Stop bugging me! : diversity in appearance of warning coloration

In nature, many animals use body coloration to communicate with each other. For example, colorations can be used as signals between individuals of the same species, but also to recognise individuals of other species, and if they may comprise a threat or not. Many animals use protective coloration to avoid predation. The two most common strategies of protective coloration are camouflage and aposematism. Camouflaged animals have coloration that minimises detection, usually by matching colours or structures in the background. Aposematic animals, on the other hand, signal to predators that they are defended. The defence can be physical structures, such as spikes and hairs, or chemical compounds that make the animal distasteful or even deadly toxic. In order for the warning signal to be effective, the predator has to recognise it as such. Studies have shown that birds for example, that are important visual predators on insects, learn to recognise and avoid unpalatable prey faster if they contrast the background or have large internal contrasts. Typical examples of aposematic species have conspicuous colours like yellow, orange or red, often in combination with black. My thesis focuses on the appearance and function of aposematic colour patterns. Even though researchers have studied aposematism for over a century, there is still a lot we do not know about the phenomenon. For example, as it is crucial that the predators recognise a warning signal, aposematic colorations should assumingly evolve homogeneously and be selected for maximal conspicuousness. Instead, there is an extensive variation of colours and patterns among warning colorations, and it is not uncommon to find typical cryptic colours, such as green and brown in aposematic colour patterns. One hypothesis to this variation is that an aposematic coloration does not have to be maximally signalling in order to be effective, instead it is sufficient to have distinct features that can be easily distinguished from edible prey. To be maximally conspicuous is one way to achieve this, but not the only way. Another hypothesis is that aposematic prey that do not exhibit maximal conspicuousness can exploit both camouflage and aposematism in a distance-dependent fashion, by being signalling when seen close up but camouflaged at a distance. Many prey animals also make use of both strategies by shifting colour at different ecological conditions such as seasonal variations, fluctuations in food resources or between life stages. Yet another explanation for the variation may be that prey animals are usually exposed to several predator species that vary in visual perception and tolerance towards various toxins. The aim with this thesis is, by studying their functions, to understand why aposematic warning signals vary in appearance, specifically in the level of conspicuousness, and if warning coloration can be combined with camouflage. In paper I, I investigated if the colour pattern of the aposematic larva of the Apollo butterfly (Parnassius apollo) can switch function with viewing distance, and be signalling at close range but camouflaged at a distance, by comparing detection time between different colour variants and distances. The results show that the natural coloration has a dual distance-dependent function. Moreover, the study shows that an aposematic coloration does not have to be selected for maximal conspicuousness. A prey animal can optimise its coloration primarily by avoiding detection, but also by investing in a secondary defence, which presence can be signalled if detected. In paper II, I studied how easily detected the coloration of the firebug (Pyrrhocoris apterus), a typical aposematic species, is at different distances against different natural backgrounds, by comparing detection time between different colour variants. Here, I found no distance-dependent switch in function. Instead, the results show that the coloration of the firebug is selected for maximal conspicuousness. One explanation for this is that the firebug is more mobile than the butterfly larva in study I, and movement is often incompatible with efficient camouflage. In paper III, I investigated if a seasonal related colour change in the chemically defended striated shieldbug (Graphosoma lineatum) is an adaptation to optimise a protective coloration by shifting from camouflage to aposematism between two seasons. The results confirm the hypothesis that the coloration expressed in the late summer has a camouflage function, blending in with the background. Further, I investigated if the internal pattern as such increased the effectiveness of the camouflage. Again, the results are in accordance with the hypothesis, as the patterned coloration was more difficult to detect than colorations lacking an internal pattern. This study shows how an aposematic species can optimise its defence by shifting from camouflage to aposematism, but in a different fashion than studied in paper I. The aim with study IV was to study the selection on aposematic signals by identifying characteristics that are common for colorations of aposematic species, and that distinguish them from colorations of other species. I compared contrast, pattern element size and colour proportion between a group of defended species and a group of undefended species. In contrast to my prediction, the results show no significant differences between the two groups in any of the analyses. One explanation for the non-significant results could be that there are no universal characteristics common for aposematic species. Instead, the selection pressures acting on defended species vary, and therefore affect their appearance differently. Another explanation is that all defended species may not have been selected for a conspicuous aposematic warning coloration. Taken together, my thesis shows that having a conspicuous warning coloration is not the only way to be aposematic. Also, aposematism and camouflage is not two mutually exclusive opposites, as there are prey species that exploit both strategies. It is also important to understand that prey animals are exposed to various selection pressures and trade-offs that affect their appearance, and determines what an optimal coloration is for each species or environment. In conclusion, I hold that the variation among warning colorations is larger and coloration properties that have been considered as archetypically aposematic may not be as widespread and representative as previously assumed.

