Brown algal phlorotannins: Improving and applying chemical methods

Phlorotannins are the least studied group of tannins and are found only in brown algae. Hitherto the roles of phlorotannins, e.g. in plant-herbivore interactions, have been studied by quantifying the total contents of the soluble phlorotannins with a variety of methods. Little attention has been given to either quantitative variation in cell-wall-bound and exuded phlorotannins or to qualitative variation in individual compounds. A quantification procedure was developed to measure the amount of cell-wall-bound phlorotannins. The quantification of soluble phlorotannins was adjusted for both large- and small-scale samples and used to estimate the amounts of exuded phlorotannins using bladder wrack (Fucus vesiculosus) as a model species. In addition, separation of individual soluble phlorotannins to produce a phlorotannin profile from the phenolic crude extract was achieved by high-performance liquid chromatography (HPLC). Along with these methodological studies, attention was focused on the factors in the procedure which generated variation in the yield of phlorotannins. The objective was to enhance the efficiency of the sample preparation procedure. To resolve the problem of rapid oxidation of phlorotannins in HPLC analyses, ascorbic acid was added to the extractant. The widely used colourimetric method was found to produce a variation in the yield that was dependent upon the pH and concentration of the sample. Using these developed, adjusted and modified methods, the phenotypic plasticity of phlorotannins was studied with respect to nutrient availability and herbivory. An increase in nutrients decreased the total amount of soluble phlorotannins but did not affect the cell-wall-bound phlorotannins, the exudation of phlorotannins or the phlorotannin profile achieved with HPLC. The presence of the snail Thedoxus fluviatilis on the thallus induced production of soluble phlorotannins, and grazing by the herbivorous isopod Idotea baltica increased the exudation of phlorotannins. To study whether the among-population variations in phlorotannin contents arise from the genetic divergence or from the plastic response of algae, or both, algae from separate populations were reared in a common garden. Genetic variation among local populations was found in both the phlorotannin profile and the content of total phlorotannins. Phlorotannins were also genetically variable within populations. This suggests that local algal populations have diverged in their contents of phlorotannins, and that they may respond to natural selection and evolve both quantitatively and qualitatively.

The application of cDNA and tissue microarray methods in the study of human carcinomas

Currently, numerous high-throughput technologies are available for the study of human carcinomas. In literature, many variations of these techniques have been described. The common denominator for these methodologies is the high amount of data obtained in a single experiment, in a short time period, and at a fairly low cost. However, these methods have also been described with several problems and limitations. The purpose of this study was to test the applicability of two selected high-throughput methods, cDNA and tissue microarrays (TMA), in cancer research. Two common human malignancies, breast and colorectal cancer, were used as examples. This thesis aims to present some practical considerations that need to be addressed when applying these techniques. cDNA microarrays were applied to screen aberrant gene expression in breast and colon cancers. Immunohistochemistry was used to validate the results and to evaluate the association of selected novel tumour markers with the outcome of the patients. The type of histological material used in immunohistochemistry was evaluated especially considering the applicability of whole tissue sections and different types of TMAs. Special attention was put on the methodological details in the cDNA microarray and TMA experiments. In conclusion, many potential tumour markers were identified in the cDNA microarray analyses. Immunohistochemistry could be applied to validate the observed gene expression changes of selected markers and to associate their expression change with patient outcome. In the current experiments, both TMAs and whole tissue sections could be used for this purpose. This study showed for the first time that securin and p120 catenin protein expression predict breast cancer outcome and the immunopositivity of carbonic anhydrase IX associates with the outcome of rectal cancer. The predictive value of these proteins was statistically evident also in multivariate analyses with up to a 13.1- fold risk for cancer specific death in a specific subgroup of patients.

Multidata inverse problems and Bayesian solution methods in astronomy

Statistical analyses of measurements that can be described by statistical models are of essence in astronomy and in scientific inquiry in general. The sensitivity of such analyses, modelling approaches, and the consequent predictions, is sometimes highly dependent on the exact techniques applied, and improvements therein can result in significantly better understanding of the observed system of interest. Particularly, optimising the sensitivity of statistical techniques in detecting the faint signatures of low-mass planets orbiting the nearby stars is, together with improvements in instrumentation, essential in estimating the properties of the population of such planets, and in the race to detect Earth-analogs, i.e. planets that could support liquid water and, perhaps, life on their surfaces. We review the developments in Bayesian statistical techniques applicable to detections planets orbiting nearby stars and astronomical data analysis problems in general. We also discuss these techniques and demonstrate their usefulness by using various examples and detailed descriptions of the respective mathematics involved. We demonstrate the practical aspects of Bayesian statistical techniques by describing several algorithms and numerical techniques, as well as theoretical constructions, in the estimation of model parameters and in hypothesis testing. We also apply these algorithms to Doppler measurements of nearby stars to show how they can be used in practice to obtain as much information from the noisy data as possible. Bayesian statistical techniques are powerful tools in analysing and interpreting noisy data and should be preferred in practice whenever computational limitations are not too restrictive.

