Made available in DSpace on 2014-12-17T14:47:04Z (GMT). No. of bitstreams: 1 MariaICDF_DISSERT.pdf: 1911495 bytes, checksum: b33fd53ec7c7e2b2d0bd96f5e0ea12e9 (MD5) Previous issue date: 2014-01-27 / Trasnversal study, with the objective of evaluating the accuracy of clinical indicators of nursing diagnosis excessive fluid volume in patients undergoing hemodialysis. The study occurred in two stages, the first consisted of the evaluation of the diagnostic indicators in study; and the second, the diagnostic inference conducted by nurse diagnosticians. The first stage occurred from december 2012 to april 2013, in a University Hospital and a Hemodialysis Clinic in Northeastern of Brazil, with a sample of 100 chronic renal failure patients on hemodialysis. The data were selected through an interview form and a physical examination, organized into spreadsheets and analyzed as to the presence or absence of the indicators of diagnosis excessive fluid volume. In the second step, the spreadsheets were sent to three nurses diagnosticians, who judged the presence or absence of diagnosis in the clientele searched. This step was conducted from july to september 2013. For analysis of the data, we used descriptive and inferential statistics. In the descriptive analysis, we used measures of central tendency and dispersion. In inferential analysis, we used the tests Chi- square, Fisher and prevalence ratios. The accuracy of the clinical indicators pertaining to the diagnosis were measured as to the specificity, sensitivity, predictive values, likelihood ratios and Diagnostic Odds Ratio. Also developed a logistic regression. The results were organized in tables and discussed with literature. This study was approved by the Ethics Committee in Research of the Federal University of Rio Grande do Norte, with Presentation Certificate for Ethics Appreciation n? 08696212.7.0000.5537. The results revealed that the diagnosis studied was present in 82% of patients. The characteristics with prevalence above 50 % that stood out were: azotemia, decreased hematocrit, electrolyte imbalance, intake exceeds output, anxiety, edema, decreased hemoglobin, oliguria and blood pressure changes. Eight defining characteristics were presented statistically significant association with the nursing diagnosis investigated: pulmonary congestion, intake exceeds output, electrolytes imbalance, jugular vein distension, edema, weight gain over short period of time, agitation and adventitious breath sounds. Among these, the 10 characteristics which showed higher prevalence ratios were: edema and weight gain over short period of time. The features with the highest sensitivity were edema, electrolytes imbalance and intake exceeds output and the standing out with greater specificity were: anasarca, weight gain over short period of time, change in respiratory pattern, adventitious breath sounds, pulmonary congestion, agitation and jugular vein distension. The indicators jugular vein distension, electrolytes imbalance, intake exceeds output, increased central venous pressure and edema, together, were identified in the logistic regression model as the most significant predictors. It is concluded that the identification of accurate clinical indicators allow a good prediction of the nursing diagnosis of excessive fluid volume in patients undergoing hemodialysis in order to assist the nurse in the inference process, which will contribute to the success of patient care. In addition, nurses will consider for diagnostic inference not only his clinical experience, but also scientific evidence of the occurrence of excessive fluid volume, contributing to the control of volemia in these patients / Estudo transversal, com o objetivo de avaliar a acur?cia dos indicadores cl?nicos do diagn?stico de enfermagem Volume de l?quidos excessivo em pacientes submetidos ? hemodi?lise. Ocorreu em duas etapas, a primeira composta pela avalia??o dos