Anabolic drugs consumption of high school students participating in school championships of physical education

This paper aims to analyse the use of anabolic drugs among Greek students participating in school championships of physical education (PE). In order to do it, a survey was conducted during the 2008 to 2009 academic year in suburban, urban and metropolitan areas in Greece. The sample was 2,535 high school students from the 10 to 12th grade, participating in the school physical education championships. The results showed that 9.6% of boys and 3.7% of girls reported that they had used anabolic drugs sometime in the past whereas 11.2% boys and 4.8% girls reported that they would intend to use them in the future. This confirms that anabolic steroids are an important problem among adolescents, and educational programs should increase their knowledge about these drugs. Information should come not only from the state, but also from coaches, teachers, trainers and parents.

La coordinación motriz en la Adolescencia y su relación con el IMC, hábitos de práctica y motivación en E.F: Estudio transversal y longitudinal

En las últimas décadas, ha aumentado el interés de la investigación sobre el desarrollo de la coordinación motriz en la adolescencia por ser una etapa sensible, crítica y crucial para la adquisición de hábitos y conductas saludables de vida. Estos estudios han mostrado que la adquisición de unos niveles óptimos de coordinación y competencia motriz van a ser determinantes para el bienestar del adolescente y van a estar relacionados e influidos por otras dimensiones del desarrollo de la persona. Recientes investigaciones han sacado a la luz datos alarmantes sobre el aumento de problemas de coordinación motriz en la población infantil y adolescente (Cantell, Smyth y Ahonen, 1994; Gómez, 2004; Ruiz, Graupera, Gutiérrez y Miyahara, 2003; Sudgen y Chambers, 2005) donde abrocharse los botones de una camisa o correr de forma armónica puede ser todo un mundo lleno de dificultades y consecuencias sobre otras dimensiones del desarrollo (Ramón-Otero y Ruiz, 2015). Estos problemas han sido tratados por investigadores como una “dificultad oculta” (Gómez, Ruiz y Mata, 2006), cuya manifestación está presente en las actividades de la vida cotidiana, en contextos deportivos, en juegos y/o en la clase de Educación Física (Ruiz, 2004). La preocupación por estas dificultades se ha extendido a nivel internacional, creando todo un campo de investigación que estudia el diagnóstico de éstos problemas, conocido bajo las siglas DCD (Developmental Coordination Disorder). El presente estudio se centra en la etapa adolescente, periodo de transición entre la etapa infantil y adulta, caracterizada por numerosos cambios biológicos, cognitivos y socioemocionales (Santrock, 2005), que van a determinar la adaptación con el entorno (Gallahue, Ozmun y Goodway, 2011; Gómez, Ruiz, y Mata, 2006). El propósito principal del estudio es analizar el desarrollo de la coordinación motriz en la etapa adolescente investigando las diferencias de género y de edad en relación con variables psicosociales, los hábitos de práctica y las variables antropométricas. El diseño de la investigación se estructura en dos estudios. El primero de ellos, de carácter transversal, analizó una muestra representativa de 1.966 adolescentes de 1º a 4º de la ESO. El segundo, de naturaleza longitudinal, utilizó un grupo de 89 adolescentes del estudio transversal los cuales fueron estudiados durante 4 años, desde los 12 a los 15 años. Los mismos instrumentos fueron utilizados en ambos estudios: el Test Sportcomp para la evaluación de la coordinación motriz, el test AMPET4 para valorar la motivación de logro para el aprendizaje en Educación Física, el inventario HBSC para conocer los hábitos saludables sobre la práctica de actividad física y, por ultimo, se utilizó un estadiómetro para obtener el peso y la altura y así calcular el índice de masa corporal (IMC). La toma de datos del Estudio Transversal se realizó en 2 cursos académicos (2011/12 - 2012/13), en la cual se requirieron 3 sesiones coincidiendo con la clase de Educación Física. En la primera sesión, se evaluó la coordinación motriz. En la segunda se aplicaron los cuestionarios (AMPET4 y HBSC) y, en la última sesión se midió el peso y la altura en un espacio reservado al estadiómetro. El análisis de datos fue descriptivo y diferencial de cada una de las variables estudiadas: motoras, psicosociales, de hábitos de práctica de actividad física y antropométricas. Asimismo, se llevaron a cabo pruebas de análisis univariante y multivariante, calculando el valor-p y las pruebas de efecto. Respecto al Estudio Longitudinal, la toma de datos se llevó cabo durante 4 años desde el 2011 al 2014. La evaluación de la coordinación motriz se realizó en cada uno de los 4 años. Sin embargo, los 2 cuestionarios y las medidas antropométricas fueron evaluadas en el primer y cuarto año. Los análisis de datos fueron descriptivos y comparativos entre las variables analizadas. En el caso de la coordinación motriz, se realizaron las pruebas de medidas repetidas y, en el caso de las demás variables analizadas, se realizaron Prueba T para muestras relacionadas. Los resultados globales mostraron que el índice motor en el Estudio Transversal fue progresivo en el conjunto de chicos. Sin embargo, en las chicas, el rendimiento se estabiliza a partir de los 13 años. En el caso del Estudio Longitudinal, este índice se estabiliza en los 3 primeros años y a la edad de los 14, es cuando comienzan a acusarse las diferencias de género. En el caso de los hombres el rendimiento mejora y, por el contrario, en las mujeres empeora. En el Estudio Transversal, el análisis de varianza mostraron diferencias en función de la edad [F(7, 1958) = 220.70, p < .001; η2 = .101], del género [F(7, 1958) = 29.76, p < .001; η2 = .044], así como en la interacción entre ambos [F(7, 1958)= 11.90, p < .001; η2 = .018]. Únicamente aparecieron diferencias significativas con la edad en todos los grupos de hombres, excepto entre 14 y 15 años. En el Longitudinal, los contrastes multivariados