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.

Quantification of Proteins and Cells: Luminometric Nonspecific Particle-Based Methods

New luminometric particle-based methods were developed to quantify protein and to count cells. The developed methods rely on the interaction of the sample with nano- or microparticles and different principles of detection. In fluorescence quenching, timeresolved luminescence resonance energy transfer (TR-LRET), and two-photon excitation fluorescence (TPX) methods, the sample prevents the adsorption of labeled protein to the particles. Depending on the system, the addition of the analyte increases or decreases the luminescence. In the dissociation method, the adsorbed protein protects the Eu(III) chelate on the surface of the particles from dissociation at a low pH. The experimental setups are user-friendly and rapid and do not require hazardous test compounds and elevated temperatures. The sensitivity of the quantification of protein (from 40 to 500 pg bovine serum albumin in a sample) was 20-500-fold better than in most sensitive commercial methods. The quenching method exhibited low protein-to-protein variability and the dissociation method insensitivity to the assay contaminants commonly found in biological samples. Less than ten eukaryotic cells were detected and quantified with all the developed methods under optimized assay conditions. Furthermore, two applications, the method for detection of the aggregation of protein and the cell viability test, were developed by utilizing the TR-LRET method. The detection of the aggregation of protein was allowed at a more than 10,000 times lower concentration, 30 μg/L, compared to the known methods of UV240 absorbance and dynamic light scattering. The TR-LRET method was combined with a nucleic acid assay with cell-impermeable dye to measure the percentage of dead cells in a single tube test with cell counts below 1000 cells/tube.

Multifrequency VLBI Observations of Selected Active Galactic Nuclei

In this dissertation, active galactic nuclei (AGN) are discussed, as they are seen with the high-resolution radio-astronomical technique called Very Long Baseline Interferometry (VLBI). This observational technique provides very high angular resolution (_ 10−300 = 1 milliarcsecond). VLBI observations, performed at different radio frequencies (multi-frequency VLBI), allow to penetrate deep into the core of an AGN to reveal an otherwise obscured inner part of the jet and the vicinity of the AGN’s central engine. Multi-frequency VLBI data are used to scrutinize the structure and evolution of the jet, as well as the distribution of the polarized emission. These data can help to derive the properties of the plasma and the magnetic field, and to provide constraints to the jet composition and the parameters of emission mechanisms. Also VLBI data can be used for testing the possible physical processes in the jet by comparing observational results with results of numerical simulations. The work presented in this thesis contributes to different aspects of AGN physics studies, as well as to the methodology of VLBI data reduction. In particular, Paper I reports evidence of optical and radio emission of AGN coming from the same region in the inner jet. This result was obtained via simultaneous observations of linear polarization in the optical and in radio using VLBI technique of a sample of AGN. Papers II and III describe, in detail, the jet kinematics of the blazar 0716+714, based on multi-frequency data, and reveal a peculiar kinematic pattern: plasma in the inner jet appears to move substantially faster that that in the large-scale jet. This peculiarity is explained by the jet bending, in Paper III. Also, Paper III presents a test of the new imaging technique for VLBI data, the Generalized Maximum Entropy Method (GMEM), with the observed (not simulated) data and compares its results with the conventional imaging. Papers IV and V report the results of observations of the circularly polarized (CP) emission in AGN at small spatial scales. In particular, Paper IV presents values of the core CP for 41 AGN at 15, 22 and 43 GHz, obtained with the help of the standard Gain transfer (GT) method, which was previously developed by D. Homan and J.Wardle for the calibration of multi-source VLBI observations. This method was developed for long multi-source observations, when many AGN are observed in a single VLBI run. In contrast, in Paper V, an attempt is made to apply the GT method to single-source VLBI observations. In such observations, the object list would include only a few sources: a target source and two or three calibrators, and it lasts much shorter than the multi-source experiment. For the CP calibration of a single-source observation, it is necessary to have a source with zero or known CP as one of the calibrators. If the archival observations included such a source to the list of calibrators, the GT could also be used for the archival data, increasing a list of known AGN with the CP at small spatial scale. Paper V contains also calculation of contributions of different sourced of errors to the uncertainty of the final result, and presents the first results for the blazar 0716+714.

