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(2020)Dog activities (urination and defecation) can be a source of nitrogen accumulation in the snowpack of urban parks. Urban parks are commonly visited by urban residents and excessive buildup of nitrogen in snowpack can be a health concern. This accumulation of nitrogen in snowpack may wash away during spring with the runoff of melting snow, which can cause the eutrophication and deterioration of lake ecosystems. In this study, I investigated the concentration of some nitrogen-related compounds along with a few physical parameters in the snowpack of urban parks and compared these between areas with high dog activities and areas with no dog activities. I hypothesized that, nitrogen concentrations will be higher in areas with high dog activities, like snowpack beside paths than in areas with no dog activities. The study was performed in 10 parks of two cities in Finland; Helsinki and Lahti. In these parks, samples were collected from snowpack immediately next to walking paths (path edge), and in control areas 8 m away from these paths, where dogs were unlikely to visit. Samples were collected from February to April of 2019. The concentration nitrogen from snow samples of both parts of parks were compared. The study showed that, the concentration of most nitrogen species, e.g. Total Nitrogen (TN), ammonium (NH4+), Total Organic Nitrogen (TON), and electrical conductivity followed a similar pattern of having higher values in snow from path edges where dog activities were the highest. However, the concentration of NO3 – and pH values were both slightly lower in the path edge samples compared to control areas undisturbed by dogs, but differences were not statistically significant. Information from this study can be useful for understanding the connection between dog activities and nitrogen build up in snowpack of urban parks, and can also be helpful for designing urban parks by considering environmental and health effects of nitrogen accumulation in the snow from dog activities. My findings can also be useful to urban waterbody related studies, e.g. eutrophication, and the accumulation of nutrients in lakes.
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(2019)Proteostasis is used by cells to maintain proteome health and understanding the biological mechanisms underpinning proteostasis is important. Despite many studies on small molecule-mediated inhibition of Sec61-dependent protein translocation, a knowledge gap exists in the quality control pathway(s) of pharmacologically-displaced secretory polypeptides. Genetic screens can be used to discover proteostasis regulators of pharmacologically-displaced secretory polypeptides. Near-haploid human HAP1 cells with an inducible Tet-on GFP reporter (reporter HAP1 cells) can be used as a cellular tool to screen for human host factors pertinent to proteostasis of secretory polypeptides. The isolation of haploid-enriched reporter HAP1 cells ensures that the inability to efficiently recover bi-allelic gene trap mutants is avoided. The use of haploid-enriched cells is a prerequisite to gene trap mutagenesis screens. Here, I present data on the isolation of haploid-enriched reporter HAP1 cells that could be used as a cellular tool in gene trap mutagenesis screens. A workflow for the isolation of haploid-enriched reporter HAP1 cells was optimised using a diploid reporter HAP1 cell line as a control. Both DNA content analysis and karyotyping showed that the isolated HAP1 reporter cell lines were haploid. In the haploid-enriched HAP1 reporter cells, the GFP-reporter compartmentalised in the ER, and a Sec61-translocon inhibitor CT8 could inhibit the GFP-mediated fluorescence. The haploid reporter HAP1 cell lines produced in this study are suitable for future gene-trap mutagenesis experiments.
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(2022)While there is no current consensus on what characterizes a heathy gut microbiome, there are evidence supporting the association of high microbial diversity with health stability in the host species. A growing literature suggests that microbial communities can withstand short-term anthropogenic changes with resilience, however enduring long-term changes might have a negative impact on the natural composition of host microbiome. Parasitism and anthropogenic disturbances resulting in habitat degradation might represent two of such long-term challenges to the invisible diversity of microbial communities, with consequences for the fitness of their host species. In this study, I tested whether parasitism by a specialist parasitoid and ecological changes due to anthropogenic habitat degradation affected the microbial community associated with the diapausing larvae of the Glanville Fritillary butterfly (Melitaea cinxia). The larval samples were collected in September 2012 from seven different communes across the Åland islands, from two extreme types of habitat patches (i.e., highly human-impacted habitats versus natural meadows). The parasitoid wasp Hyposoter horticola naturally parasitize approximately a third of the M. cinxia larvae in Åland, giving an opportunity to further investigate how parasitism might affect the larvae-associated bacterial community in this system. The DNA from the gut of each larva was extracted, and the bacterial V5-V6 region of the 16S gene was amplified and sequenced at the Finnish Institute for Molecular Medicine. The sequencing data was prepared for analysis by processing it through Mothur and QIIME2 data cleaning protocols. According to alpha- and beta-diversity metrics analyses, anthropogenic degradation of the habitat did not cause significant changes in the bacterial composition of the microbiota associated with diapausing larvae of M. cinxia, however, parasitism with the parasitoid wasp has a significant negative effect on the beta diversity of the microbial communities associated with the diapausing larvae.