mostraron que no hubo diferencias sgnificativas en el tiempo [F(3,85) = .05, p = .987, η2= .002] mostrando un nivel de coordinación estable a lo largo de los años, aunque existieron diferencias entre chicos y chicas [F(3,85) = 4.64 p = .005] con un tamaño de efecto destacable (η2 = .141). En cuanto a la motivación de logro para prender en Educación Física, en ambos estudios, los chicos fueron los que obtuvieron puntuaciones más elevadas en todas las dimensiones positivas del test (compromiso de aprendizaje, competencia autopercibida y comparada). Sin embargo, en la dimensión negativa del test, la referida a la ansiedad y al agobio ante el fracaso, fueron las chicas las que puntuaron más alto. En el Estudio Transversal, los resultados mostraron diferencias significativas en todas las dimensiones del AMPET4 en función del nivel de coordinación motriz: compromiso con el aprendizaje [F(2, 1644) = 8.66, p < .001; η2 = .010], competencia autopercibida [F(2, 1644) = 50.94, p < .001; η2 = .048], competencia comparada [F(2, 1644) = 41.56, p < .001, η2 = .020] y ansiedad [F(2, 1644) = 16.67, p < .001, η2 = .058]. En este sentido, los grupos de mejor nivel de coordinación motriz, fueron los que mayor puntuación obtuvieron en las dimensiones positivas y los que menor, en la negativa. En el Estudio Longitudinal, también se encontraron diferencias entre el primer y cuarto año de estudio en todas las dimensiones, excepto en competencia motriz autopercibida. Estas diferencias se tradujeron en una disminución en las 3 variables significativas del primer al cuarto año. Respecto al inventario HBSC, en el Estudio Longitudinal, la prueba T mostró únicamente la existencia de diferencias significativas entre el primer y cuarto año en 2 de los 11 ítems: percepción de la forma física (p = .006) y percepción de la salud (p = .047), los cuales disminuyeron en el intervalo de tiempo del estudio. En el Transversal, las diferencias se observaron en función del género (p < .001) y de la edad (p < .001). Asimismo, se mostraron diferencias significativas en todos los ítems respecto al nivel de coordinación motriz, excepto en 2 de ellos: frecuencia tiempo libre con los amigos fuera del colegio (p = .580) y facilidad para hacer amigos en el centro escolar (p = .098). Por último, en las variables antropométricas, los resultados del Estudio Transversal y Longitudinal coinciden tanto en la estatura como en el peso, apuntando, que en ambos estudios, se produce un aumento progresivo tanto en chicos como en chicas a medida que se avanza en edad. Concretamente en el Transversal, estas diferencias en la edad se encuentran en todos los grupos en ambos géneros, excepto en el conjunto de chicas entre los 14 y los 15 años. Asimismo, ambos estudios coincidieron en que tanto las ganancias en cm y kg, como las puntuaciones medias, fueron mayores en los chicos que en las chicas. Respecto al IMC, los 2 estudios coincidieron en que la evolución es paralela, y tal y como apuntan los resultados del Transversal, no se encontraron diferencias ni en la edad (p = 792) ni en el género (p = 284). No obstante, el Longitudinal apuntó únicamente diferencias significativas entre el primer y cuarto año en el conjunto de los hombres [t(41) = -4.01, p < .001]. Finalmente, y en relación con los niveles de coordinación motriz, hubo diferencias significativas en relación con el IMC (p = .012), mostrando como el grupo de peso normal coincide con puntuaciones óptimas de coordinación motriz. A modo de conclusiones, el presente estudio revela cómo la adquisición de un nivel de coordinación óptimo va a ser fundamental para el desarrollo psicosocial, para el desarrollo de hábitos saludables de práctica y para mantener un IMC dentro de la normalidad para el género y la edad. De esta manera, el desarrollo de la coordinación motriz será un aspecto fundamental para lograr un estado de bienestar físico y mental, y unos hábitos favorables para la práctica de actividad física. ABSTRACT In the past couple of decades, adolescence stage in motor coordination gained significant interest in research especially due to its sensitive and critical importance to achieving a healthy life style. These studies observed how to acquire optimum levels of coordination and motor competence, which proved crucial to the quality of the adolescent stage in addition to being influenced by other dimensions of development for each individual. Recent research shed light to an alarming set of data, which showed increased motor coordination problems in children and adolescents (Cantell, Smyth & Ahonen, 1994; Gómez, 2004; Ruiz, Graupera, Gutierrez & Miyahara, 2003; Sugden & Chambers, 2005). For instance, even to the extent that buttoning a shirt or running in a harmonic form can lead to a whole set of consequences and difficulties on the development stage. Researchers have addressed such problems in various studies such as “dificultad oculta” (Gomez, Ruiz & Mata, 2006), which literally translates as “hidden trouble”. The studies are evidently present in the activities of daily life, sporting contexts, games and/or Physical Education (Ruiz, 2004). Concern about these difficulties spread internationally, creating a whole framework research studying the diagnosis of these problems, known under the acronym DCD (Developmental Coordination Disorder). The study focuses on the adolescent stage, transition period between childhood and adulthood characterized by numerous biological, cognitive and socio-emotional changes (Santrock, 2005), which interestingly determines an individual´s adaptation to the environment (Gallahue, Ozmun & Goodway, 2011; Gomez, Ruiz & Mata, 2006). The main purpose of the study is to analyse the development of motor coordination in the adolescent stage investigating gender differences and age in relation to psychological variables, physical activity habits and anthropometric variables. The research design is structured in two studies. The first (transversal nature), analyses a representative sample of 1,966 adolescents from 1st to 4th of Secondary Education School. The second (longitudinal nature) used a group of 89 teenagers