Luminescence-based assay methods in drug discovery

The drug discovery process is facing new challenges in the evaluation process of the lead compounds as the number of new compounds synthesized is increasing. The potentiality of test compounds is most frequently assayed through the binding of the test compound to the target molecule or receptor, or measuring functional secondary effects caused by the test compound in the target model cells, tissues or organism. Modern homogeneous high-throughput-screening (HTS) assays for purified estrogen receptors (ER) utilize various luminescence based detection methods. Fluorescence polarization (FP) is a standard method for ER ligand binding assay. It was used to demonstrate the performance of two-photon excitation of fluorescence (TPFE) vs. the conventional one-photon excitation method. As result, the TPFE method showed improved dynamics and was found to be comparable with the conventional method. It also held potential for efficient miniaturization. Other luminescence based ER assays utilize energy transfer from a long-lifetime luminescent label e.g. lanthanide chelates (Eu, Tb) to a prompt luminescent label, the signal being read in a time-resolved mode. As an alternative to this method, a new single-label (Eu) time-resolved detection method was developed, based on the quenching of the label by a soluble quencher molecule when displaced from the receptor to the solution phase by an unlabeled competing ligand. The new method was paralleled with the standard FP method. It was shown to yield comparable results with the FP method and found to hold a significantly higher signal-tobackground ratio than FP. Cell-based functional assays for determining the extent of cell surface adhesion molecule (CAM) expression combined with microscopy analysis of the target molecules would provide improved information content, compared to an expression level assay alone. In this work, immune response was simulated by exposing endothelial cells to cytokine stimulation and the resulting increase in the level of adhesion molecule expression was analyzed on fixed cells by means of immunocytochemistry utilizing specific long-lifetime luminophore labeled antibodies against chosen adhesion molecules. Results showed that the method was capable of use in amulti-parametric assay for protein expression levels of several CAMs simultaneously, combined with analysis of the cellular localization of the chosen adhesion molecules through time-resolved luminescence microscopy inspection.

CFD Modelling of Direct Contact Condensation in Suppression Pools by Applying Condensation Models of Separated Flow

The condensation rate has to be high in the safety pressure suppression pool systems of Boiling Water Reactors (BWR) in order to fulfill their safety function. The phenomena due to such a high direct contact condensation (DCC) rate turn out to be very challenging to be analysed either with experiments or numerical simulations. In this thesis, the suppression pool experiments carried out in the POOLEX facility of Lappeenranta University of Technology were simulated. Two different condensation modes were modelled by using the 2-phase CFD codes NEPTUNE CFD and TransAT. The DCC models applied were the typical ones to be used for separated flows in channels, and their applicability to the rapidly condensing flow in the condensation pool context had not been tested earlier. A low Reynolds number case was the first to be simulated. The POOLEX experiment STB-31 was operated near the conditions between the ’quasi-steady oscillatory interface condensation’ mode and the ’condensation within the blowdown pipe’ mode. The condensation models of Lakehal et al. and Coste & Lavi´eville predicted the condensation rate quite accurately, while the other tested ones overestimated it. It was possible to get the direct phase change solution to settle near to the measured values, but a very high resolution of calculation grid was needed. Secondly, a high Reynolds number case corresponding to the ’chugging’ mode was simulated. The POOLEX experiment STB-28 was chosen, because various standard and highspeed video samples of bubbles were recorded during it. In order to extract numerical information from the video material, a pattern recognition procedure was programmed. The bubble size distributions and the frequencies of chugging were calculated with this procedure. With the statistical data of the bubble sizes and temporal data of the bubble/jet appearance, it was possible to compare the condensation rates between the experiment and the CFD simulations. In the chugging simulations, a spherically curvilinear calculation grid at the blowdown pipe exit improved the convergence and decreased the required cell count. The compressible flow solver with complete steam-tables was beneficial for the numerical success of the simulations. The Hughes-Duffey model and, to some extent, the Coste & Lavi´eville model produced realistic chugging behavior. The initial level of the steam/water interface was an important factor to determine the initiation of the chugging. If the interface was initialized with a water level high enough inside the blowdown pipe, the vigorous penetration of a water plug into the pool created a turbulent wake which invoked the chugging that was self-sustaining. A 3D simulation with a suitable DCC model produced qualitatively very realistic shapes of the chugging bubbles and jets. The comparative FFT analysis of the bubble size data and the pool bottom pressure data gave useful information to distinguish the eigenmodes of chugging, bubbling, and pool structure oscillations.

