Microarray based real-time analysis of nucleic acid hybridization kinetics and thermodynamics

Ziel dieser Dissertation ist die experimentelle Charakterisierung und quantitative Beschreibung der Hybridisierung von komplementären Nukleinsäuresträngen mit oberflächengebundenen Fängermolekülen für die Entwicklung von integrierten Biosensoren. Im Gegensatz zu lösungsbasierten Verfahren ist mit Microarray Substraten die Untersuchung vieler Nukleinsäurekombinationen parallel möglich. Als biologisch relevantes Evaluierungssystem wurde das in Eukaryoten universell exprimierte Actin Gen aus unterschiedlichen Pflanzenspezies verwendet. Dieses Testsystem ermöglicht es, nahe verwandte Pflanzenarten auf Grund von geringen Unterschieden in der Gen-Sequenz (SNPs) zu charakterisieren. Aufbauend auf dieses gut studierte Modell eines House-Keeping Genes wurde ein umfassendes Microarray System, bestehend aus kurzen und langen Oligonukleotiden (mit eingebauten LNA-Molekülen), cDNAs sowie DNA und RNA Targets realisiert. Damit konnte ein für online Messung optimiertes Testsystem mit hohen Signalstärken entwickelt werden. Basierend auf den Ergebnissen wurde der gesamte Signalpfad von Nukleinsärekonzentration bis zum digitalen Wert modelliert. Die aus der Entwicklung und den Experimenten gewonnen Erkenntnisse über die Kinetik und Thermodynamik von Hybridisierung sind in drei Publikationen zusammengefasst die das Rückgrat dieser Dissertation bilden. Die erste Publikation beschreibt die Verbesserung der Reproduzierbarkeit und Spezifizität von Microarray Ergebnissen durch online Messung von Kinetik und Thermodynamik gegenüber endpunktbasierten Messungen mit Standard Microarrays. Für die Auswertung der riesigen Datenmengen wurden zwei Algorithmen entwickelt, eine reaktionskinetische Modellierung der Isothermen und ein auf der Fermi-Dirac Statistik beruhende Beschreibung des Schmelzüberganges. Diese Algorithmen werden in der zweiten Publikation beschrieben. Durch die Realisierung von gleichen Sequenzen in den chemisch unterschiedlichen Nukleinsäuren (DNA, RNA und LNA) ist es möglich, definierte Unterschiede in der Konformation des Riboserings und der C5-Methylgruppe der Pyrimidine zu untersuchen. Die kompetitive Wechselwirkung dieser unterschiedlichen Nukleinsäuren gleicher Sequenz und die Auswirkungen auf Kinetik und Thermodynamik ist das Thema der dritten Publikation. Neben der molekularbiologischen und technologischen Entwicklung im Bereich der Sensorik von Hybridisierungsreaktionen oberflächengebundener Nukleinsäuremolekülen, der automatisierten Auswertung und Modellierung der anfallenden Datenmengen und der damit verbundenen besseren quantitativen Beschreibung von Kinetik und Thermodynamik dieser Reaktionen tragen die Ergebnisse zum besseren Verständnis der physikalisch-chemischen Struktur des elementarsten biologischen Moleküls und seiner nach wie vor nicht vollständig verstandenen Spezifizität bei.

An analysis of variance type method to describe and compare steady states in clinical data

Identifying and comparing different steady states is an important task for clinical decision making. Data from unequal sources, comprising diverse patient status information, have to be interpreted. In order to compare results an expressive representation is the key. In this contribution we suggest a criterion to calculate a context-sensitive value based on variance analysis and discuss its advantages and limitations referring to a clinical data example obtained during anesthesia. Different drug plasma target levels of the anesthetic propofol were preset to reach and maintain clinically desirable steady state conditions with target controlled infusion (TCI). At the same time systolic blood pressure was monitored, depth of anesthesia was recorded using the bispectral index (BIS) and propofol plasma concentrations were determined in venous blood samples. The presented analysis of variance (ANOVA) is used to quantify how accurately steady states can be monitored and compared using the three methods of measurement.

Hypoxia up-regulates expression of Eph receptors and ephrins in mouse skin.

