Optimization Of Nuclear Magnetic Resonance Spectroscopy Methods For Measuring Protein Hydration In Reverse Micelles

Water is fundamental to all aspects of protein function including folding, stability, catalysis, and dynamics. The unique characteristics of water make it the ideal solvent for supporting life but also make it incredibly difficult to study. While much has been learned about the role of water in protein function, a site resolved understanding of these interactions has remained elusive thereby leaving a large hole in the biophysical puzzle. Experimental techniques that provide a site-resolved view of protein hydration without mutation of the protein are necessary to understand the thermodynamic role of water on protein function. It has been shown that the combination of Nuclear Magnetic Resonance (NMR) spectroscopy and encapsulation of proteins within the core of reverse micelles can satisfy these requirements. The goal of this thesis is to apply and expand upon currently established techniques to make it generally applicable to all protein systems. First, previously established methods were used to examine the internal hydration patterns of staphylococcal nuclease variants with internal ionizable groups. The results demonstrate that water penetrates the hydrophobic core to stabilize buried ionizable groups. This study illustrates the utility of NMR detected hydration measurements for longstanding biophysical questions. Next, two methods to reduce data collection times were implemented for hydration dynamics measurements. This necessary time savings provides a platform for assessing the reproducibility and precision of NMR derived hydration measurements. A new data fitting method that allows for quantitative hydration dynamics measurements of protein regions generally contaminated by hydrogen exchange is introduced. Finally, one of the remaining artifacts associated with hydration dynamics measurements detected by NMR is addressed: hydrogen exchange relayed artifacts. This was accomplished by developing experiments to decouple the relaying spin and applying a new data fitting method. These methods allow for the first site and time resolved study of protein hydration in the absence of artifact is presented. These techniques introduced were applied to Ubiquitin encapsulated in AOT reverse micelles. The majority of slowed waters reside in concave regions of the protein surface. This suggests surface curvature is one of the contributing factors for the slowing of hydration waters. The experiments presented demonstrate the utility of using NMR for measuring protein-water interactions. The work presented expands and improves upon existing methodologies and provides a framework for artifact free site resolved measurement of protein water-interactions in all protein systems

Opening pervasive computing to the masses using the SEAP middleware

Abstract — The increasing availability of sensing devices has made the possibility of context-aware pervasive computing ap-plications real. However, constructing this software requires extensive knowledge about the devices and specialized program-ming languages for interacting with them. While the nature of pervasive computing lends users to demand individualized ap-plications, complexities render programming embedded devices unapproachable. In this paper we introduce the SEAP (Sensor Enablement for the Average Programmer) middleware which applies existing technologies developed for web programming to the task of collecting and using sensor data. We show how this approach can be used to create new applications and to update existing web applications to accept sensor data. I

Opening - Welcome and Instructions

By American University Washington College of Law, Texas A&M University School of Law, and University of Utah S.J. Quinney College of Law

Case studies: Raman implementation for process lifecycle management in fermentation based processes

Existing metabolite analyzers for several inline fermentation based vaccine processes are becoming obsolete in upcoming years. Analyzers are used to support classified parameters and attributes as well as characterization of the fermentation processes. A PAT solution is preferred as a replacement as it would allow for enhanced process understanding and control. Raman spectroscopy has been aligned as a core technology for implementation with varying challenges based on media, organism, processing parameters and attributes being measured. Implementation requires a highly collaborative approach across functions and sites to ensure effective implementation with no interruption to supply. Using a standardized approach to Raman model development and validation, robust models have been developed for 2 product lines with implementation scheduled over the next three years

Physical abuse of minors under 15: epidemiological study in a city in the south of Brazil