from cross-sectional study, which were studied for four years, from 12 to 15 years. The same instruments were used in both studies, namely; “Sportcomp Test” used to evaluate of motor coordination; “AMPET4 Test” which assesses the motivational achievement of learning Physical Education; “HBSC Inventory” to find out the healthy habits gained from physical activities; And finally a “stadiometer” was used to obtain the weight and height and thus calculate the body mass index (BMI). The data collection of the cross-sectional Study was conducted in two academic years (2011/12 - 2012/13), in which 3 sessions coinciding with the Physical Education level are required. In the first session, motor coordination was evaluated; questionnaires were applied in the second session (AMPET4 and HBSC); and in the last session the weight and height were measured in a reserved space for the “stadiometer”. Notably, data analysis was descriptive and differential in each of the variable studies: motor, psychological, practical and anthropometric habits of physical activity. Thus the tests were conducted in a univariate and multivariate analysis, calculating the p-value and effect tests. Regarding the Longitudinal Study, data collection was carried out during four years from 2011 to 2014 inclusively. The assessment of motor coordination was performed on each of the four years, however, the 2 questionnaires and anthropometric measures were evaluated in the first and fourth year. Data analyses were also descriptive and comparative among the variables that were put to the test. In the case of motor coordination tests, they were done on repeated measures, whilst, in the case of other variables analysed, they were accomplished through T Tests under comparable samples. The overall results showed that the engine Motor Index in Study 1 was progressive in all male gender studies, however in the females the performance remained constant after reaching 13 years of age. For the Longitudinal Study, this index is stabilized in the first 3 years and at the age of 14 is when the gender differences take place. In the case of males, the performance improves, however, in females worsens. The cross-sectional Study, analysis of variance showed differences in terms of age [F(7, 1958) = 220.70, p < .001; η2 = .101], gender [F(7, 1958) = 29.76, p <.001; η2 = .044], as well as their interaction [F(7, 1958) = 11.90, p <.001; η2 = .018]. They only show significant differences in respect to age in the male set sample, in all groups except between 14 and 15 years old. In the Longitudinal, the multivariate contrasts showed no significant differences in time [F(3,85) = 0.05, p = 0.987, η2 = 0.002] showing a stable level of coordination over the years, but if there were differences between both genders [F(3,85) = 4.64, p = .005] it took place with a noteworthy effect size (η2 = .141). In regards, to the Motivational Achievement for learning Physical Education, in both studies the male sample administered obtained higher scores on all the positive dimensions of the test (commitment to learning, self-assessed competence, and comparable competence). However, on the negative assessment side, namely, anxiety and fear of failure, the female sample scored higher than the male one. In Study 1, the multivariate analysis showed significant differences between the psychosocial dimensions and levels of motor coordination with moderate to significant effect [Lambada de Wilks = .931, F(8, 3282) = 14.99; p = <0.001; η2 = .035]. By the same token, the groups with the best level of motor coordination were the highest scoring ones in the positive dimensions, whilst the lower performing ones, performed better in the negative dimension. In the longitudinal study, there is also differences were also found between the first and fourth years of study in all dimensions, except in self-perceived motor competition. These differences resulted in a significant decrease in the 3 variables from first to fourth year. Regarding, the “HBSC Inventory”, the T test in the longitudinal study showed uniquely the existence of significant differences between the first and fourth year in 2 of the 11 items: perception of physical fitness (p = .006) and perceived health (p = 047), which diminished in the interval time of the study. In the Cross-sectional study, the se differences were also observed in gender (p < .001) and age (p < .001). Similarly, they showed significant differences in all items in respect to the motor coordination level, except in 2 of them; frequency of free time with friends outside of school (p = .580) and the ease to make friends at the educational centre (p = 098). And last but not least, the anthropometric variables, both the results of the Transversal and Longitudinal Study matched both height and weight, pointing out that in both studies a gradual increase in both genders, as they grow older. Notably in the Cross-sectional, these differences in age are found in all groups in both genders, except for the set of girls between 14 and 15 years. Thus both studies concluded that both gains in cm and kg and the mean scores were higher amongst males compared to females. Regarding BMI, the 2 studies concluded that the evolution is parallel, and as pointed cross-sectional study there isn’t differences found in age (p = 792) or in gender (p = 284). However, the Longitudinal study uniquely shows significant difference between the first and fourth year for male set sample [t (41) = -4.01, p < .001]. Finally, in relation to levels of motor coordination, there were significant differences in relation to BMI (p = .012), showing how the “normal weight group” matches the optimal scores of motor coordination. In conclusion, this study reveals how the acquisition of an optimal level of coordination is vital for psychological development, to develop and practice healthy habits, and to maintain a BMI within the normal range for age and gender. Therefore, the development of motor coordination is fundamental to achieving a state of physical and mental wellbeing, and preferable habits to pursuing physical activity.