Eph receptor tyrosine kinases and their ligands (ephrins) are key players during the development of the embryonic vasculature; however, their role and regulation in adult angiogenesis remain to be defined. Both receptors and ligands have been shown to be up-regulated in a variety of tumors. To address the hypothesis that hypoxia is an important regulator of Ephs/ephrins expression, we developed a mouse skin flap model of hypoxia. We demonstrate that our model truly represents segmental skin hypoxia by applying four independent methods: continuous measurement of partial cutaneous oxygen tension, monitoring of tissue lactate/pyruvate ratio, time course of hypoxia-inducible factor-1alpha (HIF-1alpha) induction, and localization of stabilized HIF-1alpha by immunofluorescence in the hypoxic skin flap. Our experiments indicate that hypoxia up-regulates not only HIF-1alpha and vascular endothelial growth factor (VEGF) expression, but also Ephs and ephrins of both A and B subclasses in the skin. In addition, we show that in Hep3B and PC-3 cells, the hypoxia-induced up-regulation of Ephs and ephrins is abrogated by small interfering RNA-mediated down-regulation of HIF-1alpha. These novel findings shed light on the role of this versatile receptor/ligand family in adult angiogenesis. Furthermore, our model offers considerable potential for analyzing distinct mechanisms of neovascularization in gene-targeted mice.

Medida de la conductividad térmica con el método de la aguja térmica, basado en la fuente lineal de calor transitorio, para su aplicación en los cerramientos de adobes y bloques de tierra comprimida

Existe un creciente interés internacional por el ahorro energético y la sostenibilidad en la edificación con importantes repercusiones en la Arquitectura. La inercia térmica es un parámetro fundamental para poder valorar energéticamente un edificio en condiciones reales. Para ello es necesario cambiar el enfoque tradicional de transmisión de calor en régimen estacionario por otro en régimen dinámico en el que se analizan las ondas térmicas y el flujo de calor oscilante que atraviesan los cerramientos. Los parámetros que definen la inercia térmica son: el espesor, la difusividad y el ciclo térmico. A su vez la difusividad está determinada por la conductividad térmica, la densidad y el calor específico del material. De estos parámetros la conductividad es el más complejo, variable y difícil de medir, especialmente en los cerramientos de tierra debido a su heterogeneidad y complejidad higrotérmica. En general, los métodos de medida de la conductividad o transmitancias en los paramentos presentan inconvenientes a la hora de medir un edificio construido con tierra: dificultades de implementación, el elevado coste o la fiabilidad de los resultados, principalmente. El Método de la Aguja Térmica (MAT) se basa en el principio de la evolución en el tiempo del calor emitido por una fuente lineal al insertarse en el seno de un material. Se ha escogido este método porque resulta práctico, de bajo coste y de fácil aplicación a gran escala pero tiene serios problemas de fiabilidad y exactitud. En esta tesis se desarrolla un método de medida de la conductividad térmica para Piezas de Albañilería de Tierra Cruda en laboratorio basado en el MAT, se mejora su fiabilidad, se analiza su incertidumbre, se compara con otros métodos de referencia y se aplica en adobes, Bloques de Tierra Comprimida y probetas de tierra estabilizada con distintas proporciones de paja. Este método servirá de base a una posterior aplicación in situ. Finalmente se proponen modelos matemáticos para mejorar la exactitud del dispositivo utilizado y para la estimación de la conductividad de cerramientos de tierra en función de su densidad. Con los resultados obtenidos se analizan las posibilidades de amortiguación y retardo de las ondas térmicas y capacidad de almacenaje de energía de los cerramientos en función de su densidad y humedad. There is growing international interest in energy saving and sustainability in buildings with significant impact on Architecture. Thermal inertia is a key parameter to assess energy in buildings in real conditions. This requires changing the traditional approach to heat transfer in steady state by another in dynamic regime which analyzes the thermal waves and oscillating heat flux passing through the external walls. The parameters defining the thermal inertia are: the thickness, the diffusivity and the thermal cycle. In turn, the diffusivity is determined by the thermal conductivity, density and specific heat of the material. Of these parameters, thermal conductivity is the most complex, variable and difficult to measure, especially in earth walls due to their heterogeneity and hygrothermal complexity. In general, the methods of measurement of conductivity and transmittance in walls have drawbacks when measuring a building with earth: implementation difficulties, high cost, or reliability of the results, mainly. The Thermal Needle Procedure (TNP) is based on the principle of evolution in time of heat from a line source when inserted within a material. This method was chosen because it is a practical, low cost and easy to implement on a large scale but has serious problems of reliability and accuracy. This thesis develops a laboratory method for measuring the thermal conductivity of Masonry Units Unfire Earth-based based on TNP, its uncertainty is analyzed, compared to other reference methods and applies in adobes, Compressed Earth Blocks and stabilized soil specimens with different proportions of straw. This method will form the basis of a subsequent application in situ. Finally, mathematical models are proposed to improve the accuracy of the device used, and to estimate the conductivity of earth enclosures depending on its density. With the results obtained earth enclosures are analyzed to estimate their possibilities of delay and buffer of termal waves and energy storage capacity according to their density and moisture.

