insatisfação perante o sistema. Alguns professores partem do próprio sentimento ao narrar todo desconforto que as instituições propiciam-lhes; outros enfatizam de modo objetivo seu cotidiano, contornando e explicitando diferenças na condição de ser estar na profissão .

The Arabidopsis thaliana AGRAVITROPIC 1 gene encodes a component of the polar-auxin-transport efflux carrier

Auxins are plant hormones that mediate many aspects of plant growth and development. In higher plants, auxins are polarly transported from sites of synthesis in the shoot apex to their sites of action in the basal regions of shoots and in roots. Polar auxin transport is an important aspect of auxin functions and is mediated by cellular influx and efflux carriers. Little is known about the molecular identity of its regulatory component, the efflux carrier [Estelle, M. (1996) Current Biol. 6, 1589–1591]. Here we show that mutations in the Arabidopsis thaliana AGRAVITROPIC 1 (AGR1) gene involved in root gravitropism confer increased root-growth sensitivity to auxin and decreased sensitivity to ethylene and an auxin transport inhibitor, and cause retention of exogenously added auxin in root tip cells. We used positional cloning to show that AGR1 encodes a putative transmembrane protein whose amino acid sequence shares homologies with bacterial transporters. When expressed in Saccharomyces cerevisiae, AGR1 promotes an increased efflux of radiolabeled IAA from the cells and confers increased resistance to fluoro-IAA, a toxic IAA-derived compound. AGR1 transcripts were localized to the root distal elongation zone, a region undergoing a curvature response upon gravistimulation. We have identified several AGR1-related genes in Arabidopsis, suggesting a global role of this gene family in the control of auxin-regulated growth and developmental processes.

The Fission Yeast Mitotic Regulator win1+ Encodes an MAP Kinase Kinase Kinase That Phosphorylates and Activates Wis1 MAP Kinase Kinase in Response to High Osmolarity

The Schizosaccharomyces pombe win1-1 mutant has a defect in the G2-M transition of the cell cycle. Although the defect is suppressed by wis1+ and wis4+, which are components of a stress-activated MAP kinase pathway that links stress response and cell cycle control, the molecular identity of Win1 has not been known. We show here that win1+ encodes a polypeptide of 1436 residues with an apparent molecular size of 180 kDa and demonstrate that Win1 is a MAP kinase kinase kinase that phosphorylates and activates Wis1. Despite extensive similarities between Win1 and Wis4, the two MAP kinase kinase kinases have distinct functions. Wis4 is able to compensate for loss of Win1 only under unstressed conditions to maintain basal Wis1 activity, but it fails to suppress the osmosignaling defect conferred by win1 mutations. The win1-1 mutation is a spontaneous duplication of 16 nucleotides, which leads to a frameshift and production of a truncated protein lacking the kinase domain. We discuss the cell cycle phenotype of the win1-1 cdc25-22 wee1-50 mutant and its suppression by wis genes.

Pituitary Ets-1 and GABP bind to the growth factor regulatory sites of the rat prolactin promoter

