Nanoparticle-Assisted Immunoassays for Point-of-Care Testing - With Specific Interest in Minimally Interference-Prone Assays for Cardiac Troponin I

Cardiac troponins (cTn) I and T are the current golden standard biochemical markers in the diagnosis and risk stratification of patients with suspected acute coronary syndrome. During the past few years, novel assays capable of detecting cTn‐concentrations in >50% of apparently healthy individuals have become readily available. With the emerging of these high sensitivity cTn assays, reductions in the assay specificity have caused elevations in the measured cTn levels that do not correlate with the clinical picture of the patient. The increased assay sensitivity may reveal that various analytical interference mechanisms exist. This doctoral thesis focused on developing nanoparticle‐assisted immunometric assays that could possibly be applied to an automated point‐of‐care system. The main objective was to develop minimally interference‐prone assays for cTnI by employing recombinant antibody fragments. Fast 5‐ and 15‐minute assays for cTnI and D‐dimer, a degradation product of fibrin, based on intrinsically fluorescent nanoparticles were introduced, thus highlighting the versatility of nanoparticles as universally applicable labels. The utilization of antibody fragments in different versions of the developed cTnI‐assay enabled decreases in the used antibody amounts without sacrificing assay sensitivity. In addition, the utilization of recombinant antibody fragments was shown to significantly decrease the measured cTnI concentrations in an apparently healthy population, as well as in samples containing known amounts of potentially interfering factors: triglycerides, bilirubin, rheumatoid factors, or human anti‐mouse antibodies. When determining the specificity of four commercially available antibodies for cTnI, two out of the four cross‐reacted with skeletal troponin I, but caused crossreactivity issues in patient samples only when paired together. In conclusion, the results of this thesis emphasize the importance of careful antibody selection when developing cTnI assays. The results with different recombinant antibody fragments suggest that the utilization of antibody fragments should strongly be encouraged in the immunoassay field, especially with analytes such as cTnI that require highly sensitive assay approaches.

The effect of focal point parameters in fiber laser welding of structural steel

In this thesis the effect of focal point parameters in fiber laser welding of structural steel is studied. The goal is to establish relations between laser power, focal point diameter and focal point position with the resulting quality, weld-bead geometry and hardness of the welds. In the laboratory experiments, AB AH36 shipbuilding steel was welded in an I-butt joint configuration using IPG YLS-10000 continuous wave fiber laser. The quality of the welds produced were evaluated based on standard SFS-EN ISO 13919-1. The weld-bead geometry was defined from the weld cross-sections and Vickers hardness test was used to measure hardness's from the middle of the cross-sections. It was shown that all the studied focal point parameters have an effect on the quality, weld-bead geometry and hardness of the welds produced.

Trailing end-point phenotype antibiotic-sensitive strains of Candida albicans produce different amounts of aspartyl peptidases

Candida albicans is an opportunistic fungal pathogen that causes severe systemic infections in immunosuppressed individuals. C. albicans resistance to antifungal drugs is a severe problem in patients receiving prolonged therapy. Moreover, trailing yeast growth, which is defined as a resistant MIC after 48 h of incubation with triazole antifungal agents but a susceptible MIC after 24 h, has been noted in tests of antifungal susceptibility against some C. albicans isolates. In this context, we recently noticed this phenomenon in our routine susceptibility tests with fluconazole/itraconazole and C. albicans clinical isolates. In the present study, we investigated the production of cell-associated and secreted aspartyl peptidases (Saps) in six trailing clinical isolates of C. albicans, since this class of hydrolytic enzymes is a well-known virulence factor expressed by this fungal pathogen. Sap2, which is the best-studied member of the Sap family, was detected by flow cytometry on the cell surface of yeasts and as a 43-kDa polypeptide in the culture supernatant, as demonstrated by Western blotting assay using an anti-Sap1-3 polyclonal antibody. Released aspartyl peptidase activity was measured with BSA hydrolysis and inhibited by pepstatin A, showing distinct amounts of proteolytic activity ranging from 5.7 (strain 44B) to 133.2 (strain 11) arbitrary units. Taken together, our results showed that trailing clinical isolates of C. albicans produced different amounts of both cellular and secreted aspartyl-type peptidases, suggesting that this phenotypic feature did not generate a regular pattern regarding the expression of Sap.