Motores de reacción modernos por turbinas de gas

This Bachelor’s thesis investigates the different types of jet engines used nowadays, their performance and applications. The thesis includes a general study of dynamics of fly, engine thermodynamics and contamination.

Sotilaslentäjän fyysinen suorituskyky sekä työperäiset tuki- ja liikuntaelinoireet

Kirjallisuudesta välittyy useitten vuosikymmenten ajalta tietämys sotilaslentämisen fyysisestä kuormittavuudesta. G-voimista aiheutuva kuormittuminen näyttää johtavan joko akuutisti tai pitkäaikaisesti lentäjän tuki- ja liikuntaelimistön toimintakykyä alentaviin ongelmiin. Erityisesti on selvitetty niskan alueen työperäisten ongelmien syntyä, jolloin on havaittu lentotoiminnan fyysisen kuormittavuuden johtavan ennenaikaiseen rakenteelliseen rappeumaan, haittaa aiheuttavan oireen lisäksi. Kansainvälisen kirjallisuuden mukaan ammatista johtuvista eli työperäisistä oireista kärsii vähintään 2/3 kaikista sotilaslentäjistä. Tietyin edellytyksin lentäjien kaularangan alueen rappeuma on Suomessa hyväksytty ammattitaudiksi vuodesta 1995 alkaen. On arveltu, että hyvästä fyysisestä suorituskyvystä olisi apua tuki- ja liikuntaelin (TULE)-oireilun ennaltaehkäisemisessä ja toimintakyvyn ylläpitämisessä. Tutkimusnäyttö tästä on lentäjien osalta ollut toistaiseksi erittäin niukkaa. Tämän tutkimuksen tavoitteena oli selvittää suomalaisten sotilaslentäjien työperäisen TULE-oireilun esiintyvyyttä, oireista koetun haitan tasoa, lentäjien fyysisen kunnon tasoja virkauran aikana ja näitten kaikkien välisiä yhteyksiä sekä työperäisen TULE-oireen merkitystä sotilaan toimintakykyyn. Tutkimus jakautui kahteen osaan. Poikkileikkauksena lentotoimintaperäisiä TULE-oireita kartoitettiin kyselytutkimuksella, johon vastasi vuositarkastuksen yhteydessä 267 lentäjää vuosina 2004-2005. Joukosta poimittiin ne 195 lentäjää, jotka olivat suorittaneet yleissotilaalliset kuntotestit puolen vuoden sisällä kyselyyn vastaamisesta, ja mitatut testitulokset yhdistettiin kyselytutkimusaineistoon. Tässä aineistossa toteutettiin fyysisesti erilailla kuormittuvien lentäjäryhmien välisiä vertailuja fyysisen kunnon, TULE-esiintyvyyden ja koetun haitan suhteen. Poikkileikkausosassa tutkittiin myös lentäjien virkauran aikaisia tasoeroja yleissotilaallisissa kuntotesteissä (n=195) verrattuna muihin suomalaisiin sotilaisiin. Lisäksi (N=289) selvitettiin ilmailulääketieteellisen tarkastuksen yhteydessä mitattuja, ns. ammatillisia fyysisiä erityisominaisuuksia eri ikäluokissa. Pitkittäisosassa seurattiin 67:n Hawk-suihkuharjoituskoneella aloittaneen Ilmavoimien sotilaslentäjien lentouran aikaista lentotoimintaperäisten TULE-oireitten esiintyvyyttä vuosien 1996 ja 2008 välillä. Lisäksi tutkittiin lentäjien kontakteja työterveyshuoltoon, oireen aiheuttamaa lentokelvottomuusaikaa, työn kuormituksen kumulatiivista kertymää lentotuntien lisääntyessä ja TULE-oireiden esiintyvyyden kannalta kriittisiä ajankohtia lentouran aikana. Tulokset osoittivat, että kaikki seurannassa olleet suomalaiset sotilaslentäjät kokivat jonkinasteisen