Reduction and Analysis Methods of Indigo

Throughout history indigo was derived from various plants for example Dyer’s Woad (Isatis tinctoria L.) in Europe. In the 19th century were the synthetic dyes developed and nowadays indigo is mainly synthesized from by-products of fossil fuels. Indigo is a so-called vat dye, which means that it needs to be reduced to its water soluble leucoform before dyeing. Nowadays, most of the industrial reduction is performed chemically by sodium dithionite. However, this is considered environmentally unfavourable because of waste waters contaminating degradation products. Therefore there has been interest to find new possibilities to reduce indigo. Possible alternatives for the application of dithionite as the reducing agent are biologically induced reduction and electrochemical reduction. Glucose and other reducing sugars have recently been suggested as possible environmentally friendly alternatives as reducing agents for sulphur dyes and there have also been interest in using glucose to reduce indigo. In spite of the development of several types of processes, very little is known about the mechanism and kinetics associated with the reduction of indigo. This study aims at investigating the reduction and electrochemical analysis methods of indigo and give insight on the reduction mechanism of indigo. Anthraquinone as well as it’s derivative 1,8-dihydroxyanthraquinone were discovered to act as catalysts for the glucose induced reduction of indigo. Anthraquinone introduces a strong catalytic effect which is explained by invoking a molecular “wedge effect” during co-intercalation of Na+ and anthraquinone into the layered indigo crystal. The study includes also research on the extraction of plant-derived indigo from woad and the examination of the effect of this method to the yield and purity of indigo. The purity has been conventionally studied spectrophotometrically and a new hydrodynamic electrode system is introduced in this study. A vibrating probe is used in following electrochemically the leuco-indigo formation with glucose as a reducing agent.

Food Safety Testing: Rapid Molecular Methods for Chemical and Biological Hazards

The central goal of food safety policy in the European Union (EU) is to protect consumer health by guaranteeing a high level of food safety throughout the food chain. This goal can in part be achieved by testing foodstuffs for the presence of various chemical and biological hazards. The aim of this study was to facilitate food safety testing by providing rapid and user-friendly methods for the detection of particular food-related hazards. Heterogeneous competitive time-resolved fluoroimmunoassays were developed for the detection of selected veterinary residues, that is coccidiostat residues, in eggs and chicken liver. After a simplified sample preparation procedure, the immunoassays were performed either in manual format with dissociation-enhanced measurement or in automated format with pre-dried assay reagents and surface measurement. Although the assays were primarily designed for screening purposes providing only qualitative results, they could also be used in a quantitative mode. All the developed assays had good performance characteristics enabling reliable screening of samples at concentration levels required by the authorities. A novel polymerase chain reaction (PCR)-based assay system was developed for the detection of Salmonella spp. in food. The sample preparation included a short non-selective pre-enrichment step, after which the target cells were collected with immunomagnetic beads and applied to PCR reaction vessels containing all the reagents required for the assay in dry form. The homogeneous PCR assay was performed with a novel instrument platform, GenomEra^™, and the qualitative assay results were automatically interpreted based on end-point time-resolved fluorescence measurements and cut-off values. The assay was validated using various food matrices spiked with sub-lethally injured Salmonella cells at levels of 1-10 colony forming units (CFU)/25 g of food. The main advantage of the system was the exceptionally short time to result; the entire process starting from the pre-enrichment and ending with the PCR result could be completed in eight hours. In conclusion, molecular methods using state-of-the-art assay techniques were developed for food safety testing. The combination of time-resolved fluorescence detection and ready-to-use reagents enabled sensitive assays easily amenable to automation. Consequently, together with the simplified sample preparation, these methods could prove to be applicable in routine testing.