Effect of point mutations on Herbaspirillum seropedicae NifA activity

NifA is the transcriptional activator of the nif genes in Proteobacteria. It is usually regulated by nitrogen and oxygen, allowing biological nitrogen fixation to occur under appropriate conditions. NifA proteins have a typical three-domain structure, including a regulatory N-terminal GAF domain, which is involved in control by fixed nitrogen and not strictly required for activity, a catalytic AAA+ central domain, which catalyzes open complex formation, and a C-terminal domain involved in DNA-binding. In Herbaspirillum seropedicae, a β-proteobacterium capable of colonizing Graminae of agricultural importance, NifA regulation by ammonium involves its N-terminal GAF domain and the signal transduction protein GlnK. When the GAF domain is removed, the protein can still activate nif genes transcription; however, ammonium regulation is lost. In this work, we generated eight constructs resulting in point mutations in H. seropedicae NifA and analyzed their effect on nifH transcription in Escherichia coli and H. seropedicae. Mutations K22V, T160E, M161V, L172R, and A215D resulted in inactive proteins. Mutations Q216I and S220I produced partially active proteins with activity control similar to wild-type NifA. However, mutation G25E, located in the GAF domain, resulted in an active protein that did not require GlnK for activity and was partially sensitive to ammonium. This suggested that G25E may affect the negative interaction between the N-terminal GAF domain and the catalytic central domain under high ammonium concentrations, thus rendering the protein constitutively active, or that G25E could lead to a conformational change comparable with that when GlnK interacts with the GAF domain.

Spectrally Tuned Lanthanide Photoluminescence for Point-of-Care Testing

Point-of-care (POC) –diagnostics is a field with rapidly growing market share. As these applications become more widely used, there is an increasing pressure to improve their performance to match the one of a central laboratory tests. Lanthanide luminescence has been widely utilized in diagnostics because of the numerous advantages gained by the utilization of time-resolved or anti-Stokes detection. So far the use of lanthanide labels in POC has been scarce due to limitations set by the instrumentation required for their detection and the shortcomings, e.g. low brightness, of these labels. Along with the advances in the research of lanthanide luminescence, and in the field of semiconductors, these materials are becoming a feasible alternative for the signal generation also in the future POC assays. The aim of this thesis was to explore ways of utilizing time-resolved detection or anti-Stokes detection in POC applications. The long-lived fluorescence for the time-resolved measurement can be produced with lanthanide chelates. The ultraviolet (UV) excitation required by these chelates is cumbersome to produce with POC compatible fluorescence readers. In this thesis the use of a novel light-harvesting ligand was studied. This molecule can be used to excite Eu(III)-ions at wavelengths extending up to visible part of the spectrum. An enhancement solution based on this ligand showed a good performance in a proof-of-concept -bioaffinity assay and produced a bright signal upon 365 nm excitation thanks to the high molar absorptivity of the chelate. These features are crucial when developing miniaturized readers for the time-resolved detection of fluorescence. Upconverting phosphors (UCPs) were studied as an internal light source in glucose-sensing dry chemistry test strips and ways of utilizing their various emission wavelengths and near-infrared excitation were explored. The use of nanosized NaYF :Yb3+,Tm3+-particles enabled the replacement of an external UV-light source with a NIR-laser and gave an additional degree of freedom in the optical setup of the detector instrument. The new method enabled a blood glucose measurement with results comparable to a current standard method of measuring reflectance. Microsized visible emitting UCPs were used in a similar manner, but with a broad absorbing indicator compound filtering the excitation and emission wavelengths of the UCP. This approach resulted in a novel way of benefitting from the non-linear relationship between the excitation power and emission intensity of the UCPs, and enabled the amplification of the signal response from the indicator dye.

