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Not much attention is paid on the conservation of invertebrates despite their importance to the ecosystems in general and their benefits and ecosystem services to us, humans. This study is part of a project aiming to start the Sampled Red List Index (SRLI) for spiders. The IUCN Red List Index (RLI) is used for measuring the overall extinction risk of groups of species and the sampled approach is a way to evaluate the trajectory towards extinction of megadiverse groups without the need to assess every species of the whole group of interest. A random sample of 200 spider species were selected from the global checklist and assessed according to IUCN Red List Categories and Criteria. Given the availability of data, I was able to calculate the extent of occurrence and area of occupancy for 80 species, of which 70 species had an EOO above 20000 km² and 75 species an AOO above 2000 km², the thresholds below which species can be considered threatened. The trends in EOO and AOO were assumed to be stable for most species (49 species) given the inexistence of monitoring data for any taxon. Evidence of decline was found for only 10 species, usually inferred from habitat loss. Habitat data was collected for 118 species. The most common habitat type was forest (73 species), followed by grasslands (24 species) and artificial habitats (22 species). For 44 species the habitat trend was inferred to be stable, only declining, according to available knowledge, for 14 species and increasing for one species. For the remaining 141 species the habitat trend could not be inferred and was thus assumed to be unknown. The most commonly mentioned threat types were agriculture (11 species), fires (7 species) and logging (6 species). For 39 species there were no known threats and for the rest of the 132 species the threats were unknown. Conservation actions in place were observed for 104 species, most commonly site and area protection (100 species) and resource and habitat protection (88 species). Conservation actions such as education and awareness (8 species), resource and habitat protection (7 species) and site and area management (6 species) were to take into consideration. All the 200 species were estimated to be in need of further basic research especially on threats (143 species and distribution (140 species), but also on life history and ecology (135 species). Due to several knowledge shortfalls, including the Wallacean (distribution of species), Prestonian (population trends) and Hutchinsonian (response to environmental change), no threat category could be reached for the vast majority of the species. The results show that an IUCN category could be reached for only 59 species, of which 55 were assessed as Least Concern and a threatened category was reached for only 4 species (t as Critically Endangered and one as Vulnerable). The baseline SRLI at this first point in time was 0.95 (in a 0-1 scale, where 0 means all species are extinct and 1 for all species are Least Concern). We hypothesize however that among the 141 Data Deficient species there should be a higher proportion of threatened species than among the 59 evaluated. This would be due to two reasons. First, the scarcity of information on many species might in part be due to their rarity. Second, widespread species were often the only for which an assessment could be reached, creating a bias in the dataset towards a large base SRLI value. The strategy currently imposed by IUCN is therefore clearly inadequate for taxa with scarce information, which represent the vast majority of species. I propose the future use of a different, non-random, approach to the selection of species in the SRLI and its adoption for other taxa which represent in fact most extant and threatened species.

Urban greenspaces are valuable resources for humans and are an integral part of the urban landscape. They are also popular sites for various social and cultural activities, such as dog walking. Dog ownership rates are increasing globally at a rapid rate, hence increasing the dog populations within urban cities. The increasing number of dogs could potentially have negative consequences to the quality of urban parks. Furthermore, there is a lack of research on the impacts of dogs on urban environments. The aim of this study is to gain a better understanding of the impacts of dogs on path-side soils in urban greenspaces. Soil samples were collected from both Helsinki and Lahti. The impact of dog urine was studied by comparing path-side soil samples between areas with high dog activity (i.e., parks) and similar areas with low to no dog activity (i.e., cemeteries). Soil samples were collected three times during the growing season in 2021 at three different treatments: close by 1) an evergreen tree, 2) a lamppost, and 3) a lawn area. The samples were analyzed for soil pH, electrical conductivity, organic matter, soil moisture, NO3--N, and NH4+-N. Net N mineralization and net nitrification rates were also calculated. The results indicated that dog urine is a significant contributor to the input of nitrogen in urban parks. Soil NO3-N and NH4+-N concentrations were significantly higher in parks than in cemeteries and exhibited clear seasonal trends. Soil pH was generally lower and electrical conductivity higher in parks than in cemeteries. Poles and trees exhibited the greatest concentrations, suggesting that the impact of dog urine is highly localized around path-side objects. None of the treatments in cemeteries showed little to no changes throughout the growing season. Lastly, the net nitrification rates were significantly higher in parks during spring and summer. In contrast, net mineralization rates were generally higher in cemeteries than parks. These results confirm that urban planners need to take into consideration the impact of dogs to mitigate any potential negative consequences to the quality of urban parks.

