Frequency-Domain and Wide-Pulse Time-Domain Measurements of Lanthanide Luminescence and Lanthanide-Based Resonance Energy Transfer

Resonance energy transfer (RET) is a non-radiative transfer of the excitation energy from the initially excited luminescent donor to an acceptor. The requirements for the resonance energy transfer are: i) the spectral overlap between the donor emission spectrum and the acceptor absorption spectrum, ii) the close proximity of the donor and the acceptor, and iii) the suitable relative orientations of the donor emission and the acceptor absorption transition dipoles. As a result of the RET process the donor luminescence intensity and the donor lifetime are decreased. If the acceptor is luminescent, a sensitized acceptor emission appears. The rate of RET depends strongly on the donor–acceptor distance (r) and is inversely proportional to r6. The distance dependence of RET is utilized in binding assays. The proximity requirement and the selective detection of the RET-modified emission signal allow homogeneous separation free assays. The term lanthanide-based RET is used when luminescent lanthanide compounds are used as donors. The long luminescence lifetimes, the large Stokes’ shifts and the intense, sharply-spiked emission spectra of the lanthanide donors offer advantages over the conventional organic donor molecules. Both the organic lanthanide chelates and the inorganic up-converting phosphor (UCP) particles have been used as donor labels in the RET based binding assays. In the present work lanthanide luminescence and lanthanide-based resonance energy transfer phenomena were studied. Luminescence lifetime measurements had an essential role in the research. Modular frequency-domain and time-domain luminometers were assembled and used successfully in the lifetime measurements. The frequency-domain luminometer operated in the low frequency domain ( 100 kHz) and utilized a novel dual-phase lock-in detection of the luminescence. One of the studied phenomena was the recently discovered non-overlapping fluorescence resonance energy transfer (nFRET). The studied properties were the distance and temperature dependences of nFRET. The distance dependence was found to deviate from the Förster theory and a clear temperature dependence was observed whereas conventional RET was completely independent of the temperature. Based on the experimental results two thermally activated mechanisms were proposed for the nFRET process. The work with the UCP particles involved the measurement of the luminescence properties of the UCP particles synthesized in our laboratory. The goal of the UCP particle research is to develop UCP donor labels for binding assays. In the present work the effect of the dopant concentrations and the core–shell structure on the total up-conversion luminescence intensity, the red–green emission ratio, and the luminescence lifetime was studied. Also the non-radiative nature of the energy transfer from the UCP particle donors to organic acceptors was demonstrated for the first time in aqueous environment and with a controlled donor–acceptor distance.

Luminescent Lanthanide Reporters: New Concepts for Use in Bioanalytical Applications

Lanthanides represent the chemical elements from lanthanum to lutetium. They intrinsically exhibit some very exciting photophysical properties, which can be further enhanced by incorporating the lanthanide ion into organic or inorganic sensitizing structures. A very popular approach is to conjugate the lanthanide ion to an organic chromophore structure forming lanthanide chelates. Another approach, which has quickly gained interest, is to incorporate the lanthanide ions into nanoparticle structures, thus attaining improved specific activity and binding capacity. The lanthanide-based reporters usually express strong luminescence emission, multiple narrow emission lines covering a wide wavelength range, and exceptionally long excited state lifetimes enabling timeresolved detection. Because of these properties, the lanthanide-based reporters have found widespread applications in various fields of life. This study focuses on the field of bioanalytical applications. The aim of the study was to demonstrate the utility of different lanthanide-based reporters in homogeneous Förster resonance energy transfer (FRET)-based bioaffinity assays. Several different model assays were constructed. One was a competitive bioaffinity assay that utilized energy transfer from lanthanide chelate donors to fluorescent protein acceptors. In addition to the conventional FRET phenomenon, a recently discovered non-overlapping FRET (nFRET) phenomenon was demonstrated for the first time for fluorescent proteins. The lack of spectral overlap in the nFRET mechanism provides sensitivity and versatility to energy transfer-based assays. The distance and temperature dependence of these phenomena were further studied in a DNA-hybridization assay. The distance dependence of nFRET deviated from that of FRET, and unlike FRET, nFRET demonstrated clear temperature dependence. Based on these results, a possible excitation mechanism operating in nFRET was proposed. In the study, two enzyme activity assays for caspase-3 were also constructed. One of these was a fluorescence quenching-based enzyme activity assay that utilized novel inorganic particulate reporters called upconverting phosphors (UCPs) as donors. The use of UCPs enabled the construction of a simple, rather inexpensive, and easily automated assay format that had a high throughput rate. The other enzyme activity assay took advantage of another novel reporter class, the lanthanidebinding peptides (LBPs). In this assay, energy was transferred from a LBP to a green fluorescent protein (GFP). Using the LBPs it was possible to avoid the rather laborious, often poorly repeatable, and randomly positioned chemical labeling. In most of the constructed assays, time-resolved detection was used to eliminate the interfering background signal caused by autofluorescence. The improved signal-to-background ratios resulted in increased assay sensitivity, often unobtainable in homogeneous assay formats using conventional organic fluorophores. The anti-Stokes luminescence of the UCPs, however, enabled the elimination of autofluorescence even without time-gating, thus simplifying the instrument setup. Together, the studied reporters and assay formats pave the way for increasingly sensitive, simple, and easily automated bioanalytical applications.

