A novel method for rapid and selective extraction of male DNA from rape kits using alkaline lysis and pressure cycling technology (PCT)

There is an increasing demand for DNA analysis because of the sensitivity of the method and the ability to uniquely identify and distinguish individuals with a high degree of certainty. But this demand has led to huge backlogs in evidence lockers since the current DNA extraction protocols require long processing time. The DNA analysis procedure becomes more complicated when analyzing sexual assault casework samples where the evidence contains more than one contributor. Additional processing to separate different cell types in order to simplify the final data interpretation further contributes to the existing cumbersome protocols. The goal of the present project is to develop a rapid and efficient extraction method that permits selective digestion of mixtures. ^ Selective recovery of male DNA was achieved with as little as 15 minutes lysis time upon exposure to high pressure under alkaline conditions. Pressure cycling technology (PCT) is carried out in a barocycler that has a small footprint and is semi-automated. Typically less than 10% male DNA is recovered using the standard extraction protocol for rape kits, almost seven times more male DNA was recovered from swabs using this novel method. Various parameters including instrument setting and buffer composition were optimized to achieve selective recovery of sperm DNA. Some developmental validation studies were also done to determine the efficiency of this method in processing samples exposed to various conditions that can affect the quality of the extraction and the final DNA profile. ^ Easy to use interface, minimal manual interference and the ability to achieve high yields with simple reagents in a relatively short time make this an ideal method for potential application in analyzing sexual assault samples.^

Simulação da destilação molecular de filme descendente para o petróleo

The heavy part of the oil can be used for numerous purposes, e.g. to obtain lubricating oils. In this context, many researchers have been studying alternatives such separation of crude oil components, among which may be mentioned molecular distillation. Molecular distillation is a forced evaporation technique different from other conventional processes in the literature. This process can be classified as a special distillation case under high vacuum with pressures that reach extremely low ranges of the order of 0.1 Pascal. The evaporation and condensation surfaces must have a distance from each other of the magnitude order of mean free path of the evaporated molecules, that is, molecules evaporated easily reach the condenser, because they find a route without obstacles, what is desirable. Thus, the main contribution of this work is the simulation of the falling-film molecular distillation for crude oil mixtures. The crude oil was characterized using UniSim® Design and R430 Aspen HYSYS® V8.5. The results of this characterization were performed in spreadsheets of Microsoft® Excel®, calculations of the physicochemical properties of the waste of an oil sample, i.e., thermodynamic and transport. Based on this estimated properties and boundary conditions suggested by the literature, equations of temperature and concentration profiles were resolved through the implicit finite difference method using the programming language Visual Basic® (VBA) for Excel®. The result of the temperature profile showed consistent with the reproduced by literature, having in their initial values a slight distortion as a result of the nature of the studied oil is lighter than the literature, since the results of the concentration profiles were effective allowing realize that the concentration of the more volatile decreases and of the less volatile increases due to the length of the evaporator. According to the transport phenomena present in the process, the velocity profile tends to increase to a peak and then decreases, and the film thickness decreases, both as a function of the evaporator length. It is concluded that the simulation code in Visual Basic® language (VBA) is a final product of the work that allows application to molecular distillation of petroleum and other similar mixtures.

Gravidade do olho seco em pacientes internados em unidade de terapia intensiva: análise de conceito e construção de definições