Offshore Measurements of Ocean Waves using Stereo Vision Systems

In recent years, remote sensing imaging systems for the measurement of oceanic sea states have attracted renovated attention. Imaging technology is economical, non-invasive and enables a better understanding of the space-time dynamics of ocean waves over an area rather than at selected point locations of previous monitoring methods (buoys, wave gauges, etc.). We present recent progress in space-time measurement of ocean waves using stereo vision systems on offshore platforms, which focus on sea states with wavelengths in the range of 0.01 m to 10 m. Classical epipolar techniques and modern variational methods are reviewed to reconstruct the sea surface from the stereo pairs sequentially in time. The statistical and spectral properties of the resulting observed waves are analyzed. Current improvements of the variational methods are discussed as future lines of research.

In situ surveillance of capacitive pressure transmitter dynamic poles

In this work, a methodology is proposed to find the dynamic poles of a capacitive pressure transmitter in order to enhance and extend the online surveillance of this type of sensor based on the response time measurement by applying noise analysis techniques and the dynamic data system procedure. Several measurements taken from a pressurized water reactor have been analyzed. The methodology proposes an autoregressive fit whose order is determined by the sensor dynamic poles. Nevertheless, the signals that have been analyzed could not be filtered properly in order to remove the plant noise; thus, this was considered as an additional pair of complex conjugate poles. With this methodology we have come up with the numerical value of the sensor second real pole in spite of its low influence on the sensor dynamic response. This opens up a more accurate online sensor surveillance since the previous methods were achieved by considering one real pole only.

AR modelling and DDS for in situ surveillance of capacitive pressure transmitters

In this work, a methodology is proposed to find the dynamics poles of a capacitive pressure transmitter in order to enhance and extend the on line surveillance of this type of sensors based on the response time measurement by applying noise analysis techniques and the Dynamic Data System. Several measurements have been analyzed taken from a Pressurized Water Reactor. The methodology proposes an autoregressive fit whose order is determined by the sensor dynamics poles. Nevertheless, the signals that have been analyzed, could not be filtered properly in order to remove the plant noise, thus, this was considered as an additional pair of complex conjugate poles. With this methodology we have come up with the numerical value of the sensor second real pole in spite of its low influence on the sensor dynamic response. This opens up a more accurate on line sensor surveillance since the previous methods were achieved by considering one real pole only.

Offshore remote sensing of the ocean by stereo vision systems

In recent years, remote sensing imaging systems for the measurement of oceanic sea states have attracted renovated attention. Imaging technology is economical, non-invasive and enables a better understanding of the space-time dynamics of ocean waves over an area rather than at selected point locations of previous monitoring methods (buoys, wave gauges, etc.). We present recent progress in space-time measurement of ocean waves using stereo vision systems on offshore platforms, which focus on sea states with wavelengths in the range of 0.01 m to 1 m. Both traditional disparity-based systems and modern elevation-based ones are presented in a variational optimization framework: the main idea is to pose the stereoscopic reconstruction problem of the surface of the ocean in a variational setting and design an energy functional whose minimizer is the desired temporal sequence of wave heights. The functional combines photometric observations as well as spatial and temporal smoothness priors. Disparity methods estimate the disparity between images as an intermediate step toward retrieving the depth of the waves with respect to the cameras, whereas elevation methods estimate the ocean surface displacements directly in 3-D space. Both techniques are used to measure ocean waves from real data collected at offshore platforms in the Black Sea (Crimean Peninsula, Ukraine) and the Northern Adriatic Sea (Venice coast, Italy). Then, the statistical and spectral properties of the resulting observed waves are analyzed. We show the advantages and disadvantages of the presented stereo vision systems and discuss future lines of research to improve their performance in critical issues such as the robustness of the camera calibration in spite of undesired variations of the camera parameters or the processing time that it takes to retrieve ocean wave measurements from the stereo videos, which are very large datasets that need to be processed efficiently to be of practical usage. Multiresolution and short-time approaches would improve efficiency and scalability of the techniques so that wave displacements are obtained in feasible times.