indicadores do diagn?stico em estudo; e a segunda, pela infer?ncia diagn?stica realizada por enfermeiros diagnosticadores. A primeira etapa aconteceu nos meses de dezembro de 2012 a abril de 2013, em um Hospital Universit?rio e em uma Cl?nica de Hemodi?lise do Nordeste do Brasil, com uma amostra de 100 pacientes renais cr?nicos submetidos ? hemodi?lise. Os dados foram obtidos mediante formul?rio de entrevista e exame f?sico, organizados em planilhas eletr?nicas e analisados quanto ? presen?a ou aus?ncia dos indicadores do diagn?stico Volume de l?quidos excessivo. Na segunda etapa, as planilhas foram encaminhadas a tr?s diagnosticadores, que julgaram a presen?a ou aus?ncia do diagn?stico na clientela pesquisada. Essa etapa desenvolveu-se nos meses de julho a setembro de 2013. Para a an?lise dos dados, utilizou-se a estat?stica descritiva e inferencial. Na an?lise descritiva, utilizaram-se medidas de tend?ncia central e de dispers?o. Na inferencial, utilizaram-se os testes de Qui-quadrado, Fisher e as raz?es de preval?ncia. Mensurou-se a acur?cia dos indicadores cl?nicos do diagn?stico estudado por meio da especificidade, da sensibilidade, dos valores preditivos, das raz?es de verossimilhan?a e da Odds Ratio Diagn?stica. Desenvolveu-se tamb?m uma regress?o log?stica. Os resultados foram organizados em tabelas e discutidos com literatura pertinente. O estudo foi aprovado pelo Comit? de ?tica da Universidade Federal do Rio Grande do Norte, com Certificado de Apresenta??o para Aprecia??o ?tica n? 08696212.7.0000.5537. Os resultados revelaram que o diagn?stico de enfermagem estudado esteve presente em 82% dos pacientes. Dentre as caracter?sticas com preval?ncia acima de 50% destacaram-se: azotemia, hemat?crito diminu?do, eletr?litos alterados, ingest?o maior que o d?bito, ansiedade, edema, hemoglobina diminu?da, olig?ria e mudan?a na press?o arterial. 8 Apresentaram associa??o estatisticamente significativa com o diagn?stico de enfermagem investigado oito caracter?sticas definidoras: congest?o pulmonar, ingesta maior que o d?bito, eletr?litos alterados, distens?o da jugular, edema, ganho de peso em um curto per?odo de tempo, agita??o e ru?dos advent?cios. Dentre estas, as que apresentaram maiores raz?es de preval?ncia foram edema e ganho de peso em um curto per?odo de tempo. A caracter?stica com maior sensibilidade foi o edema e sobressa?ram-se com maior especificidade ru?dos advent?cios, congest?o pulmonar e agita??o. Os indicadores distens?o da jugular, eletr?litos alterados, ingesta maior que o d?bito, press?o venosa central aumentada e edema, em conjunto, foram identificados no modelo da regress?o como as caracter?sticas preditoras mais significantes. Conclui-se que a identifica??o de indicadores cl?nicos acurados permite uma boa predi??o do diagn?stico de enfermagem Volume de l?quidos excessivo em pacientes submetidos ? hemodi?lise, de modo a auxiliar no processo de infer?ncia do enfermeiro, o que contribuir? no sucesso do cuidado prestado ao paciente. Al?m disso, os enfermeiros considerar?o para a infer?ncia diagn?stica n?o apenas sua experi?ncia cl?nica, mas tamb?m evid?ncias cient?ficas da ocorr?ncia do Volume de l?quidos excessivo, contribuindo para o controle da volemia nesses pacientes

Submitted by Pedro Silva Filho (pfsilva@ipen.br) on 2017-10-09T19:31:24Z No. of bitstreams: 0 / Made available in DSpace on 2017-10-09T19:31:24Z (GMT). No. of bitstreams: 0 / A marcação por feixe laser em superfícies é utilizada para assegurar a identificação e a rastreabilidade em biomateriais. A texturização via feixe laser confere maior aderência às superfícies dos dispositivos médicos implantáveis. Este trabalho teve o objetivo de avaliar o comportamento da superfície do aço inoxidável austenítico ABNT NBR ISO 5832-1 submetido à marcação e texturização com feixe laser de fibra óptica, alterando-se a frequência dos pulsos; frente ao seu comportamento biomecânico, por meio de: ensaios de resistência à tração, fadiga e desgaste; verificar a susceptibilidade à corrosão localizada, por ensaios eletroquímicos em solução que simula os fluidos corpóreos; e caracterizar a sua microestrutura. Os tratamentos alteraram a rugosidade e a dureza dos biomateriais em função do aumento da frequência dos pulsos. A microestrutura e composição química das superfícies sofreram mudanças que afetaram diretamente a camada passiva dos aços inoxidáveis. Este efeito foi comprovado com o uso de SVET, XPS e caracterização de propriedades eletrônicas do filme passivo. Os dois tipos de tratamentos implicaram em aumento de susceptibilidade magnética das superfícies. Os parâmetros utilizados para as marcações e texturizações não causaram diminuição na viabilidade celular, de modo que não apresentou citotoxicidade mesmo após incubação prolongada. Este biomaterial mostrou-se adequado perante os ensaios biomecânicos, uma vez que os tratamentos a laser, nas condições utilizadas, não induziram a formação de tensões superficiais de magnitude capaz de levar à fratura por fadiga, indicando vida em fadiga infinita; tampouco se pôde relacionar a região de fratura por tração com as marcações a laser. O volume de desgaste diminuiu em função do aumento da dureza produzido pela elevação da frequência do pulso nas texturizações. O caráter visual das marcações e texturizações por feixe laser foi assegurado após a maioria dos ensaios realizados. / Tese (Doutorado em Tecnologia Nuclear) / IPEN/T / Instituto de Pesquisas Energéticas e Nucleares - IPEN-CNEN/SP

Des recherches en sciences judiciaires ont montré récemment une possible corrélation entre le temps d’entreposage d’échantillons de fluides corporels et la dégradation de l’ARN dans ceux-ci. Le moment où une tache a été déposée sur une scène de crime peut être important pour déterminer la pertinence d’un échantillon dans une enquête. Dans ce mémoire, nous rapportons les profils de dégradation de quatre ARN différents mesurés par RT-qPCR, soit l’ARN ribosomique 18S et les ARNm de la β-actine, de la glyceraldehyde-3-phosphate déhydrogénase et de la cyclophiline A, obtenus de taches de sang, de salive et de sperme, entreposés à la température de la pièce ou au congélateur à -80°C sur une période de 6 mois. Nos résultats montrent une faible variation interindividuelle pour le sang et le sperme, mais une différence importante entre les donneurs pour la salive. De plus, le profil de dégradation est semblable pour tous les transcrits, mais diffère entre les fluides. La congélation des échantillons stabilise les ARN avant leur analyse. Finalement, la quantité d’ARN détecté est en relation avec le temps d’entreposage et pourrait être utilisée afin d’estimer l’âge des échantillons lorsque l’impact des conditions d’entreposage sur la dégradation de l’ARN sera mieux connu. / Recent studies in forensic science have shown a possible correlation between the degradation rate of some RNA transcripts and the age of bloodstains. The time of deposition of a stain can be of major importance to determine the relevance of a sample in a forensic investigation. In this thesis, we describe the degradation profiles of the 18S ribosomal RNA and the β-actin, glyceraldehyde-3-phosphate dehydrogenase and cyclophilin A mRNAs, measured by RT-qPCR and obtained from dried blood, semen and saliva stains stored at room temperature or frozen at -80°C up to 6 months. Our results showed low inter-individual variation for blood and semen stains, but a high variation was observed between donors for saliva. Moreover, degradation profile of each transcripts was similar, but differed between fluids. Freezing samples prevented RNA degradation over time. Finally, RNA quantity was in relation with the time of storage and could be used to estimate the time since deposition of a stain when the effects of various storage conditions on RNA degradation profiles will be better documented.