OBJETIVO: apresentar o perfil de casos notificados de violência física contra menores de 15 anos em Londrina, Paraná, no ano de 2006. MÉTODO: Estudo transversal, com coleta retrospectiva nos prontuários dos Conselhos Tutelares e serviços de atendimento do município. Os dados foram processados e tabulados pelo programa Epi Info. RESULTADOS: Foram estudados 479 casos de violência por força corporal e 9 casos de violência por outros meios (7 por instrumentos, 1 por objeto cortante e 1 por substância corrosiva). Na violência por força corporal, predominaram vítimas do sexo feminino (53,4 por cento ) e maior risco na idade de seis anos (12,2 por 1.000). O pai foi o agressor mais frequente (48,8 por cento ) e o alcoolismo esteve presente em 64,0 por cento dos casos. A violência por instrumentos foi praticada através de cinta (42,9 por cento ), fio (28,6 por cento ), ferro (14,3 por cento ) e instrumento de cozinha (14,3 por cento ), com vítimas do sexo feminino (85,7 por cento ), na faixa etária de doze anos (33,3 por cento ), sendo o pai (71,4 por cento ) e a mãe (28,6 por cento ) os únicos agressores, com o alcoolismo presente em 57,1 por cento destas situações. A vítima de violência por objeto cortante era do sexo masculino, 13 anos e o agressor, desconhecido, tinha de 15 a 19 anos. A violência por substância corrosiva teve como vítima um adolescente de 13 anos, do sexo masculino, cujo agressor foi o pai, sendo o alcoolismo a situação presente. CONCLUSÕES: Os resultados apontam para a importância epidemiológica do abuso físico contra crianças e adolescentes e podem contribuir para a elaboração de estratégias de prevenção e acompanhamento das vítimasOBJECTIVES: to build the epidemiological profile of deliberate violence against minors under 15 years of age in Londrina, Paraná in 2006. METHODS: cross-sectional study with retrospective data collected from the records of Tutelary Councils and care services in the city. Data were processed and tabulated with Epi Info software. RESULTS: 479 cases of violence by use of physical force and 9 cases of violence by others means (7 by instruments, 1 by cutting object and 1 by corrosive substance) were studied. Most victims of physical force were female (53.4%) and the highest risk age was 6 (12.2 per 1,000). The father was the main aggressor (48.8%) and alcoholism was present in 64.0% of cases. Violence by instrument was inflicted with belt (42.9%), wire (28.6%), iron (14.3%), or kitchen tool (14.3%), mainly with 85.7% female victims in the 12 year-old age group (33.3%). Father (71.4%) and mother (28.6%) were the only aggressors, with alcoholism present in 57.1% of the situations. The victim of violence with a cutting device was a 13 year-old male, the aggressor was unknown and between 15 and 19 years of age. Violence by a corrosive substance victimized a male adolescent aged 13. The aggressor was the alcoholic father. CONCLUSION: outcomes point toward the epidemiological importance of physical abuse of children and adolescents, and can contribute to structure strategies to prevent child abuse and follow-up victim

Maltrato infantil: un rescate de la historia y de las políticas de protección

O estudo apresenta uma revisão bibliográfica, cujo objetivo foi conhecer e divulgar a evolução histórica da violência contra a criança, bem como as políticas desenvolvidas na atenção à violência contra menores, além de discutir a importância da prevenção e da atuação dos profissionais de saúde. A pesquisa bibliográfica foi realizada na base de dados MEDLINE, LILACS e SciELO, selecionando-se os estudos com os descritores: Violência, Agressão, Maus-Tratos, Síndrome da Criança Maltratada, Pediatria, não se fazendo restrição aos idiomas espanhol, inglês e francês. A literatura mostra a importância de ações preventivas e a necessidade de discussões e reflexões entre os diferentes setores que possam culminar em políticas e estratégias preventivas, diagnósticas e terapêuticas, além da relevância de incluir o tema na formação dos profissionais de saúde para que possam contribuir para o diagnóstico, tratamento e profilaxia do abuso infantil, rompendo as cadeias de determinação e fatalidade.The study presents a literature research of violence against children, intended to better understand this phenomenon and present the results to the public knowledge; the same objectives were pursued with the policies developed to deal with the violence against children, at the same time discussing the importance of the prevention and the interventions of the health professionals. The literature research was conducted on the data banks MEDLINE, LILACS and SciELO, selecting the studies with the keywords: Violence, Aggression, Child Abuse, Shaken Child Syndrome, and Pediatrics; the search was not restricted to the Spanish, English and French languages. The literature showed the importance of preventive measures and the need to discuss and reflect, among different sectors, in order to obtain policies and strategies leading to prevention, diagnostics and therapeutics. It was demonstrated the relevance of including these aspects in the training of the health professionals, which will allow them to contribute to diagnosis, treatment and prevention of child abuse, breaking the chains of predetermined fate and destiny.El estudio tuvo por objetivo realizar una revisión bibliográfica, con la finalidad de conocer y divulgar la evolución histórica de la violencia contra los niños, así como las políticas desarrolladas en la atención a la violencia contra menores; además se discute la importancia de la prevención y de la actuación de los profesionales de la salud. La investigación bibliográfica fue realizada en la base de datos MEDLINE, LILACS y SciELO, seleccionando los estudios con las palabras claves: Violencia, Agresión, Maltrato, Síndrome del Niño Maltratado y, Pediatría; no se hizo restricción a los idiomas castellano, inglés y francés. La literatura muestra la importancia de las acciones preventivas y la necesidad de discutir y reflexionar, entre los diferentes sectores, para que culminen en políticas y estrategias preventivas, diagnósticas y terapéuticas. Es relevante incluir el tema en la formación de los profesionales de la salud para que puedan contribuir para el diagnóstico, tratamiento y profilaxis del abuso infantil, rompiendo las cadenas de determinación y fatalidad