Dry eye syndrome is a multifactorial disease of the tear film, resulting from the instability of the lacrimal functional unit that produces volume change, up or tear distribution. In patients in intensive care the cause is enhanced due to various risk factors, such as mechanical ventilation, sedation, lagophthalmos, low temperatures, among others. The study's purpose is to build an assessment tool of Dry Eye Severity in patients in intensive care units based on the systematization of nursing care and their classification systems. The aim of this study is to build an assessment tool of Dry Eye Severity in hospitalized patients in Care Unit Intensiva.Trata is a methodological study conducted in three stages, namely: context analysis, concept analysis, construction of operational definitions and magnitudes of nursing outcome. For the first step we used the methodological framework for Hinds, Chaves and Cypress (1992). For the second step we used the model of Walker and Avant and an integrative review Whitemore seconds, Knalf (2005). This step enabled the identification of the concept of attributes, background and consequent ground and the construction of the settings for the result of nursing severity of dry eye. For the construction of settings and operational magnitudes, it was used Psicometria proposed by Pasquali (1999). As a result of context analysis, visualized from the reflection that the matter should be discussed and that nursing needs to pay attention to the problem of eye injury, so minimizing strategies are created this event with a high prevalence. With the integrative review were located from the crosses 19 853 titles, selected 215, and from the abstracts 96 articles were read in full. From reading 10 were excluded culminating in the sample of 86 articles that were used to analyze the concept and construction of settings. Selected articles were found in greater numbers in the Scopus database (55.82%), performed in the United States (39.53%), and published mainly in the last five years (48.82). Regarding the concept of analysis were identified as antecedents: age, lagophthalmos, environmental factors, medication use, systemic diseases, mechanical ventilation and ophthalmic surgery. As attributes: TBUT <10s, Schimer I test <5 mm in Schimer II test <10mm, reduced osmolarity. As consequential: the ocular surface damage, ocular discomfort, visual instability. The settings were built and added indicators such as: decreased blink mechanism and eyestrain.

Qualidade da água de um reservatório e do solo da zona ripária sob diferentes usos na região semiárida do Rio Grande do Norte

Reservoirs are artificial ecosystems intermediate between rivers and lakes widely used in the Brazilian semiarid region as a way to provide water supply due to the said region’s water scarcity. The use of water from these supply sources for multiple uses, along with occupation and utilization of its riparian zone without proper management, directly influences the increased nutrient flow into aquatic environments, there with contributing to the acceleration of eutrophication. The semi-arid region is characterized by peculiar weather conditions, such as severe evaporation, high temperatures with little variation throughout the year and long water residence time, making it susceptible to prolonged drought occurrence, which tends to concentrate the nutrients in reservoirs, which favors the development of eutrophic conditions. Moreover, it is common soil use and occupation by carrying out activities with potential environmental impact on natural resources such as agriculture, livestock farming and lack of sanitation. The aim of this study is both to evaluate the water quality of the Cruzeta Reservoir, located in the semiarid region of Rio Grande do Norte, during a prolonged drought period, and assess the quality of its riparian zone soil under different uses, by monitoring physical-chemical variables. Along the prolonged drought, high levels of turbidity, suspended solids, nutrients and chlorophyll a were verified as present, therefore featuring low water quality. In the riparian zone of Cruzeta Reservoir, the areas under use of agriculture and livestock farming appeared as one of the main diffuse sources of nutrients to the said reservoir, featuring the highest levels of phosphorus and nitrogen in the soil, originated from decomposition of animal excreta and from the use of fertilizers, creating a tendency to increased eutrophication of such water supply source. The indicators of water and soil quality are useful for monitoring and evaluating the conservation status of natural resources, allowing the control and mitigation of the reservoir eutrophication process. This study confirmed the hypothesis that the reduction of water level, resulting from prolonged drought event, aggravates the symptoms of eutrophication; and also that using the soil under severalways modifies the physic chemical properties of the soil, having livestock farming and agriculture as the usages with greatest potential towards yielding P and N to the aquatic environment.

A dinâmica geográfica da construção de um território para moradias sociais: estudo testemunhal do empreendimento Vitória Brasil em Uberlândia (MG)

This work describes and analyzes the situation planned with the current, according to the housing development project style of social interest (Gleba 9), named Victory Brazil at Shopping Park II neighborhood, located in the southern sector of Uberlândia (MG), built with Program resources Minha Casa, Minha Vida (PMCMV). The specific objectives are: to present the housing policy of social interest directed to the Minha Casa, Minha Vida in the city of Uberlândia, from its history to its current information; trace the environmental characterization of the housing Vitoria Brazil through the development of thematic maps; identify the impacts generated by the housing projects of social interest; and show through interviews with 25 residents of Vitoria Brazil, the structural problems of the houses. Therefore, the research became quali-quantitative with descriptive approach and use of semi- structured interviews to collect data. In addition, the iconographic sources, ie photographs of the problems raised by the residents of the whole Victory Brazil and maps of the study area, enlarged the qualitative and dialogic nature of this study, as there is opportunity to contextualize specific way what was reported the residents interviewed. We understand that the city is intrinsically related to the design of agglomeration, characterized by the production of the possessor market capitalism a public administration system that is guided by the ideals of consumption and utilization of space, and its inhabitants are high class with the construction of equipment and adequate social services. One of the important biases to solving the problems of urbanization is by the intervention of the government through public policies through the master plan so that would allow prospects to enable the population of low-income housing in locations that offer housing and decent urbanization conditions.