Distance Transforms on Gray-Level Surfaces

This thesis studies gray-level distance transforms, particularly the Distance Transform on Curved Space (DTOCS). The transform is produced by calculating distances on a gray-level surface. The DTOCS is improved by definingmore accurate local distances, and developing a faster transformation algorithm. The Optimal DTOCS enhances the locally Euclidean Weighted DTOCS (WDTOCS) with local distance coefficients, which minimize the maximum error from the Euclideandistance in the image plane, and produce more accurate global distance values.Convergence properties of the traditional mask operation, or sequential localtransformation, and the ordered propagation approach are analyzed, and compared to the new efficient priority pixel queue algorithm. The Route DTOCS algorithmdeveloped in this work can be used to find and visualize shortest routes between two points, or two point sets, along a varying height surface. In a digital image, there can be several paths sharing the same minimal length, and the Route DTOCS visualizes them all. A single optimal path can be extracted from the route set using a simple backtracking algorithm. A new extension of the priority pixel queue algorithm produces the nearest neighbor transform, or Voronoi or Dirichlet tessellation, simultaneously with the distance map. The transformation divides the image into regions so that each pixel belongs to the region surrounding the reference point, which is nearest according to the distance definition used. Applications and application ideas for the DTOCS and its extensions are presented, including obstacle avoidance, image compression and surface roughness evaluation.

New Distance Transforms for Gray Level Image Compression

This thesis deals with distance transforms which are a fundamental issue in image processing and computer vision. In this thesis, two new distance transforms for gray level images are presented. As a new application for distance transforms, they are applied to gray level image compression. The new distance transforms are both new extensions of the well known distance transform algorithm developed by Rosenfeld, Pfaltz and Lay. With some modification their algorithm which calculates a distance transform on binary images with a chosen kernel has been made to calculate a chessboard like distance transform with integer numbers (DTOCS) and a real value distance transform (EDTOCS) on gray level images. Both distance transforms, the DTOCS and EDTOCS, require only two passes over the graylevel image and are extremely simple to implement. Only two image buffers are needed: The original gray level image and the binary image which defines the region(s) of calculation. No other image buffers are needed even if more than one iteration round is performed. For large neighborhoods and complicated images the two pass distance algorithm has to be applied to the image more than once, typically 3 10 times. Different types of kernels can be adopted. It is important to notice that no other existing transform calculates the same kind of distance map as the DTOCS. All the other gray weighted distance function, GRAYMAT etc. algorithms find the minimum path joining two points by the smallest sum of gray levels or weighting the distance values directly by the gray levels in some manner. The DTOCS does not weight them that way. The DTOCS gives a weighted version of the chessboard distance map. The weights are not constant, but gray value differences of the original image. The difference between the DTOCS map and other distance transforms for gray level images is shown. The difference between the DTOCS and EDTOCS is that the EDTOCS calculates these gray level differences in a different way. It propagates local Euclidean distances inside a kernel. Analytical derivations of some results concerning the DTOCS and the EDTOCS are presented. Commonly distance transforms are used for feature extraction in pattern recognition and learning. Their use in image compression is very rare. This thesis introduces a new application area for distance transforms. Three new image compression algorithms based on the DTOCS and one based on the EDTOCS are presented. Control points, i.e. points that are considered fundamental for the reconstruction of the image, are selected from the gray level image using the DTOCS and the EDTOCS. The first group of methods select the maximas of the distance image to new control points and the second group of methods compare the DTOCS distance to binary image chessboard distance. The effect of applying threshold masks of different sizes along the threshold boundaries is studied. The time complexity of the compression algorithms is analyzed both analytically and experimentally. It is shown that the time complexity of the algorithms is independent of the number of control points, i.e. the compression ratio. Also a new morphological image decompression scheme is presented, the 8 kernels' method. Several decompressed images are presented. The best results are obtained using the Delaunay triangulation. The obtained image quality equals that of the DCT images with a 4 x 4