Methods of genetic diversity creation and functional display for directed evolution experiments

Protein engineering aims to improve the properties of enzymes and affinity reagents by genetic changes. Typical engineered properties are affinity, specificity, stability, expression, and solubility. Because proteins are complex biomolecules, the effects of specific genetic changes are seldom predictable. Consequently, a popular strategy in protein engineering is to create a library of genetic variants of the target molecule, and render the population in a selection process to sort the variants by the desired property. This technique, called directed evolution, is a central tool for trimming protein-based products used in a wide range of applications from laundry detergents to anti-cancer drugs. New methods are continuously needed to generate larger gene repertoires and compatible selection platforms to shorten the development timeline for new biochemicals. In the first study of this thesis, primer extension mutagenesis was revisited to establish higher quality gene variant libraries in Escherichia coli cells. In the second study, recombination was explored as a method to expand the number of screenable enzyme variants. A selection platform was developed to improve antigen binding fragment (Fab) display on filamentous phages in the third article and, in the fourth study, novel design concepts were tested by two differentially randomized recombinant antibody libraries. Finally, in the last study, the performance of the same antibody repertoire was compared in phage display selections as a genetic fusion to different phage capsid proteins and in different antibody formats, Fab vs. single chain variable fragment (ScFv), in order to find out the most suitable display platform for the library at hand. As a result of the studies, a novel gene library construction method, termed selective rolling circle amplification (sRCA), was developed. The method increases mutagenesis frequency close to 100% in the final library and the number of transformants over 100-fold compared to traditional primer extension mutagenesis. In the second study, Cre/loxP recombination was found to be an appropriate tool to resolve the DNA concatemer resulting from error-prone RCA (epRCA) mutagenesis into monomeric circular DNA units for higher efficiency transformation into E. coli. Library selections against antigens of various size in the fourth study demonstrated that diversity placed closer to the antigen binding site of antibodies supports generation of antibodies against haptens and peptides, whereas diversity at more peripheral locations is better suited for targeting proteins. The conclusion from a comparison of the display formats was that truncated capsid protein three (p3Δ) of filamentous phage was superior to the full-length p3 and protein nine (p9) in obtaining a high number of uniquely specific clones. Especially for digoxigenin, a difficult hapten target, the antibody repertoire as ScFv-p3Δ provided the clones with the highest affinity for binding. This thesis on the construction, design, and selection of gene variant libraries contributes to the practical know-how in directed evolution and contains useful information for scientists in the field to support their undertakings.

Scientific creativity and diversity of physical work environment: framework development

The aim of this research was to develop a framework to analyze how physical environment influences scientific creativity. Due to the relative novelty of this topic, there is still a gap in the unified method to study connection between physical environment and creativity. Therefore, in order to study this issue deeply, the qualitative method was used (interviews and qualitative questionnaire). Scientists (PhD students and senior researchers) of Graduate School of Management were interviewed to build the model and one expert interview was conducted to assess its validity. The model highlights several dimensions via which physical environment can influence scientific creativity: Comfort, Instruments and Diversity. Comfort and Instruments are considered to be related mostly to productivity, an initial requirement for creativity, while Diversity is the factor responsible for supporting all the stages of scientific creative process. Thus, creative physical environment is not one place by its nature, but an aggregative phenomenon. Due to two levels of analysis, the model is named the two-level model of creative physical environment.

Prediction of market switching and delisting events from OMX First North Nordic multilateral stock exchange

This thesis studies the predictability of market switching and delisting events from OMX First North Nordic multilateral stock exchange by using financial statement information and market information from 2007 to 2012. This study was conducted by using a three stage process. In first stage relevant theoretical framework and initial variable pool were constructed. Then, explanatory analysis of the initial variable pool was done in order to further limit and identify relevant variables. The explanatory analysis was conducted by using self-organizing map methodology. In the third stage, the predictive modeling was carried out with random forests and support vector machine methodologies. It was found that the explanatory analysis was able to identify relevant variables. The results indicate that the market switching and delisting events can be predicted in some extent. The empirical results also support the usability of financial statement and market information in the prediction of market switching and delisting events.