The role of import intermediary companies in the Russian imports from the EU A case study of a Russian industrial company’s point of view on import intermediaries

The literature on agency suggests different implications for the use of export intermediaries. However, only few studies provide a view on import intermediaries. This thesis tries for its part to fill this research gap by studying the import intermediaries in the EU–Russia trade from a Russian industrial company’s point of view. The aim is to describe import intermediation and explain the need for import intermediary companies in the EU–Russia trade. The theoretical framework of this thesis originates from an article by Peng and York (2001), in which they study the performance of export intermediaries. This thesis applies resource-based theory, transaction cost theory and agency cost theory, following the idea of Peng and York. The resource-based theory approach is utilised for describing an ideal import intermediary company, and transaction cost theory provides a basis for understanding the benefits of using the services of import intermediary companies, while agency cost theory is applied in order to understand the risks the Russian industrial company faces when it decides to use the services of import intermediaries. The study is performed in the form of a case interview with a representative of a major Russian metallurgy company. The results of the study suggest that an ideal intermediary has the skills required specifically for the imports process, in order to save time and money of the principal company. The intermediary company helps reducing the amount of time the managers and the staff of the principal company use to make imports possible, thus reducing the salary costs and providing the possibility to concentrate on the company’s core competencies. The benefits of using the services of import intermediary companies are the reduced transaction costs, especially salary costs that are minimised because of the effectiveness and specialisation of import intermediaries. Intermediaries are specialised in the imports process and thus need less time and resources to organise the imports. They also help to reduce the fixed salary costs, because their services can be used only when needed. The risks of being misled by intermediaries are minimised by the competition on the import intermediary market. In case an intermediary attempts fraud, it gets replaced by its rival.

Effect of the implementation of the Hazard Analysis Critical Control Point (HACCP) prerequisite program in an institutional foodservice unit in Southern Brazil

The aims of this study were to investigate the hygienic practices in the food production of an institutional foodservice unit in Southern Brazil and to evaluate the effect of implementing good food handling practices and standard operational procedures using microbiological hygiene indicators. An initial survey of the general operating conditions classified the unit as regular in terms of compliance with State safety guidelines for food service establishments. An action plan that incorporated the correction of noncompliance issues and the training of food handlers in good food handling practices and standard operational procedures were then implemented. The results of the microbiological analysis of utensils, preparation surfaces, food handlers' hands, water, and ambient air were recorded before and after the implementation of the action plan. The results showed that the implementation of this type of practice leads to the production of safer foods.

Hygienic-sanitary working practices and implementation of a Hazard Analysis and Critical Control Point (HACCP) plan in lobster processing industries

This study aimed to verify the hygienic-sanitary working practices and to create and implement a Hazard Analysis Critical Control Point (HACCP) in two lobster processing industries in Pernambuco State, Brazil. The industries studied process frozen whole lobsters, frozen whole cooked lobsters, and frozen lobster tails for exportation. The application of the hygienic-sanitary checklist in the industries analyzed achieved conformity rates over 96% to the aspects evaluated. The use of the Hazard Analysis Critical Control Point (HACCP) plan resulted in the detection of two critical control points (CCPs) including the receiving and classification steps in the processing of frozen lobster and frozen lobster tails, and an additional critical control point (CCP) was detected during the cooking step of processing of the whole frozen cooked lobster. The proper implementation of the Hazard Analysis Critical Control Point (HACCP) plan in the lobster processing industries studied proved to be the safest and most cost-effective method to monitor each critical control point (CCP) hazards.