Regulación del movimiento y anticipación en el resto del primer servicio en tenis

Aunque se han logrado importantes avances en estudios de laboratorio con diseños experimentales poco representativos (e.g., Farrow y Reid, 2012; Nieminen, Piirainen, Salmi, y Linnamo, 2013), a día de hoy, todavía se desconoce a cabalidad cómo los jugadores de tenis de diferente nivel de pericia calibran o ajustan sus movimientos a las demandas espacio-temporales presentes en la tarea de resto de un primer servicio. ! Escasos trabajos se han llevado a cabo in situ y a la mayoría se les puede cuestionar algún aspecto de la metodología empleada. Así pues, en varios estudios la frecuencia de grabación ha sido limitada (e.g., a 50 Hz en Jackson y Gudgeon, 2004; Triolet, Benguigui, Le Runigo y Williams, 2013), o la velocidad del saque ha sido visiblemente inferior a la habitual (cf. Carboch, Süss y Kocib, 2014; Williams, Singer y Weigelt, 1998). También, en algunos estudios los participantes experimentados no han sido jugadores de nivel internacional (e.g., Avilés, Ruiz, Sanz y Navia, 2014), y el tamaño muestral ha sido muy pequeño (e.g., Gillet, Leroy, Thouvarecq, Mégrot y Stein, 2010). ! Además, en los diferentes trabajos se han utilizado una diversidad de métodos e instrumentos de medida y los criterios de codificación del inicio de los movimientos y de las respuestas han diferido; como consecuencia el lapso visomotor de respuesta (LVMr) ha sido muy dispar variando considerablemente de 198 a 410 ms. Considerando los inconvenientes señalados anteriormente, el presente estudio tuvo como objetivo determinar un modelo técnico de regulación temporal de los movimientos y de la respuesta del restador, tomando en cuenta el flujo continuo de información proporcionado por el sacador. Para ello, se realizó un análisis cronométrico de los restos de doce jugadores de diferente nivel deportivo (seis internacionales y seis nacionales) que respondieron de forma natural enviando sus devoluciones hacia las dianas. Se grabaron las acciones de los restadores y sacadores con una cámara Casio Exilim Pro Ex-F1 de alta velocidad (300 Hz) y luego se realizó un análisis imagen por imagen cada 3.33 ms. Una vez obtenidos los datos de los vídeos se realizaron análisis con las pruebas de ANOVA de un factor, ANCOVA con la velocidad del saque como covariable, U de Mann-Whitney y Chi-cuadrado de Pearson. En cuanto a la regulación del movimiento hasta el momento del despegue, los jugadores internacionales iniciaron sus acciones antes que los jugadores nacionales lo que podría indicar una mejor preparación al ejecutar los movimientos como reflejo del nivel de pericia. Los jugadores internacionales iniciaron la elevación del pie posterior a -293 ms y los jugadores nacionales a -202 ms. Todas estas acciones se fueron enlazando unas con otras y fue en el momento del impacto del sacador donde los restadores demostraron una remarcable coordinación perceptivo-motriz. Por consiguiente, los jugadores internacionales despegaron e iniciaron el vuelo a tan solo -6.5 ms del impacto y los jugadores nacionales lo hicieron más tarde a +19.5 ms. A lo largo de la secuencia temporal, todo parece indicar que las informaciones que utilizan los restadores interactúan entre sí; información más temprana y menos fiable para anticipar o moverse antes e información más tardía y más fiable para regular la temporalización de las acciones. Los restadores de nivel internacional y nacional anticiparon a nivel espacial en un bajo porcentaje (7.7% vs. 13.6%) y en tiempos similares (-127 vs. -118 ms) sugiriendo que la utilización de variables ópticas tempranas y menos fiables solo se produce en contadas ocasiones. Por otra parte, estos datos se relacionan con una gran precisión en la respuesta ya que tanto los jugadores internacionales como los nacionales demostraron un alto porcentaje de acierto al responder (95.4% vs. 96.7%). Se había señalado que los jugadores internacionales y nacionales se diferenciarían en el tiempo de caída (i.e., aterrizaje) del primer pie del salto preparatorio, sin embargo ese efecto no fue encontrado (128 vs. 135 