Development and Characterization of Monovalent and Bivalent RNA Aptamers Targeting the Common Pathway of Coagulation

Anticoagulant agents are commonly used drugs to reduce blood coagulation in acute and chronic clinical settings. Many of these drugs target the common pathway of coagulation because it is critical for thrombin generation and disruption of this portion of the pathway has profound effects on the hemostatic process. Currently available drugs for these indications struggle with balancing desired activity with immunogenicity and poor reversibility or irreversibility in the event of hemorrhage. While improvements are being made with the current drugs, new drugs with better therapeutic indices are needed for surgical intervention and chronic indications to prevent thrombosis from occurring.

A class of therapeutics known as aptamers may be able to meet the need for safer anticoagulant agents. Aptamer are short single-stranded RNA oligonucleotides that adopt specific secondary and tertiary structures based upon their sequence. They can be generated to both enzymes and cofactors because they derive their inhibitory activity by blocking protein-protein interactions, rather than active site inhibition. They inhibit their target proteins with a high level of specificity and bind with high affinity to their target. Additionally, they can be reversed using two different antidote approaches, specific oligonucleotide antidotes, or with cationic, “universal” antidotes. The reversal of their activity is both rapid and durable.

The ability of aptamers to be generated to cofactors has been conclusively proven by generating an aptamer targeting the common pathway coagulation cofactor, Factor V (FV). We developed two aptamers with anticoagulant ability that bind to both FV and FVa, the active cofactor. Both aptamers were truncated to smaller functional sizes and had specific point mutant aptamers developed for use as controls. The anticoagulant activity of both aptamer-mutant pairs was characterized using plasma-based clotting assays and whole blood assays. The mechanism of action resulting in anticoagulant activity was assessed for one aptamer. The aptamer was found to block FVa docking to membrane surfaces, a mechanism not previously observed in any of our other anticoagulant aptamers.

To explore development of aptamers as anticoagulant agents targeting the common pathway for surgical interventions, we fused two anticoagulant aptamers targeting Factor X and prothrombin into a single molecule. The bivalent aptamer was truncated to a minimal size while maintaining robust anticoagulant activity. Characterization of the bivalent aptamer in plasma-based clotting assays indicated we had generated a very robust anticoagulant therapeutic. Furthermore, we were able to simultaneously reverse the activity of both aptamers with a single oligonucleotide antidote. This rapid and complete reversal of anticoagulant activity is not available in the antithrombotic agents currently used in surgery.

PENTAZOCINE VERSUS PENTAZOCINE WITH RECTAL DICLOFENAC FOR POSTOPERATIVE PAIN RELIEF AFTER CESAREAN SECTION- A DOUBLE BLIND RANDOMIZED PLACEBO CONTROLLED TRIAL IN A LOW RESOURCE AREA.

BACKGROUND: The unimodal approach of using pentazocine as post-cesarean section pain relief is inadequate, hence the need for a safer, easily available and more effective multimodal approach. AIM: To evaluate the effectiveness of rectal diclofenac combined with intramuscular pentazocine for postoperative pain following cesarean section. METHODS: In this double blind clinical trial, 130 pregnant women scheduled for cesarean section under spinal anesthesia were randomly assigned to two groups. Group A received 100mg diclofenac suppository and group B received placebo suppository immediately following surgery, 12 and 24h later. Both groups also received intramuscular pentazocine 30mg immediately following surgery and 6 hourly postoperatively in the first 24 h. Postoperative pain was assessed by visual analogue scale at end of surgery and 2, 12 and 24 h after surgery. Patient satisfaction scores were also assessed. RESULTS: One hundred and sixteen patients completed the study. Combining diclofenac and pentazocine had statistically significant reduction in pain intensity at 2, 12, and 24 hours postoperatively compared to pentazocine alone (p <0.05). No significant side effects were noted in both groups. The combined group also had significantly better patient satisfaction scores. CONCLUSION: The addition of diclofenac suppository to intramuscular pentazocine provides better pain relief after cesarean section and increased patient satisfaction.