The role of import intermediary companies in the Russian imports from the EU A case study of a Russian industrial company’s point of view on import intermediaries

The literature on agency suggests different implications for the use of export intermediaries. However, only few studies provide a view on import intermediaries. This thesis tries for its part to fill this research gap by studying the import intermediaries in the EU–Russia trade from a Russian industrial company’s point of view. The aim is to describe import intermediation and explain the need for import intermediary companies in the EU–Russia trade. The theoretical framework of this thesis originates from an article by Peng and York (2001), in which they study the performance of export intermediaries. This thesis applies resource-based theory, transaction cost theory and agency cost theory, following the idea of Peng and York. The resource-based theory approach is utilised for describing an ideal import intermediary company, and transaction cost theory provides a basis for understanding the benefits of using the services of import intermediary companies, while agency cost theory is applied in order to understand the risks the Russian industrial company faces when it decides to use the services of import intermediaries. The study is performed in the form of a case interview with a representative of a major Russian metallurgy company. The results of the study suggest that an ideal intermediary has the skills required specifically for the imports process, in order to save time and money of the principal company. The intermediary company helps reducing the amount of time the managers and the staff of the principal company use to make imports possible, thus reducing the salary costs and providing the possibility to concentrate on the company’s core competencies. The benefits of using the services of import intermediary companies are the reduced transaction costs, especially salary costs that are minimised because of the effectiveness and specialisation of import intermediaries. Intermediaries are specialised in the imports process and thus need less time and resources to organise the imports. They also help to reduce the fixed salary costs, because their services can be used only when needed. The risks of being misled by intermediaries are minimised by the competition on the import intermediary market. In case an intermediary attempts fraud, it gets replaced by its rival.

ATTITUDES TOWARD RUSSIAN CUSTOMERS IN THE FINNISH RETAIL TRADE SECTOR – A salesperson’s point of view

The amount of Russian tourists in Finland has increased significantly in the past years. The impact of Russian tourism to the Finnish retail trade sector is enormous, since Russian tourists often spend a lot of money particularly on shopping. Shopping tourism is mainly focused in the near border cities, such as Imatra and Lappeenranta, and in addition in Helsinki metropolitan area. The purpose of this study is to map the attitudes and perceptions of the sales personnel who are working in the Finnish retail trade sector towards Russian customers and to discover which elements affect these attitudes. The theories in this study are based on cultural elements and elements related to sales behavior and performance. Cultural differences between Finland and Russia, cultural distance and cultural intelligence form the cultural aspect of this study. Customer orientation vs. sales orientation (SOCO), adaptive selling, selling skills and job competency, salesperson’s affect and empathy toward customers, and job autonomy form the elements concerning sales behavior and performance. Furthermore, the attitude – behavior link, based on social psychology is addressed. A survey was conducted in two retail trade chains operating in Finland. These retail companies have stores and department stores in different geographical areas in Finland and the survey was conducted in altogether 19 cities. In addition to the theories that were discussed, two expert interviews were conducted in order to get a deeper understanding of the phenomenon at hand. Moreover the interviews helped in the formulation of the hypotheses and the questionnaire design. The questionnaires were sent directly to the stores, where they were placed so that they were available for the sales personnel. Altogether 487 usable responses were collected. The returned questionnaires were analyzed with IBM SPSS 21 statistics program. The results of this study indicated that the attitudes toward Russian customers are more negative compared to other foreign customers. However, the respondents’ attitudes toward and perceptions of Russian customers varied a lot. From the background variables age, education level, length of employment in current workplace, and length of experience in customer service had an effect on the attitudes of the respondents. In addition, the perceptions of Russian customers were more positive in the Eastern Finland compared to Helsinki metropolitan area. The cultural elements; cultural knowledge, cultural distance and cultural intelligence all affected the attitudes of the respondents. From the elements related to sales behavior and performance customer orientation, salesperson’s affect and empathy toward customers, and perceived job autonomy had an effect on the attitudes