Methods for assessing the sustainability of integrated municipal waste management and

The general striving to bring down the number of municipal landfills and to increase the reuse and recycling of waste-derived materials across the EU supports the debates concerning the feasibility and rationality of waste management systems. Substantial decrease in the volume and mass of landfill-disposed waste flows can be achieved by directing suitable waste fractions to energy recovery. Global fossil energy supplies are becoming more and more valuable and expensive energy sources for the mankind, and efforts to save fossil fuels have been made. Waste-derived fuels offer one potential partial solution to two different problems. First, waste that cannot be feasibly re-used or recycled is utilized in the energy conversion process according to EU’s Waste Hierarchy. Second, fossil fuels can be saved for other purposes than energy, mainly as transport fuels. This thesis presents the principles of assessing the most sustainable system solution for an integrated municipal waste management and energy system. The assessment process includes: · formation of a SISMan (Simple Integrated System Management) model of an integrated system including mass, energy and financial flows, and · formation of a MEFLO (Mass, Energy, Financial, Legislational, Other decisionsupport data) decision matrix according to the selected decision criteria, including essential and optional decision criteria. The methods are described and theoretical examples of the utilization of the methods are presented in the thesis. The assessment process involves the selection of different system alternatives (process alternatives for treatment of different waste fractions) and comparison between the alternatives. The first of the two novelty values of the utilization of the presented methods is the perspective selected for the formation of the SISMan model. Normally waste management and energy systems are operated separately according to the targets and principles set for each system. In the thesis the waste management and energy supply systems are considered as one larger integrated system with one primary target of serving the customers, i.e. citizens, as efficiently as possible in the spirit of sustainable development, including the following requirements: · reasonable overall costs, including waste management costs and energy costs; · minimum environmental burdens caused by the integrated waste management and energy system, taking into account the requirement above; and · social acceptance of the selected waste treatment and energy production methods. The integrated waste management and energy system is described by forming a SISMan model including three different flows of the system: energy, mass and financial flows. By defining the three types of flows for an integrated system, the selected factor results needed in the decision-making process of the selection of waste management treatment processes for different waste fractions can be calculated. The model and its results form a transparent description of the integrated system under discussion. The MEFLO decision matrix has been formed from the results of the SISMan model, combined with additional data, including e.g. environmental restrictions and regional aspects. System alternatives which do not meet the requirements set by legislation can be deleted from the comparisons before any closer numerical considerations. The second novelty value of this thesis is the three-level ranking method for combining the factor results of the MEFLO decision matrix. As a result of the MEFLO decision matrix, a transparent ranking of different system alternatives, including selection of treatment processes for different waste fractions, is achieved. SISMan and MEFLO are methods meant to be utilized in municipal decision-making processes concerning waste management and energy supply as simple, transparent and easyto- understand tools. The methods can be utilized in the assessment of existing systems, and particularly in the planning processes of future regional integrated systems. The principles of SISMan and MEFLO can be utilized also in other environments, where synergies of integrating two (or more) systems can be obtained. The SISMan flow model and the MEFLO decision matrix can be formed with or without any applicable commercial or free-of-charge tool/software. SISMan and MEFLO are not bound to any libraries or data-bases including process information, such as different emission data libraries utilized in life cycle assessments.

Bayesian methods for estimation, optimization and experimental design

Mathematical models often contain parameters that need to be calibrated from measured data. The emergence of efficient Markov Chain Monte Carlo (MCMC) methods has made the Bayesian approach a standard tool in quantifying the uncertainty in the parameters. With MCMC, the parameter estimation problem can be solved in a fully statistical manner, and the whole distribution of the parameters can be explored, instead of obtaining point estimates and using, e.g., Gaussian approximations. In this thesis, MCMC methods are applied to parameter estimation problems in chemical reaction engineering, population ecology, and climate modeling. Motivated by the climate model experiments, the methods are developed further to make them more suitable for problems where the model is computationally intensive. After the parameters are estimated, one can start to use the model for various tasks. Two such tasks are studied in this thesis: optimal design of experiments, where the task is to design the next measurements so that the parameter uncertainty is minimized, and model-based optimization, where a model-based quantity, such as the product yield in a chemical reaction model, is optimized. In this thesis, novel ways to perform these tasks are developed, based on the output of MCMC parameter estimation. A separate topic is dynamical state estimation, where the task is to estimate the dynamically changing model state, instead of static parameters. For example, in numerical weather prediction, an estimate of the state of the atmosphere must constantly be updated based on the recently obtained measurements. In this thesis, a novel hybrid state estimation method is developed, which combines elements from deterministic and random sampling methods.

