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Secondary growth in plant vasculature tissues originate from meristematic tissue vascular cambium. Vascular cambium produces xylem inwards and phloem outwards in the plant root and stem. Vascular cambium regulatory mechanisms are not completely known, and while plants and especially trees are dependent on the mechanical support and transportation systems the vasculature provide, increased knowledge of the vascular cambium and especially xylem development is valuable. In this thesis, I explored Arabidopsis thaliana single-cell RNA-sequencing data from secondary growth stage via cloning transcriptional reporter lines, and later utilized the transcriptomics data to find potential xylem development regulators. By loss-of-function mutant and inducible overexpression line screening, I found two genes in a family of MYB transcription factors to be potential regulators in xylem development. Loss-of-function mutant did not show significant phenotype, but overexpression lines under constitutive 35S promoter showed extreme halt of growth, and a stain-filled vessel phenotype in a fixed and resin-embedded Arabidopsis cross-section. Under developing xylem -specific promoters, inducible overexpression lines showed the same phenotype, and while grown to the secondary growth stage, I detected some patterning defects. Thus, these results indicate, that these MYBs expressed in the xylem parenchyma in the single-cell transcriptomics data, might have a function in xylem development.

In recent years, the science community has increasingly emphasized the influence of behavior, lifestyles, and culture on greenhouse gas emissions. Finland’s emissions per capita (~10 tCO2/yr.) are amongst the highest in the world, and it has been calculated that to comply with the Paris Agreement and to limit global warming to well below 2 °C, Finnish households should reduce their carbon footprint by 70 % by 2030. This requires drastic lifestyle changes, especially regarding mobility, housing, and nutrition. However, knowing this is not enough to change behaviors. Human behavior is influenced by several different internal and external factors such as knowledge, values, and social structures. This master’s thesis is a case study conducted in the Environmental Change and Global Sustainability (ECGS) master’s program at the University of Helsinki with the aim to enrich theories about pro-environmental behavior by providing insights on the future aspirations of sustainability science students, who have both extensive knowledge and experience in the field of sustainability. The data for this study was collected through an anonymous, online questionnaire with four open-ended questions. In total, 61 out of 274 ECGS students answered the questionnaire, after which qualitative content analysis with both inductive and deductive elements was applied to analyze the results. The results confirm that ECGS students are indeed well aware of the most impactful lifestyle changes for reducing personal carbon footprint. Despite their extensive knowledge on sustainability, the students’ future aspirations still come quite close to the dominant cultural vision of a good life in Finland. The implications of these findings once again confirm the importance of facilitating sustainable ways of living through changes in societal structures, discourses, and norms.

Diabetes mellitus (DM) is a metabolic disorder, which in 2021 alone affected approximately 537 million adults. DM is a multi-organ disease with several comorbidities, one of which is chronic kidney disease (CKD), which often leads to renal impairment and kidney damage. While current treatment strategies have improved, they fail to protect the kidneys efficiently, which is why further understanding and renoprotective strategies are required. Podocytes are terminally differentiated cells central to the proper function of the glomerular filtration barrier (GFB) in the kidneys, and their injury can lead to the leakage of protein into the primary urine, which is a hallmark of CKD. One of the potential causes of podocyte injury in DM is hyperfiltration induced increase in fluid flow shear stress (FFSS). Podocyte responses to FFSS are still, however, relatively unknown. We exposed cultured human podocytes in vitro to FFSS at 2 dyne/cm2 for 2 hours via a novel flow chamber system. From the FFSS experiments, we studied podocyte motility from live cell imaging, protein expression levels by Western blotting and finally did immunofluorescent labelling to identify protein localizations in the cells. We discovered that podocytes express different modes of motility upon FFSS exposure, notably bleb-like motility previously only described in tumor and embryonic cells. In addition, we observed that podocytes significantly increased the phosphorylation of both AMPK and Ezrin, indicating the activation of pro-survival signalling as well as formation of bleb-like protrusions in response to FFSS stimuli. However, we did not observe significant podocyte loss, indicating that podocytes are capable of withstanding increased FFSS for short exposures such as 60 minutes. We believe that upon FFSS exposure, podocytes activate pro-survival mechanisms such as increased phosphorylation of AMPK and changes in motility in order to better withstand the increased shear stress. However, increased FFSS in for example DM patients is persistent, making it potentially a key factor in the development of podocyte injury and ultimately kidney damage.  

Multiple sclerosis (MS) is an immune-cell mediated demyelinating disease with one of the World’s highest prevalence in Finland. Although the central nervous system (CNS) has a remyelinating response towards acute demyelination, in MS it often fails and fibrotic scarring occurs. A subtype of perivascular cells, fibroblasts (FBs), have shown to offer a favorable environment for fibrosis and promote fibrotic scar formation, which hampers neuronal recovery. The process is driven by immune response, including macrophages that can express phenotypes between pro- (M1) and anti-inflammatory (M2) profiles. This thesis aims to represent an opening exploration to the role of macrophages in meningeal fibroblasts’ (mFBs) proliferation, survival and fibronectin expression (scarring activity). Thus, mFBs were exposed to soluble factors derived from M1, M2 and unpolarized M0 macrophages. While immunocytochemistry (ICC) against Ki-67 revealed that M1-derived soluble factors induce a transient delay in mFBs’ proliferation, this condition also increases mFBs’ fibronectin expression. On the other hand, ICC showed that M1-derived soluble factors do not alter the proportion of active caspase-3 positive apoptotic mFBs and, in consistence with this, lactate dehydrogenase (LDH) assay shows no changes in LDH release, both approaches indicating no effects in cell survival. These findings suggest a potential profibrotic role of M1-derived soluble factors in fibrotic scarring that may counteract remyelination in MS.