lentotoimintaperäisen TULE-oireen uransa aikana. Niskan ammattitautiluokituksen tasoisen ongelman esiintyvyys oli 4 % koko lentäjäpopulaatiosta ja 10 % suihkuharjoituskonevaiheen jo läpäisseistä, mutta vastaavanlaisia TULE-ongelmia, ilman riittävää näyttöä ammattitaudista, esiintyi lähes joka kolmannella sotilaslentäjällä. Alaselän osalta lentäjät oireilivat lähes samassa määrin, mutta näitä oireita ei toistaiseksi ole mahdollista määrittää ammattitaudiksi. Lentäjät kävivät varsin vähän valittamassa oireistaan työterveyshuoltoon, jossa käytäneen vasta silloin, kun oire jo selvästi heikentää työtehtävissä vaadittavaa toimintakykyä. Merkittävin lentotoimintaperäisten oireitten esiintymisen kasvu ajoittui 200 Hawk-lentotunnin kohdalle, jolloin koneella saavutetaan eräänlainen optimaalinen G-indeksi eli taktisen liikehtelyn G-tasoylitysten vaihtelu. Tämän jälkeen lentäjät ovat erityisen alttiina akuuteille lennonaikaisille TULE-ongelmille. Oireitten esiintyminen kasvoi eksponentiaalisesti noin 600 lentotuntiin asti. Monimuuttujamallien mukaan työperäisen TULE-oireen esiintyvyysriskiä vähensivät alaraajojen hyvä motoriikka, korkeat valintapisteet ja korkea kaulan fleksion voimataso maksimaalisessa isometrisessä testissä. Yleissotilaallisilla kuntotasoilla ei ollut yhteyttä oireiluun, mutta lihaskunnoltaan voimakkaimmat lentäjät kärsivät tilastollisesti merkittävästi vähemmän haittaa lentotoimintaperäisistä TULE-oireistaan. Yleissotilaallisissa kuntotesteissä lentäjät olivat parempia kuin muut suomalaiset sotilaat. Aktiivisimman lentouran aikana, 30-40-vuotiaina, lentäjien fyysinen suorituskyky oli normaaliväestöön nähden vain keskimääräinen ja urheilijoihin nähden keskimääräistä heikompi. Käytännössä lentäjät eivät kyenneet ylläpitämään valintavaiheen fyysistä suorituskykyään edes kadettivaiheen loppuun asti. Huomattavaa oli lisäksi, että aktiivisen lentouran päätyttyä fyysinen kunto näytti jossain määrin palautuvan kohti lähtötasoa lentäjien ikääntymisestä huolimatta. Lentäjien valintavaiheen aikana mitatun fyysisen suorituskyvyn tason säilyminen aktiivisen lentopalveluksen loppuun asti vaatisi lentäjien fyysisen toimintakyvyn ylläpidon ja kehittämisen tehostamista koulutuksen ja työuran eri vaihessa. Tähän tavoitteeseen nähden Ilmavoimien fyysisen kasvatuksen järjestelyt vaikuttivat alimitoitetuilta. Operatiivisesti huolestuttavaa oli Ilmavoimien ohjaajien fyysisen suorituskyvyn heikentyminen silloin, kun heidän taitojensa puolesta olisi pitänyt olla suorituskykyisimpiä taistelutehtäviinsä. Myös lentäjän terveyttä ja toimintakykyä pitäisi pystyä reaaliaikaisemmin seuraamaan koko lentouran aikana. Ilmavoimille suositellaan moniammatillista lähestymistä sotilaslentäjien toimintakyvyn ylläpitämiseen ja terveysriskien hallintaan yhdessä liikunnan, työterveyshuollon, lentoturvallisuusalan ja operatiivisen suunnittelun asiantuntijoitten kanssa. Lisäksi suositellaan avoimempaa ja eettisesti kestävämpää suhtautumista ammattiin liittyvien terveysongelmien kuvaamiseen sekä fyysisen kunnon kysymyksiin jo lentäjien rekrytointivaiheessa.