Do retail investors use technical analysis? Evidence from Stuttgart stock exchange.

Tutkielma käyttää automaattista kuviontunnistusalgoritmia ja yleisiä kahden liukuvan keskiarvon leikkauspiste –sääntöjä selittääkseen Stuttgartin pörssissä toimivien yksityissijoittajien myynti-osto –epätasapainoa ja siten vastatakseen kysymykseen ”käyttävätkö yksityissijoittajat teknisen analyysin menetelmiä kaupankäyntipäätöstensä perustana?” Perusolettama sijoittajien käyttäytymisestä ja teknisen analyysin tuottavuudesta tehtyjen tutkimusten perusteella oli, että yksityissijoittajat käyttäisivät teknisen analyysin metodeja. Empiirinen tutkimus, jonka aineistona on DAX30 yhtiöiden data vuosilta 2009 – 2013, ei tuottanut riittävän selkeää vastausta tutkimuskysymykseen. Heikko todistusaineisto näyttää kuitenkin osoittavan, että yksityissijoittajat muuttavat kaupankäyntikäyttäytymistänsä eräiden kuvioiden ja leikkauspistesääntöjen ohjastamaan suuntaan.

Oxygen Photoreduction in Cyanobacteria

Cyanobacteria are well-known for their role in the global production of O2 via photosynthetic water oxidation. However, with the use of light energy, cyanobacteria can also reduce O2. In my thesis work, I have investigated the impact of O2 photoreduction on protection of the photosynthetic apparatus as well as the N2-fixing machinery. Photosynthetic light reactions produce intermediate radicals and reduced electron carriers, which can easily react with O2 to generate various reactive oxygen species. To avoid prolonged reduction of photosynthetic components, cyanobacteria use “electron valves” that dissipate excess electrons from the photosynthetic electron transfer chain in a harmless way. In Synechocystis sp. PCC 6803, flavodiiron proteins Flv1 and Flv3 comprise a powerful electron sink redirecting electrons from the acceptor side of Photosystem I to O2 and reducing it directly to water. In this work, I demonstrate that upon Ci-depletion Flv1/3 can dissipate up to 60% of the electrons delivered from Photosystem II. O2 photoreduction by Flv1/3 was shown to be vital for cyanobacteria in natural aquatic environments and deletion of Flv1/3 was lethal for both Synechocystis sp. PCC 6803 and Anabaena sp. PCC 7120 under fluctuating light conditions. The lethal phenotype observed in the absence of Flv1/3 results from oxidative damage to Photosystem I, which appeared to be a primary target of reactive oxygen species produced upon sudden increases in light intensity. Importantly, cyanobacteria also possess other O2 photoreduction pathways which can protect the photosynthetic apparatus. This study demonstrates that respiratory terminal oxidases are also capable of initiating O2 photoreduction in mutant cells lacking the Flv1/3 proteins and grown under fluctuating light. Photoreduction of O2 by Rubisco was also shown in Ci-depleted cells of the mutants lacking Flv1/3, and thus provided the first evidence for active photorespiratory gas-exchange in cyanobacteria. Nevertheless, and despite the existence of other O2 photoreduction pathways, the Flv1/3 route appears to be the most robust and rapid system of photoprotection. Several groups of cyanobacteria are capable of N2 fixation. Filamentous heterocystous N2- fixing species, such as Anabaena sp. PCC 7120, are able to differentiate specialised cells called heterocysts for this purpose. In contrast to vegetative cells which perform oxygenic photosynthesis, heterocysts maintain a microoxic environment for the proper function of the nitrogenase enzyme, which is extremely sensitive to O2. The genome of Anabaena sp. PCC 7120 harbors two copies of genes encoding Flv1 and Flv3 proteins, designated as “A” and “B” forms. In this thesis work, I demonstrate that Flv1A and Flv3A are expressed only in the vegetative cells of filaments, whilst Flv1B and Flv3B are localized exclusively in heterocysts. I further revealed that the Flv3B protein is most responsible for the photoreduction of O2 in heterocysts, and that this reaction plays an important role in protection of the N2-fixing machinery and thus, the provision of filaments with fixed nitrogen. The function of the Flv1B protein remains to be elucidated; however the involvement of this protein in electron transfer reactions is feasible. Evidence provided in this thesis indicates the presence of a great diversity of O2 photoreduction reactions in cyanobacterial cells. These reactions appear to be crucial for the photoprotection of both photosynthesis and N2 fixation processes in an oxygenic environment.

