Photoinhibition of Photosystem II. Kinetics, Photoprotection and Mechanism

Photosystem II (PSII) is susceptible to light-induced damage defined as photoinhibition. In natural conditions, plants are capable of repairing the photoinhibited PSII by on-going degradation and re-synthesis of the D1 reaction centre protein of PSII. Photoinhibition is induced by both visible and ultraviolet light and photoinhibition occurs under all light intensities with the same efficiency per photon. In my thesis work, I studied the reaction kinetics and mechanism of photoinhibition of PSII, as well as photoprotection in leaves of higher plants. Action spectroscopy was used to identify photoreceptors of photoinhibition. I found that the action spectrum of photoinhibition in vivo shows resemblance to the absorption spectra of manganese model compounds of the oxygen evolving complex (OEC) suggesting a role for manganese as a photoreceptor of photoinhibition under UV and visible light. In order to study the protective effect of non-photochemical quenching, the action spectrum was measured from leaves of wild type Arabidopsis thaliana and two mutants impaired in nonphotochemical quenching of chlorophyll a excitations. The findings of action spectroscopy and simulations of chlorophyll-based photoinhibition mechanisms suggested that quenching of antenna excitations protects less efficiently than would be expected if antenna chlorophylls were the only photoreceptors of photoinhibition. The reaction kinetics of prolonged photoinhibition was studied in leaves of Cucurbita maxima and Capsicum annuum. The results indicated that photoinhibitory decrease in both the oxygen evolution activity and ratio of variable to maximum fluorescence follows firstorder kinetics in vivo. The persistence of first-order kinetics suggests that already photoinhibited reaction centres do not protect against photoinhibition and that the mechanism of photoinhibition does not have a reversible intermediate. When Cucurbita maxima leaves were photoinhibited with saturating single-turnover flashes and continuous light, the light response curve of photoinhibition was found to be essentially a straight line with both types of illumination, suggesting that similar photoinhibition mechanisms might function during illumination with continuous light and during illumination with short flashes.

Supermatrix-hankkeen vahvuudet, heikkoudet, mahdollisuudet ja uhat

Supermatrix on Finnet-liitto ry:n jäsenyritysten hanke, joka tuo nopeiden valokuituyhteyksien avulla erilaisia sähköisiä palveluita kuluttajien koteihin ja yritysten toimipisteisiin. Näitä palveluita ovat mm. virtualisoidut henkilö-kohtaiset tietokoneet, tiedon tallennuspalvelut ja multimediasisältö, kuten teräväpiirtotelevisio. Tässä diplomityössä arvioidaan Supermatrixin palveluiden toteutumis-mahdollisuuksia sekä teknologian, että markkinoiden kannalta. Työn tarkoituksena oli myös laskea virtualisoitujen tietokoneiden avulla tuotetun työpöytäpalvelun tuotantokustannus. Kustannuslaskentaa varten testattiin useiden valmistajien teknologioita, ja parhaimmista vaihtoehdoista laskettiin tuotantokustannukset. Tulevia Supermatrix-operaattoreita varten kehitettiin työkalu, jolla työpöytäpalvelun vaatimien investointien kannattavuus saadaan nopeasti selville. Tuotantohintojen laskelmat osoittavat, että työpöytäpalvelua on taloudelli-sesti mahdollista tuottaa luomaan lisäarvoa nopeille laajakaistayhteyksille. Yrityksissä työpöytien virtualisointi on jo tämän päivän teknologiaa, mutta kuluttaja-asiakkaiden innostuminen palvelusta vaatii uuden markkinan luomisen. Uhkana on, että kuluttajien ostokäyttäytymisen muuttaminen ei onnistu, koska Supermatrix ei tuota fyysisen tuotteen ostamisen tarjoamaa mielihyvää.