Nuevos desarrollos en la medida de descargas parciales y técnicas avanzadas de análisis para el diagnóstico del estado de los aislamientos en sistemas eléctricos de alta tensión

En esta tesis se desarrolla una investigación sobre las técnicas de medida de descargas parciales (DP) que se aplican en medidas on-line, en condiciones normales de operación de las instalaciones eléctricas de alta tensión (AT). También se realiza un estudio de técnicas avanzadas de procesado y análisis de las señales medidas, que permiten realizar diagnósticos precisos del estado de los aislamientos eléctricos de AT. Uno de los objetivos fundamentales de la tesis ha sido disponer de un procedimiento eficaz de medida y procesado de las señales de DP, para la realización de medidas on-line tanto de forma itinerante, como mediante monitorización temporal o permanente. La implementación del nuevo procedimiento de medida permite obtener resultados satisfactorios en la detección, identificación y localización de defectos de aislamiento. Se ha dedicado especial interés al desarrollo de un método de clasificación de señales, que permite separar pulsos de ruido y diferentes fuentes de DP, presentes de forma simultánea en las instalaciones de AT. El estudio de la clasificación de señales se ha completado con la aplicación de un método para la detección de manera asistida, de los diferentes grupos de pulsos de ruido y de DP. La aplicación de este método de detección de grupos de pulsos, facilita la labor de los técnicos especialistas a la hora de diagnosticar el estado de los elementos aislantes. Al efecto de verificar de forma práctica las aportaciones de la tesis, se han realizado medidas de DP tanto en laboratorio como en campo. Las medidas experimentales en laboratorio se han efectuado en el Laboratorio de Alta Tensión de la Escuela Técnica Superior de Ingeniería y Diseño Industrial (LAT-UPM), de la Universidad Politécnica de Madrid. Por otro lado, las medidas experimentales en campo se han llevado a cabo en instalaciones de AT propiedad de compañías de transporte y distribución de energía eléctrica. La realización de ensayos de DP en estas instalaciones ha sido posible, gracias a los proyectos de investigación llevados a cabo por el grupo de trabajo del LAT-UPM, con diferentes empresas del sector durante los diez últimos años. ABSTRACT This thesis develops techniques for measuring partial discharges (PD) that are applied in on-line measurements, under normal operating conditions of the high voltage (HV) electrical installations. In addition there are studied advanced techniques for the processing and analysis of the measured signals, that permit precise diagnostics of the state of HV electrical insulation systems. One of the fundamental objectives of the thesis is to make available an effective procedure for measuring and processing PD signals, for making on-line measurements, either in an itinerant way or in temporary or permanent monitoring. The implementation of the new measurement procedure yields satisfactory results in the detection, identification and localization of insulation defects. Special attention has been devoted to the development of a method for classifying signals, that separates noise pulses and various PD sources present simultaneously in the HV installations. The study of the classification of signals has been completed by the application of a method for detecting, in a user assisted manner, the different groups of noise pulses and of PD. The application of this method for detecting groups of pulses facilitates the work of the specialist technicians to diagnose the condition of the insulation elements. To demonstrate the practical value of the thesis, PD measurements were made in laboratory as well as in field installations. The experimental measurements in laboratory were made in the High Voltage Laboratory (LAT-UPM) of the High Technical School of Engineering and Industrial Design, of the Polytechnic University of Madrid. Field measurements were realized in the HV installations of companies providing electrical energy transport and distribution. The realization of PD tests in these facilities was possible thanks to the research projects carried out by the working group of the LAT-UPM during the last ten years, with different companies operating in the sector.