ms). Tampoco se hallaron diferencias en el porcentaje de caída con el pie contrario a la dirección de la pelota (58% vs. 62%). Donde sí ambos grupos se diferenciaron fue en el tiempo de caída del segundo pie (147 vs. 168 ms). Esta diferencia de 21 ms fue crucial y fue una prueba de la mayor rapidez de los jugadores internacionales; sugiriendo que ésta acción se podría relacionar con el momento del inicio de la respuesta. Aunque los jugadores internacionales hayan demostrado ser más rápidos en relación con sus capacidades funcionales, ambos grupos no se diferenciaron en todas las variables relacionadas con el LVMr. Ellos no utilizaron esos valiosos milisegundos ganados en el instante de la caída del segundo pie para responder más pronto, ya que el LVMr del miembro superior fue el mismo para ambos grupos (179 vs. 174 ms). Es como si hubiesen tenido todo el tiempo del mundo para seguir ajustando sus acciones hasta el propio golpeo. Además, estos tiempos largos sugieren que en la gran mayoría de los restos la información clave que determinó la respuesta fue detectada (extraída) en momentos cercanos al golpeo del sacador y en la primera parte del vuelo de la pelota. Asimismo, se constató que en general el LVMr se ve influenciado por el tipo de información utilizada. De esta manera, cuando se tomaron en cuenta los ensayos en los que hubo anticipación espacial reflejados en el LVMr del cuerpo entero los tiempos disminuyeron (152 vs. 136 ms). Por otra parte, existieron ocasiones (13%) en los que tanto los jugadores internacionales como los nacionales respondieron tarde recibiendo saques directos (208 vs. 195 ms). Es muy posible que en estos casos los jugadores hayan tenido problemas para detectar la información respondiendo fuera de los márgenes temporales de acción lo que mermó su rendimiento. Lo mismo pudo haber ocurrido cuando ambos grupos de jugadores corrigieron el movimiento del miembro superior tras el impacto (17% vs. 10%) lo que aumentó el tiempo en responder al redirigir la respuesta hacia el lado correcto (208 vs. 205 ms). Además, los jugadores internacionales obtuvieron tiempos de movimiento menores que el de los jugadores nacionales (509 vs. 531 ms) lo que se reflejó en un tiempo total de actuación menor (683 vs. 703 ms). Por último, en cuanto al rendimiento del resto, los jugadores internacionales obtuvieron valores superiores a los jugadores nacionales (1.3 vs. 0.9). ABSTRACT Although there have been significant advances in laboratory studies with unrepresentative experimental designs (e.g., Farrow y Reid, 2012; Nieminen, Piirainen, Salmi, y Linnamo, 2013), today it is still unknown to full extent how tennis players of different levels of expertise calibrate or adjust their movements to the spatial-temporal demands present in the return of a first serve. Few studies have been carried out in situ and some aspects of the methodology most of them used can be questioned. Thus, in several studies the recording frequency has been limited (e.g., a 50 Hz en Jackson y Gudgeon, 2004; Triolet, Benguigui, Le Runigo y Williams, 2013), or serve speed was visibly lower than the usual one (cf. Carboch, Süss y Kocib, 2014; Williams, Singer y Weigelt, 1998). Also, in some studies, experienced participants have not played at international level (e.g., Avilés, Ruiz, Sanz y Navia, 2014), and the sample size has been very small (e.g., Gillet, Leroy, Thouvarecq, Mégrot y Stein, 2010). Furthermore, different works have used a variety of methods and measurement instruments and coding criteria of the onset of movements and responses have differed; due to this, visuomotor response delay (LVMr) has been very uneven, varying considerably from 198-410 ms. Considering the drawbacks mentioned above, this study aimed to determine a technical model of temporal regulation of movements and returner’s response, taking into account the continuous flow of information provided by the server. For this, a chronometric analysis of the returns of twelve players (six international and six national) of different sports level, that naturally responded by hitting their returns towards the targets, was