ToPI - uma abordagem online para identificar locais de interesse utilizando fotografias geo-referenciadas

Due to the growing use of social networks people no longer just consume data, they also produce and share it. Geo-tagged information, i.e., data with geographical location, have been used in many attempts to identify popular places and help tourists that will visit unfamiliar cities. This Master Thesis presents an online strategy that uses geo-tagged photos and their metadata in order to identify places of interest inside a given geographical area and retrieve relevant related information. The whole process runs automatically in real time, returning updated information about places. The proposed strategy takes into account the inherent dynamism of social media, and thus is robust under inconsistencies and/or outdated information, a common issue in solutions that rely on previously stored data. The analysis of the results showed that our approach is very promising, returning places that present high agreement with those from a popular travel website.

Team reflexivity and innovation:the moderating role of team context

We developed and tested a team level contingency model of innovation, integrating theories regarding work demands, team reflexivity - the extent to which teams collectively reflect upon their working methods and functioning -, and team innovation. We argued that highly reflexive teams will be more innovative than teams low in reflexivity when facing a demanding work environment. The relationships between team reflexivity, a demanding work environment (i.e. quality of the physical work environment and work load) and team innovation was examined among 98 primary health care teams (PHCTs) in the UK, comprised of 1137 individuals. Results showed that team reflexivity is positively related to team innovation, and that there is an interaction between team reflexivity, team level workload, and team innovation, such that when team level workload is high, combined with a high level of team reflexivity, team innovation is also higher. The complementary interaction between team reflexivity, quality of physical work environment, and team innovation, showed that when the quality of the work environment is low, combined with a high level of team reflexivity, team innovation was also higher. These results are discussed in the context of the need for team reflexivity and team innovation among teams at work facing high work demands.

The sound of whiteness: early music vocal performance practice in Britain

American Musicological Society annual meeting, San Francisco, 10 Nov. 2011

Leader's reflections on knowledge, knowing and not knowing: an analysis of change over time

One commonality across the leadership and knowledge related literature is the apparent neglect of the leaders own knowledge. This thesis sought to address this issue through conducting exploratory research into the content of leader’s personal knowledge and the process of knowing it. The empirical inquiry adopted a longitudinal approach, with interviews conducted at two separate time periods with an extended time-interval between each. The findings from this research contrast with images of leadership which suggest leaders are in control of what they know, that they own their own knowledge. The picture that emerges is one of individuals struggling to keep abreast of the knowledge required to deal with the dynamics and uncertainties of organisational life. Much knowledge is tacit, provisional and perishable and the related process of knowing more organic, evolutionary and informal than any structured or orchestrated approach. The collective nature of knowing is a central feature, with these leaders embedded in networks of uncontrollable relationships. In view of the indeterminate nature of knowing, the boundary between what is known and what one needs to know is both amorphous and ephemeral, and the likelihood of knowledge-absences is escalated. A significant finding in this regard is the identification of two critical points where not-knowing is most likely (entry and exit from role) and the differing implications of each. Overtime the knowledge that is legitimised or prioritised is significantly altered as these leaders replace the dogmas that were previously held in high esteem with the lessons from their own experience. This experience brings increased self-knowledge and a deeper appreciation of the values and morals instilled in their early lives. In view of the above findings, this study makes theoretical contribution to a number of core literatures: authentic leadership, role transition and knowledge-absences. In terms of leadership development, the findings point to the necessity to prepare leaders for the challenges they will encounter at the pivotal stages of the leadership role.