ZigBee-verkot ja evaluointilaitteen suunnittelu

In this thesis a closer look at ZigBee wireless network protocol was made and a ZigBee evaluation device was designed and built for testing it. ZigBee is a digital wireless network technology published in 2004, which was developed for wireless control and automation systems. It is based on the IEEE 802.15.4-2003 standard, and its aim was to create a reliable wireless networking protocol with secure data transmissions, low power consumption and low manufacturing costs. The low power consumption requirement has also led to a slow data rate and a short range, but in a ZigBee network individual network devices can route messages to each other, so short range is not a problem. In automation and control systems a fast data rate is not usually an important requirement either. ZigBee network can form automatically and one ZigBee network can theoretically include thousands of devices, all of which can communicate with each other. ZigBee network is also self-healing, so an individual device dropping out of the network, like breaking down, does not affect functionality of the network. Evaluation device was designed and implemented in four different versions, which were tested with six different antenna designs. All antennas were tested at a range of 100 meters in an open terrain using two different frequencies and transmission powers. Also a range test was performed for selected antennas using transmission power of 4,5 dBm, which was maximum power of the ZigBee chip used in the evaluation device. Range test resulted in a measured range of several hundred meters, which can be considered as a good result for a short-range technology.

RF disturbances produced by high-power photovoltaic solar plants

In this thesis, analysis of electromagnetic compatibility of high-power photovoltaic solar plant is made. Current standards suitable for photovoltaic applications are given. Measurements of antenna factor for experimental setup are shown. Also, measurements of common mode disturbance voltages in high-power solar plant are given. Importance of DC-side filter is shown. In the last part of the work, electromagnetic simulations are made. These simulations show influence of several factors to EMC of power plant. Based on these simulations and measurements recommendations are given.

Liiketoiminnan kehittäminen verkostoitumalla, tutkimus sähkö- ja teleasennustoimialalta

Tällä tutkimuksella annetaan työkaluja pienille mikrotason asennusyrityksille heidän tiellään tuottavampaan ja tehokkaampaan yritystoimintaan. Kehitysmenetelmäksi on valittu verkostoituminen, joka tapahtuu alueellisten toimijoiden kesken ja on lähinnä horisontaalista. Tutkimuksen kohderyhmänä on sähkö-, tele- ja antenniasennusyrittäjät tai alalle aikovat uudet yrittäjät. Luotu malli on sovelias lähes kaikille kiinteistöjen teknisen kunnossapidon alalla toimiville mikroyrityksille. Tutkimuksen teoreettisessa osassa käsitellään verkostoyrittäjyyden perusteita ja syventävästi esitetyn case-mallin mukaista verkostotoiminnan erityispiirteitä. Empiirinen osa koostuu sähköisesti toteutetusta internetpohjaisesta kysely- tutkimuksesta, jonka kohteena on pienet sähkö-, tele- ja antenniasennusyritykset. Kyselytutkimus on toteutettu yhteistyössä alan urakointiliittojen (STUL ry ja SANT ry) kanssa. Kysely noudattaa samoja aihealueita kuin professori Asko Miettisen sähköurakointialalle 1988 toteuttaman raportin SÄHKÖURAKOINTI 2000 kyselylomake pidemmän tarkastelujänteen saamiseksi. Kysely selvittää yritysten nykytilaa ja näkemyksiä alan lähitulevaisuudesta. Empiiristä osaa tuetaan voimakkaasti case- esityksin, jotka pohjautuvat kirjoittajan 40 vuoden yrittäjäkokemukseen tällä alalla, joka on ollut voimakkaastikin lähinnä vertikaalisesti verkottunut. Tutkimus osoittaa, että alan taloudellinen ja toiminnallinenkin kehitys on polkenut paikallaan viimeiset vuosikymmenet, vaikka alan teknologiakehitys on ollut tarkasteluajanjaksona varsin merkittävää. Alan yritysten heikohkojen resurssien vuoks entistä voimakkaampi alueellinen yhteistyö verkostoitumalla on mahdollisuus tuottavampaan yritystoimintaan ja arvontuottoon yritykselle sekä luo mahdollisuuden myös suuntautumisen uusille kehittyneimmille teknologian alueille.