Future prospects of short distance rail based passenger transport in Northern Europe

The worlds’ population is increasing and cities have become more crowded with people and vehicles. Communities in the fringe of metropolitans’ increase the traffic done with private cars, but also increase the need for public transportation. People have typically needs traveling to work located in city centers during the morning time, and return to suburbs in the afternoon or evening. Rail based passenger transport is environmentally friendly transport mode with high capacity to transport large volume of people. Railways have been regulated markets with national incumbent having monopoly position. Opening the market for competition is believed to have a positive effect by increasing the efficiency of the industry. National passenger railway market is opened for competition only in few countries, where as international traffic in EU countries was deregulated in 2010. The objective of this study is to examine the passenger railway market of three North European countries, Sweden, Denmark and Estonia. The interest was also to get an understanding of the current situation and how the deregulation has proceeded. Theory of deregulation is unfolded with literature analyses and empirical part of the study is constructed from two parts. Customer satisfaction survey was chosen as a method to collect real life experiences from the passengers and measure their knowledge of the market situation and possible changes appeared. Interviews of experts from the industry and labor unions give more insights and able better understanding for example of social consequences caused from opening the market for competition. Expert interviews were conducted by using semi-structured theme interview. Based on the results of this study, deregulation has proceeded quite differently in the three countries researched. Sweden is the most advanced country, where the passenger railway market is open for new entrants. Denmark and Estonia are lagging behind. Opening the market is considered positive among passengers and most of the experts interviewed. Common for the interviews were the labour unions negative perspective concerning deregulation. Despite the fact deregulation is considered positive among the respondents of the customer satisfaction survey, they could not name railway undertakings operating in their country. Generally respondents were satisfied with the commuter trains. Ticket price, punctuality of trains and itinerary affect the most to customer satisfaction.

Essays on auditor independence

Auditor independence is a cornerstone of the auditing profession and the basic principle that underpins the reputation of the auditing profession in the public eye. Indeed, it is the attribute most demanded from auditors by the public. Therefore, the sustainability of the auditing profession depends on how auditors can protect this principle. This dissertation consists of four interrelated essays concerned with auditor independence. Specifically, it examines situations that can threaten and impair auditor independence. In addition, this dissertation also examines several variables that may enhance and protect auditor independence. The first essay aims to examine the impact of social pressures occurring within audit firms on auditors’ judgment in the setting of a society with “high power distance” and “low individualism” cultural dimensions. The social pressures consisted of obedience pressure exerted by an auditor’s superior and conformity pressure exerted by an auditor’s colleague. Moreover, two moderating variables—a multi-dimensional professional commitment and locus of control—were included as moderator variables in the relationship between the social pressures faced by auditors and their judgment. The findings show that obedience and conformity pressures influence auditor judgment. Auditors who face the social pressures will make a judgment that may be even diametrically opposite to the independence principle. The findings also indicate that a multi-dimensional professional commitment and locus of control may potentially influence auditor judgment in a situation with social pressures. The second essay aims to investigate the association of advocacy and familiarity threats caused by auditor fee dependence and auditor tenure on auditor independence based on Finnish data, law, and auditing environment. This essay was motivated by the Green Paper on Audit Policy, published by the European Commission in 2010 that questions whether the maximum fee collected from a client should be regulated and whether consecutive assignments should be limited, among others. Contrary to popular belief, this essay does not find evidence that audit fees and long auditor tenure will jeopardise auditor independence. Therefore, the findings do not support policies to regulate auditor fees or limit auditor tenure in Finland. The third essay aims to examine the effect of client intimidation on auditor independence in an audit-client conflict situation. Intimidation threat is one of five independence threats that are explicitly referenced in the IFAC’s independence framework. Client intimidation was manifested in the client threatening to replace the auditor if the auditor did not adopt the client’s position. In addition, this essay examines the role of auditor’s perceived pressure and multi-dimensions of professional commitment as moderator variables. The findings suggest that auditors who experience client intimidation in an audit conflict situation are more likely to have their independence impaired than those who are in a similar situation but without client intimidation. Moreover, auditors who experience client intimidation perceive higher pressure than those who do not experience intimidation. Finally, auditors’ affective and continuance professional commitment dimensions moderate the relationship between auditors’ perceived pressures and auditor independence. The aim of the fourth essay is twofold. First, it aims to develop a scale for measuring auditors’ reputation awareness. Second, it aims to examine the correlation between the levels of auditor reputation awareness and auditor independence. A seven-item scale was developed as the reputation awareness scale. The findings indicate that the scale consists of one dimension. It also has a level of satisfactory reliability and a high level of validity. The findings show that there is a positive correlation between the level of auditors’ reputation awareness and auditor independence.