A Novel Method for Rapid and Selective Extraction of Male DNA from Rape Kits using Alkaline Lysis and Pressure Cycling Technology (PCT)

There is an increasing demand for DNA analysis because of the sensitivity of the method and the ability to uniquely identify and distinguish individuals with a high degree of certainty. But this demand has led to huge backlogs in evidence lockers since the current DNA extraction protocols require long processing time. The DNA analysis procedure becomes more complicated when analyzing sexual assault casework samples where the evidence contains more than one contributor. Additional processing to separate different cell types in order to simplify the final data interpretation further contributes to the existing cumbersome protocols. The goal of the present project is to develop a rapid and efficient extraction method that permits selective digestion of mixtures. Selective recovery of male DNA was achieved with as little as 15 minutes lysis time upon exposure to high pressure under alkaline conditions. Pressure cycling technology (PCT) is carried out in a barocycler that has a small footprint and is semi-automated. Typically less than 10% male DNA is recovered using the standard extraction protocol for rape kits, almost seven times more male DNA was recovered from swabs using this novel method. Various parameters including instrument setting and buffer composition were optimized to achieve selective recovery of sperm DNA. Some developmental validation studies were also done to determine the efficiency of this method in processing samples exposed to various conditions that can affect the quality of the extraction and the final DNA profile. Easy to use interface, minimal manual interference and the ability to achieve high yields with simple reagents in a relatively short time make this an ideal method for potential application in analyzing sexual assault samples.

Estudio sobre la variación perfecto simple y perfecto compuesto en los discursos presidenciales de Evo Morales : Marcas del contacto lingüístico

El trabajo presenta un análisis cualitativo-cuantitativo del uso variable del perfecto simple y perfecto compuesto en los discursos presidenciales de Evo Morales, de acuerdo con la perspectiva Etnopragmática desarrollada por García 1990, 1995; Martínez 1995, 2000; Mauder 2000. Se considera la singularidad de una variedad en situación de contacto lingüístico: la del español en contacto con el aymara y el quechua, y se indaga sobre las transformaciones que pueden producirse cuando diferentes lenguas -diferentes visiones de mundo- entran en contacto. En relación con esto, veremos que la presencia, tanto en la lengua quechua como en la lengua aymara, de un sistema evidencial gramaticalizado podría influir en la variación del perfecto simple y compuesto en los discursos de Morales, en particular, que la presencia de la forma del perfecto compuesto podría funcionar como una marca de evidencia directa y su uso contribuiría a la estrategia discursiva del orador de mostrar ciertos hechos de la historia de Bolivia como cercanos a su vivencia y experiencia personales, marcando un mayor grado compromiso con respecto a la veracidad de lo que enuncia. De esta manera, el orador consigue aunar su voz con la de los pueblos originarios, formar parte de los grupos discriminados y excluidos de la historia de su país y así, contar con su apoyo y acompañamiento durante su mandato.

Estudio sobre la variación perfecto simple y perfecto compuesto en los discursos presidenciales de Evo Morales : Marcas del contacto lingüístico

El trabajo presenta un análisis cualitativo-cuantitativo del uso variable del perfecto simple y perfecto compuesto en los discursos presidenciales de Evo Morales, de acuerdo con la perspectiva Etnopragmática desarrollada por García 1990, 1995; Martínez 1995, 2000; Mauder 2000. Se considera la singularidad de una variedad en situación de contacto lingüístico: la del español en contacto con el aymara y el quechua, y se indaga sobre las transformaciones que pueden producirse cuando diferentes lenguas -diferentes visiones de mundo- entran en contacto. En relación con esto, veremos que la presencia, tanto en la lengua quechua como en la lengua aymara, de un sistema evidencial gramaticalizado podría influir en la variación del perfecto simple y compuesto en los discursos de Morales, en particular, que la presencia de la forma del perfecto compuesto podría funcionar como una marca de evidencia directa y su uso contribuiría a la estrategia discursiva del orador de mostrar ciertos hechos de la historia de Bolivia como cercanos a su vivencia y experiencia personales, marcando un mayor grado compromiso con respecto a la veracidad de lo que enuncia. De esta manera, el orador consigue aunar su voz con la de los pueblos originarios, formar parte de los grupos discriminados y excluidos de la historia de su país y así, contar con su apoyo y acompañamiento durante su mandato.