Biocompatibility Assessment of Biosorbable Polymer Coated Nitinol Alloys

Owing to an increased risk of aging population and a higher incidence of coronary artery disease (CAD), there is a need for more reliable and safer treatments. Numerous varieties of durable polymer-coated drug eluting stents (DES) are available in the market in order to mitigate in-stent restenosis. However, there are certain issues regarding their usage such as delayed arterial healing, thrombosis, inflammation, toxic corrosion by-products, mechanical stability and degradation. As a result, significant amount of research has to be devoted to the improvement of biodegradable polymer-coated implant materials in an effort to enhance their bioactive response. In this investigation, magneto-electropolished (MEP) and a novel biodegradable polymer coated ternary Nitinol alloys, NiTiTa and NiTiCr were prepared to study their bio and hemocompatibility properties. The initial interaction of a biomaterial with its surroundings is dependent on its surface characteristics such as, composition, corrosion resistance, work of adhesion and morphology. In-vitro corrosion tests such as potentiodynamic polarization and electrochemical impedance spectroscopic (EIS) studies were conducted to determine the coating stability and longevity. In-vitro hemocompatibility studies and HUVEC cell growth was performed to determine their thrombogenic and biocompatibility properties. Critical delamination load of the polymer coated Nitinol alloys was determined using Nano-scratch analysis. Sulforhodamine B (SRB) assays were performed to elucidate the effect of metal ions leached from Nitinol alloys on the viability of HUVEC cells. Scanning Electron Microscopy (SEM), X-ray Photoelectron Spectroscopy (XPS), contact angle meter and X-ray diffraction (XRD) were used to characterize the surface of the alloys. MEP treated and polymer coated (PC) Nitinol alloys displayed a corrosion resistant polymer coating as compared to uncoated alloys. MEP and PC has resulted in reduced Ni and Cr ion leaching from NiTi5Cr and subsequently low cytotoxicity. Thrombogenicity tests revealed significantly less platelet adhesion and confluent endothelial cell growth on polymer coated and uncoated ternary MEP Nitinol alloys. Finally, this research addresses the bio and hemocompatibility of MEP + PC ternary Nitinol alloys that could be used to manufacture blood contacting devices such as stents and vascular implants which can lead to lower U.S. healthcare spending.

Terrain Controls and Landscape-Scale Susceptibility Modelling of Active-Layer Detachments, Sabine Peninsula, Melville Island, Nunavut

Modelling the susceptibility of permafrost slopes to disturbance can identify areas at risk to future disturbance and result in safer infrastructure and resource development in the Arctic. In this study, we use terrain attributes derived from a digital elevation model, an inventory of permafrost slope disturbances known as active-layer detachments (ALDs) and generalised additive modelling to produce a map of permafrost slope disturbance susceptibility for an area on northern Melville Island, in the Canadian High Arctic. By examining terrain variables and their relative importance, we identified factors important for initiating slope disturbance. The model was calibrated and validated using 70 and 30 per cent of a data-set of 760 mapped ALDs, including disturbed and randomised undisturbed samples. The generalised additive model calibrated and validated very well, with areas under the receiver operating characteristic curve of 0.89 and 0.81, respectively, demonstrating its effectiveness at predicting disturbed and undisturbed samples. ALDs were most likely to occur below the marine limit on slope angles between 3 and 10° and in areas with low values of potential incoming solar radiation (north-facing slopes).

The Economic Case for Electric Mining Equipment and Technical Considerations Relating to their Implementation