performed. Actions of servers and returners were recorded with a Casio Exilim Pro Ex-F1 high speed camera (300 Hz) and then every 3.33 ms analysis was made frame by frame. Once the data of the videos were obtained, analyses were performed using one factor ANOVA test, ANCOVA with the speed of the serve as a covariate, U of Mann- Whitney and Pearson’s Chi-square test. As for the regulation of movement until the moment of serve, international players began their actions before national players, which could indicate that they were better prepared to execute movements reflecting the level of their expertise. International players began raising the rear foot at -293 ms and national players at -202 ms. All these actions were being linked to each other and it was at the moment of impact of the server when the receivers demonstrated a remarkable perceptual-motor coordination. Therefore, international players took off and started their flight just -6.5 ms before the serve and national players did the same somewhat later: +19.5 ms after the serve. Along the timeline, everything seems to indicate that the information used by returners interact with each other; early information which is less reliable to anticipate or move before, and later information more reliable appears to regulate the timing of actions. Returners of international and national levels anticipated at spatial level in a low percentage (7.7% vs. 13.6%) and in similar times (-127 vs. -118 ms) suggesting that the use of early and less reliable optical variables is only produced on rare occasions. Moreover, these data relate to a precise response as both international and national players showed a high percentage of success in responding (95.4% vs. 96.7%). It had been noted that international and national players would differ in the time the fall (i.e., landing) of the first foot of the split-step, however, this effect was not found (128 vs. 135 ms). No differences in the percentage of fall with the opposite foot to the direction of the ball (58% vs. 62%) were found. Where the two groups differed was in the time of the fall of the second foot (147 vs. 168 ms). This difference of 21 ms was crucial and it was a proof of mayor speed of international players; suggesting that this action could be related to the onset time of response. Although international players have proven to be faster in relation to their functional capabilities, both groups did not differ in all variables related to LVMr. They did not use those precious milliseconds earned at the time of the fall of the second foot to respond as soon, since the LVMr of the upper limb was the same for both groups (179 vs. 174 ms). It is as if they had all the time in the world to continue to adjust their actions until the return itself. Furthermore, these long times suggest that in the vast majority of the returns, key information that determined the response was detected (pick-up) in moments close to the hit of the server and in the first part of the ball flight. It was also found that in general the LVMr is influenced by the type of information used. Thus, when taking into account the trials during which there was spatial anticipation, reflected in LVMr of the whole body, the times decreased (152 vs. 136 ms). On the other hand, there were occasions (13%) where both international and national players responded late, thus receiving aces (208 vs. 195 ms). It is quite possible that in these cases the players have had trouble to pick-up information, responding out of temporary margins of action, which affected their performance. The same could have occurred when both groups of players corrected upper limb movement after impact (17% vs. 10%), which increased the time to respond and to redirect the return towards the right side (208 vs. 205 ms). Moreover, international players scored lower movement times than the national players (509 vs. 531 ms), which was reflected in a shorter total response time (683 vs. 703 ms). Finally, as far as the performance of return is concerned, international players scored above the national players values (1.3 vs. 0.9).