Manifestation of the pairing symmetry in the vortex core structure in iron-based superconductors

The superconducting gap is a basic character of a superconductor. While the cuprates and conventional phonon-mediated superconductors are characterized by distinct d- and s-wave pairing symmetries with nodal and nodeless gap distributions respectively, the superconducting gap distributions in iron-based superconductors are rather diversified. While nodeless gap distributions have been directly observed in Ba1–xKxFe2As2, BaFe2–xCoxAs2, LiFeAs, KxFe2–ySe2, and FeTe1–xSex, the signatures of a nodal superconducting gap have been reported in LaOFeP, LiFeP, FeSe, KFe2As2, BaFe2–xRuxAs2, and BaFe2(As1–xPx)2. Due to the multiplicity of the Fermi surface in these compounds s± and d pairing states can be both nodeless and nodal. A nontrivial orbital structure of the order parameter, in particular the presence of the gap nodes, leads to effects in which the disorder is much richer in dx2–y2-wave superconductors than in conventional materials. In contrast to the s-wave case, the Anderson theorem does not work, and nonmagnetic impurities exhibit a strong pair-breaking influence. In addition, a finite concentration of disorder produces a nonzero density of quasiparticle states at zero energy, which results in a considerable modification of the thermodynamic and transport properties at low temperatures. The influence of order parameter symmetry on the vortex core structure in iron-based pnictide and chalcogenide superconductors has been investigated in the framework of quasiclassical Eilenberger equations. The main results of the thesis are as follows. The vortex core characteristics, such as, cutoff parameter, ξh, and core size, ξ2, determined as the distance at which density of the vortex supercurrent reaches its maximum, are calculated in wide temperature, impurity scattering rate, and magnetic field ranges. The cutoff parameter, ξh(B; T; Г), determines the form factor of the flux-line lattice, which can be obtained in _SR, NMR, and SANS experiments. A comparison among the applied pairing symmetries is done. In contrast to s-wave systems, in dx2–y2-wave superconductors, ξh/ξc2 always increases with the scattering rate Г. Field dependence of the cutoff parameter affects strongly on the second moment of the magnetic field distributions, resulting in a significant difference with nonlocal London theory. It is found that normalized ξ2/ξc2(B/Bc2) dependence is increasing with pair-breaking impurity scattering (interband scattering for s±-wave and intraband impurity scattering for d-wave superconductors). Here, ξc2 is the Ginzburg-Landau coherence length determined from the upper critical field Bc2 = Φ0/2πξ2 c2, where Φ0 is a flux quantum. Two types of ξ2/ξc2 magnetic field dependences are obtained for s± superconductors. It has a minimum at low temperatures and small impurity scattering transforming in monotonously decreasing function at strong scattering and high temperatures. The second kind of this dependence has been also found for d-wave superconductors at intermediate and high temperatures. In contrast, impurity scattering results in decreasing of ξ2/ξc2(B/Bc2) dependence in s++ superconductors. A reasonable agreement between calculated ξh/ξc2 values and those obtained experimentally in nonstoichiometric BaFe2–xCoxAs2 (μSR) and stoichiometric LiFeAs (SANS) was found. The values of ξh/ξc2 are much less than one in case of the first compound and much more than one for the other compound. This is explained by different influence of two factors: the value of impurity scattering rate and pairing symmetry.

Cultural distance and negotiation behavior : Perception of entrepreneurs operating in Turku

