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Antimicrobial resistance is an emerging concern at the global scale, threatening the effectiveness of antibiotics in treating bacterial infections. Among anthropogenically impacted environments, wastewater treatment plants have been indicated as possible reservoirs of antibiotic resistance genes, putative hotspots for their horizontal gene transfer, and a source of their dissemination to the environment. Generally, the abundance of antibiotic resistance genes is reduced during the wastewater treatment process. However, some genes were shown to be enriched in purified effluent water and dried sludge, which are then released to the environment, compared to influent water. Also, the taxonomy of the hosts carrying antibiotic resistance genes could change as a result of horizontal gene transfer events. The aim of this study was to analyse and compare the host range of a series of antibiotic resistance genes in influent water, effluent water and dried sludge collected from the Viikinmäki wastewater treatment plant in Helsinki, Finland, by applying Emulsion, Paired Isolation and Concatenation PCR (epicPCR). EpicPCR is a method that can link a gene of interest to the 16S rRNA gene from the genome of the host bacterium, without any cultivation step. The abundance of the hosts was also evaluated by sequencing the 16S rRNA gene from the whole bacterial community. In several cases, the target antibiotic resistance genes (blaIMP, blaNDM, ermB, ermF, sul1 and strB) were carried in effluent water and dried sludge by taxa that were not hosting them in influent water, suggesting that horizontal gene transfer events might have occurred during the treatment. All the examined genes were detected both in abundant and in rare taxa, including genera that also comprise pathogenic species, such as Arcobacter and Acinetobacter. Some of the detected hosts were not previously known to show resistant phenotypes, namely members of the family Methylophilaceae. These results corroborate the idea that wastewater treatment plants might be hotspots for the horizontal gene transfer of resistance determinants, and potentially disseminate antibiotic resistant pathogens to the environment. However, in order to ensure the accuracy of the results, the limits of epicPCR as a method need to be identified and addressed.

Ants are among the most successful organisms in the world. They can be found almost anywhere on the planet and due to their high degree of sociality and complex societies they have become some of the most abundant creatures in most terrestrial ecosystems. Although sociality has benefits in the form of more efficient foraging, brood care, reproduction and protection from predators, it has costs too. Ants live in high densities in their nests and have frequent contact between them which can facilitate an efficient transmission of pathogens within the nest. Ants have become highly successful in spite of their potentially high susceptibility to pathogens. They share the same innate immune responses of other arthropods and have unique adaptations for coping with pathogens. In extension to physiological strategies for coping with pathogens, ants engage in behavioural strategies as well. Ants and other eusocial insects can also harness the structure and behaviour of the colony to prevent and cope with pathogen infections through social immunity. Ants can also engage in self-medication behaviour to combat disease. Self-medication is a behavioural strategy where individuals respond to pathogen infections by seeking out and using biologically active compounds to alleviate the effects of pathogens in a way that would be detrimental for uninfected individuals. The behaviour can be either therapeutic of prophylactic depending on when the compounds are used in relation to encountering the pathogen, and it can be extended beyond the self to other kin. While ants have been proven to medicate themselves with reactive oxygen species (ROS) in laboratory conditions, it remains unknown how they do it in the wild. In my thesis, I studied how the ant Lasius platythorax self-medicate in a natural setting by developing a multi-trophic system of ant – pathogen – aphid – plant interactions. In this system, the ants infected with a fungal pathogen (Beauveria bassiana) had the opportunity to forage on the nectar produced by the extrafloral nectaries (EFNs) of a broad bean plant (Vicia faba) infested by vetch aphids (Megoura viciae). Plants that are stressed by aphids react with a systemic production of ROS, which ants are known to use for self-medication, and ROS could therefore be present in the EFN nectar as well, along with other potentially medicinal compounds. The aphids themselves could present the ants with both ROS, if it accumulates in the aphids due to the immune responses of the plant, and protein if eaten. In my thesis I found out that infected ants increase their foraging on EFN nectar during the first three days after infection compared to healthy ants. This immediate response to a pathogen infection shown by the infected ants fits in a self-medication context as well as the infection cycle of the pathogen, making this a strong case for self-medication. The change in foraging by the infected ants did not reflect on the changes in ROS content in the ants, possibly due to a lack of ROS in the nectar, but instead were likely to be caused by self-generation of ROS in the infected ants. The aphids feeding on the plant contained a higher ROS content compared to the ants, but I found no evidence of ants preying on the aphids, possibly due to the M. viciae being unpalatable for the ants or the ants finding medicinal compounds in the EFN nectar. The result of my thesis is a first step in to identifying natural ways for ants to obtain and use medicinal compounds from their environments and opens up new avenues of research in the topic of self-medication. The result also highlights the importance of biodiversity for the conservation efforts for ants and other insects. Insects are facing a drastic decline in both abundance and diversity due to human impact on their environments, including the prevalence in pathogens. By understanding the full extent of the immune strategies that insects use, including self-medication, we can develop more efficient methods of conservation to help them.