Ets factors play a critical role in oncogenic Ras- and growth factor-mediated regulation of the proximal rat prolactin (rPRL) promoter in pituitary cells. The rPRL promoter contains two key functional Ets binding sites (EBS): a composite EBS/Pit-1 element located at –212 and an EBS that co-localizes with the basal transcription element (BTE, or A-site) located at –96. Oncogenic Ras exclusively signals to the –212 site, which we have named the Ras response element (RRE); whereas the response of multiple growth factors (FGFs, EGF, IGF, insulin and TRH) maps to both EBSs. Although Ets-1 and GA binding protein (GABP) have been implicated in the Ras and insulin responses, respectively, the precise identity of the pituitary Ets factors that specifically bind to the RRE and BTE sites remains unknown. In order to identify the Ets factor(s) present in GH4 and GH3 nuclear extracts (GH4NE and GH3NE) that bind to the EBSs contained in the RRE and BTE, we used EBS-RRE and BTE oligonucleotides in electrophoretic mobility shift assays (EMSAs), antibody supershift assays, western blot analysis of partially purified fractions and UV-crosslinking studies. EMSAs, using either the BTE or EBS-RRE probes, identified a specific protein–DNA complex, designated complex A, which contains an Ets factor as determined by oligonucleotide competition studies. Using western blot analysis of GH3 nuclear proteins that bind to heparin–Sepharose, we have shown that Ets-1 and GABP, which are MAP kinase substrates, co-purify with complex A, and supershift analysis with specific antisera revealed that complex A contains Ets-1, GABPα and GABPβ1. In addition, we show that recombinant full-length Ets-1 binds equivalently to BTE and EBS-RRE probes, while recombinant GABPα/β preferentially binds to the BTE probe. Furthermore, comparing the DNA binding of GH4NE containing both Ets-1 and GABP and HeLa nuclear extracts devoid of Ets-1 but containing GABP, we were able to show that the EBS-RRE preferentially binds Ets-1, while the BTE binds both GABP and Ets-1. Finally, UV-crosslinking experiments with radiolabeled EBS-RRE and BTE oligonucleotides showed that these probes specifically bind to a protein of ∼64 kDa, which is consistent with binding to Ets-1 (54 kDa) and/or the DNA binding subunit of GABP, GABPα (57 kDa). These studies show that endogenous, pituitary-derived GABP and Ets-1 bind to the BTE, whereas Ets-1 preferentially binds to the EBS-RRE. Taken together, these data provide important insights into the mechanisms by which the combination of distinct Ets members and EBSs transduce differential growth factor responses.

Identification of the Gene Encoding the Tryptophan Synthase β-Subunit from Chlamydomonas reinhardtii1

We report the isolation of a Chlamydomonas reinhardtii cDNA that encodes the β-subunit of tryptophan synthase (TSB). This cDNA was cloned by functional complementation of a trp-operon-deleted strain of Escherichia coli. Hybridization analysis indicated that the gene exists in a single copy. The predicted amino acid sequence showed the greatest identity to TSB polypeptides from other photosynthetic organisms. With the goal of identifying mutations in the gene encoding this enzyme, we isolated 11 recessive and 1 dominant single-gene mutation that conferred resistance to 5-fluoroindole. These mutations fell into three complementation groups, MAA2, MAA7, and TAR1. In vitro assays showed that mutations at each of these loci affected TSB activity. Restriction fragment-length polymorphism analysis suggested that MAA7 encodes TSB. MAA2 and TAR1 may act to regulate the activity of MAA7 or its protein product.

Differences in water release for the binding of EcoRI to specific and nonspecific DNA sequences.

The free energy difference between complexes of the restriction nuclease EcoRI with nonspecific DNA and with the enzyme's recognition sequence is linearly dependent on the water chemical potential of the solution, set using several very different solutes, ranging from glycine and glycerol to triethylene glycol and sucrose. This osmotic dependence indicates that the nonspecific complex sequesters some 110 waters more than the specific complex with the recognition sequence. The insensitivity of the difference in number of waters released to the solute identity further indicates that this water is sequestered in a space that is sterically inaccessible to solutes, most likely at the protein-DNA interface of the nonspecific complex. Calculations based on the structure of the specific complex suggest that the apposing DNA and protein surfaces in the nonspecific complex retain approximately a full hydration layer of water.

Extraordinary divergence and positive Darwinian selection in a fusagenic protein coating the acrosomal process of abalone spermatozoa.

During fertilization in marine invertebrates, fusion between sperm and egg cell membranes occurs at the tip of the sperm acrosomal process. In abalone sperm the acrosomal process is coated with an 18-kDa protein. In situ, this protein has no effect on the egg vitelline envelope, but in vitro it is a potent fusagen of liposomes. Thus, the 18-kDa protein may mediate membrane fusion between the gametes, a step in gamete recognition known to restrict heterospecific fertilization in other species. The cDNA and deduced amino acid sequences of the 18-kDa protein were determined for five species of California abalone. The deduced amino acid sequences exhibit extraordinary divergence; the percent identity varies from 27% to 87%. Analysis of nucleotide substitution shows extremely high frequencies of amino acid-altering substitution compared to silent substitution, demonstrating that positive Darwinian selection promotes the divergence of this protein. However, amino acid replacement is conservative with respect to size and polarity of residue. The data support the developing idea that in free-spawning marine invertebrates, the proteins mediating fertilization may be subjected to intense, and as yet unknown, selective forces. The extraordinary divergence of fertilization proteins may be related to the establishment of barriers to heterospecific fertilization.