Underground hardrock mining can be very energy intensive and in large part this can be attributed to the power consumption of underground ventilation systems. In general, the power consumed by a mine’s ventilation system and its overall scale are closely related to the amount of diesel power in operation. This is because diesel exhaust is a major source of underground air pollution, including diesel particulate matter (DPM), NO2 and heat, and because regulations tie air volumes to diesel engines. Furthermore, assuming the size of airways remains constant, the power consumption of the main system increases exponentially with the volume of air supplied to the mine. Therefore large diesel fleets lead to increased energy consumption and can also necessitate large capital expenditures on ventilation infrastructure in order to manage power requirements. Meeting ventilation requirements for equipment in a heading can result in a similar scenario with the biggest pieces leading to higher energy consumption and potentially necessitating larger ventilation tubing and taller drifts. Depending on the climate where the mine is located, large volumes of air can have a third impact on ventilation costs if heating or cooling the air is necessary. Annual heating and cooling costs, as well as the cost of the associated infrastructure, are directly related to the volume of air sent underground. This thesis considers electric mining equipment as a means for reducing the intensity and cost of energy consumption at underground, hardrock mines. Potentially, electric equipment could greatly reduce the volume of air needed to ventilate an entire mine as well as individual headings because they do not emit many of the contaminants found in diesel exhaust and because regulations do not connect air volumes to electric motors. Because of the exponential relationship between power consumption and air volumes, this could greatly reduce the amount of power required for mine ventilation as well as the capital cost of ventilation infrastructure. As heating and cooling costs are also directly linked to air volumes, the cost and energy intensity of heating and cooling the air would also be significantly reduced. A further incentive is that powering equipment from the grid is substantially cheaper than fuelling them with diesel and can also produce far fewer GHGs. Therefore, by eliminating diesel from the underground workers will enjoy safer working conditions and operators and society at large will gain from a smaller impact on the environment. Despite their significant potential, in order to produce a credible economic assessment of electric mining equipment their impact on underground systems must be understood and considered in their evaluation. Accordingly, a good deal of this thesis reviews technical considerations related to the use of electric mining equipment, especially ones that impact the economics of their implementation. The goal of this thesis will then be to present the economic potential of implementing the equipment, as well as to outline the key inputs which are necessary to support an evaluation and to provide a model and an approach which can be used by others if the relevant information is available and acceptable assumptions can be made.

‘Every Coward’s Choice’? Political Objection to Military Service in Apartheid South Africa as Sexual Citizenship

Sexuality was articulated by the apartheid state as a means of disciplining the white population and marginalizing white opponents of apartheid. As such, homophobia was a recurrent feature of political and legal discourse. The End Conscription Campaign (ECC) opposed compulsory conscription for all white men in the apartheid era South African Defence Force (SADF). Its challenge was a potentially radical and profoundly destabilizing one and it articulated a competing definition of citizenship to that offered by the state. The pro‐ and anti‐conscription discourse was inherently gendered and overtly sexualized. The South African government regularly associated men who objected to military service with effeminacy, cowardice and sexual ‘deviance’. The case of Dr Ivan Toms' objection, a gay objector who wished to cite his sexuality as a primary motivation for his objection, reveals the unwillingness of the ECC to engage in sexual politics. Using Shane Phelan's and Zygmunt Bauman's concept of friends, enemies and strangers, this paper investigates the construction of both white gay men and white people who opposed apartheid as ‘strangers’ and suggests that the deployment of homophobia by the state was a stigmatizing discourse aimed at purging the ECC's political message from the public realm. In this context the ECC adopted an assimilatory discursive strategy, whereby they attempted to be ‘respectable whites’, negotiating over shared republican territory. This populist strategy, arguably safer in the short term, avoided issues of sexuality and the fundamental conflation of sexuality and citizenship in apartheid South Africa. The ECC thus circumscribed its radical and deconstructive political potential and did not offer a ‘radical democratic’ message in opposition to apartheid.

