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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.

Kaurapohjaiset elintarvikkeet ovat terveellisiä. Monet niiden terveyshyödyt johtuvat kauran liukoisen ravintokuidun suuresta β-glukaanipitoisuudesta. β-glukaanin terveysvaikutukset ovat riippuvaisia sen molekyylipainosta ja viskositeetista. Viskositeetilla on myös muuta merkitystä kaurapohjaisissa elintarvikkeissa kuten kaurajugurteissa ja kauramaidoissa. Aiemmassa Folafibre-tutkimusprojektissa oli tutkittu kaurakuidun folaattipitoisuuden (B9 vitamiini) kasvattamista fermentoimalla sitä eri mikrobeilla. Hyvin folaattia tuottaneet mikrobit myös alensivat kaurakuituvalmisteen viskositeettia, ja erittivät glykosyylihydrolaaseja ja/tai proteaasia. Oli kuitenkin jäänyt epäselväksi, olivatko entsyymit ainut syy viskositeetin alenemiseen, ja kuinka paljon kullakin niistä oli vaikutusta viskositeettiin. Lisäksi tämän alan kirjallisuudessa on pidetty epäselvänä, onko esim. tärkkelyksen ja β-glukaanin välillä jotain interaktioita jotka mm. nostavat niiden viskositeettia yli yksittäisten polymeerien viskositeetin summan. Näitä voitaisiin selvittää puhdistamalla viskositeettia alentaneista mikrobikannoista kyseiset entsyymit ja tutkimalla niiden vaikutusta kaurakuidun viskositeettiin. Yhdeksi mikrobiksi valittiin Exiguobacterium sp. RB3 kanta, koska em. syiden lisäksi tässä bakteerisuvussa esiintyy psykrofiilejä ja mikrobeja jotka voivat kasvaa korkeassa pH:ssa, jolloin sen erittämät entsyymit saattaisivat olla aktiivisia matalissa lämpötiloissa ja korkeassa pH:ssa, ja olla siten teollisesti kiinnostavia. Toiseksi mikrobiksi valittiin Bacillus sp. ABM5119, koska sitä oli käytetty monissa Folafibre-projektin tutkimuksissa. Työn tavoitteena oli puhdistaa Bacillus sp. ABM5119:n endo-β-1,4-glukanaasi, ja Exiguobacterium sp. RB3:n α-amylaasi ainakin siinä määrin, että ne ovat muista endoaktiivisista glykosyylihydrolaaseista ja proteaaseista puhtaita, ja sitten mitata puhdistettuja entsyymejä ja proteaasia käyttäen niiden vaikutus yhdessä ja erikseen keitetyn kaurakuituvalmisteen viskositeettiin. Lisäksi tavoitteena oli karakterisoida Exiguobacterium sp. RB3 amylaasin olennaisimmat biokemialliset ominaisuudet, sekvensoida sen geenin amylaasia koodaava alue, ja selittää sen ominaisuuksia myös sekvenssistä johdettavissa olevan tiedon perusteella. Keitetyn kaurakuitupreparaatin viskositeettia alentavat eniten α-amylaasi ja endo-β-1,4-glukanaasi. Proteaasi ei vaikuta kaurakuitupreparaatin viskositeettiin, kun kuitupreparaatti on keitetty. Endo-β-1,4-glukanaasi alentaa viskositeettia yhtä hyvin kuin β-1,3-1,4-glukanaasi. Synergiaa viskositeetin alentamisessa α-amylaasilla ja endo-β-1,4-glukanaasilla ei havaittu, mutta havaittiin että β-glukaani saattaa estää tärkkelyksen retrogradaatiota. Hyvin pienetkin entsyymiaktiivisuudet vaikuttavat viskositeettiin. Jos halutaan β-glukaanin viskositeetin ja siten terveysvaikutusten säilyvän, täytyy esim. kauramaidon valmistuksessa käytettävien entsyymivalmisteiden olla β-glukanaaseista hyvin puhtaita. Mikrobifermentaatioissa tapahtuvat viskositeetin alenemat johtuvat lähinnä mikrobien erittämistä glykosyylihydrolaaseista, ei niinkään esim. niiden metabolian sivutuotteista kuten happiradikaaleista. Exiguobacterium sp. RB3 α-amylaasi on rakenteeltaan Bacillus licheniformis α-amylaasin kaltainen glykosyylihydrolaasiryhmän 13 entsyymi. Se sitoo rakenteeseensa kolme kalsiumiatomia, ja kalsiumpitoisuus vaikuttaa sen aktiivisuuteen. Se on aktiivisimmillan pH alueella 5,0 – 7,5. Se sietää detergenttejä, toisin kuin eräs aiemmin karakterisoitu Exiguobacterium-α-amylaasi. RB3 α-amylaasin turnover number oli korkea, 29000 1/s. Exiguobacterium-suvun psykrofiilisessä haarassa esiintyy kahta eri α-amylaasia, joista yksi on tässä karakterisoitu, ja toinen on selvästi erilainen rakenteeltaan ja biokemiallisilta ominaisuuksiltaan.

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.