Value at Risk in Foreign Exchange Risk Management

This thesis examines the suitability of VaR in foreign exchange rate risk management from the perspective of a European investor. The suitability of four different VaR models is evaluated in respect to have insight if VaR is a valuable tool in managing foreign exchange rate risk. The models evaluated are historical method, historical bootstrap method, variance-covariance method and Monte Carlo simulation. The data evaluated are divided into emerging and developed market currencies to have more intriguing analysis. The foreign exchange rate data in this thesis is from 31st January 2000 to 30th April 2014. The results show that the previously mentioned VaR models performance in foreign exchange risk management is not to be considered as a single tool in foreign exchange rate risk management. The variance-covariance method and Monte Carlo simulation performs poorest in both currency portfolios. Both historical methods performed better but should also be considered as an additional tool along with other more sophisticated analysis tools. A comparative study of VaR estimates and forward prices is also included in the thesis. The study reveals that regardless of the expensive hedging cost of emerging market currencies the risk captured by VaR is more expensive and thus FX forward hedging is recommended

From genotype to phenotype : diversity and population structure of Flavobacterium psychrophilum

Flavobacterium psychrophilum is the etiological agent of bacterial cold-water disease (BCWD) causing high fish mortalities and significant economic losses to the freshwater salmonid aquaculture industry around the world. Today BCWD outbreaks are mainly treated with environmentally hazardous antimicrobial agents and alternative preventative measures are urgently needed in order to ensure the well-being of animals and the sustainability of aquaculture. The diversity of pathogenic bacteria challenges the development of universal control strategies and in many cases the pathogen population structure, i.e. the total genetic diversity of the species must be taken into account. This work integrates the tools of modern molecular biology and conventional phenotypic microbiology to gain knowledge about the diversity and population structure of F. psychrophilum. The present work includes genetic characterization of a large collection of isolates collected from diverse origins and years, from aquaculture in a whole region including different countries, and provides the first international validation of a universal multilocus sequence typing (MLST) approach for unambiguous genetic typing of F. psychrophilum. Population structure analyses showed that the global F. psychrophilum population is subdivided into pathogenic species-specific clones, of which one particular genetic lineage, clonal complex CC-ST2, has been responsible for the majority of BCWD outbreaks in rainbow trout (Oncorhynchus mykiss) in European aquaculture facilities over several decades. Genotypic and phenotypic population heterogeneity affecting antimicrobial resistance in F. psychrophilum within BCWD outbreaks was discovered. Specific genotypes were associated with severe infections in farmed rainbow trout and Atlantic salmon (Salmo salar), and in addition to high adherence, antimicrobial resistance was strongly associated with outbreak strains. The study brought additional support for the hypothesis of an epidemic F. psychrophilum population structure, where recombination is an important force for the generation and maintenance of genetic diversity, and a significant contribution towards mapping the genetic diversity of this important fish pathogen. Evidence indicating dissemination of virulent strains with commercial movement of fish and fish products was strengthened.

Genetic diversity and phenotypic variability of phytoplankton populations in the Baltic Sea