Los más pequeños toman la palabra: La Escuela Infantil que a ellos les gustaría

El derecho a ser escuchado y participar en aquellos asuntos que afectan a los niños es un derecho fundamental de la infancia escasamente respetado por el adulto, quien tiende a silenciar su voz, infravalorando su visión y posible aportación. Ante esta realidad, la Escuela Infantil se erige como un marco privilegiado para la escucha y la participación de la infancia en aquellos temas que les afectan, contribuyendo a la formación de ciudadanos democráticos. A partir de un marco de escucha donde se concibe al pequeño como un sujeto activo, competente y experto en su propia vida, se ha llevado a cabo una investigación que ha adoptado el formato de estudio de caso, en la que se ha recogido a través de las técnicas del dibujo y de las conversaciones con niños, la visión que poseen los más pequeños sobre la Escuela Infantil, analizando el bienestar del niño en el contexto escolar, el sentido y finalidad que los niños otorgan a la escuela; el grado de satisfacción con la misma; sus gustos y preferencias en el contexto escolar; la autonomía y libertad que dicen poseer en la escuela; su visión sobre los aspectos organizativos que rigen el funcionamiento del centro y su perspectiva sobre cómo es y cómo les gustaría que fuera la participación de las familias en la escuela. Finalmente, se recogen sus propuestas de mejora para hacer de la Escuela Infantil a la que asisten su escuela ideal. Los resultados del estudio ponen de manifiesto la demanda de los niños de una Escuela Infantil diferente a la que conocen y la existencia de un pensamiento divergente del pensamiento adulto que es necesario escuchar y tener en consideración, siendo sus aportaciones de gran valía para poder iniciar procesos de innovación y mejora educativa.

Doing Relationships and Sexuality Education with Young People in State Care

Background: Existing literature indicates that young people in state carehave particular sexual health needs that include addressing their social andemotional well-being, yet little has been published as to how thesecomponents of sex education are actually delivered by service-providers.Objective: To analyse the processes involved in delivering relationship andsexuality education to young people in state care from the perspectives ofa sample of service-providers with a role in sexual health care delivery.Design: Qualitative methodological strategy.Setting: Service-delivery sites at urban and rural locations in Ireland.Method: Twenty-two service-providers were interviewed in depth, and datawere analysed using a qualitative analytical strategy resembling modifiedanalytical induction.Findings: Participants proffered their perceptions and examples of theirpractices of sex education in relation to the following themes: (1)acknowledging the multi-dimensional nature of sexual health in the case ofyoung people in care; (2) personal and emotional development educationto address poor self-esteem, emotional disconnectedness and an inabilityto recognise and express emotions; (3) social skills’ education as part of arepertoire of competencies needed to negotiate relationships and safer sex;(4) the application of positive social skills embedded in everyday socialsituations; and (5) factual sexuality education.Conclusion: Insights into service providers’ perceptions of the multidimensionalnature of the sexual health needs of young people in statecare, and the ways in which these service-providers justified their practicemake visible the complex character of sex education and the degree of skillrequired to deliver it to those in state care.

Gas Solubility and Vapor Pressure of Electrolytes Based on Alkylcarbonates Containing Litfsi, Lifap, and LiPF6 Lithium Salt: Measurement and Prediction

Most liquid electrolytes used in commercial lithium-ion batteries are composed by alkylcarbonate mixture containing lithium salt. The decomposition of these solvents by oxidation or reduction during cycling of the cell, induce generation of gases (CO2, CH4, C2H4, CO …) increasing of pressure in the sealed cell, which causes a safety problem [1]. The prior understanding of parameters, such as structure and nature of salt, temperature pressure, concentration, salting effects and solvation parameters, which influence gas solubility and vapor pressure of electrolytes is required to formulate safer and suitable electrolytes especially at high temperature.

We present in this work the CO2, CH4, C2H4, CO solubility in different pure alkyl-carbonate solvents (PC, DMC, EMC, DEC) and their binary or ternary mixtures as well as the effect of temperature and lithium salt LiX (X = LiPF6, LiTFSI or LiFAP) structure and concentration on these properties. Furthermore, in order to understand parameters that influence the choice of the structure of the solvents and their ability to dissolve gas through the addition of a salt, we firstly analyzed experimentally the transport properties (Self diffusion coefficient (D), fluidity (h-1), and conductivity (s) and lithium transport number (tLi) using the Stock-Einstein, and extended Jones-Dole equations [2]. Furthermore, measured data for the of CO2, C2H4, CH4 and CO solubility in pure alkylcarbonates and their mixtures containing LiPF6; LiFAP; LiTFSI salt, are reported as a function of temperature and concentration in salt. Based on experimental solubility data, the Henry’s law constant of gases in these solvents and electrolytes was then deduced and compared with values predicted by using COSMO-RS methodology within COSMOthermX software. From these results, the molar thermodynamic functions of dissolution such as the standard Gibbs energy, the enthalpy, and the entropy, as well as the mixing enthalpy of the solvents and electrolytes with the gases in its hypothetical liquid state were calculated and discussed [3]. Finally, the analysis of the CO2 solubility variations with the salt addition was then evaluated by determining specific ion parameters Hi by using the Setchenov coefficients in solution. This study showed that the gas solubility is entropy driven and can been influenced by the shape, charge density, and size of the anions in lithium salt.