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.

The effects of welding heat input on the usability of high strength steels in welded structures

High strength steel (HSS) has been in use in workshops since the 1980s. At that time, the significance of the term HSS differed from the modern conception as the maximum yield strength of HSSs has increased nearly every year. There are three different ways to make HSS. The first and oldest method is QT (quenched and tempered) followed by the TMCP (thermomechanical controlled process) and DQ (direct quenching) methods. This thesis consists of two parts, the first of which part introduces the research topic and discusses welded HSS structures by characterizing the most important variables. In the second part of the thesis, the usability of welded HSS structures is examined through a set of laboratory tests. The results of this study explain the differences in the usability of the welded HSSs made by the three different methods. The results additionally indicate that usage of different HSSs in the welded structures presumes that manufacturers know what kind of HSS they are welding. As manufacturers use greater strength HSSs in welded structures, the demands for welding rise as well. Therefore, during the manufacturing process, factors such as heat input, cooling time, weld quality, and more must be under careful observation.

Solids removal in pyrolysis

Pyrolysis is a process for turning biomass into liquid fuel. The process consists of heating the biomass in inert conditions and quenching the resulting vapors into oil. The oil has many potential uses, such as heating fuel in peak heating plants. In order to broaden the application base and improve the quality of the oil, solids removal has to be addressed. The solids may also increase the probability of plugging in downstream equipment. The purpose of this research was to gain an understanding of the formation of solids in the pyrolysis process and to assess options for reducing the solid content of the oil. From literature it is known that the solids can be removed either by hot vapor filtration, liquid treatment or multiple cyclones. Hot vapor filtration decreases yield, but improves the stability of the oil while simultaneously removing solids and ash. Liquid treatment techniques are good for removing large particles but involve losses of pyrolysis liquid. Cyclones are a traditional robust technique used regularly in pyrolysis. In the experimental part of this thesis, a 2 MWfuel pyrolysis setup with 2 cyclones in series was operated and monitored. Solid and liquid samples were collected from various parts of the process for further examination. Sampling and sample treatment techniques were developed. The chemical properties of the pyrolysis char were also analyzed and assessed as a function of reactor temperature and fluidizing velocity. By measuring the particle size distributions it was noticed that there were much smaller particles collected from the second cyclone than fed into pyrolysis. The solids in the pyrolysis oil were even smaller. This was most likely caused by attrition and shrinkage. Due to better separation efficiency of the cyclones in large particles, excess attrition should be avoided.

Mahdollisuudet sisäilmaoireiden poistamiseen huonekohtaisen ilmanpuhdistimen avulla eri aikakauden rakennuksissa

Tämän diplomityön tavoitteena on selvittää sisäilmaongelmien aiheuttajia ja keinoja niiden poistamiseen. Kokeellisessa osuudessa on tavoitteena selvittää kenttämittauksin sisäilman epäpuhtauspitoisuuksia ja ilmanpuhdistimien vaikutusta sisäilman laatuun. Työ toteutetaan kirjallisuustutkimuksen, sisäilmamittauksien sekä asiantuntija- ja sisäilmakyselyjen avulla. Tutkimukseen valittiin viisi erilaisia puhdistustekniikoita käyttäviä ilmanpuhdistimia. Valitut ilmanpuhdistimet olivat viikon ajan viidessä eri koululuokassa, joissa tehtiin sisäilmamittauksia ennen puhdistimen käynnistystä ja sen jälkeen. Luokista kerättiin ilmanäytteitä Andersen keräimellä ja VOC-pumpulla sekä pölynäytteitä pintasivelynä. Tutkimuskohteissa ei havaittu merkittäviä sisäilmaongelmia ja ilmanäytteiden pitoisuudet olivat pääasiassa alhaisia. Poikkeavia sienisukuja ei myöskään esiintynyt valtalajeina. Rakennetutkimukset ja materiaalinäytteet ovat ensisijaisia tutkimusmenetelmiä ongelmakohteissa. Syytä sisäilman aiheuttamaan oireiluun ei vielä tarkalleen tiedetä. Suunnitteluun, valvontaan ja kunnossapitoon tulee panostaa enemmän. Mittausmenetelmiä tulee myös kehittää lisää ja huolehtia ilmanvaihdon toimivuudesta. Tutkimuksia tuleekin tehdä aina kokonaisvaltaisesti.

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.

Ejektoripumpun suorituskyvyn kokeellinen testaus

Tässä kandidaatintyössä tutkitaan ejektoripumpun suorituskykyä teoreettisesti tutkimukseen pohjautuen ja kokeellisesti testilaitteistolla. Suorituskyvyn tarkastelun lähtökohtana on selvittää ejektorin käyttömahdollisuuksia mikro-ORC-energiamuuntimessa esi- ja pääsyöttöpumpun kavitoinnin estämiseksi ja koko prosessin huollettavuuden parantamiseksi. Ejektorin suorituskyvyn tarkastelu suoritetaan kuitenkin yleisesti sovelluskohteesta riippumatta tehdyt rajaukset huomioiden. Työn tarkoituksena on esitellä testaukseen rakennettava testilaitteisto ominaisuuksineen sekä määrittää kokeellisesti rakennettavalla laitteistolla ejektorin suorituskyvyn tunnuslukuja teoreettisen ja kokeellisen tarkastelun välisten korrelaatioiden selvittämiseksi sekä tilanteeseen sopivan ejektorin valitsemiseksi.