Indiana University-Purdue University Indianapolis (IUPUI) / Articular cartilage is subjected to dynamic mechanical loading during normal daily activities. This complex mechanical loading, including cell deformation and interstitial fluid flow, affects chondrocyte mechano-chemical signaling and subsequent cartilage homeostasis and remodeling. Focal adhesion kinase (FAK) and Src are known to be main mechanotransduction proteins, but little is known about the effect of mechanical loading on FAK and Src under its varying magnitudes and types. In this study, we addressed two questions using C28/I2 chondrocytes subjected to the different types and magnitudes of mechanical loading: Does a magnitude of the mechanical loading affect activities of FAK and Src? Does a type of the mechanical loading also affect their activities? Using fluorescence resonance energy transfer (FRET)-based FAK and Src biosensor in live C28/I2 chondrocytes, we monitored the effects of interstitial fluid flow and combined effects of cell deformation/interstitial fluid flow on FAK and Src activities. The results revealed that both FAK and Src activities in C28/I2 chondrocytes were dependent on the different magnitudes of the applied fluid flow. On the other hand, the type of mechanical loading differently affected FAK and Src activities. Although FAK and Src displayed similar activities in response to interstitial fluid flow only, simultaneous application of cell deformation and interstitial fluid flow induced differential FAK and Src activities possibly due to the additive effects of cell deformation and interstitial fluid flow on Src, but not on FAK. Collectively, the data suggest that the intensities and types of mechanical loading are critical in regulating FAK and Src activities in chondrocytes.

Dreiwertige Actinide (An(III)) und Lanthanide (Ln(III)) stellen im Falle ihrer Inkorporation eine ernste Gefahr für die Gesundheit des Menschen dar. An(III) sind künstlich erzeugte, stark radioaktive Elemente, die insbesondere bei der nuklearen Energiegewinnung in Kernkraftwerken entstehen. Durch Störfälle oder nicht fachgerechte Lagerung radioaktiven Abfalls können sie in die Umwelt und die Nahrungskette des Menschen gelangen. Ln(III) sind hingegen nicht radioaktive Elemente, die natürlicherweise vorkommen und für vielfältige Anwendungen in Technik und Medizin abgebaut werden. Folglich kann der Mensch sowohl mit An(III) als auch Ln(III) in Kontakt kommen bzw. sie inkorporieren. Es ist daher von enormer Wichtigkeit, das Verhalten dieser Elemente im menschlichen Körper aufzuklären. Während makroskopische Vorgänge wie Verteilung, Anreicherung und Ausscheidung bereits sehr gut untersucht sind, ist das Wissen hinsichtlich der chemischen Bindungsform (Speziation) von An(III) und Ln(III) in Körperflüssigkeiten noch sehr lückenhaft. In der vorliegenden Arbeit wurde daher erstmals die chemische Bindungsform von Cm(III) und Eu(III) in natürlichem menschlichem Urin (in vitro) spektroskopisch aufgeklärt und die gebildeten Komplexe identifiziert. Hierzu wurden auch grundlegende Untersuchungen zur Komplexierung von Cm(III) und Eu(III) in synthetischem Modellurin sowie mit den urinrelevanten organischen Modellliganden Harnstoff, Alanin, Phenylalanin, Threonin und Citrat durchgeführt und die noch unbekannten Komplexbildungskonstanten bestimmt. Abschließend wurden alle experimentellen Ergebnisse mit Literaturdaten und Vorherberechnungen mittels thermodynamischer Modellierung verglichen. Auf Grund der hervorragenden Lumineszenzeigenschaften von Cm(III) und Eu(III) konnte insbesondere auch die Eignung der zeitaufgelösten laserinduzierten Fluoreszenzspektroskopie (TRLFS) als Methode zur Untersuchung dieser Metallionen in unbehandelten, komplexen biologischen Flüssigkeiten demonstriert werden. Die Ergebnisse dieser Arbeit liefern damit neue Erkenntnisse zu den biochemischen Reaktionen von An(III) und Ln(III) in Körperflüssigkeiten auf molekularer Ebene und tragen zu einem besseren Verständnis der bekannten, makroskopischen Effekte dieser Elemente bei. Darüber hinaus sind sie die Grundlage weiterführender in-vivo-Untersuchungen.:1 Motivation und Zielstellung 2 Speziationsbestimmung exogener Schwermetalle in Biofluiden 2.1 Actinide und Lanthanide 2.2 Biochemisches Verhalten exogener Schwermetalle im Menschen 2.3 Speziationsbestimmung von Metallen 3 Komplexbildung von Curium(III) und Europium(III) mit organischen Modellliganden 3.1 Lumineszenzspektroskopische Eigenschaften von Curium(III) und Europium(III) in Wasser 3.2 Harnstoff – Hauptbestandteil des menschlichen Urins 3.3 Citronensäure – ubiquitäres