Exercise-induced hyperinsulinism (EIHI) is a pathological condition characterized by aberrant insulin secretion triggered by physical exercise or pyruvate exposure. The monocarboxylate transporter protein (MCT1), encoded by SLC16A1, is ubiquitously expressed in almost all cell types except pancreatic islet cells. In patients with EIHI, mutations in the regulatory regions of the SLC16A1 gene are thought to lead to the unwanted expression of MCT1 on the beta cell membrane, allowing the influx of elevated lactate and pyruvate blood levels during exercise. These substrates feed into the Krebs cycle, increasing insulin release. This excessive insulin secretion can lead to hypoglycemia during exercise, causing weakness, syncope, and confusion. Since EIHI has never been studied using a human stem cell-derived islets, this thesis aims to establish a robust model in which to investigate the disease mechanism in detail. To achieve this, we reprogrammed EIHI patients’ fibroblasts into a stable pluripotent state and further differentiated them into functional pancreatic stem cell-derived islets (SC-islets) in vitro. These SC-islets were then matured further in vivo (in immunocompromised mice) and compared to healthy SC-islet controls. Rigorous quality control measures were implemented throughout the differentiation process to ensure its efficacy and the expression regulation of SLC16A1 was studied during SC-islet development. Extensive phenotypic characterization was conducted using immunohistochemistry, quantitative gene expression level analysis, and insulin secretion assays with glucose and pyruvate. Contrary to expectations, the results of this study demonstrated that despite the SLC16A1 promoter mutation, the expression of SLC16A1 was downregulated similarly to the control cell line during development in vitro, resulting in similar pyruvate-stimulated insulin secretion to the control cells. Interestingly, immunohistochemical analysis of in vivo implanted SC-islets showed a clear phenotype with an increased number of MCT1-positive cells only in the mutant grafts, some of which were endocrine cells. In conclusion, the phenotypic manifestations of EIHI were not visible in the setting of in vitro modeling, which was attributed to the similar expression levels of MCT1 in both the mutant and control cell lines. However, following in vivo implantation, there was a noticeable increase in MCT1 expression exclusively in the mutant cells. This finding suggests a distinct regulatory mechanism of MCT1 expression, which might be impacted by the in vivo surroundings and the maturation state of human islets.

Flat-footed flies (Diptera: Platypezidae) are a group of small (3-6 mm) flies classified into four subfamilies based on diagnostic morphological characteristics. There are approximately 277 species worldwide. One of the subfamilies, the cosmopolitan Microsaniinae or the smoke-flies, is of particular interest. Smoke-flies are pyrophilic i.e. they are attracted to sources of smoke, such as burning wood or forest debris where they swarm. Smoke-flies are frequently observed in the presence of smoke; however, their biology remains entirely unknown. While the adult and larval habitat is understood for most other Palaearctic flat-footed flies, this information remains elusive for smoke-flies. Three different approaches are explored to increase knowledge on the biology of smoke-flies. Controlled burns organized by Metsähallitus are visited to observe the behavior of smoke-flies in a natural setting that mimics forest fires. Molecular and morphological identification is employed to identify the phoretic mites carried by smoke-flies, which can help to deduce the ecology and habitat preferences of the mites and consequently infer the habitats of smoke-fly larvae. Additionally, shotgun metagenomic sequencing of entire smoke-fly samples is utilized to determine if it’s possible to identify a molecular signature of the fungi in the genomic reads, which could provide insights into the ecological aspects of their habitats. The results confirm that the reproduction of smoke-flies is directly linked to forest fires. The use of an emergence trap in freshly burned forest demonstrated that smoke-fly larvae likely reside within the soil. These findings are supported by the taxonomic classification of fungal reads obtained through shotgun metagenomic sequencing, which identified soil-based arbuscular mycorrhizal fungi in adult smoke-fly samples. Although the analysis of phoretic mites provided no additional information on the biology of smoke-flies, the presence of mites in nearly all smoke-flies captured by the emergence trap confirm the ecological linkage between the habitat of mites and smoke-fly larvae. Integration of the results obtained from this study provides key information on the biology of smoke-flies and the specific habitat of the unknown larvae.