En av naturens mest grundläggande aspekter är den enorma mängd av variation som existerar mellan arter. Denna variation har lett oss till att klassificera olika organismer på basis av morfologiska skillnader och på senare tid till att jämföra genetiska skillnader på individens nivå. Den marina kiselalgen Skeletonema marinoi är en av de vanligaste växtplanktonarter i Östersjön under vårblomningen och anses viktig för den årliga produktionen. En av mina främsta målsättningar var att beskriva den intra-specifika diversiteten hos denna art längs med miljögradienter i Östersjön. Ett annat mål var att klargöra de faktorer som eventuellt är involverade i konfigurationen av genetisk diversitet och differentiering. Med hjälp av genetiska markörer visade jag att den genetiska diversiteten hos S. marinoi populationer i Östersjön är lägre jämfört med populationer i östra delen av Nordsjön. Arten är genetiskt uppdelad så att en utpräglad population förekommer i Östersjön och en annan, genetiskt åtskild population förekommer norr om de Danska sunden. Resultaten visar att de genetiskt åtskilda populationerna är anpassade till lokala salinitetsförhållanden. Genflödet mellan populationerna korrelerade kraftigt med havströmmar i området. Mina studier avslöjade även omfattande variation av fenotypiska, ekologiskt vikitga särdrag hos olika kloner. Djurplankton som äter kiselalger kunde modifiera den klonala mångfalden av fenotypiskt variabla S. marinoi populationer. En ökad klonal mångfald ledde till högre prestationsförmåga i fråga om primär produktion och stabiliserade ekofysiologiska funktioner. Som visas i denna avhandling består en art allt som oftast av åtskilliga genetiska varianter med fenotypiska skillnader. Kunskap om sådana intra-specifika skillnader är en förutsättning för att vi skall kunna förstå var och varför arter förekommer. Denna kunskap utgör även en grund för prognoser som siktar på att förutspå huruvida arter kan anpassa sig till framtida miljöförhållanden. ------------------------------------------------------ Suunnaton määrä variaatioita eliölajien välillä on perustavanlaatuinen ominaisuus luonnossa. Perinteisesti tätä monimuotoisuutta on käytetty organismien luokittelemiseen eri lajeihin niiden morfologisten eroavaisuuksien perusteella. Hiljattain myös geneettisten erojen huomioimista yksilötasolla on hyödynnetty lajien luokittelemisessa. Merialueilla esiintyvä piilevä, Skeletonema marinoi on yksi Itämeren tavallisimmista kasviplanktonlajeista kevätkukinnan aikana. Tavoitteenani oli selventää geneettistä ja fenotyyppistä monimuotoisuutta pitkin Itämeren ympäristögradienttejä. Geneettisen monimuotoisuuteen ja erkaantumiseen vaikuttavien tekijöiden selvittäminen oli tärkeä aspekti väitöstutkimuksessani. Geneettisiä markkereita käyttämällä pystyin toteamaan, että S. marinoi levän geneettinen monimuotoisuus on Itämeressä merkittävästi alhaisempi kuin läheisessä Pohjanmeren itäosassa. Tutkittu laji jakautuu geneettisesti yhteen erilliseen populaatioon Itämeressä ja toiseen selvästi erottuvaan populaatioon Tanskan salmien pohjoispuolella. Kokeellisten tulosten perusteella nämä geneettisesti erilaistuneet populaatiot ovat kumpikin sopeutuneet paikalliseen veden suolapitoisuuteen. Populaatioiden välisen geenivirran ja merivirtojen luoman yhteyden välillä havaittiin vahva korrelaatio. Tutkimukseni paljastivat myös laajaa vaihtelua Skeletonema-kloonien ekologisesti tärkeissä ominaisuuksissa. Kokeellisten tutkimusteni perusteella laiduntajat pystyivät muuttamaan geneettisten kloonien lukumäärää monimuotoisissa S. marinoi populaatioissa. Lisääntynyt kloonien lukumäärä paransi perustuotantokykyä ja vakautti ekofysiologisia toimintoja. Kuten tässä väitöstutkimuksessa osoitetaan, lajit koostuvat useimmiten lukuisista geneettisistä muunnelmista, jotka eroavat usein fenotyypeiltään. Ymmärtääksemme missä tietyt lajit esiintyvät ja miksi, tarvitsemme tietoa lajien sisäisistä vaihteluista. Tämä tieto on tarpeellista, jotta voimme ennustaa lajien sopeutumista tuleviin ympäristönmuutoksiin.

Making home rules for mother tongues : the legal implications of linguistic diversity in the design of autonomy regimes

Principen om nationalismen där det politiska och det nationella är samspelt kan vara av markant betydelse för uppbyggande av autonomiska regimer. Likaså tillåter decentralicering och delegering av befogenheter för språk och utbildning (officiellt erkännande av språk, standardisering av språk, undervisningsspråk och relaterade läroplaner) formning av identiteter inom dessa autonomiska regimer. Resultatet är en ofullkomlig cirkulär relation där språk, samfund och politiska institutioner ömsesidigt och kontinuerligt formar varandra: lingvistiskt mångfald prägar och formger autonomiska ordningar och vice-versa. De juridiska implikationerna av territoriella och icke-territoriella former av autonomi är dock av en annan art. Emedan territoriell autonomi bygger på idéen om ett eventuellt inkluderande hemland för lingvistiska grupper, vars vistelseort är avgörande, förstärker den icke-territoriella autonomin idéen om ett exclusivt samfund bestående av själv-identifierade medlemmar som är kapabla till självstyre oavsett territoriella gränser. Denna avhandling utgör an analys av sådana juridiska implikationer genom komparativa och institutionella analyser. Avhandlingen föreslår som resultat en serie av normativa och pragmatiska rekommendationer inriktade på att främja demokratiseringsprocesser i linje med principer om multikulturalism.