Regulation of photosynthetic electron transfer in plant chloroplasts

This thesis focuses on the molecular mechanisms regulating the photosynthetic electron transfer reactions upon changes in light intensity. To investigate these mechanisms, I used mutants of the model plant Arabidopsis thaliana impaired in various aspects of regulation of the photosynthetic light reactions. These included mutants of photosystem II (PSII) and light harvesting complex II (LHCII) phosphorylation (stn7 and stn8), mutants of energy-dependent non-photochemical quenching (NPQ) (npq1 and npq4) and of regulation of photosynthetic electron transfer (pgr5). All of these processes have been extensively investigated during the past decades, mainly on plants growing under steady-state conditions, and therefore many aspects of acclimation processes may have been neglected. In this study, plants were grown under fluctuating light, i.e. the alternation of low and high intensities of light, in order to maximally challenge the photosynthetic regulatory mechanisms. In pgr5 and stn7 mutants, the growth in fluctuating light condition mainly damaged PSI while PSII was rather unaffected. It is shown that the PGR5 protein regulates the linear electron transfer: it is essential for the induction of transthylakoid ΔpH that, in turn, activates energy-dependent NPQ and downregulates the activity of cytochrome b6f. This regulation was shown to be essential for the photoprotection of PSI under fluctuations in light intensity. The stn7 mutants were able to acclimate under constant growth light conditions by modulating the PSII/PSI ratio, while under fluctuating growth light they failed in implementing this acclimation strategy. LHCII phosphorylation ensures the balance of the excitation energy distribution between PSII and PSI by increasing the probability for excitons to be trapped by PSI. LHCII can be phosphorylated over all of the thylakoid membrane (grana cores as well as stroma lamellae) and when phosphorylated it constitutes a common antenna for PSII and PSI. Moreover, LHCII was shown to work as a functional bridge that allows the energy transfer between PSII units in grana cores and between PSII and PSI centers in grana margins. Consequently, PSI can function as a quencher of excitation energy. Eventually, the LHCII phosphorylation, NPQ and the photosynthetic control of linear electron transfer via cytochrome b6f work in concert to maintain the redox poise of the electron transfer chain. This is a prerequisite for successful plant growth upon changing natural light conditions, both in short- and long-term.

VIRVE-tukiaseman peittoalueen mallintaminen

Tekniikan kehityksen myötä reaalimaailman ilmiöitä voidaan mallintaa yhä tarkemmin. Tässä työssä tutkitaan VIRVE-radioverkon tukiaseman peittoalueen mallintamista. Työn tavoitteena oli mallintaa VIRVE-tukiaseman peittoalue mahdollisimman tarkasti. Tavoitteena oli myös löytää VIRVE-tukiasemaan peittoalueen kannalta nykyistä paremmat antenniratkaisut. Tutkimuksen teoreettisessa osuudessa selvitettiin tukiaseman peittoalueen mallintamiseen liittyviä tekijöitä, kuten radioaaltojen ominaisuuksia ja etenemismalleja sekä antennien teoriaa. Eri etenemismalleilla laskettuja tukiasemien peittoennusteita verrattiin empiirisesti toteutettuihin tukiasemien peittoalueiden mittaustuloksiin. Työn tuloksena löydettiin radioaaltojen etenemismalli, jolla mallinnettavien tukiasemien peittoalue-ennusteet vastasivat hyvin empiirisesti mitattuja tuloksia. Lisäksi löydettiin nykyistä paremmat antenniratkaisut VIRVE-tukiasemiin.