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.

Histamine receptors are known to be expressed throughout the peripheral nervous system and are involved in regulating the gut and immune system. The gut-brain axis, which consists of bidirectional signaling between the central nervous system and gastrointestinal tract, links gut functions to emotional and cognitive controls in the brain. Many animal models are known to express histamine receptors in their gut and brain tissue which can be altered by a compromised gut-brain axis like stress. Histamine receptors also play an important role in many gastric and intestinal disorders. However, the precise expression pattern of histamine receptors in zebrafish gut tissue is unknown, as is whether their expression levels also change with stress. Here, I show that zebrafish gut contains several histamine receptors, but their role involving stress within the gut remains unknown. I found that histamine receptors hrh1 and hrh3 as well as the enzyme that synthesizes histamine, histidine decarboxylase (hdc), are expressed in zebrafish gut and brain in wildtype and hdc knockout adult zebrafish using in situ hybridization. Stress induction on wildtype male zebrafish through chronic social defeat and analysis of histamine receptor and hdc mRNA levels using quantitative real time PCR showed no differences in subordinate, dominate, or control fish. However, it did provide quantitative data that hrh1, hrh2, and hdc mRNA expresses in the adult gut. My results demonstrate the first data to suggest histamine receptors are expressed in zebrafish gut, and that even though stress can alter the gut-brain axis, it may not do so through the regulation of these receptors.

White matter (WM) structural connectivity alterations have been linked to depression. This study aimed to identify structural connectivity metrics associated with Major depressive disorder (MDD) and predictive of different symptom phenotypes. The study sample included N=29 control and N=86 subjects with MDD who underwent a clinical interview, Mini-International Neuropsychiatric Interview (MINI), and assessment of depression symptoms severity. Using a 3T MRI scanner, diffusion tensor imaging (DTI) was employed to capture WM connectivity markers at baseline. While no distinct differences between control and MDD groups were observed at the whole-brain network level, significant alterations were evident at the node level. Clinical group demonstrated enhanced connectivity, particularly in the DefaultB and LimbicB subsystems, as evidenced by measures such as eigenvector centrality. Furthermore, notable differences were observed in clustering coefficient and local efficiency, predominantly in DefaultB, LimbicB, and VisPeri networks, with MDD patients showing higher connectivity. Analysis of the association between WM structural connectivity measures, both global (e.g. global efficiency) and local (e.g. clustering coefficient) with MDD symptom scores and related symptoms, revealed no significant correlation at the whole-brain level, both at baseline and post-intervention. Distinct patterns were identified when evaluating node-level metrics averaged across networks, which together with group differences, point to MDD patients exhibiting characteristics consistent with regular networks. Hierarchical clustering based on standardized baseline DTI structural connectivity within the clinical cohort revealed three distinct clusters of MDD patients, with the first cluster exhibiting a higher WHO-5 score, indicating a potential association with better well-being. These findings provide insight into MDD-specific brain regions’ structural alterations and underscore the heterogeneity of depression symptom profiles. Further research is needed, including a higher sample size and control for confounding factors.

Despite the advances in the management of breast cancer, discovery of novel and targeted drugs remains a challenge. It has been suggested that drug failure rates in clinical trials might be diminished by improving the predictive potential of preclinical cancer models. Three-dimensional (3D) scaffold-based cell culture has emerged as an attractive platform for mimicking tissue-like microenvironment, since it is well-known that cells respond to the cues in the extracellular matrix (ECM). The aim of this thesis was to develop fibrin-based hydrogels and evaluate their performance in 3D cell culture of breast cancer cells. The fibrin gel formulation was first optimized by testing the effect of different buffers on gel properties. Structural properties were examined with scanning electron microscopy and mechanical properties measured with oscillatory rheometry. Three different fibrin concentrations of the optimized formulation were then used as scaffolds for DU4475 breast cancer cells. After seven days of culture, the morphology, phenotype and proliferation of the resulting cell structures were assessed by using techniques such as light microscopy, immunofluorescent confocal microscopy and Western blot analysis. The desired properties for 3D cell culture were obtained by preparing fibrin gels at high pH in the absence of calcium. The main finding of the thesis was that fibrin concentration strongly affected the phenotype of DU4475 cells, with cells cultured in the softest gel retaining their original characteristics to the greatest extent. In the future, the developed scaffold could possibly be used in drug discovery and personalized medicine by culturing tumor explants from patients. However, the methods used in the study must be further optimized and the results validated with other breast cancer cell lines and with primary tissues.