References

[1] S.A. Freunberger, Y. Chen, Z. Peng, J.M. Griffin, L.J. Hardwick, F. Bardé, P. Novák, P.G. Bruce, Journal of the American Chemical Society 133 (2011) 8040-8047.

[2] P. Porion, Y.R. Dougassa, C. Tessier, L. El Ouatani, J. Jacquemin, M. Anouti, Electrochimica Acta 114 (2013) 95-104.

[3] Y.R. Dougassa, C. Tessier, L. El Ouatani, M. Anouti, J. Jacquemin, The Journal of Chemical Thermodynamics 61 (2013) 32-44.

Synthesis of 2-Alkynoates by Palladium(II)-Catalyzed Oxidative Carbonylation of Terminal Alkynes and Alcohols

A homogeneous PdII catalyst, utilizing a simple and inexpensive amine ligand (TMEDA), allows 2-alkynoates to be prepared in high yields by an oxidative carbonylation of terminal alkynes and alcohols. The catalyst system overcomes many of the limitations of previous palladium carbonylation catalysts. It has an increased substrate scope, avoids large excesses of alcohol substrate and uses a desirable solvent. The catalyst employs oxygen as the terminal oxidant and can be operated under safer gas mixtures.

Quantitative Comparison of SensibleThings and Dweet.io

The objective of this paper is to perform a quantitative comparison of Dweet.io and SensibleThings from different aspects. With the fast development of internet of things, the platforms for internet-of-things face bigger challenges. This paper will evaluate both systems in four parts. The first part shows the general comparison of input ways and output functions provided by the platforms. The second part shows the security comparison, which focuses on the protocol types of the packets and the stability during the communication. The third part shows the scalability comparison when the value becomes bigger. The fourth part shows the scalability comparison when speeding up the processes. After the comparisons, I concluded that Dweet.io is more easy to use on devices and supports more programming languages. Dweet.io realizes visualization and it can be shared. Dweet.io is safer and more stable than SensibleThings. SensibleThings provides more openness. SensibleThings has better scalability in handling big values and quick speed.

Pilot Study on Improving the Efficiency of Transportation Projects Using Laser Scanning, 2003

Laser scanning is a terrestrial laser-imaging system that creates highly accurate three-dimensional images of objects for use in standard computer-aided design software packages. This report describes results of a pilot study to investigate the use of laser scanning for transportation applications in Iowa. After an initial training period on the use of the scanner and Cyclone software, pilot tests were performed on the following projects: intersection and railroad bridge for training purposes; section of highway to determine elevation accuracy and pair of bridges to determine level of detail that can be captured; new concrete pavement to determine smoothness; bridge beams to determine camber for deck-loading calculations; stockpile to determine volume; and borrow pit to determine volume. Results show that it is possible to obtain 2-6 mm precision with the laser scanner as claimed by the manufacturer compared to approximately one-inch precision with aerial photogrammetry using a helicopter. A cost comparison between helicopter photogrammetry and laser scanning showed that laser scanning was approximately 30 percent higher in cost depending on assumptions. Laser scanning can become more competitive to helicopter photogrammetry by elevating the scanner on a boom truck and capturing both sides of a divided roadway at the same time. Two- and three-dimensional drawings were created in MicroStation for one of the scanned highway bridges. It was demonstrated that it is possible to create such drawings within the accuracy of this technology. It was discovered that a significant amount of time is necessary to convert point cloud images into drawings. As this technology matures, this task should become less time consuming. It appears that laser scanning technology does indeed have a place in the Iowa Department of Transportation design and construction toolbox. Based on results from this study, laser scanning can be used cost effectively for preliminary surveys to develop TIN meshes of roadway surfaces. It also appears that this technique can be used quite effectively to measure bridge beam camber in a safer and quicker fashion compared to conventional approaches. Volume calculations are also possible using laser scanning. It seems that measuring quantities of rock could be an area where this technology would be quite beneficial since accuracy is more important with this material compared to soil. Other applications for laser scanning could include developing as-built drawings of historical structures such as the bridges of Madison County. This technology could also be useful where safety is a concern such as accurately measuring the surface of a highway active with traffic or scanning the underside of a bridge damaged by a truck. It is recommended that the Iowa Department of Transportation initially rent the scanner when it is needed and purchase the software. With time, it may be cost justifiable to purchase the scanner as well. Laser scanning consultants can be hired as well but at a higher cost.