1.6 School of Law, Lincoln University

Another dilemma also had to be dealt with; Lloyd Gaines was determined to attend law school, not just anywhere but at the University of Missouri. Shortly after the Supreme Court decision, Lloyd Gaines left his civil service job in Michigan and returned home to St. Louis, arriving on New Year’s Eve, 1938. In the meantime, to pay his bills, he took a job as a filling station attendant. On January 9, 1939, Gaines spoke to the St. Louis chapter of the NAACP. He told them he stood “ready, willing, and able to enroll at MU.” Gaines later quit his gas station job. He explained to his family that the station owner substituted inferior gas and that he could not, in good conscience, continue to work there. In the meantime, the state Supreme Court sent the Gaines case back to Boone County to determine whether the new law school at Lincoln would comply with the US Supreme Court’s requirement of “substantial equality.”

ESCOLA PÚBLICA DE TEMPO INTEGRAL: UM ESTUDO DE CASO DAS REPRESENTAÇÕES SOCIAIS DOS PROFESSORES SOBRE OS PROCESSOS FORMATIVOS

O presente trabalho analisa as origens do modelo de Escola Pública de Tempo Integral, a partir de uma abordagem histórica que compara dois modelos: o da Escola-parque e a proposta implantada em 2006, no Estado de São Paulo. Investiga a questão com base na Teoria das Representações Sociais de Moscovici com o intuito de desvelar as representações sociais de professores, para aprofundar a reflexão sobre a sua inserção e preparação para a atuação neste modelo de escola. Prioriza, também, uma discussão sobre uma divisão que se percebe no contexto da Escola Pública de Tempo Integral, que se refere a uma distinção entre a proposta de atividades desenvolvidas sobre as atividades do currículo normal (consideradas mais importantes) e as atividades de extensão do currículo oficinas (consideradas menos importantes), cujas diferenças são geradas pela ausência de preparação destes profissionais nos processos formativos do projeto.(AU)

ESCOLA PÚBLICA DE TEMPO INTEGRAL: UM ESTUDO DE CASO DAS REPRESENTAÇÕES SOCIAIS DOS PROFESSORES SOBRE OS PROCESSOS FORMATIVOS

O presente trabalho analisa as origens do modelo de Escola Pública de Tempo Integral, a partir de uma abordagem histórica que compara dois modelos: o da Escola-parque e a proposta implantada em 2006, no Estado de São Paulo. Investiga a questão com base na Teoria das Representações Sociais de Moscovici com o intuito de desvelar as representações sociais de professores, para aprofundar a reflexão sobre a sua inserção e preparação para a atuação neste modelo de escola. Prioriza, também, uma discussão sobre uma divisão que se percebe no contexto da Escola Pública de Tempo Integral, que se refere a uma distinção entre a proposta de atividades desenvolvidas sobre as atividades do currículo normal (consideradas mais importantes) e as atividades de extensão do currículo oficinas (consideradas menos importantes), cujas diferenças são geradas pela ausência de preparação destes profissionais nos processos formativos do projeto.(AU)

Temporal distribution of the ganglion cell volleys in the normal rat optic nerve

We describe experiments on behaving rats with electrodes implanted on the cornea, in the optic chiasm, and on the visual cortex; in addition, two red light-emitting diodes (LED) are permanently attached to the skull over the left eye. Recordings timelocked to the LED flashes reveal both the local events at each electrode site and the orderly transfer of visual information from retina to cortex. The major finding is that every stimulus, regardless of its luminance, duration, or the state of retinal light adaptation, elicits an optic nerve volley with a latency of about 10 ms and a duration of about 300 ms. This phenomenon has not been reported previously, so far as we are aware. We conclude that the retina, which originates from the forebrain of the developing embryo, behaves like a typical brain structure: it translates, within a few hundred milliseconds, the chemical information in each pattern of bleached photoreceptors into a corresponding pattern of ganglion cell neuronal information that leaves via the optic nerve. The attributes of each rat ganglion cell appear to include whether the retinal neuropile calls on it to leave after a stimulus and, if so when, within a 300-ms poststimulus epoch. The resulting retinal analysis of the scene, on arrival at the cortical level, is presumed to participate importantly in the creation of visual perceptual experiences.