Active Magnetic Bearing System Identification Methods and Rotor Model Updating

Active magnetic bearing is a type of bearing which uses magnetic field to levitate the rotor. These bearings require continuous control of the currents in electromagnets and data from position of the rotor and the measured current from electromagnets. Because of this different identification methods can be implemented with no additional hardware. In this thesis the focus was to implement and test identification methods for active magnetic bearing system and to update the rotor model. Magnetic center calibration is a method used to locate the magnetic center of the rotor. Rotor model identification is an identification method used to identify the rotor model. Rotor model update is a method used to update the rotor model based on identification data. These methods were implemented and tested with a real machine where rotor was levitated with active magnetic bearings and the functionality of the methods was ensured. Methods were developed with further extension in mind and also with the possibility to apply them for different machines with ease.

Methods of Telecommunication Technology Forecasting

This master's thesis coversthe concepts of knowledge discovery, data mining and technology forecasting methods in telecommunications. It covers the various aspects of knowledge discoveryin data bases and discusses in detail the methods of data mining and technologyforecasting methods that are used in telecommunications. Main concern in the overall process of this thesis is to emphasize the methods that are being used in technology forecasting for telecommunications and data mining. It tries to answer to some extent to the question of do forecasts create a future? It also describes few difficulties that arise in technology forecasting. This thesis was done as part of my master's studies in Lappeenranta University of Technology.

Multivariable regression analysis and hot-spot stress approach into fatigue life estimations of welded joints

Kolmen eri hitsausliitoksen väsymisikä arvio on analysoitu monimuuttuja regressio analyysin avulla. Regression perustana on laaja S-N tietokanta joka on kerätty kirjallisuudesta. Tarkastellut liitokset ovat tasalevy liitos, krusiformi liitos ja pitkittäisripa levyssä. Muuttujina ovat jännitysvaihtelu, kuormitetun levyn paksuus ja kuormitus tapa. Paksuus effekti on käsitelty uudelleen kaikkia kolmea liitosta ajatellen. Uudelleen käsittelyn avulla on varmistettu paksuus effektin olemassa olo ennen monimuuttuja regressioon siirtymistä. Lineaariset väsymisikä yhtalöt on ajettu kolmelle hitsausliitokselle ottaen huomioon kuormitetun levyn paksuus sekä kuormitus tapa. Väsymisikä yhtalöitä on verrattu ja keskusteltu testitulosten valossa, jotka on kerätty kirjallisuudesta. Neljä tutkimustaon tehty kerättyjen väsymistestien joukosta ja erilaisia väsymisikä arvio metodeja on käytetty väsymisiän arviointiin. Tuloksia on tarkasteltu ja niistä keskusteltu oikeiden testien valossa. Tutkimuksissa on katsottu 2mm ja 6mm symmetristäpitkittäisripaa levyssä, 12.7mm epäsymmetristä pitkittäisripaa, 38mm symmetristä pitkittäisripaa vääntökuormituksessa ja 25mm/38mm kuorman kantavaa krusiformi liitosta vääntökuormituksessa. Mallinnus on tehty niin lähelle testi liitosta kuin mahdollista. Väsymisikä arviointi metodit sisältävät hot-spot metodin jossa hot-spot jännitys on laskettu kahta lineaarista ja epälineaarista ekstrapolointiakäyttäen sekä paksuuden läpi integrointia käyttäen. Lovijännitys ja murtumismekaniikka metodeja on käytetty krusiformi liitosta laskiessa.

Project controlling and risk management in German machine and plant engineering industry

Tutkielman tavoitteena on määritellä projektikontrolloinnin ja - riskijohtamisen roolit ja toiminnot saksalaisissa kone- ja tehdassuunnitteluteollisuusyrityksissä. Tämä on kvalitatiivinen tutkielma, jossa käytetään voimakkaasti kuvailevia metodeita. Materiaali tutkimuksen empiiriseen osaan kerättiin kyselykaavakkeen avulla. Kyselykaavakkeiden tulokset käsiteltiin Microsoft Office Access- ohjelmalla ja analysoitiin Microsoft Office Excel- ohjelmalla ja Pivot table- työkalun avulla. Tutkimustulokset osoittavat, että asianmukaisessa projektikontrollointi- ja riskijohtamismetodien käytössä ja käyttötiheydessä esiintyy puutteita saksalaisissa kone- ja tehdassuunnitteluteollisuusyrityksissä. Tehostamalla ja keskittymällä enemmän projektikontrollointi- ja riskijohtamismetodeihin ja prosesseihin sekä projektien että yritysten suorituskyky paranisi.