This thesis aims to illuminate expert dissensus on the policy instruments for reducing the carbon footprint of household consumption. In this thesis, I focus specifically on the policy instruments for addressing Finnish households’ consumption, proposed in the policy Delphi study by Salo et al. (2023), and experts’ responses regarding the measures’ importance. Exploring expert dissensus regarding consumption policy instruments is essential since identifying the measures that elicit expert dissensus and unveiling experts’ opinions on these measures promotes transparency in policymaking, helps to identify underlying assumptions, inconsistencies, and potential blind spots within the proposed measures, and therefore, contributes to better policy formulation. Two research questions are answered in this thesis to explore expert dissensus regarding consumption policy instruments: 1) What policy instruments to reduce the carbon footprint of household consumption elicit the most dissensus among experts? and 2) On what grounds do experts agree or disagree on the instruments exhibiting expert dissensus? A quantitative analysis involving calculating the interquartile range of experts’ responses was done to reveal which policy instruments exhibited expert dissensus. As a result, 26.8% of all proposed policy instruments were identified to exhibit expert dissensus. Subsequently, experts’ open comments regarding these policy instruments were examined through a thematic analysis to understand the grounds for expert dissensus. As a result of this research, seven themes emerging from experts’ open comments were identified: 1) Effectiveness, 2) Feasibility and practicality, 3) Content of policy mix, 4) Content of policy measure, 5) Consideration of other factors guiding consumption, 6) Consideration of possible side effects of utilizing the measure, and 7) Normative claims. Of these themes, the most prevalent was effectiveness, and the least prevalent were normative claims. As previous literature points out, the evaluation of policy instruments involves a blend of normative and empirical aspects. While it is unclear to what extent experts resorted to implicit normative assumptions within their evaluation process, this thesis concludes that information and uncertainty-related aspects, especially regarding effectiveness, require further consideration in sustainable consumption policymaking and research regarding policy measures exhibiting expert dissensus.