Biomolekül0 3.4 Aminosäuren – Grundbausteine des Lebens 4 Speziation von Curium(III) und Europium(III) in menschlichen Urinproben 4.1 Charakterisierung und Analyse der natürlichen menschlichen Urinproben 4.2 Bestimmung der Speziation von Curium(III) und Europium(III) in Modellurin 4.3 Bestimmung der Speziation von Curium(III) und Europium(III) in menschlichem Urin 5 Diskussion 5.1 Vergleich der Komplexbildungseigenschaften von Curium(III) und Europium(III) 5.2 Thermodynamische Modellierung der Speziation von Curium(III) und Europium(III) in menschlichem Urin 5.3 Ausblick 6 Experimentelles / In case of incorporation, trivalent actinides (An(III)) and lanthanides (Ln(III)) pose a serious health risk to humans. An(III) are artificial, highly radioactive elements which are mainly produced during the nuclear fuel cycle in nuclear power plants. Via hazardous accidents or nonprofessional storage of radioactive waste, they can be released in the environment and enter the human food chain. In contrast, Ln(III) are nonradioactive, naturally occurring elements with multiple applications in technique and medicine. Consequently it is possible that humans get in contact and incorporate both, An(III) and Ln(III). Therefore, it is of particular importance to elucidate the behaviour of these elements in the human body. While macroscopic processes such as distribution, accumulation and excretion are studied quite well, knowledge about the chemical binding form (speciation) of An(III) and Ln(III) in various body fluids is still sparse. In the present work, for the first time, the speciation of Cm(III) and Eu(III) in natural human urine (in vitro) has been investigated spectroscopically and the formed complex identified. For this purpose, also basic investigations on the complex formation of Cm(III) and Eu(III) in synthetic model urine as well as with the urinary relevant, organic model ligands urea, alanine, phenylalanine, threonine and citrate have been performed and the previously unknown complex stability constants determined. Finally, all experimental results were compared to literature data and predictions calculated by thermodynamic modelling. Since both, Cm(III) and Eu(III), exhibit unique luminescence properties, particularly the suitability of time-resolved laser-induced fluorescence spectroscopy (TRLFS) could be demonstrated as a method to investigate these metal ions in untreated, complex biofluids. The results of this work provide new scientific findings on the biochemical reactions of An(III) and Ln(III) in human body fluids on a molecular scale and contribute to a better understanding of the known macroscopic effects of these elements. Furthermore, they are the basis of subsequent in vivo investigations.:1 Motivation und Zielstellung 2 Speziationsbestimmung exogener Schwermetalle in Biofluiden 2.1 Actinide und Lanthanide 2.2 Biochemisches Verhalten exogener Schwermetalle im Menschen 2.3 Speziationsbestimmung von Metallen 3 Komplexbildung von Curium(III) und Europium(III) mit organischen Modellliganden 3.1 Lumineszenzspektroskopische Eigenschaften von Curium(III) und Europium(III) in Wasser 3.2 Harnstoff – Hauptbestandteil des menschlichen Urins 3.3 Citronensäure – ubiquitäres Biomolekül0 3.4 Aminosäuren – Grundbausteine des Lebens 4 Speziation von Curium(III) und Europium(III) in menschlichen Urinproben 4.1 Charakterisierung und Analyse der natürlichen menschlichen Urinproben 4.2 Bestimmung der Speziation von Curium(III) und Europium(III) in Modellurin 4.3 Bestimmung der Speziation von Curium(III) und Europium(III) in menschlichem Urin 5 Diskussion 5.1 Vergleich der Komplexbildungseigenschaften von Curium(III) und Europium(III) 5.2 Thermodynamische Modellierung der Speziation von Curium(III) und Europium(III) in menschlichem Urin 5.3 Ausblick 6 Experimentelles

Do metal particles (including particles of pure metals, alloys, metal oxides and compounds) pose a hazard or risk to human health? In the light of this question, this thesis summarizes results from research conducted on metal particles, and describes the elaboration and implementation of an in vitro test methodology to study metal release from particles through corrosion and dissolution processes in synthetic biological media relevant for human exposure through inhalation/ingestion and dermal contact. Bioaccessible metals are defined as the pool of released metals from particles that potentially could be made available for absorption by humans or other organisms. Studies of bioaccessible metals from different metal particles within this thesis have shown that the metal release process is