Design and Material Selection of High-Speed Rotating Electrical Machines

The increasing emphasis on energy efficiency is starting to yield results in the reduction in greenhouse gas emissions; however, the effort is still far from sufficient. Therefore, new technical solutions that will enhance the efficiency of power generation systems are required to maintain the sustainable growth rate, without spoiling the environment. A reduction in greenhouse gas emissions is only possible with new low-carbon technologies, which enable high efficiencies. The role of the rotating electrical machine development is significant in the reduction of global emissions. A high proportion of the produced and consumed electrical energy is related to electrical machines. One of the technical solutions that enables high system efficiency on both the energy production and consumption sides is high-speed electrical machines. This type of electrical machines has a high system overall efficiency, a small footprint, and a high power density compared with conventional machines. Therefore, high-speed electrical machines are favoured by the manufacturers producing, for example, microturbines, compressors, gas compression applications, and air blowers. High-speed machine technology is challenging from the design point of view, and a lot of research is in progress both in academia and industry regarding the solution development. The solid technical basis is of importance in order to make an impact in the industry considering the climate change. This work describes the multidisciplinary design principles and material development in high-speed electrical machines. First, high-speed permanent magnet synchronous machines with six slots, two poles, and tooth-coil windings are discussed in this doctoral dissertation. These machines have unique features, which help in solving rotordynamic problems and reducing the manufacturing costs. Second, the materials for the high-speed machines are discussed in this work. The materials are among the key limiting factors in electrical machines, and to overcome this limit, an in-depth analysis of the material properties and behavior is required. Moreover, high-speed machines are sometimes operating in a harsh environment because they need to be as close as possible to the rotating tool and fully exploit their advantages. This sets extra requirements for the materials applied.

Crystal growth, characterization and point contact spectroscopy on CuxTiSe₂

Various s e t s of single c rys t a l s and poly c rys t a l s of Cux TiSe2 were grown. X- r ay diffraction and ene rgy dispersive spe c t ros copy r e sul t s verified tha t the c rys t a l s were the cor r e c t compos i t ion and c rys t a l s t ruc tur e . Re s i s t ivi ty me a sur ement s and magne t i c sus c ept ibi l i ty me a sur ement s de t e rmined the supe r conduc t ing t r ans i t ion t empe r a tur e s for the c rys t a l s . The c rys t a l s in each growth had various supe r conduc t ing t r ans i t ion t empe r a tur e s . Also, the me a sur ement s indi c a t ed tha t the c rys t a l s were inhomogeneous. Point cont a c t spe c t ros copy expe r iment s were employed on various single c rys t a l s . Inspe c t ion of the da t a indi c a t ed tha t the ma t e r i a l ha s a single ene rgy gap . A progr am was bui l t ut i l i z ing the Levenbe rg-Ma rqua rdt me thod and theory on point cont a c t spe c t ros copy to de t e rmine the supe r conduc t ing ene rgy gap. Plot s of the supe r conduc t ing ene rgy gap a t various t empe r a tur e s were in di s agr e ement wi th wha t was expe c t ed for a convent iona l supe r conduc tor .

Glacial stratigraphy and micromorphology of the Pine Point Pb-­‐Zn mining district, Northwest Territories

This investigation aims to gain a better understanding of the glacial history of the Pine Point Mining district, Northwest Territories, by examining the sedimentological properties of the glacial sediments including, geochemical analysis, heavy mineral concentrate analysis, clast macro-­‐fabrics, pebble lithologies, and micromorphological investigation. Four till units were identified, and three were associated with identified erosional bedrock features and streamlined landforms in the area, indicating a minimum of three ice flow directions. Sedimentological properties suggest that these units were all Type-­B tectomict/mélange till, emplaced as part of a soft subglacial deformable bed. The lack of ice-­‐marginal advance and retreat sequences within the studied till, suggests the Middle Wisconsinan Laurentide Ice margin was likely north and west of the Pine Point area, as opposed to along the margin of the Canadian Shield and Western Sedimentary Basin where it has been suggested to have existed.

Notice from the Crown Lands Department of Ottawa that the lands on Long Point, Lake Erie would be offered at public auction on May 4, 1866

Notice from the Crown Lands Department of Ottawa that the lands on Long Point, Lake Erie would be offered at public auction on May 4, 1866. This is attached to a small newspaper clipping with the same information, dated April 20, 1866.

Flyer entitled Long Point Company which was written by Edward Harris

Flyer entitled Long Point Company which was written by Edward Harris (2 printed pages). There are 2 copies of this flyer. One is addressed to S.D. Woodruff and the other is addressed to Joseph Woodruff, May 28, 1879.