Animal welfare is a multifaceted concept that encompasses the overall well-being of animals, considering their physical and psychological health, behaviour, social interactions, and ability to engage in species-specific behaviours. Though there isn't a single universally accepted definition, the World Organisation for Animal Health (OIE) provides a widely recognized definition. According to the OIE, animal welfare refers to how well an animal copes with its living conditions. An animal is considered to have good welfare if it is healthy, comfortable, well-nourished, safe, able to express innate behaviour, and not suffering from pain, fear, or distress. In my thesis, the main objective is to identify knowledge gaps in recognizing factors that influence welfare among animal caretakers and to gather essential information that can serve as a foundation for future efforts to improve animal welfare in Korkeasaari Zoo. The project was conducted in collaboration with the zoo and involved studying and assessing the welfare of four different species. The assessments were based on scientific knowledge obtained through extensive literature research. The findings were then compared and analyzed by using a general linear model (GLM) with the zookeepers' own assessments. The results revealed that, overall, zookeepers tended to assess animal welfare more positively than the knowledge-based evaluation, although the extent of this difference varied among species. Notably, the results brought to light that certain species, especially the Asiatic lion, have specific welfare requirements that might not be fully recognized by the keepers. This implies there are opportunities for enhancing animal welfare in these cases. Furthermore, assessments varied depending on the type of question posed, with a significant observation being that nutritional requirements consistently received the highest evaluation across all species, regardless of the evaluator. This underscores the heightened attention that zookeepers pay to the nutritional well-being of the animals. This likely stems from the availability of well-documented nutritional information, in contrast to other facets of welfare in the studied species that might still be less comprehensively understood. My research is practical in nature, focusing on four specific species within one zoo. It may not primarily contribute to theoretical advancements but rather offer practical insights and applications. This marks just the starting point and indicates the need for further exploration and advancement. Shedding light on the various factors that shape animal welfare is pivotal for refining the operational protocols within the zoo setting. Such insights can pave the way for constructive measures that contribute to the betterment of the animals' well-being within the zoo environment. For instance, the notable difference in the evaluation of Asiatic lions, where the zookeepers scored them higher than the literature-based assessment, emphasizes the importance of incorporating scientific knowledge into zoo management practices. By bridging this gap and aligning assessments with well-established scientific data, we can make substantial strides in improving animal welfare in Zoos.

Plants are vital to all terrestrial ecosystems by providing ecosystem services through photosynthesis- derived compounds. Throughout the millennia, plant metabolism has diversified in the form of all plant secondary metabolites, ranging from metabolite groups such as terpenes to alkaloids to flavonoids. Many of these secondary metabolites are economically valued for their chemical, pharmaceutical and physical properties. The flavonoids are one of the largest groups and are known to provide competitional advantages and increase of survival of many plant species in extreme environments. One of the critical enzymes in the whole biosynthesis pathway of flavonoids is the dihydroflavonol 4- reductase (DFR). DFR regulates the formation of leucoanthocyanidins, predecessors of colourful anthocyanins. Anthocyanins are an economically significant group of molecules, especially for horticulturists and plant breeders, but also for nutritional and health scientists due to their potential health benefits. Dihydroflavonol 4-reductase is a much-studied enzyme due to its significant role in flavonoid biosynthesis and the economic interests of plant breeders and alike. Previous studies have expanded the knowledge of flavonoid biosynthesis and have identified several amino acid residues in the DFR structure affecting the substrate specificity of the enzyme and, consequently, the flower colours. However, only a single crystal structure model of the dihydroflavonol 4-reductase has been solved so far, originating from the grapevine Vitis vinifera. Although a single crystal structure can facilitate further structure-to-function studies associated with dihydroflavonol 4-reductase, further studies need to be carried out to shine a light on the functional basis of the enzyme. Therefore, this study aims to resolve petunia and gerbera dihydroflavonol 4-reductase crystal structures, expanding the knowledge of structural variations within the uncharted families of angiosperms, Solanaceae and Asteraceae. Several recombinant protein expression systems were utilised in my attempts to solve the crystal structure of the DFRs. These systems ranged from the bacterium Escherichia coli to yeast species such as Saccharomyces cerevisiae and Pichia pastoris, as well as the tobacco plant Nicotiana benthamiana. The genes encoding for Petunia wildtype DFRA, three mutants, and three Gerbera DFR variants were cloned to several expression vectors. Their presence and expression were identified using various genetic methodologies and enzymological assays. The expression of DFRs using an E. coli-based expression system was verified. However, the trials with E. coli were deemed unsuccessful due to the majority of the protein ending in inclusion bodies with no detectable activity. An alternative system using agroinfiltration of N. benthamiana was later utilised, as significant amounts were detected in the plant tissue extracts following the agrobacterial infiltration. Although the proteins were expressed in high quantities, no purification procedures have been established to provide plant tissue-extracted protein in crystallography-grade purity. With the protein supplied by a plant-based system and several small- scale purification steps, purified DFR enzymes could be utilised in crystallisation studies. Due to significant contamination by RuBisCO in the protein samples, alternative systems based on S. cerevisiae and Pichia pastoris were investigated, and a successful Pichia-based expression was established. Several sets of plasmids with variable expression systems were constructed in this study, facilitating future experiments into the dynamics and structure of dihydroflavonol 4-reductases. Ground-breaking techniques based on computational modelling were utilised to hypothesise the role of prior determined amino acid residues in enzyme catalysis and substrate recognition. Possible crystallisation-related issues originating from protein structure were approached using the same techniques, opening new windows and possibilities into determining the structure of Petunia hybrida and Gerbera hybrida dihydroflavonol 4-reductase structures using tools of protein engineering.