Direct torque control of permanent magnet synchronous machines - analysis and implementation

Thedirect torque control (DTC) has become an accepted vector control method besidethe current vector control. The DTC was first applied to asynchronous machines,and has later been applied also to synchronous machines. This thesis analyses the application of the DTC to permanent magnet synchronous machines (PMSM). In order to take the full advantage of the DTC, the PMSM has to be properly dimensioned. Therefore the effect of the motor parameters is analysed taking the control principle into account. Based on the analysis, a parameter selection procedure is presented. The analysis and the selection procedure utilize nonlinear optimization methods. The key element of a direct torque controlled drive is the estimation of the stator flux linkage. Different estimation methods - a combination of current and voltage models and improved integration methods - are analysed. The effect of an incorrect measured rotor angle in the current model is analysed andan error detection and compensation method is presented. The dynamic performance of an earlier presented sensorless flux estimation method is made better by improving the dynamic performance of the low-pass filter used and by adapting the correction of the flux linkage to torque changes. A method for the estimation ofthe initial angle of the rotor is presented. The method is based on measuring the inductance of the machine in several directions and fitting the measurements into a model. The model is nonlinear with respect to the rotor angle and therefore a nonlinear least squares optimization method is needed in the procedure. A commonly used current vector control scheme is the minimum current control. In the DTC the stator flux linkage reference is usually kept constant. Achieving the minimum current requires the control of the reference. An on-line method to perform the minimization of the current by controlling the stator flux linkage reference is presented. Also, the control of the reference above the base speed is considered. A new estimation flux linkage is introduced for the estimation of the parameters of the machine model. In order to utilize the flux linkage estimates in off-line parameter estimation, the integration methods are improved. An adaptive correction is used in the same way as in the estimation of the controller stator flux linkage. The presented parameter estimation methods are then used in aself-commissioning scheme. The proposed methods are tested with a laboratory drive, which consists of a commercial inverter hardware with a modified software and several prototype PMSMs.

Theoretical and numerical study of thermomagnetic convection in magnetic fluids

In this thesis, the magnetic field control of convection instabilities and heat and mass transfer processesin magnetic fluids have been investigated by numerical simulations and theoretical considerations. Simulation models based on finite element and finite volume methods have been developed. In addition to standard conservation equations, themagnetic field inside the simulation domain is calculated from Maxwell equations and the necessary terms to take into account for the magnetic body force and magnetic dissipation have been added to the equations governing the fluid motion.Numerical simulations of magnetic fluid convection near the threshold supportedexperimental observations qualitatively. Near the onset of convection the competitive action of thermal and concentration density gradients leads to mostly spatiotemporally chaotic convection with oscillatory and travelling wave regimes, previously observed in binary mixtures and nematic liquid crystals. In many applications of magnetic fluids, the heat and mass transfer processes including the effects of external magnetic fields are of great importance. In addition to magnetic fluids, the concepts and the simulation models used in this study may be applied also to the studies of convective instabilities in ordinary fluids as well as in other binary mixtures and complex fluids.

Critical flux and fouling in ultrafiltration of proteins

In this thesis different parameters influencing critical flux in protein ultrafiltration and membrane foul-ing were studied. Short reviews of proteins, cross-flow ultrafiltration, flux decline and criticalflux and the basic theory of Partial Least Square analysis (PLS) are given at the beginning. The experiments were mainly performed using dilute solutions of globular proteins, commercial polymeric membranes and laboratory scale apparatuses. Fouling was studied by flux, streaming potential and FTIR-ATR measurements. Critical flux was evaluated by different kinds of stepwise procedures and by both con-stant pressure and constant flux methods. The critical flux was affected by transmembrane pressure, flow velocity, protein concentration, mem-brane hydrophobicity and protein and membrane charges. Generally, the lowest critical fluxes were obtained at the isoelectric points of the protein and the highest in the presence of electrostatic repulsion between the membrane surface and the protein molecules. In the laminar flow regime the critical flux increased with flow velocity, but not any more above this region. An increase in concentration de-creased the critical flux. Hydrophobic membranes showed fouling in all charge conditionsand, furthermore, especially at the beginning of the experiment even at very low transmembrane pressures. Fouling of these membranes was thought to be due to protein adsorption by hydrophobic interactions. The hydrophilic membranes used suffered more from reversible fouling and concentration polarisation than from irreversible foul-ing. They became fouled at higher transmembrane pressures becauseof pore blocking. In this thesis some new aspects on critical flux are presented that are important for ultrafiltration and fractionation of proteins.