Switchable lanthanide fluorescence probes in homogenous molecular diagnostics

The number of molecular diagnostic assays has increased tremendously in recent years.Nucleic acid diagnostic assays have been developed, especially for the detection of human pathogenic microbes and genetic markers predisposing to certain diseases. Closed-tube methods are preferred because they are usually faster and easier to perform than heterogenous methods and in addition, target nucleic acids are commonly amplified leading to risk of contamination of the following reactions by the amplification product if the reactions are opened. The present study introduces a new closed-tube switchable complementation probes based PCR assay concept where two non-fluorescent probes form a fluorescent lanthanide chelate complex in the presence of the target DNA. In this dual-probe PCR assay method one oligonucleotide probe carries a non-fluorescent lanthanide chelate and another probe a light absorbing antenna ligand. The fluorescent lanthanide chelate complex is formed only when the non-fluorescent probes are hybridized to adjacent positions into the target DNA bringing the reporter moieties in close proximity. The complex is formed by self-assembled lanthanide chelate complementation where the antenna ligand is coordinated to the lanthanide ion captured in the chelate. The complementation probes based assays with time-resolved fluorescence measurement showed low background signal level and hence, relatively high nucleic acid detection sensitivity (low picomolar target concentration). Different lanthanide chelate structures were explored and a new cyclic seven dentate lanthanide chelate was found suitable for complementation probe method. It was also found to resist relatively high PCR reaction temperatures, which was essential for the PCR assay applications. A seven-dentate chelate with two unoccupied coordination sites must be used instead of a more stable eight- or nine-dentate chelate because the antenna ligand needs to be coordinated to the free coordination sites of the lanthanide ion. The previously used linear seven-dentate lanthanide chelate was found to be unstable in PCR conditions and hence, the new cyclic chelate was needed. The complementation probe PCR assay method showed high signal-to-background ratio up to 300 due to a low background fluorescence level and the results (threshold cycles) in real-time PCR were reached approximately 6 amplification cycles earlier compared to the commonly used FRET-based closed-tube PCR method. The suitability of the complementation probe method for different nucleic acid assay applications was studied. 1) A duplex complementation probe C. trachomatis PCR assay with a simple 10-minute urine sample preparation was developed to study suitability of the method for clinical diagnostics. The performance of the C. trachomatis assay was equal to the commercial C. trachomatis nucleic acid amplification assay containing more complex sample preparation based on DNA extraction. 2) A PCR assay for the detection of HLA-DQA1*05 allele, that is used to predict the risk of type 1 diabetes, was developed to study the performance of the method in genotyping. A simple blood sample preparation was used where the nucleic acids were released from dried blood sample punches using high temperature and alkaline reaction conditions. The complementation probe HLA-DQA1*05 PCR assay showed good genotyping performance correlating 100% with the routinely used heterogenous reference assay. 3) To study the suitability of the complementation probe method for direct measurement of the target organism, e.g., in the culture media, the complementation probes were applied to amplificationfree closed-tube bacteriophage quantification by measuring M13 bacteriophage ssDNA. A low picomolar bacteriophage concentration was detected in a rapid 20- minute assay. The assay provides a quick and reliable alternative to the commonly used and relatively unreliable UV-photometry and time-consuming culture based bacteriophage detection methods and indicates that the method could also be used for direct measurement of other micro-organisms. The complementation probe PCR method has a low background signal level leading to a high signal-to-background ratio and relatively sensitive nucleic acid detection. The method is compatible with simple sample preparation and it was shown to tolerate residues of urine, blood, bacteria and bacterial culture media. The common trend in nucleic acid diagnostics is to create easy-to-use assays suitable for rapid near patient analysis. The complementation probe PCR assays with a brief sample preparation should be relatively easy to automate and hence, would allow the development of highperformance nucleic acid amplification assays with a short overall assay time.