Estudio sobre la variación perfecto simple y perfecto compuesto en los discursos presidenciales de Evo Morales : Marcas del contacto lingüístico

El trabajo presenta un análisis cualitativo-cuantitativo del uso variable del perfecto simple y perfecto compuesto en los discursos presidenciales de Evo Morales, de acuerdo con la perspectiva Etnopragmática desarrollada por García 1990, 1995; Martínez 1995, 2000; Mauder 2000. Se considera la singularidad de una variedad en situación de contacto lingüístico: la del español en contacto con el aymara y el quechua, y se indaga sobre las transformaciones que pueden producirse cuando diferentes lenguas -diferentes visiones de mundo- entran en contacto. En relación con esto, veremos que la presencia, tanto en la lengua quechua como en la lengua aymara, de un sistema evidencial gramaticalizado podría influir en la variación del perfecto simple y compuesto en los discursos de Morales, en particular, que la presencia de la forma del perfecto compuesto podría funcionar como una marca de evidencia directa y su uso contribuiría a la estrategia discursiva del orador de mostrar ciertos hechos de la historia de Bolivia como cercanos a su vivencia y experiencia personales, marcando un mayor grado compromiso con respecto a la veracidad de lo que enuncia. De esta manera, el orador consigue aunar su voz con la de los pueblos originarios, formar parte de los grupos discriminados y excluidos de la historia de su país y así, contar con su apoyo y acompañamiento durante su mandato.

Seawater carbonate chemistry, growth rate and hatching processes of Symsagittifera roscoffensis during experiments, 2012

As a consequence of anthropogenic CO2 emissions, oceans are becoming more acidic, a phenomenon known as ocean acidification. Many marine species predicted to be sensitive to this stressor are photosymbiotic, including corals and foraminifera. However, the direct impact of ocean acidification on the relationship between the photosynthetic and nonphotosynthetic organism remains unclear and is complicated by other physiological processes known to be sensitive to ocean acidification (e.g. calcification and feeding). We have studied the impact of extreme pH decrease/pCO2 increase on the complete life cycle of the photosymbiotic, non-calcifying and pure autotrophic acoel worm, Symsagittifera roscoffensis. Our results show that this species is resistant to high pCO2 with no negative or even positive effects on fitness (survival, growth, fertility) and/or photosymbiotic relationship till pCO2 up to 54 K µatm. Some sub-lethal bleaching is only observed at pCO2 up to 270 K µatm when seawater is saturated by CO2. This indicates that photosymbiosis can be resistant to high pCO2. If such a finding would be confirmed in other photosymbiotic species, we could then hypothesize that negative impact of high pCO2 observed on other photosymbiotic species such as corals and foraminifera could occur through indirect impacts at other levels (calcification, feeding).

Process Monitoring and Defect Detection of Additive Manufacturing Using Inline Coherent Imaging

With applications ranging from aerospace to biomedicine, additive manufacturing (AM) has been revolutionizing the manufacturing industry. The ability of additive techniques, such as selective laser melting (SLM), to create fully functional, geometrically complex, and unique parts out of high strength materials is of great interest. Unfortunately, despite numerous advantages afforded by this technology, its widespread adoption is hindered by a lack of on-line, real time feedback control and quality assurance techniques. In this thesis, inline coherent imaging (ICI), a broadband, spatially coherent imaging technique, is used to observe the SLM process in 15 - 45 $\mu m$ 316L stainless steel. Imaging of both single and multilayer builds is performed at a rate of 200 $kHz$, with a resolution of tens of microns, and a high dynamic range rendering it impervious to blinding from the process beam. This allows imaging before, during, and after laser processing to observe changes in the morphology and stability of the melt. Galvanometer-based scanning of the imaging beam relative to the process beam during the creation of single tracks is used to gain a unique perspective of the SLM process that has been so far unobservable by other monitoring techniques. Single track processing is also used to investigate the possibility of a preliminary feedback control parameter based on the process beam power, through imaging with both coaxial and 100 $\mu m$ offset alignment with respect to the process beam. The 100 $\mu m$ offset improved imaging by increasing the number of bright A-lines (i.e. with signal greater than the 10 $dB$ noise floor) by 300\%. The overlap between adjacent tracks in a single layer is imaged to detect characteristic fault signatures. Full multilayer builds are carried out and the resultant ICI images are used to detect defects in the finished part and improve upon the initial design of the build system. Damage to the recoater blade is assessed using powder layer scans acquired during a 3D build. The ability of ICI to monitor SLM processes at such high rates with high resolution offers extraordinary potential for future advances in on-line feedback control of additive manufacturing.