This thesis investigates the influence of cultural distance on entrepreneurs’ negotiation behaviour. For this purpose, Turku was chosen as the unit of analysis due to the exponential demographic change experienced during the last two decades that has derived in a more diversified local environment. The research aim set for this study was to identify to what extent entrepreneurs face cultural distance, how cultural distance influences the entrepreneur’s negotiation behaviour and how can it be addressed in order to turn dissimilarities into opportunities. This study presented the relation and apparent dichotomy of cultural distance and global culture, including the component of diversity. The impact of cultural distance in the entrepreneurial mindset and its consequent effect in negotiation behaviour was presented too. Addressing questions about the way individuals perceive, behave and interact allowed the use of interviews for this qualitative research study. In the empirical part of this study it was found that negotiation behaviour differed in terms of how congenial entrepreneurs felt when managing cultural distance, encompassing their performance. It was also acknowledged that after time and effort, some of the personal traits were enhanced while others reduced, allowing for more flexibility and adaptation. Furthermore, depending on the level of trust and shared interests, entrepreneurs determined their attitudinal approach, being adaptive or reactive subject to situational aspects. Additionally, it was found that the acquisition of cultural savvy not necessarily conveyed to more creativity. This experiential learning capability led to the proposition of new ways of behaviour. Likewise, it was proposed that growing cultural intelligence bridge distances, reducing mistrusts and misunderstandings. The capability of building more collaborative relationships allows entrepreneurs to see cultural distance as a cultural perspective instead of as a threat. Therefore it was recommended to focus on proximity rather than distance to better identify and exploit untapped opportunities and better perform when negotiating in whichever cultural conditions.

Optimal distance of the light-curing tip for light transmission through the resin composite

Hammaslääketieteessä käytetettävien komposiittien valonläpäisevyys vaihtelee. Samoin LED-valokovettimet eroavat toisistaan valotehonsa ja muotoilunsa perusteella. On yleisesti tiedossa, että valokovettimesta tulevan valon intensiteetti pinta-alayksikköä kohden heikkenee, kun kovettimen etäisyys kasvaa. Toisaalta ei ole tiedossa, miten valokovetettavan kohteen ja valokovettimen kärjen väliin sijoitettu materiaali tarkalleenottaen vaikuttaa valon intensiteettiin eri etäisyyksiä käytettäessä. Tämän tutkimuksen tarkoituksena on selvittää, miten valokovetettavan kohteen ja valokovettimen kärjen väliin asetettava etukäteen polymerisoitu materiaali vaikuttaa valon intensiteettiin eri etäisyyksillä. Tutkimus suoritettiin käyttämällä kahta eri valokovetinta. Jotta etäisyyden vaikutusta valotustehoon voitiin demonstroida, vaihdettiin kovettimen etäisyyttä sensorista 0,2,4,6,8,10mm välillä. Valotehot rekisteröitiin MARC resin calibrator -laitteella. Sensorin ja valokovettimen kärjen väliin asetettavat erilaiset komposiittilevyt olivat valmiiksi kovetettuja,1mm paksuisia, filleripitoisuuksiltaan neljää erilaista muovia. Valotehot rekisteröitiin jokaiselta etäisyydeltä komposiitin ollessa sensorin päällä. Rinnakkaisesti verrattiin myös etäisyyden vaikutusta valotehoon ilman esikovetettua materiaalia kovettimen kärjen ja valoa mittaavan sensorin välissä. Vertailun suorittamiseksi laskettiin intensiteettisuhdeluku muovillisen ja muovittoman arvon välillä aina tietyllä etäisyydellä Valokovettimen kärjen etäisyyden kasvattaminen sensorista (eli valokovetettavasta kohteesta) odotusten mukaisesti pienensi valotehoa. Laittamalla sensorin ja kovettimen väliin komposiittilevy, valoteho pieneni odotetusti vielä enemmän. Tutkittaessa intensiteettisuhdetta (valoteho muovin kanssa : valoteho ilman muovia) kuitenkin huomattiin, että 4-6mm:n kohdalla suhdeluku oli suurempi kuin 0,2,8 ja 10mm kohdalla. Johtopäätöksenä oli, että suurin mahdollinen valokovetusteho saavutetan laittamalla kovetuskärki mahdollisimman lähelle kohdetta. Jos valokovetettavan kohteen ja valokovettimen kärjen välissä oli kiinteä komposiittipalanen, suurin mahdollinen valokovetusteho kohteeseen saavutetaan edelleen laittamalla kovetuskärki kiinni muoviin. Jos etäisyyttä muovin pinnasta sen sijaan kasvatettiin, valokovetusteho ei laskenutkaan niin nopeasti kuin oli odotettu. Tämä voi liittyä siihen, että tehokkaan valokeilan halkaisijan koko on suurempi verrattuna komposiitin sekä sensorin halkaisian kokoon. Toiseksi on arvioitu, että resiinikomposiitin täyteaineet voisivat fokusoida läpi kulkevaa valoa sensoriin. Se, pitääkö tämä ilmiö paikkansa, vaatii kuitenkin enemmän tutkimusta