Follicle-stimulating hormone (FSH) regulates mammalian reproduction. The hormonal function of FSH is exerted through its receptor, FSHR. Hormone binding leads to signal transduction by the synthesis of the second messenger (intracellular cAMP) and the activation of other downstream signaling pathways. FSH receptors are mainly found in ovaries and testes where FSH stimulates e.g. follicular growth and spermatogenesis, respectively. It has been previously shown that FSHR mutations are linked to infertility through abnormalities in the receptor function. For example, inactivating FSHR mutation (Ala189Val) leads to arrest of follicular development in females and reduced sperm counts in males. FSH action seems to be especially critical for folliculogenesis and essential for female fertility. The development of gonads and dysfunctions affecting reproduction are the focus of our research group. Studies of recent years have demonstrated extragonadal FSHR expression, including endothelial cells of female reproductive tract and developing placenta. The physiological relevance and function of this extragonadal FSHR is still not well known, especially during embryogenesis. In addition, the expression or function of the mutated FSHR has not been studied in any cells endogenously expressing the receptor. As a consequence, the purpose of this thesis was to study FSHR expression and function in human pluripotent stem cells (hPSCs) as a model for early human development. The study was conducted in a human embryonic stem cell line (hESC H9, 46, XX) and two human induced pluripotent stem cell lines (hiPSC HEL127.6 and HEL128.5; 46, XX). iPSCs were obtained from female patients carrying the A189V mutation in the FSHR gene. Cells were differentiated for 8/12 days using two in vitro cell culturing protocols for distinct differentiation attempts. The first protocol (protocol D) was designed for embryonal cell differentiation, the other (trophoblast protocol) for extraembryonal differentiation. Cells at different stages of differentiation were studied with qRT-PCR and immunocytochemistry. At d8, cells were stimulated with FSH for qPCR studies and for cAMP assay. Control cells (H9) differentiated by protocol D endogenously expressed functional FSHR. These cells responded dose-dependently to FSH stimulation by substantially increasing cAMP production, downregulating FSHR mRNA expression and altering inhibin gene expression. Patient-derived iPSCs carrying the mutation expressed FSHR as well but the receptors were non-functional as expected. Both H9 and patient-derived cells that differentiated into trophoblast-like cells with the other protocol, also expressed FSHR at low level but did not respond to FSH stimulation. Preliminary results with protocol D indicate that the cells resemble extraembryonic cell types. The study has revealed novel information and insight about extragonadal FSHR expression and function during differentiation. However, the exact identity and the biological role of these cells is yet to be confirmed.

In this thesis I focus on a novel disaster response and preparedness mechanism called forecast-based financing. The mechanism is linked to the changing paradigm of humanitarian response that calls for more localized and more resilience building solutions to addressing and preventing humanitarian crisis. It is also in the core of the anticipation agenda which argues that waiting for disasters to happen is not a sustainable option and that forecast data and pre-agreed triggers and actions should be used in order to prevent both loss of lives and mitigate the cost and impact of disasters. Main hypothesis is that climate related hazards to livelihoods and food security seems to be the sector where forecast-based financing could have most potential for increasing resilience and sustainability. Slow onset crises with long lead-time allow for better targeting and more variety of actions. As the lifetime of the action is longer, there is less chance of action which is in vain. Furthermore, the actions which are more localized, for example direct support to farmers, can decrease their vulnerabilities. I aim at taking a critical approach to assessing this potentiality associated with the forecast-based financing mechanism through case study. The three cases (Mongolia, Kenya, Zimbabwe) were selected from pilots implemented by the main actors: the Red Cross, World Food Programme (WFP) and Food and Agriculture Organization (FAO) and Start Network. This thesis uses a combination of evaluative and heuristic approach to qualitative case study analysis. To answer the first research question, 1) is the forecast-based financing mechanism successful in prioritization of actions in a way that best address the needs and resources of vulnerable populations, I aim at finding out if mechanism is effective (or potentially effective) in delivering impact. For the second research question, 2) are the actions sustainable and do they bring socio-economic benefits that go beyond meeting acute humanitarian needs, I will see if new pathways are found for confirming the defined hypothesis. I am using heuristic approach in terms of finding new links e.g. between actions and needs of either donors, actors or beneficiaries. I asses and analyse available reports and evaluations (secondary data) of the selected operations. I conducted eleven (11) semi-structured key informant interviews (primary data) using practitioner’s perspective for retrieving qualitative data, for further understanding and for triangulation. All key informants were affiliated to the cases. My analysis show that the potentiality for development impacts and long-term transformation of the forecast-based financing is there but it is not utilized in the cases reviewed nor is it perceived in a same way across practitioners of different backgrounds. Currently the mechanism is used more for effective response, not for addressing the root causes of vulnerability. In general, the entitlement or empowering of a person who is affected by disaster currently does not go beyond securing bridge over lean season, avoiding negative coping mechanisms or e.g. better yield or survival of livestock. Sustainability potential of the forecast-based financing seems to be currently underutilized and international funding envelopes do not offer an alternative to the humanitarian funding launched case-by-case. Most of the practitioners interviewed were clearly in favour of linking and using forecast-based financing in some way to long-term programming, thinking outside of the framework of humanitarian response, extending lead time significantly and adding positive reinforcement inputs. I argue that with a lead time that goes long in advance, towards development actions, the mechanism needs to be reframed for the donors and the sources of funding might need to be reconsidered. To implement meaningful resilience actions in slow onset cases, triggers need to be early enough and actions in two phases: 1) anticipatory and benefiting from forecast and 2) early response. At beneficiary level the actions should be geared up to better address underlying socio-economic vulnerabilities and take advantage of the long lead time.