The supermassive binary black hole system OJ287

Abstract This doctoral thesis concerns the active galactic nucleus (AGN) most often referred to with the catalogue number OJ287. The publications in the thesis present new discoveries of the system in the context of a supermassive binary black hole model. In addition, the introduction discusses general characteristics of the OJ287 system and the physical fundamentals behind these characteristics. The place of OJ287 in the hierarchy of known types of AGN is also discussed. The introduction presents a large selection of fundamental physics required to have a basic understanding of active galactic nuclei, binary black holes, relativistic jets and accretion disks. Particularly the general relativistic nature of the orbits of close binaries of supermassive black holes is explored with some detail. Analytic estimates of some of the general relativistic effects in such a binary are presented, as well as numerical methods to calculate the effects more precisely. It is also shown how these results can be applied to the OJ287 system. The binary orbit model forms the basis for models of the recurring optical outbursts in the OJ287 system. In the introduction, two physical outburst models are presented in some detail and compared. The radiation hydrodynamics of the outbursts are discussed and optical light curve predictions are derived. The precursor outbursts studied in Paper III are also presented, and tied into the model of OJ287. To complete the discussion of the observable features of OJ287, the nature of the relativistic jets in the system, and in active galactic nuclei in general, is discussed. Basic physics of relativistic jets are presented, with additional detail added in the form of helical jet models. The results of Papers II, IV and V concerning the jet of OJ287 are presented, and their relation to other facets of the binary black hole model is discussed. As a whole, the introduction serves as a guide, though terse, for the physics and numerical methods required to successfully understand and simulate a close binary of supermassive black holes. For this purpose, the introduction necessarily combines a large number of both fundamental and specific results from broad disciplines like general relativity and radiation hydrodynamics. With the material included in the introduction, the publications of the thesis, which present new results with a much narrower focus, can be readily understood. Of the publications, Paper I presents newly discovered optical data points for OJ287, detected on archival astronomical plates from the Harvard College Observatory. These data points show the 1900 outburst of OJ287 for the first time. In addition, new data points covering the 1913 outburst allowed the determination of the start of the outburst with more precision than was possible before. These outbursts were then successfully numerically modelled with an N-body simulation of the OJ287 binary and accretion disc. In Paper II, mechanisms for the spin-up of the secondary black hole in OJ287 via interaction with the primary accretion disc and the magnetic fields in the system are discussed. Timescales for spin-up and alignment via both processes are estimated. It is found that the secondary black hole likely has a high spin. Paper III reports a new outburst of OJ287 in March 2013. The outburst was found to be rather similar to the ones reported in 1993 and 2004. All these outbursts happened just before the main outburst season, and are called precursor outbursts. In this paper, a mechanism was proposed for the precursor outbursts, where the secondary black hole collides with a gas cloud in the primary accretion disc corona. From this, estimates of brightness and timescales for the precursor were derived, as well as a prediction of the timing of the next precursor outburst. In Paper IV, observations from the 2004–2006 OJ287 observing program are used to investigate the existence of short periodicities in OJ287. The existence of a _50 day quasiperiodic component is confirmed. In addition, statistically significant 250 day and 3.5 day periods are found. Primary black hole accretion of a spiral density wave in the accretion disc is proposed as the source of the 50 day period, with numerical simulations supporting these results. Lorentz contracted jet re-emission is then proposed as the reason for the 3.5 day timescale. Paper V fits optical observations and mm and cm radio observations of OJ287 with a helical jet model. The jet is found to have a spine–sheath structure, with the sheath having a much lower Lorentz gamma factor than the spine. The sheath opening angle and Lorentz factor, as well as the helical wavelength of the jet are reported for the first time. Tiivistelmä Tässä väitöskirjatutkimuksessa on keskitytty tutkimaan aktiivista