The demand for CO2-free electricity will grow in the future, which is expected to lead to an increase in onshore wind power capacity. Like all energy forms, wind power can have negative impacts on the environment, which can pose different risks for wind power companies. Several companies that have wind power in their portfolio have recently committed to biodiversity targets, typically aiming for no net loss or net positive impact on biodiversity. These targets are expected to be a possible avenue for competitive advantage, however, there is a research gap on whether biodiversity considerations can bring competitive advantage to companies. In my research, executed as a paid assignment for Fortum Renewables Oy, I examine wind power investors’ perceptions of biodiversity targets and whether they perceive that the targets can bring competitive advantage to wind power companies. In this thesis, I aim to provide answers to three primary research questions: 1) How do investors manage their investments’ biodiversity impacts? 2) Can setting biodiversity targets bring competitive advantage to wind power companies? and 3) What kind of biodiversity targets do investors value? For this research I interviewed investors known to invest in onshore wind power in Finland. I conducted seven interviews with eight informants, including both finance and sustainability professionals, representing seven different investors. I analysed the interview data using inductive thematic analysis provided by Braun and Clarke (2006). My research found that investors identify several possibilities for both risk mitigation and competitive advantage, which can be achieved through good biodiversity management. Thus, investors see value in biodiversity considerations, such as biodiversity targets. Simultaneously, investors recognised several issues regarding these targets, which hinder achieving competitive advantage through them. The findings of my research may prove useful both for companies that have or aim to set biodiversity targets and investors wanting to manage their investments’ biodiversity impacts.

Social media influencers are called the opinion leaders of our time, as they have a lot of power over what peo- ple think and consume. Various sustainability actors expect that influencers take a strong role in the transfor- mation towards sustainability. In my thesis I examine, how do the reactions of the influencers’ followers and the perceived sense of responsibility and power of the influencer affect the way influencers share content on sustainability. In my research I also consider how different influencers receive differing responses to content related to sustainability. My analysis consists of interviews with six Finnish social media influencers. I conducted 5 semi-structured in- terviews, and received one written answer to the interview questions. I used thematic analysis as the analysis method in my research. The themes that arose from the interview data and from previous research are relation- ality between the influencer and their followers, connection between followers’ reactions and influencers’ sus- tainability content and the perceived sense of power and responsibility of the influencer. Three influencer profiles were created based on the interview data. The profiles describe different types of in- fluencers with different, specific audiences. The profiles demonstrate how different audiences respond to con- tent related to sustainability in different ways, how different influencers share sustainability content in different ways, and how influencers perceive their responsibility in different ways. My research shows that both the reac- tions of the followers and the perceived sense of responsibility of the influencer affect how an influencer shares content on sustainability. According to my research, the influencer's perceived power does not affect content shared on sustainability. This study contributes to the understanding of the issues that affect influencers’ behavior in sharing sustainabil- ity-related content. With this knowledge, it is possible to move closer to understanding whether influencers can be part of the change towards a more sustainable future. shared on sustainability. This study contributes to the understanding of the issues that affect influencers’ behavior in sharing sustainabil- ity-related content. With this knowledge, it is possible to move closer to understanding whether influencers can be part of the change towards a more sustainable future.