Switchable Lanthanide Luminescence for Detection of Biomolecules

Binary probes are oligonucleotide probe pairs that hybridize adjacently to a complementary target nucleic acid. In order to detect this hybridization, the two probes can be modified with, for example, fluorescent molecules, chemically reactive groups or nucleic acid enzymes. The benefit of this kind of binary probe based approach is that the hybridization elicits a detectable signal which is distinguishable from background noise even though unbound probes are not removed by washing before measurement. In addition, the requirement of two simultaneous binding events increases specificity. Similarly to binary oligonucleotide probes, also certain enzymes and fluorescent proteins can be divided into two parts and used in separation-free assays. Split enzyme and fluorescent protein reporters have practical applications among others as tools to investigate protein-protein interactions within living cells. In this study, a novel label technology, switchable lanthanide luminescence, was introduced and used successfully in model assays for nucleic acid and protein detection. This label technology is based on a luminescent lanthanide chelate divided into two inherently non-luminescent moieties, an ion carrier chelate and a light harvesting antenna ligand. These form a highly luminescent complex when brought into close proximity; i.e., the label moieties switch from a dark state to a luminescent state. This kind of mixed lanthanide complex has the same beneficial photophysical properties as the more typical lanthanide chelates and cryptates - sharp emission peaks, long emission lifetime enabling time-resolved measurement, and large Stokes’ shift, which minimize the background signal. Furthermore, the switchable lanthanide luminescence technique enables a homogeneous assay set-up. Here, switchable lanthanide luminescence label technology was first applied to sensitive, homogeneous, single-target nucleic acid and protein assays with picomolar detection limits and high signal to background ratios. Thereafter, a homogeneous four-plex nucleic acid array-based assay was developed. Finally, the label technology was shown to be effective in discrimination of single nucleotide mismatched targets from fully matched targets and the luminescent complex formation was analyzed more thoroughly. In conclusion, this study demonstrates that the switchable lanthanide luminescencebased label technology can be used in various homogeneous bioanalytical assays.

Veden desinfiointi UV-LED-valolla

Veden riittämätön puhdistus aiheuttaa riskin veden käyttäjille. Miljoonia kuolemia vuosittain aiheuttavien vesiteitse leviävien sairauksien ehkäisemiseksi vaaditaan tehokkaita juomaveden desinfiointimenetelmiä. Kuivuuden ja väestönkasvun myötä veden tarve on lisääntynyt ja vedenkulutus tulee yhä kasvamaan. Tästä syystä mahdollisuus kierrättää vettä hyödyntäen sitä esimerkiksi kasteluun on saanut yhä enemmän huomiota. Kierrätettävä vesi on kuitenkin käsiteltävä huolellisesti sen sisältämän mikrobiologisen kontaminaatioriskin vuoksi. Ultraviolettisäteily luokitellaan fysikaaliseksi desinfiointimenetelmäksi. Sen tehokkuus perustuu mikro-organismien absorboimaan UV-säteilyyn, jonka aiheuttamien DNA:ssa tai RNA:ssa tapahtuvien muutoksien seurauksena mikro-organismi inaktivoituu ja estyy lisääntymästä. UV-desinfioinnissa on tyypillisesti käytetty elohopeahöyrylamppuja. Vaihtoehtoinen UV-säteilyn lähde ovat LEDit eli valoa emittoivat diodit. Matalapaine-elohopeahöyrylamppujen emittoima säteily on aallonpituudella 254 nm ja keskipaine-elohopeahöyrylamppujen emittoima säteily on laajakaistaista säteilyä. Energiatehokkuuden lisäksi LEDien etuna on, että niillä voidaan tuottaa kapeakaista säteilyä aallonpituudella, joka parhaiten absorboituu DNA:han. Tämän diplomityön tarkoituksena oli tutkia, onko UVC-alueen aallonpituuksien yhdistelmillä synergistisiä etuja LEDien desinfiointitehokkuuteen, kun desinfioidaan virtaavaa vettä useilla säteilyannoksilla ja indikaattorimikrobina käytetään kolibakteeria. Tavoitteena oli myös tutkia tällä hetkellä saatavissa olevien LEDien desinfiointitehokkuutta energiatehokkuuden näkökulmasta. Yksittäisistä aallonpituuksista desinfiointitehokkuudeltaan parhaimmaksi osoittautui 260 nm, aallonpituuksien yhdistelmistä tehokkain oli 265 nm:n ja 260 nm:n yhdistelmä. Muilla aallonpituuksien yhdistelmillä ei saavutettu odotettua parempaa desinfiointitehokkuutta. Optiselta teholtaan parhaimmat LEDit, 265 nm, 270 nm ja 275 nm olivat kokeiden perusteella myös energiatehokkuuden kannalta tarkasteltuina parhaimmat sekä yksittäin että yhdistelminä. UVC-aallonpituuksia emittoivien LEDien optisen tehokkuuden paraneminen on edellytys LEDien hyödyntämiselle desinfioinnissa.