Immucillin H, a powerful transition-state analog inhibitor of purine nucleoside phosphorylase, selectively inhibits human T lymphocytes

Transition-state theory has led to the design of Immucillin-H (Imm-H), a picomolar inhibitor of purine nucleoside phosphorylase (PNP). In humans, PNP is the only route for degradation of deoxyguanosine, and genetic deficiency of this enzyme leads to profound T cell-mediated immunosuppression. This study reports the biological effects and mechanism of action of Imm-H on malignant T cell lines and on normal activated human peripheral T cells. Imm-H inhibits the growth of malignant T cell leukemia lines with the induction of apoptosis. Imm-H also inhibits activated normal human T cells after antigenic stimulation in vitro. However, Imm-H did not inhibit malignant B cells, colon cancer cell lines, or normal human nonstimulated T cells, demonstrating the selective activity of Imm-H. The effects on leukemia cells were mediated by the cellular phosphorylation of deoxyguanosine and the accumulation of dGTP, an inhibitor of ribonucleotide diphosphate reductase. Cells were protected from the toxic effects of Imm-H when deoxyguanosine was absent or when deoxycytidine was present. Guanosine incorporation into nucleic acids was selectively blocked by Imm-H with no effect on guanine, adenine, adenosine, or deoxycytidine incorporation. Imm-H may have clinical potential for treatment of human T cell leukemia and lymphoma and for other diseases characterized by abnormal activation of T lymphocytes. The design of Imm-H from an enzymatic transition-state analysis exemplifies a powerful approach for developing high-affinity enzyme inhibitors with pharmacologic activity.

The pKa of His-24 in the folding transition state of apomyoglobin

In native apomyoglobin, His-24 cannot be protonated, although at pH 4 the native protein forms a molten globule folding intermediate in which the histidine residues are readily protonated. The inability to protonate His-24 in the native protein dramatically affects the unfolding/refolding kinetics, as demonstrated by simulations for a simple model. Kinetic data for wild type and for a mutant lacking His-24 are analyzed. The pKa values of histidine residues in native apomyoglobin are known from earlier studies, and the average histidine pKa in the molten globule is determined from the pH dependence of the equilibrium between the native and molten globule forms. Analysis of the pH-dependent unfolding/refolding kinetics reveals that the average pKa of the histidine residues, including His-24, is closely similar in the folding transition state to the value found in the molten globule intermediate. Consequently, protonation of His-24 is not a barrier to refolding of the molten globule to the native protein. Instead, the normal pKa of His-24 in the transition state, coupled with its inaccessibility in the native state, promotes fast unfolding at low pH. The analysis of the wild-type results is confirmed and extended by using the wild-type parameters to fit the unfolding kinetics of a mutant lacking His-24.

Tetraploid State Induces p53-dependent Arrest of Nontransformed Mammalian Cells in G1

A “spindle assembly” checkpoint has been described that arrests cells in G1 following inappropriate exit from mitosis in the presence of microtubule inhibitors. We have here addressed the question of whether the resulting tetraploid state itself, rather than failure of spindle function or induction of spindle damage, acts as a checkpoint to arrest cells in G1. Dihydrocytochalasin B induces cleavage failure in cells where spindle function and chromatid segregation are both normal. Notably, we show here that nontransformed REF-52 cells arrest indefinitely in tetraploid G1 following cleavage failure. The spindle assembly checkpoint and the tetraploidization checkpoint that we describe here are likely to be equivalent. Both involve arrest in G1 with inactive cdk2 kinase, hypophosphorylated retinoblastoma protein, and elevated levels of p21WAF1 and cyclin E. Furthermore, both require p53. We show that failure to arrest in G1 following tetraploidization rapidly results in aneuploidy. Similar tetraploid G1 arrest results have been obtained with mouse NIH3T3 and human IMR-90 cells. Thus, we propose that a general checkpoint control acts in G1 to recognize tetraploid cells and induce their arrest and thereby prevents the propagation of errors of late mitosis and the generation of aneuploidy. As such, the tetraploidy checkpoint may be a critical activity of p53 in its role of ensuring genomic integrity.

Glutamate is required to maintain the steady-state potassium pool in Salmonella typhimurium.