Permafrost peatlands have the capacity to store significant amounts of carbon, and thus they act as important controllers of the climate. Approximately 14% of the world’s soil organic carbon pool is stored in permafrost peatlands, which are sensitive to climatic fluctuations due to their location in the high latitudes of the subarctic zone. Permafrost peatlands also act as a habitat for a large number of moisture-sensitive organisms, such as bryophytes and testate amoebae, which can be used to study how the hydrology of peatlands has changed and will continue to change throughout time, giving us an opportunity to predict the future of peatlands under a changing climate. In this Master’s Thesis I examined the testate amoebae composition and used these species as indicators to study hydrological fluctuations from three subarctic permafrost peatland cores extracted from Taavavuoma and Abisko in northern Sweden. The species compositions were combined with radiocarbon (14C) and lead (210Pb) dates to reconstruct the past water table levels for the late Holocene, spanning four climatic periods. The reconstructions were then compared to past studies on testate amoebae to understand how permafrost peatlands and their species assemblages respond to changes in the hydrology of the active layer of the peat. Out of the study sites only the Taavavuoma cores spanned the Dark Age Cold Period (DACP) and Medieval Climate Anomaly (MCA). Species compositions in both cores indicated fluctuating water tables during the DACP, but during the MCA the results began to contradict with one site showing a wetter, and the other a drier MCA. Two out of three study sites indicated a wetter Little Ice Age and a drier Post-Industrial Warming, supporting past studies indicating similar results, whereas one study site gave opposite results. The results indicated large variability in testate amoebae assemblages throughout time, indicating that the hydrology of peatlands can change very abruptly and vary considerably even on a local scale. Modelling is however complicated by the poorly known ecology of testate amoebae, which is why a multi-proxy approach is essential to reliably predict the future fate of permafrost peatlands.

The crops, pollinator species and the agricultural conditions in the Northern agricultural district differ from those in most other areas. For example, in Finland the importance of insect pollination hasn't been researched very broadly. The oil plant widely cultivated in Finland, turnip rape (Brassica rapa subsp. Oleifera) has been discovered to benefit from insect pollination. The aim of this study is to examine whether the yield is different with turnip rapes where insect pollination is free or prevented, and how pollinator species and the number of flower visits affect the turnip rape yield. The study was mainly conducted in July 2017 on field parcels in Uusimaa. In the turnip rape fields, different pollinators and their visits in turnip rape flowers were monitored. To prevent insect pollination, the turnip rapes were covered with gauze bags before flowering began so that several plants came into one bag. This allowed cross-pollination. The total number of flower visits by pollinators was linked to the number of racemes and seeds of the plant: insect pollination decreased the number of racemes and increased the number of seeds per plant. The honeybees and hoverflies were a major group of pollinators explaining the turnip rape yield. Openly pollinated turnip rapes contained fewer racemes and more siliques and seeds per silique than enclosed plants. In addition, the weight of the seeds was higher In this study, it was discovered that there are differences between diverse pollinator species groups when it comes to pollination efficiency. It is still not clear how exactly the diversity of pollinator species affects pollination. Several studies show that after a certain threshold, flower visits by honey bees no more have an increasing effect on crop yields. Thus, other pollinators are also needed for optimal pollination. This study especially highlighted the significance of wild pollinators in pollinating crop plants. There seems to be demand for long-term monitoring of the pollination efficiency of different pollinator groups in the northern agricultural region