galaksiydintä OJ287. Väitöskirjan osana olevat tieteelliset julkaisut esittelevät OJ287-systeemistä saatuja uusia tuloksia kaksoismusta-aukkomallin kontekstissa. Väitöskirjan johdannossa käsitellään OJ287:n yleisiä ominaisuuksia ja niitä fysikaalisia perusilmiöitä, jotka näiden ominaisuuksien taustalla vaikuttavat. Johdanto selvittää myös OJ287-järjestelmän sijoittumisen aktiivisten galaksiytimien hierarkiassa. Johdannossa käydään läpi joitakin perusfysiikan tuloksia, jotka ovat tarpeen aktiivisten galaksiydinten, mustien aukkojen binäärien, relativististen suihkujen ja kertymäkiekkojen ymmärtämiseksi. Kahden toisiaan kiertävän mustan aukon keskinäisen radan suhteellisuusteoreettiset perusteet käydään läpi yksityiskohtaisemmin. Johdannossa esitetään joitakin analyyttisiä tuloksia tällaisessa binäärissä havaittavista suhteellisuusteoreettisista ilmiöistä. Myös numeerisia menetelmiä näiden ilmiöiden tarkempaan laskemiseen esitellään. Tuloksia sovelletaan OJ287-systeemiin, ja verrataan havaintoihin. OJ287:n mustien aukkojen ratamalli muodostaa pohjan systeemin toistuvien optisten purkausten malleille. Johdannossa esitellään yksityiskohtaisemmin kaksi fysikaalista purkausmallia, ja vertaillaan niitä. Purkausten säteilyhydrodynamiikka käydään läpi, ja myös ennusteet purkausten valokäyrille johdetaan. Johdannossa esitellään myös Julkaisussa III johdettu prekursoripurkausten malli, ja osoitetaan sen sopivan yhteen OJ287:n binäärimallin kanssa. Johdanto esittelee myös relativististen suihkujen fysiikkaa sekä OJ287- systeemiin liittyen että aktiivisten galaksiydinten kontekstissa yleisesti. Relativististen suihkujen perusfysiikka esitellään, kuten myös malleja kierteisistä suihkuista. Julkaisujen II, IV ja V OJ287-systeemin suihkuja koskevat tulokset esitellään binäärimallin kontekstissa. Kokonaisuutena johdanto palvelee suppeana oppaana, joka esittelee tarvittavan fysiikan ja tarpeelliset numeeriset menetelmät mustien aukkojen binäärijärjestelmän ymmärtämiseen ja simulointiin. Tätä tarkoitusta varten johdanto yhdistää sekä perustuloksia että joitakin syvällisempiä tuloksia laajoilta fysiikan osa-alueilta kuten suhteellisuusteoriasta ja säteilyhydrodynamiikasta. Johdannon sisältämän materiaalin avulla väitöskirjan julkaisut, ja niiden esittämät tulokset, ovat hyvin ymmärrettävissä. Väitöskirjan julkaisuista ensimmäinen esittelee uusia OJ287-systeemistä saatuja havaintopisteitä, jotka on paikallistettu Harvardin yliopiston observatorion arkiston valokuvauslevyiltä. OJ287:n vuonna 1900 tapahtunut purkaus nähdään ensimmäistä kertaa näissä havaintopisteissä. Uudet havaintopisteet mahdollistivat myös vuoden 1913 purkauksen alun ajoittamisen tarkemmin kuin aiemmin oli mahdollista. Havaitut purkaukset mallinnettiin onnistuneesti simuloimalla OJ287-järjestelmän mustien aukkojen paria ja kertymäkiekkoa. Julkaisussa II käsitellään mekanismeja OJ287:n sekundäärisen mustan aukon spinin kasvamiseen vuorovaikutuksessa primäärin kertymäkiekon ja systeemin magneettikenttien kanssa. Julkaisussa arvioidaan maksimispinin saavuttamisen ja spinin suunnan vakiintumisen aikaskaalat kummallakin mekanismilla. Tutkimuksessa havaitaan sekundäärin spinin olevan todennäköisesti suuri. Julkaisu III esittelee OJ287-systeemissä maaliskuussa 2013 tapahtuneen purkauksen. Purkauksen havaittiin muistuttavan vuosina 1993 ja 2004 tapahtuneita purkauksia, joita kutsutaan yhteisnimityksellä prekursoripurkaus (precursor outburst). Julkaisussa esitellään purkauksen synnylle mekanismi, jossa OJ287-systeemin sekundäärinen musta aukko osuu primäärisen mustan aukon kertymäkiekon koronassa olevaan kaasupilveen. Mekanismin avulla johdetaan arviot prekursoripurkausten kirkkaudelle ja aikaskaalalle. Julkaisussa johdetaan myös ennuste seuraavan prekursoripurkauksen ajankohdalle. Julkaisussa IV käytetään vuosina 2004–2006 kerättyjä havaintoja OJ287- systeemistä lyhyiden jaksollisuuksien etsintään. Julkaisussa varmennetaan systeemissä esiintyvä n. 50 päivän kvasiperiodisuus. Lisäksi tilastollisesti merkittävät 250 päivän ja 3,5 päivän jaksollisuudet havaitaan. Julkaisussa esitetään malli, jossa primäärisen mustan aukon kertymäkiekossa oleva spiraalitiheysaalto aiheuttaa 50 päivän jaksollisuuden. Mallista tehty numeerinen simulaatio tukee tulosta. Systeemin relativistisen suihkun emittoima aikadilatoitunut säteily esitetään aiheuttajaksi 3,5 päivän jaksollisuusaikaskaalalle. Julkaisussa V sovitetaan kierresuihkumalli OJ287-systeemistä tehtyihin optisiin havaintoihin ja millimetri- sekä senttimetriaallonpituuden radiohavaintoihin. Suihkun rakenteen havaitaan olevan kaksijakoinen ja koostuvan ytimestä ja kuoresta. Suihkun kuorella on merkittävästi pienempi Lorentzin gamma-tekijä kuin suihkun ytimellä. Kuoren avautumiskulma ja Lorentztekijä sekä suihkun kierteen aallonpituus raportoidaan julkaisussa ensimmäistä kertaa.