influenced by material properties, particle specific properties, size distribution, surface area and morphology, as well as the chemistry of synthetic biological test media simulating various human exposure scenarios. The presence of metal particles in proximity to humans and the fact that metals can be released from particles to a varying extent is the hazard referred to in the title. The bioavailable metal fraction of the released metals (the fraction available for uptake/absorption by humans through different exposure routes) is usually significantly smaller than the bioaccessible pool of released metals, and is largely related to the chemical form and state of oxidation of the released metals. Chemical speciation measurements of released chromium for instance revealed chromium to be complexed to its non-available form in simulated lung fluids. Such measurements provide an indirect measure of the potential risk for adverse health effects, when performed at relevant experimental conditions. A more direct way to assess risks is to conduct toxicological in-vitro testing of metal particles, for instance on lung cell cultures relevant for human inhalation. Induced toxicity of metal particles on lung cells includes both the effect of the particles themselves and of the released metal fraction (including bioaccessible and bioavailable metals), the latter shown to be less predominant. The toxic response was clearly influenced by various experimental conditions such as sonication treatment of particles and the presence of serum proteins. Thorough characterization of metal particles assessing parameters including chemical surface composition, degree of agglomeration in solution, size distribution, surface area and morphology was performed and discussed in relation to generated results of bioaccessibility, bioavailability and induced toxicity. One important conclusion was that neither the surface composition nor the bulk composition can be used to assess the extent of metals released from chromium-based alloy particles. These findings emphasize that information on physical-chemical properties and surface characteristics of particles is essential for an in-depth understanding of metal release processes and for further use and interpretation of bioaccessibility data to assess hazard and reduce any risks induced by human exposure to metal particles. / QC 20100803

The open nature of Cell-Free Protein Synthesis (CFPS) systems has enabled flexible design, easy manipulation, and novel applications of protein engineering in therapeutic production, biocatalysis, and biosensors. This dissertation reports on three advances in the application of CFPS systems for 1) improving biocatalysis performance in industrial applications by site-specific covalent enzyme immobilization, 2) expressing and optimizing a difficult to express a mammalian protein in bacterial-based CFPS systems and its application for cost-effective, on-demand biosensors compatible with human body fluids, and 3) streamlining the procedure of an E. coli extract with built-in compatibility with human body fluid biosensors. Site-specific covalent immobilization stabilizes enzymes and facilitates recovery and reuse of enzymes which improves the net profit margin of industrial enzymes. Yet, the suitability of a given site on the enzyme for immobilization remains a trial-and-error procedure. This dissertation reports the reliability of several design heuristics and a coarse-grain molecular simulation in predicting the optimum sites for covalent immobilization of a target enzyme, TEM-1 ?-lactamase. This work demonstrates that the design heuristics can successfully identify a subset of favorable locations for experimental validation. This approach highlights the advantages of combining coarse-grain simulation and high-throughput experimentation using CFPS to efficiently identify optimal enzyme immobilization sites. Additionally, this dissertation reports high-yield soluble expression of a difficult-to-express protein (murine RNase Inhibitor or m-RI) in E. coli-lysate-based CFPS. Several factors including reaction temperature, reaction time, redox potential, and presence of folding chaperones in CFPS reactions were altered to find suitable conditions for m-RI expression. m-RI with the highest activity and stability was used to develop a lyophilized CFPS biosensor in human body fluids which reduced the cost of biosensor test by ~90%. Moreover, an E. coli extract with RNase inhibition activity was developed and tested which further streamlines the production of CFPS biosensors compatible with human body fluids.