Modeling of fractal antennas

In this Master Thesis the characteristics of the chosen fractal microstrip antennas are investigated. For modeling has been used the structure of the square Serpinsky fractal curves. During the elaboration of this Master thesis the following steps were undertaken: 1) calculation and simulation of square microstrip antennа, 2) optimizing for obtaining the required characteristics on the frequency 2.5 GHz, 3) simulation and calculation of the second and third iteration of the Serpinsky fractal curves, 4) radiation patterns and intensity distribution of these antennas. In this Master’s Thesis the search for the optimal position of the port and fractal elements was conducted. These structures can be used in perspective for creation of antennas working at the same time in different frequency range.

Radar cross-section measurement using the Doppler effect

In this Master Thesis we discuss issues related to the measurement of the effective scattering surface, based on the Doppler Effect. Modeling of the detected signal was made. Narrowband signal filtering using low-frequency amplifier was observed. Parameters of the proposed horn antennas were studied; radar cross section charts for three different objects were received.

Determinants of Bone Strength and Predictors of Hip Fracture among Finnish Adults. Results from the Health 2000 Survey and the Mini-Finland Health Survey

The objective of this thesis was to identify the determinants of bone strength and predictors of hip fracture in representative samples of Finnish adults. A secondary objective was to construct a simple multifactorial model for hip fracture prediction over a 10-year follow-up period. The study was based on the Health 2000 Survey conducted during 2000 to 2001 (men and women aged 30 years or over, n=6 035) and the Mini-Finland Health Survey conducted during 1978 to 1980 (women aged 45 years or over, n=2 039). Study subjects participated in health interviews and comprehensive health examination. In the Health 2000 Survey, bone strength was assessed by means of calcaneal quantitative ultrasound (QUS). The follow-up information about hip fractures was drawn from the National Hospital Discharge Register. In this study, age, weight, height, serum 25-hydroxyvitamin D (S-25(OH)D), physical activity, smoking and alcohol consumption as well as menopause and eventual HRT in women were found to be associated with calcaneal broadband ultrasound attenuation (BUA) and speed of sound (SOS). Parity was associated with a decreased risk of hip fracture in postmenopausal women. Age, height, weight or waist circumference, quantitative ultrasound index (QUI), S-25(OH)D and fall-related factors, such as maximal walking speed, Parkinson’s disease, and the number of prescribed CNS active medication were significant independent predictors of hip fracture. At the population level, the incremental value of QUS appeared to be minor in hip fracture prediction when the fall-related risk factors were taken into account. A simple multifactorial model for hip fracture prediction presented in this study was based on readily available factors (age, gender, height, waist circumference, and fallrelated factors). Prospective studies are needed to test this model in patient-based study populations.

Modelling of faults in low-voltage cables

Power line modelling has become an interesting research area in recent years as a result of advances in the power line distribution network system. Extensive knowledge about the power line cable characteristics can be implemented in a software algorithm in a modern broadband power-line communication modem. In this study, a novel approach for modelling power line cables (AMCMK) based on the broadband impedance spectroscopy (BIS) and transmission line matrix (TLM) techniques is recommended in characterizing a healthy cable and the various faults associated with low-voltage cables for both open and short circuit situation. Models for different cable conditions are developed and tuned, which include six models for both healthy and faulty cables situations. The models are on the basis of impedance response analysis of the cable. The resulting spectra from the simulations are also cross-correlated to determine the degree of similarities between the healthy cable spectra and their respective faulty spectra.