Measurement of 8-hydroxy-2′-deoxyguanosine in DNA by high-performance liquid chromatography-mass spectrometry: comparison with measurement by gas chromatography-mass spectrometry

Measurement of 8-hydroxy-2′-deoxyguanosine (8-OH-dGuo) in DNA by high-performance liquid chromatography/mass spectrometry (LC/MS) was studied. A methodology was developed for separation by LC of 8-OH-dGuo from intact and modified nucleosides in DNA hydrolyzed by a combination of four enzymes: DNase I, phosphodiesterases I and II and alkaline phosphatase. The atmospheric pressure ionization-electrospray process was used for mass spectral measurements. A stable isotope-labeled analog of 8-OH-dGuo was used as an internal standard for quantification by isotope-dilution MS (IDMS). Results showed that LC/IDMS with selected ion-monitoring (SIM) is well suited for identification and quantification of 8-OH-dGuo in DNA at background levels and in damaged DNA. The sensitivity level of LC/IDMS-SIM was found to be comparable to that reported previously using LC-tandem MS (LC/MS/MS). It was found that approximately five lesions per 106 DNA bases can be detected using amounts of DNA as low as 2 µg. The results also suggest that this lesion may be quantified in DNA at levels of one lesion per 106 DNA bases, or even lower, when more DNA is used. Up to 50 µg of DNA per injection were used without adversely affecting the measurements. Gas chromatography/isotope-dilution MS with selected-ion monitoring (GC/IDMS-SIM) was also used to measure this compound in DNA following its removal from DNA by acidic hydrolysis or by hydrolysis with Escherichia coli Fpg protein. The background levels obtained by LC/IDMS-SIM and GC/IDMS-SIM were almost identical. Calf thymus DNA and DNA isolated from cultured HeLa cells were used for this purpose. This indicates that these two techniques can provide similar results in terms of the measurement of 8-OH-dGuo in DNA. In addition, DNA in buffered aqueous solution was damaged by ionizing radiation at different radiation doses and analyzed by LC/IDMS-SIM and GC/IDMS-SIM. Again, similar results were obtained by the two techniques. The sensitivity of GC/MS-SIM for 7,8-dihydro-8-oxoguanine was also examined and found to be much greater than that of LC/MS-SIM and the reported sensitivity of LC/MS/MS for 8-OH-dGuo. Taken together, the results unequivocally show that LC/IDMS-SIM is well suited for sensitive and accurate measurement of 8-OH-dGuo in DNA and that both LC/IDMS-SIM and GC/IDMS-SIM can provide similar results.

Semiconductor measurement technology:

Title varies slightly.

Development of Cookstove Emissions and Performance Testing Suite with Time-resolved Particulate Matter Analysis and Excess Air Estimation

Thesis (Master's)--University of Washington, 2016-06

Primary aldosteronism - actual epidemics or false alarm?

The prevalence of "primary aldosteronism" (PAL) cannot be precisely determined at this time, given 1) lack of a universally accepted definition, and 2) normotensive as well as normokalemic phases in the evolutionary development of a disease eventually characterized by hypertension and hypokalemia. The exception is fully genetically characterised forms such as glucocorticoid-suppressible hyperaldosteronism, the true prevalence of which could be proven today by universal screening using a single blood sample, but this is neither practical nor appropriate. Controversy has arisen regarding the rareness, or otherwise, of PAL because of 1) rediscovery in the last 12 years of the normokalemic phase described by Conn, 2) application of widely available methods for measurement of aldosterone and renin to "screening", 3) variable quality of these methods, and of their application, and 4) lack of the necessary "diagnostic", in addition to "screening", tests in some studies. PAL is significantly more common than previously thought, and a very important potentially curable form of hypertension. Early diagnosis and specific treatment avoids morbidity. The current focus on increased detection is essential, and will help to resolve the question of prevalence.

Time-Frequency Distribution Moments of Heart Rate Variability for Neonatal Seizure Detection

In this paper, we propose features extracted from the heart rate variability (HRV) based on the first and second conditional moments of time-frequency distribution (TFD) as an additional guide for seizure detection in newborn. The features of HRV in the low frequency band (LF: 0-0.07 Hz), mid frequency band (MF: 0.07-0.15 Hz), and high frequency band (HF: 0.15-0.6 Hz) have been obtained by means of the time-frequency analysis using the modified-B distribution (MBD). Results of ongoing time-frequency research are presented. Based on our preliminary results, the first conditional moment of HRV which is also known as the mean/central frequency in the LF band and the second conditional moment of HRV which is also known as the variance/instantaneous bandwidth (IB) in the HF band can be used as a good feature to discriminate the newborn seizure from the non-seizure

Ultrasonic measurement technique of burning metals in normal and reduced gravity

The in situ real time measurement of the regression rate of a melting interface (RRMI) is performed by the ultrasonic measurement system reported here. The RRMI is the rate at which a solid/liquid interface (SLI) moves along a metallic rod while burning in an oxygen-enriched atmosphere and is an important flatnmability indicator. The ultrasonic transducer and associated equipment used to drive the transducer and record the echo signal is described, along with the process that transforms the acquired signals into a RRMI value. Test rods of various metals and geometric shapes were burned at several test conditions in different test facilities. The RRMI results with quantified errors are presented and reviewed. The effect of reduced gravity on burning metals is important to space-applications and RRMI results obtained in a reduced gravity environment are also presented.