Resultados de los casos de violencia contra niños y adolescentes en el poder judicial

Objetivos: Analisar, decorridos cinco anos da notificação, o desfecho dado pela Vara da Infância e da Juventude de Londrina (PR) aos casos de violência contra crianças e adolescentes; identificar os tipos de violência com maior taxa de encaminhamento à vara da Infância e Juventude. Métodos: Estudo descritivo, quantitativo, cuja população de estudo foram crianças e adolescentes de até 15 anos de idade, residentes em Londrina (PR), vítimas de violência notificada pelos Conselhos Tutelares à Vara da Infância e da Juventude, em 2002. Resultados: Dos 230 casos, dos quais 40,0% dos processos foram arquivados; 3,9% foram arquivados enquanto ainda inquéritos; em 1,7% houve condenação do réu; 31,7% dos processos encontravam-se em andamento. Conclusão: Os achados contribuem para ampliar o conhecimento das medidas de proteção adotadas pelo poder judiciário frente aos casos de violência contra crianças e adolescentes.Objective: To analyze the decisions of the department of justice regarding cases of violence against children and teenagers for the last five years of notification. Methods: A descriptive quantitative study with 230 cases of violence against children and teenager that have been reported to the department of justice in 2002 in Londrina, PR. Results: A great number of the processes (40.0%) had been archived. A few of the processes were archived during questioning (3.9%) and only 1.7% of the processes resulted in the plaintiff's conviction. Approximately one third of the processes (31.7%) were in progress. Conclusion: Findings may contribute to awareness of the population regarding protection measures adopted by the department of justice for the cases of violence against children and teenagers.Objetivos: Analizar, después de transcurridos cinco años de la notificación, el resultado dado por el Tribunal de la Infancia y de la Juventud de Londrina (PR) a los casos de violencia contra niños y adolescentes. Métodos: Estudio descriptivo, cuantitativo, cuya población de estudio fueron niños y adolescentes de hasta 15 años de edad, residentes en Londrina (PR), víctimas de violencia notificada por los Consejos Tutelares al Tribunal de la Infancia y de la Juventud, en 2002. Resultados: De los 230 casos estudiados, 40% de los procesos fueron archivados; 3,9% fueron archivados en cuanto se encontraban en proceso de investigación; en 1,7% hubo condenación del reo; 31,7% de los procesos se encontraban en andamiento. Conclusión: Lo encontrado contribuye para ampliar el conocimiento de las medidas de protección adoptadas por el poder judicial frente a los casos de violencia contra niños y adolescentes

Case study: Raman implementation for process lifecycle management in fermentation based processes

Existing metabolite analyzers for several inline fermentation based vaccine processes are becoming obsolete in upcoming years. Analyzers are used to support classified parameters and attributes as well as characterization of the fermentation processes. A PAT solution is preferred as a replacement as it would allow for enhanced process understanding and control. Raman spectroscopy has been aligned as a core technology for implementation with varying challenges based on media, organism, processing parameters and attributes being measured. Implementation requires a highly collaborative approach across functions and sites to ensure effective implementation with no interruption to supply. Using a standardized approach to Raman model development and validation, robust models have been developed for 2 product lines with implementation scheduled over the next three years

Valid attacks in argumentation frameworks with recursive attacks

The purpose of this work is to study a generalisation of Dung’s abstract argumentation frameworks that allows representing recursive attacks, that is, a class of attacks whose targets are other attacks. We do this by developing a theory of argumentation where the classic role of attacks in defeating arguments is replaced by a subset of them, which is extension dependent and which, intuitively, represents a set of “valid attacks” with respect to the extension. The studied theory displays a conservative generalisation of Dung’s semantics (complete, preferred and stable) and also of its principles (conflictfreeness, acceptability and admissibility). Furthermore, despite its conceptual differences, we are also able to show that our theory agrees with the AFRA interpretation of recursive attacks for the complete, preferred and stable semantics