In many bacteria, accumulation of K+ at high external osmolalities is accompanied by accumulation of glutamate. To determine whether there is an obligatory relationship between glutamate and K+ pools, we studied mutant strains of Salmonella typhimurium with defects in glutamate synthesis. Enteric bacteria synthesize glutamate by the combined action of glutamine synthetase and glutamate synthase (GS/GOGAT cycle) or the action of biosynthetic glutamate dehydrogenase (GDH). Activity of the GS/GOGAT cycle is required under nitrogen-limiting conditions and is decreased at high external ammonium/ammonia ((NH4)+) concentrations by lowered synthesis of GS and a decrease in its catalytic activity due to covalent modification (adenylylation by GS adenylyltransferase). By contrast, GDH functions efficiently only at high external (NH4)+ concentrations, because it has a low affinity for (NH4)+. When grown at low concentrations of (NH4)+ (< or = 2 mM), mutant strains of S. typhimurium that lack GOGAT and therefore are dependent on GDH have a low glutamate pool and grow slowly; we now demonstrate that they have a low K+ pool. When subjected to a sudden (NH4)+ upshift, strains lacking GS adenylyltransferase drain their glutamate pool into glutamine and grow very slowly; we now find that they also drain their K+ pool. Restoration of the glutamate pool in these strains at late times after shift was accompanied by restoration of the K+ pool and a normal growth rate. Taken together, the results indicate that glutamate is required to maintain the steady-state K+ pool -- apparently no other anion can substitute as a counter-ion for free K+ -- and that K+ glutamate is required for optimal growth.

Thrombospondin 1 expression in transformed endothelial cells restores a normal phenotype and suppresses their tumorigenesis.

Murine endothelial cells are readily transformed in a single step by the polyomavirus oncogene encoding middle-sized tumor antigen. These cells (bEND.3) form tumors (hemangiomas) in mice which are lethal in newborn animals. The bEND.3 cells rapidly proliferate in culture and express little or no thrombospondin 1 (TS1). To determine the role of TS1 in regulation of endothelial cell phenotype, we stably transfected bEND.3 cells with a human TS1 expression vector. The cells expressing human TS1 were readily identified by their altered morphology and exhibited a slower growth rate and lower saturation density than the parental bEND.3 cells. The TS1-expressing cells also formed aligned cords of cells instead of clumps or cysts in Matrigel. Moreover, while the bEND.3 cells formed large tumors in nude mice within 48 hr, the TS1-expressing cells failed to form tumors even after 1 month. The TS1-transfected cells expressed transforming growth factor beta mRNA and bioactivity at levels similar to those of the parental or vector-transfected bEND.3 cells, indicating that the effects of TS1 expression are not due to the activation of transforming growth factor beta by TS1. TS1 expression resulted in a > 100-fold decrease in net fibrinolytic (urokinase-type plasminogen activator, uPA) activity due to more plasminogen-activator inhibitor 1 and less uPA secretion. TS1 thus appears to be an important regulator of endothelial cell phenotype required for maintaining the quiescent, differentiated state.

N-tert-butyl-alpha-phenylnitrone improves recovery of brain energy state in rats following transient focal ischemia.

Recent results have demonstrated that the spin trapping agent N-tert-butyl-alpha-phenylnitrone (PBN) reduces infarct size due to middle cerebral artery occlusion (MCAO), even when given after ischemia. The objective of the present study was to explore whether PBN influences recovery of energy metabolism. MCAO of 2-hr duration was induced in rats by an intraluminal filament technique. Brains were frozen in situ at the end of ischemia and after 1, 2, and 4 hr of recirculation. PBN was given 1 hr after recirculation. Neocortical focal and perifocal ("penumbra") areas were sampled for analyses of phosphocreatine (PCr), creatine, ATP, ADP, AMP, glycogen, glucose, and lactate. The penumbra showed a moderate-to-marked decrease and the focus showed a marked decrease in PCr and ATP concentrations, a decline in the sum of adenine nucleotides, near-depletion of glycogen, and an increase in lactate concentration after 2 hr of ischemia. Recirculation for 1 hr led to only a partial recovery of energy state, with little further improvement after 2 hr and signs of secondary deterioration after 4 hr, particularly in the focus. After 4 hr of recirculation, PBN-treated animals showed pronounced recovery of energy state, with ATP and lactate contents in both focus and penumbra approaching normal values. Although an effect of PBN on mitochondria cannot be excluded, the results suggest that PBN acts by preventing a gradual compromise of microcirculation. The results justify a reevaluation of current views on the pathophysiology of focal ischemic damage and suggest that a therapeutic window of many hours exists in stroke.