Eläinten hyvinvoinnin parantamiseen tarvitaan paljon tieteellistä tutkimusta tunteista, älykkyydestä ja lajikohtaisista tarpeista. Tiedon pohjalta pystytään vertaamaan ja kehittämään eläinten tarpeisiin soveltuvia olosuhteita, kuten esimerkiksi virikkeitä ja hoitomuotoja eläintarhoissa. Eläintarhaolosuhteissa eläimen hyvinvoinnin saavuttaminen vaatii kompromisseja tilan ja resurssien rajallisuuden vuoksi. Parhaimman kompromissin löytämiseen vaaditaan tieteellistä tietoa lajikohtaisesta hyvinvoinnista. Maisterintutkielman tarkoituksena oli tuottaa tieteelliseen tietoon perustuva arvio Korkeasaaren eläintarhassa valittujen lajien hyvinvoinnista, jotka olivat huulipekari (Tayassu pecari), pikkumarmosetti (Cebuella pygmaea), punaniskakenguru (Macropus rufogriseus) ja sinikurkkuara (Ara glaucogularis). Tavoitteena oli tunnistaa tekijät, jotka vaikuttavat kyseisten eläinten hyvinvointiin luonnossa, ja verrata niitä eläintarhassa olevien yksilöiden olosuhteisiin. Tämän avulla pystyttiin arvioimaan lajikohtaiset tarpeet, joita tarvitaan eläinten hyvinvoinnin saavuttamiseksi. Vertasin hyvinvointiarvioitani tutkittavien lajien hoitajien tekemiin arvioihin, minkä tarkoituksena oli ymmärtää, riippuuko hyvinvoinnin arvio arvioijan näkökulmasta, ja onko lajien välillä eroja siinä, mihin seikkoihin arvioitsijat kiinnittävät huomiota. Arviointeja vertailemalla oli myös tarkoitus löytää tekijöitä, joilla on vaikutusta lajikohtaiseen hyvinvointiin Korkeasaaressa, minkä pohjalta voidaan ymmärtää ja kehittää mahdollisia hyvinvointia edistäviä toimenpiteitä. Toteutin tutkielman eri osioissa. Ensin suoritin kirjallisuuskatsauksen, jonka tarkoituksena oli määrittää tutkittavien lajien käyttäytyminen ja hyvinvoinnin tarpeet tieteellisten tutkimuksien avulla. Seuraavaksi selvitin eläinten olosuhteet Korkeasaaressa aitauksien kuvaamisen, tarkastelun ja hoitajien haastattelujen avulla. Tämän jälkeen tein hyvinvointikartoitukset, joita verrattiin tutkittavien eläinten hoitajien vastaaviin tilastollisesti. Hoitajat arvioivat lajien hyvinvoinnin aina paremmaksi kuin minä, ja erot arvioinneissa olivat suuria. Johtopäätöksenä voidaan pitää, että suora vertailu luonnonympäristöön voi johtaa huomattavasti alhaisempiin hyvinvointiarvioihin. Tällainen on esimerkiksi trooppisen ilmaston vertailu Suomen vuodenaikaisvaihteluun, vaikka eläimillä olisi aina mahdollisuus suotuisiin lämpö- ja valaistusolosuhteisiin. Hyvinvoinnin osa-alueiden välillä ei ollut merkittäviä eroja. Huomattavaa on, että hoitajat arvioivat samalla myös paitsi eläimen olosuhteita, myös omaa onnistumistaan niiden hoidossa. Tämä voi vaikuttaa siihen, että yksikään laji ei saanut hoitajilta yhtään 1-arviota, kun taas minulla oli objektiivisempi ote arviointiin, sillä kritiikki ei kohdistunut minuun edes välillisesti.

Protein kinases are signaling molecules that regulate vital cellular and biological processes by phosphorylating cellular proteins. Kinases are linked to variety of diseases such as cancer, immune deficiencies and degenerative diseases. This thesis work aimed to identify direct substrates for protein kinases in the CMGC family, which consists of the cyclin-depended kinases (CDK), mitogen activated protein kinases (MAPK), glycogen synthase kinase-3 (GSK3) and CDC-like kinases (CLK). CMGC kinases have been identified as cancer hubs in interactome studies, but large-scale identification of direct substrates has been difficult due to the lack of efficient methods. Here, we present a heavy-labeled 18O-ATP-based kinase assay combined with LC-MS/MS analysis for direct substrate identification. In the assay, HEK and HeLa cell lysates are treated with a pan-kinase inhibitor FSBA which irreversibly blocks endogenous kinases. After the removal of FSBA, cell lysates are incubated with the kinase of interest and a heavy-labeled ATP, which contains 18O isotope at the γ-phosphate position. Resulting phosphopeptides are enriched with Ti4+- IMAC before the LC-MS/MS analysis, which distinguishes the desired phosphorylation events based on a mass shift caused by the heavy 18O. With this pipeline of methods, we managed to quantify and identify direct substrates for 26 members of CMGC kinase family. A total of 1345 substrates and 3841 interacting kinase-substrate pairs were identified in cytosolic cell lysates, from which 165 were annotated in the PhosphoSitePlus® database. To identify substrates for kinases with nuclear localization, ten kinases were tested with nuclear HEK cell lysate. We identified 194 kinase-substrate pairs, 141 of which were unique to the nuclear fraction and 27 annotated in the PhosphoSitePlus® database. Finally, kinases with outstandingly high amounts of novel substrates were subjected to gene ontology analysis. We were able to link the gene ontology classifications of novel substrates to the biological processes regulated by the kinase of interest. These results indicate that heavy-labeled 18O-ATP-based kinase assay linked LC-MS/MS is a useful tool for large-scale direct kinase substrate identification.