Molecular Mechanisms and Interactions in Regulation of Photosynthetic Reactions

In photosynthesis, light energy is converted to chemical energy, which is consumed for carbon assimilation in the Calvin-Benson-Bassham (CBB) cycle. Intensive research has significantly advanced the understanding of how photosynthesis can survive in the ever-changing light conditions. However, precise details concerning the dynamic regulation of photosynthetic processes have remained elusive. The aim of my thesis was to specify some molecular mechanisms and interactions behind the regulation of photosynthetic reactions under environmental fluctuations. A genetic approach was employed, whereby Arabidopsis thaliana mutants deficient in specific photosynthetic protein components were subjected to adverse light conditions and assessed for functional deficiencies in the photosynthetic machinery. I examined three interconnected mechanisms: (i) auxiliary functions of PsbO1 and PsbO2 isoforms in the oxygen evolving complex of photosystem II (PSII), (ii) the regulatory function of PGR5 in photosynthetic electron transfer and (iii) the involvement of the Calcium Sensing Receptor CaS in photosynthetic performance. Analysis of photosynthetic properties in psbo1 and psbo2 mutants demonstrated that PSII is sensitive to light induced damage when PsbO2, rather than PsbO1, is present in the oxygen evolving complex. PsbO1 stabilizes PSII more efficiently compared to PsbO2 under light stress. However, PsbO2 shows a higher GTPase activity compared to PsbO1, and plants may partially compensate the lack of PsbO1 by increasing the rate of the PSII repair cycle. PGR5 proved vital in the protection of photosystem I (PSI) under fluctuating light conditions. Biophysical characterization of photosynthetic electron transfer reactions revealed that PGR5 regulates linear electron transfer by controlling proton motive force, which is crucial for the induction of the photoprotective non-photochemical quenching and the control of electron flow from PSII to PSI. I conclude that PGR5 controls linear electron transfer to protect PSI against light induced oxidative damage. I also found that PGR5 physically interacts with CaS, which is not needed for photoprotection of PSII or PSI in higher plants. Rather, transcript profiling and quantitative proteomic analysis suggested that CaS is functionally connected with the CBB cycle. This conclusion was supported by lowered amounts of specific calciumregulated CBB enzymes in cas mutant chloroplasts and by slow electron flow to PSI electron acceptors when leaves were reilluminated after an extended dark period. I propose that CaS is required for calcium regulation of the CBB cycle during periods of darkness. Moreover, CaS may also have a regulatory role in the activation of chloroplast ATPase. Through their diverse interactions, components of the photosynthetic machinery ensure optimization of light-driven electron transport and efficient basic production, while minimizing the harm caused by light induced photodamage.