Due to population growth, urbanization, and increase in life expectancy, the urban population is growing, and by 2050 68% of the global population is expected to live in urban areas. Even though the air quality in urban settings has greatly improved in recent decades due to increased legislation, restriction, and monitoring, the negative health impacts associated with pollutants have not completely diminished. Air quality varies on a local scale due to urban form and function, that creates differences in experienced exposure among individuals. These exposure differences among socio-economic groups have been studied, but no clear consensus has been found, as the results have been very diverse and even contradictory. Therefore, conducting local level studies is important in order to identify local patterns of exposure and to recognize them in urban planning. However, previous literature on the topic in Finnish context is lacking. This thesis studies interpolated air pollutant exposure among one susceptible socio-economic group – the elderly – and aims to identify possible hotspots of both in Helsinki, Finland. The distribution of the elderly and air quality is assessed through Moran’s I calculations. Global Moran’s I is used to assess for spatial autocorrelation, and local Moran’s I is applied to identify local clusters. To further examine the relationship, correlation coefficients are calculated through Spearman’s rank correlation coefficient. According to the results, there is a weak positive correlation between the elderly and air quality, indicating that generally air quality tends to be worse when the number of elderly people increase. However, the relationship is stronger among younger age groups, although the differences between age groups is very small. The exposure was also assessed through overlapping high value clusters, which indicate that elderly clusters in the city center are located in areas with low air quality, while elderly clusters in Lauttasaari and Vuosaari are located in areas with good air quality. Previous studies regarding the relationship between age and air pollution exposure have been contradicting in different cities, and therefore these results provide important knowledge about the problem specifically in the context of Helsinki. Additionally, the results are be observed in the light of the larger discourse around socio-economic status and air pollution, but further studies on the topic are still needed. Including multiple socio-economic variables and the dimension of negative health outcomes would aid in identifying 1) the most important socio-economic factors in the context of negative health outcomes associated with air pollution, and 2) those areas where multiple important socio-economic factors and low air quality are overlapping and therefore might have higher risk for negative health outcomes.

The endoplasmic reticulum (ER) is an important organelle of the cell where a high number of proteins are synthesized and modified to obtain their final structure. Therefore, the ER stress, which is caused by accumulation of unfolded proteins in the ER, is not to be taken lightly since it could contribute to many diseases, such as cancer and neurodegenerative diseases. The response to the ER stress is the unfolded protein response (UPR), which is an adaptive system that helps in adjusting for increased folding needs within the ER. One of the main protein branches in the UPR is inositol requiring enzyme 1 (IRE1). IRE1 detects the status of protein folding inside the ER and initiates the UPR signaling pathway to achieve either normal folding status or cell death. The aim of this research was to express yeast IRE1 in E.coli and human IRE1 in insect cells, purify with affinity chromatography and study the IRE1’s crystal structure with a small molecule modulator that could possibly enhance its activity. The protein was expressed successfully and purified with glutathione S-transferase (GST) tag, and the activity of the pure protein was determined. The structural studies were not fully completed since the absolute purity and yield that was necessary for crystallization was not achieved due to loss of protein during gel filtration and precipitation. Based on the results it is likely that the structure of the protein could be solved and further biochemical and structural studies with F10 are possible.