Plants control the exchange of gases through the stomatal pores. Stomata are formed by guard cells and the closure of stomata are regulated via a complex signaling network in response to various biotic and abiotic stimuli, such as pathogens, elevated levels of CO2 and darkness. The leucine-rich repeat receptor-like kinase (LRR-RLK) GUARD CELL HYDROGEN PEROXIDE-RESISTANT1 (GHR1) is part of the network regulating stomatal closure. GHR1 is an inaktive pseudokinase that can activate SLOW ANION CHANNEL-ASSOCIATED1 (SLAC1), an anion channel that is crucial for stomatal closure, via interacting proteins. The exact role of GHR1 is still partly unknown, however, it has been suggested that GHR1 could function as a scaffold or as an allosteric regulator of additional components required for stomatal closure. The aim of this study was to identify novel interactors of GHR1. First stable plant lines expressing fusion proteins GHR1-YFP, GHR1W799*-YFP and plain YFP as a negative control were generated and from these lines fusion protein expression levels and the subcellular localization were studied. Next the plant lines were used for purifying GHR1 interacting proteins with the use of co-immunoprecipitation and identification of the proteins with mass spectrometry. The unlikely GHR1 interactor candidates were then filtered from the mass spectrometry data. The subcellular localization and the protein expression of the interacting proteins were studied with the use of internet databases. Literature of the GHR1 interacting proteins were studied in order to make possible connections with GHR1 and stomatal closure. In this study 38 GHR1 interactors were identified. Literature search revealed that many of the identified interactors had a known role in stomatal movements. These included proteins such as PLASMA MEMBRANE INTRINSIC PROTEIN2-1 (PIP2-1) and BETA CARBONIC ANHYDRASE 4 (BCA4), that are known to have a role in stomatal closure. Future work includes confirming the interactions with independent methods and studying the molecular mechanisms related to stomatal movements. The GHR1 interactome identified here for the first time reveals novel parts of the network regulating stomatal movements and thus increases our understanding of molecular mechanisms behind stomatal functions.

The TTN gene is composed of 364 exons (363 coding) and encodes for titin, the largest protein in nature. Pathogenic TTN variants result in a wide spectrum of skeletal muscle and cardiac disorders known as Titinopathies. These differ in inheritance patterns, onset age, disease course and severity. The biological mechanisms underlying disease-causing variants specific to titinopathy patients are still elusive. Investigating gene signatures causing the biological pathomechanisms is crucial for understanding genotype-phenotype corelations. RNA-sequencing emerges as a valuable technique for analysing transcriptomic data and exploring gene expression profiles of patient and control samples. To elucidate common pathomechanisms in titinopathies, including adult tibial muscular dystrophy (TMD) due to heterozygous FINmaj variant, and biallelic recessive titinopathies, an extensive differential gene expression (DGE) analysis was conducted using three RNA cohorts from human muscle biopsies. The cohorts involved two polyA enriched and one rRNA depleted batch-corrected cohort. Human DGE analysis identified 265 commonly upregulated genes and 147 commonly downregulated genes in the titinopathy cohorts. A significant downregulation of TTN expression levels was observed in one of the cohorts. To validate and understand the biological significance of these findings, data from a mouse model was incorporated with homozygous Ttn FINmaj variants. Common genes among all cohorts accounted for the structural integrity of the extracellular matrix. This study indicates the pathomechanisms for a skeletal muscle pathology and discusses the future steps in efficiently performing RNA-Seq for titinopathies.