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.

In Finland and in many other European countries, indoor housing of dairy cattle has been increasing at the expense of grazing. According to Finnish legislation, only dairy cows housed in tie-stalls are obligated to graze for 60 days per year between May and September. Pressure to intensify production leads to increase in herd sizes, regional concentration, lack of suitable pastureland near large barns, and automation of milking. Conversely, grazing is regarded as important for animal welfare, maintenance of biodiversity and cultural landscapes. Farmer perceptions towards grazing have not been studied in Finland comprehensively, which need to be understood better in order to provide adequate support for sustainable management of grazing. The research aims at gaining understanding of the external and internal factors driv- ing the farm-level decision making related to grazing regimes. Concept of good farming ideal is used to examine farmers’ decision-making. The data was collected from 26 cattle farms with a gradient of grazing from zero to maximum. Firstly, an online questionnaire was sent out to farmers and secondly, semi-structured inter- views were conducted with 7 dairy producers, to further explore farmer views about grazing, biodiversity, and animal welfare. The results show farmers who practice grazing to perceive it more positively than the farmers who do not graze. Some of the farmers who grazed seemed to consider it almost as an intrinsic value, whereas farmers with no grazing saw it as an inef- ficient production method, which they were afraid would become an obligation. The most im- portant internal factors driving decision-making about grazing based on the questionnaire and interviews, were animal welfare, decreased workload because of grazing, infrastructure of the farm, economic aspects, and benefits of grazing to biodiversity and landscape values. Exter- nal factors found to affect the decision of grazing were incentives, regulations, and consumer expectations. Animal welfare was found to be a basis for production for many and the most important feature valued for good farming, to which grazing was seen to contribute. The re- sults imply subsidies regarding grazing should be higher for grazing to be considered profita- ble and more enjoyable by farmers.

Biodiversity loss and ecosystem service degradation and the related economic costs are increasingly recognized as sources of financial risks. The risks are arising through physical and transition sources of risks caused by dependencies and impacts upon biodiversity and ecosystem services. Therefore, it has become increasingly important for both individual financial institutions as well as central banks and financial supervisors to better understand and manage these risks. However, biodiversity loss is associated with unique complexity and uncertainty, making it a challenging task. The purpose of this thesis was to study the linkages between biodiversity loss and financial stability in Finland. This was done by assessing the financial exposure of Finnish credit institutions to sectors dependent on ecosystem services for their production processes. A quantitative analysis was conducted in order to combine loan data obtained from the Bank of Finland and ecosystem service data obtained from the ENCORE database. The results showed that 23% of loans provided by Finnish credit institutions are exposed to high or very high biodiversity-related financial risks. The sectors associated with most value at risks were real estate and agriculture, forestry and fishing sectors. Disruptions of ecosystem services providing surface water and climate regulation were posing the highest financial risks. The findings represent a first step towards assessing the exposure of the Finnish financial system to biodiversity-related financial risks. The results and previous literature indicate that physical biodiversity-related financial risk exposures are significant for individual financial institutions and for the whole financial system, despite the prevailing methodological challenges and gaps in knowledge. In order to complete a comprehensive biodiversity-related financial risk assessment, further research is needed.

Food literacy is a noteworthy topic to be studied due to food’s considerable environmental and health effects. When food literacy and its characteristics are known, food literacy can be used as a tool to improve people’s health and the condition of the environment. Thus, this thesis aims to reveal the extent of food literacy among Finnish upper secondary school students. In this context, food literacy emphasises food-related environmental and health knowledge. Environmental knowledge is understanding of the global environmental impact of food. Health knowledge, on the other hand, is the familiarity with the relationship between excessive meat consumption and Finnish common diseases along with beliefs regarding diets and food products as a source of protein. This thesis also aims to identify to what extent does the food literacy differ based on gender, study year and living area. This study was performed as a quantitative sample survey and the data was collected using an online Typeform -questionnaire. The questionnaire reached respondents from many different Finnish localities, mainly from cities. The final data consisted of 1320 individuals and it was analysed using IMB SPSS Statistics 24 software. The following methods were used to analyse data: frequency analysis, an Independent Samples t Test, a One-way ANOVA, and a Post-hoc LSD test. Gender, study year and living area were used as grouping variables to examine the differences between groups. The results show that the students named school as the main source of food literacy. Moreover, the results indicate that awareness regarding food production, dietary health and proteins increase significantly from the first to the third study year. The students acknowledged food production causing environmental problems and that the share of food in the consumer’s climatic impact is considerable. Nevertheless, the students underestimated the climatic impact of cheese and they were unaware of the more specific characteristics of food’s environmental impacts. They also had food-related environmental misconceptions considering packaging, transportation and meat consumption. Moreover, approximately half or more of the students were aware of the connection between excessive meat consumption and the increased risk of distinct common diseases. Most of the students acknowledged a versatile vegetarian diet as being a healthy choice. The study also reveals that female students had notably higher dietary health knowledge than male students. This Master’s thesis study mainly supports the findings of previous studies on food-related knowledge. The results elucidate the extent, characteristics, gaps and misconceptions of students’ food literacy. These findings may be utilized to improve school education on food literacy, alter misconceptions and fill the gaps of knowledge in pursuit of improving the health of people and the condition of the environment.