Organisational and systems factors impacting on patient safety in acute care organisations: lessons from four multi-site research studies

Background Patient safety is concerned with preventable harm in healthcare, a subject that became a focus for study in the UK in the late 1990s. How to improve patient safety, presented both a practical and a research challenge in the early 2000s, leading to the eleven publications presented in this thesis. Research question The overarching research question was: What are the key organisational and systems factors that impact on patient safety, and how can these best be researched? Methods Research was conducted in over 40 acute care organisations in the UK and Europe between 2006 and 2013. The approaches included surveys, interviews, documentary analysis and non-participant observation. Two studies were longitudinal. Results The findings reveal the nature and extent of poor systems reliability and its effect on patient safety; the factors underpinning cases of patient harm; the cultural issues impacting on safety and quality; and the importance of a common language for quality and safety across an organisation. Across the publications, nine key organisational and systems factors emerged as important for patient safety improvement. These include leadership stability; data infrastructure; measurement capability; standardisation of clinical systems; and creating an open and fair collective culture where poor safety is challenged. Conclusions and contribution to knowledge The research presented in the publications has provided a more complete understanding of the organisation and systems factors underpinning safer healthcare. Lessons are drawn to inform methods for future research, including: how to define success in patient safety improvement studies; how to take into account external influences during longitudinal studies; and how to confirm meaning in multi-language research. Finally, recommendations for future research include assessing the support required to maintain a patient safety focus during periods of major change or austerity; the skills needed by healthcare leaders; and the implications of poor data infrastructure.

Type-Aware Programming Models for Distributed Applications

Thesis (Ph.D.)--University of Washington, 2016-08

Towards Empowerment: Narrative Study of Counselling Psychology Trainees and How They Make Sense of Their Personal and Professional Development in the Context of Their Past Experiences

The discipline of counselling psychology continues to grow and change in response to social, economic and political pressures. It has been argued that its quest for a coherent and distinct identity, which emphasises the possibility of the coexistence of multiple approaches, creates an inherently uncertain and dilemmatic training environment that may hinder the development of trainees’ professional identities. In order to gain a deeper understanding of the issue at hand, the aim was to explore how final year trainees and newly qualified counselling psychologists constructed and made sense of their emerging professional identities and what experiences, past and present, they drew upon in the context of their training to shape those identities. Applying narrative inquiry to analyse eight open-ended interviews, eight preliminary themes were originally identified in participants’ narratives, which with further refinements lead to stories of struggle and marginalisation, growth and discovery, and power and resilience. Participants’ stories of struggle and marginalisation emerged in reference to early family dynamics and stressful life experiences, which seemed to also foster a strong identification with the counselling psychology profession, while stories of growth and discovery focused on the importance of having supportive figures, who helped to instill a sense of security and create an atmosphere of openness. It was in this learning environment that participants felt it was possible to develop a more resilient, empowered professional self, which allowed them to shed an earlier sense of struggle and vulnerability. However, where more of an emphasis was placed on power and resilience, there seemed to be less room for participants to express other feelings that came into conflict with their preferred sense of professional self. While there seems to be a need for a ‘safer’ climate, in which trainees could voice and acknowledge anxieties, vulnerabilities and limitations, addressing concerns around power and vulnerability that may be contributing to the silencing of particular voices and identities may be equally important if trainees are to develop coherent and distinct counselling psychologist identities.