School Principals' Attitudes Towards Performance Based Compensation

The main objective of this study is to determine the attitudes of school principals regarding a performance based compensation system. This study identifies the attitudes towards specific factors that should be considered in the implementation of a system of performance based compensation. The data have been analyzed to determine if a principal's demographic characteristics affect his/her level of agreement with performance based compensation and the factors for implementation. In addition, this study unveils areas of concern that principals have conveyed regarding the implementation of a performance based compensation system. Data was obtained from 444 public school principals representing 444 schools and 178 districts in the state of Colorado. Measures used in the treatment of the data include descriptive statistics and one-way ANOVA. The major findings of this study were: 1. 82.4% of respondents believe that teachers, principals and administrators should be included in performance based compensation (PBC). 2. The top two indicators that respondents believed should be included in a PBC system are student achievement (88.5%) and teacher evaluations (77.6%) 3. The 3 largest obstacles to PBC that respondents identified are: a. The capacity to link student achievement to teacher evaluations (82.9%) b. Teacher Union Resistance (67.1%) c. Cost (55.9%) 4. Principals in urban, rural and suburban geographic groups disagree about the effects of performance based compensation. 5. The top 5 overall concerns regarding Performance Based Compensation were: a. Concerns regarding effectively using assessment to measure performance of all teachers/equity between teachers b. Concerns regarding evaluation (time for principals to learn, consistency from school to school, time for principals to evaluate, quality of evaluation tool). c. Not in favor of PBC due to philosophical views or concerns about lack of research. d. Concerns regarding the equity between classrooms and districts across the state due to poverty levels and unequal resources. e. Concerns that performance based compensation will result in a decline in teacher collaboration and an increase in competition between teachers. Based upon these findings, the researcher concluded that there is not a strong general acceptance of performance based compensation systems. However, urban principals in Colorado tend to view PBC somewhat more favorably than do principals in suburban or rural areas. Most importantly, systems to link student achievement to teacher evaluation must be collaboratively created to ensure PBC systems are equitable, consistent and fair.

Variabilidade intratípica de HPV 16 em relação à origem étnica e HLA de uma população de alto risco para o câncer do colo do útero

A infecção por papilomavirus é a principal causa de desenvolvimento de neoplasias intraepiteliais cervicais (NIC) e câncer do colo do útero (CCU). Estudos epidemiológicos têm demonstrado que a persistência do genoma viral encontra-se associado a variantes moleculares específicas de papilomavirus humano (HPV) de alto risco. As moléculas HLA de classe II têm um importante papel na resposta imune. Associações entre HLA e CCU ou infecção por HPV tem sido demonstrado em diferentes populações. O nosso objetivo foi verificar se a variabilidade de HLA-DRB1 e DQB1 estavam associada ao CCU e NIC III em mulheres de Belém, uma população formada pelos 3 principais grupos étnicos humanos e uma área de alto risco para o CCU no Norte do Brasil. Foi investigada a existência de diferenças na distribuição de alelos HLA entre mulheres com CCU e NIC III portadoras de diferentes variantes de HPV-16 e mulheres citologicamente normais. Os genes HLA DQB1 e DRB1 foram tipados pelo método de PCR-SSO em 95 casos e 287 controles de mulheres com citologia normal atendidas em um centro de prevenção do colo do útero na mesma cidade. As variantes de HPV-16 foram tipadas por sequenciamento de um fragmento da região controladora do genoma viral (LCR). O polimorfismo na posição 350 do gene E6 foi tipado baseado em um protocolo de hibridização em pontos, para identificar a alteração na posição 350T→G. A magnitude das associações foi estimada por odds ratio (OR) e os respectivos intervalos de confiança (IC), ajustados para potenciais fatores de confusão. Uma associação positiva foi observada entre CCU e os haplótipos DRB1* 150 l-DQB1*0602, DRB1*04-DQB1*0301 e DRB1*1602-DQB1*0301. Ao contrário, DRB1*01-DQB1*0501 mostrou um efeito protetor. Os alelos DRB1*0804, DQB1*0402 apresentaram efeito protetor contra positividade por HPV. O alelo DQB1*0502 e o grupo DRB1*15 foram positivamente associados. Os nossos resultados mostram que as associações positivas de DRB1*1501 e DRB1*1602 podem ser atribuídas a variantes asiático-americanas quando comparado a variantes européias. O risco conferido a DRB1*1501 foi encontrado associado tanto a variantes E6350G quanto a variantes E6350T, entretanto, o maior efeito foi devido às variantes E6250T. A associação positiva de DRB1*1602 foi significativa somente no grupo de mulheres positivas para E6350G. Estes resultados estão de acordo com a composição étnica da população estudada bem como um maior potencial oncogênico de certas variantes. Nossos dados sugerem que a contribuição dos alelos HLA na susceptibilidade genética ao CCU difere de acordo com a distribuição das variantes de HPV em uma dada região geográfica ou grupo étnico.