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(2022)Mutations in the KCNQ1 gene have been implicated in the onset of hypopituitarism. Regulating KCNQ1 expression would therefore enable future clinical research on the mechanism of the disease. CRISPR offers a flexible toolset for controlling genetic expression via knockout, knock-in, knockdown, and gene activation. Of these approaches, CRISPR activation (CRISPRa) is distinguished by its ability to induce gene overexpression in a cell’s native context, making it a valuable tool in the interrogation of genetic disorder pathogenesis. This thesis therefore tested the efficacy of a CRISPRa subsystem in increasing KCNQ1 expression. The CRISPRa subsystem, VPR, was chosen because of its high activation efficiency and the ease of controlling the activation system of its doxycycline-inducible mode of action. The cell line used for the experiment, HEK293, was similarly chosen because of its ease of culture and transfection. To validate the proper functioning of the activation system, expression rates of the related genes ASCL1 and GHRH were measured as positive controls. The activation system successfully upregulated the expression rates of the two genes. As the dCas9-VPR system is dependent on the Tet-ON operator for inducing activation in a controllable manner, a test for dCas9 leakage was conducted. RT-qPCR analysis showed the upregulation of ASCL1 expression in the uninduced state of the system, confirming the presence of dCas9-VPR leakage. The dCas9-VPR system finally aimed to test the expression rate of KCNQ1. Although one novel guide RNA successfully upregulated KCNQ1 expression, it did so inefficiently and its success was not shared by any of the other tested guide RNAs. Altogether, the dCas9-VPR system was successfully established in HEK293 cells, and the leakage of the inducible system was confirmed, however, KCNQ1 activation by CRISPRa requires further optimization.
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Activity and functional connectivity of key nodes in depression network : A resting state EEG study (2024)Major Depressive Disorder (MDD) is a prevalent health issue worldwide, yet one third of patients are unresponsive to first-line treatment. Repetitive Transcranial Magnetic Stimulation (rTMS) is a promising alternative treatment and better understanding of the neural network impairments associated with MDD could significantly improve its efficacy. Recent research has identified a ‘depression network’ involving key brain regions, suggesting that MDD symptoms arise from functional connectivity impairments within this network. This thesis aims to map the activity and functional connectivity of six bilateral regions of interest (ROIs) implicated in MDD: dorsolateral prefrontal cortex (dlPFC), dorsomedial prefrontal cortex (dmPFC), ventromedial prefrontal cortex (vmPFC), inferior frontal gyrus (IFG), intraparietal sulci (IPS) and pre-supplementary motor area (pre-SMA). The intention is to describe biomarkers of depression and map the depression network to identify connectivity impairments between these cortical sites. Literature on these ROIs was reviewed and rsEEG data from 24 MDD patients and 9 healthy controls was collected to analyze the activity and functional connectivity of selected ROIs. Our results suggest that MDD involves widespread connectivity impairment including in regions that have not previously been included in the research on depression, such as IFG and IPS. Most ROIs showed trends of reduced activity in delta, theta, alpha and beta bands. The effects were most noticeable in the theta band, especially in the IFG, dmPFC, and dlPFC. However, the functional connectivity impairments were more prominent and significant, particularly in the beta and alpha bands. Notably, the IPS and vmPFC stood out as key nodes with the most dysfunctional connections in MDD. These findings support the idea of a depression network characterized by connectivity abnormalities rather than localized activity impairments. This study emphasizes the importance of a network-based approach in understanding MDD. The search for biomarkers as well as novel stimulation targets should consider regions beyond the traditional dlPFC or the frontal cortex, to include regions such as IPS, IFG and vmPFC.
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(2022)A systemic change in the current modes of production and consumption, a so-called sustainability transition, is required to overcome large-scale society-transforming phenomena such as the climate change. This in turn demands changes in socio-technical systems, i.e., the networks of actors, institutions, technologies, material artefacts, and knowledge creation. In this thesis, the Finnish construction and housing sector is used as a case study, and an example of one socio-technical system. By focusing on the socio aspect of the socio-technical, I analyse how actors who are involved in the current system may inhibit or enable a sustainability transition. I seek to answer the following questions: what the relevant definitions of and foci for climate-wise action are among stakeholders in the sector in Finland; how actor-related barriers manifest themselves; and which actors could enable or speed up the transition. This thesis builds on sustainability transition theories, especially multi-level perspective and strategic niche management, to better understand actor roles and relationships. Based on a literature review, I define three actor-related barriers to transition (misaligned vision and focus, small network, and pro-regime actor resistance) and one potential enabler (intermediaries). These are then applied to the Finnish context. In this thesis, I employed stakeholder analysis as the methodology, interviewing a pre-defined set of 21 stakeholders. The results were analysed using content and social network analyses. The results suggest that although the understanding of climate-wise construction and housing is gaining a more holistic perspective, the three barriers all still manifest in the sector in Finland: all the stakeholders are engaged in energy-related topics, but hold differing foci on household choices, low-carbon materials and circularity; the network amongst actors seems relatively dense and inclusive, but improvement points emerge with closer examination; although results suggest that development has happened in the recent years, industries and incumbents are still considered too slow-moving. The importance of intermediation is also recognised by many but defining and picking potential intermediaries out of the crowd is a complex task. Overall, the sector may be moving forward in the transition, but the stakeholders create and uphold both barriers and opportunities in the process.
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(2022)This study aims to understand how cities adapt to environmentally induced hazards, like floods. Extreme floods are interesting firstly, because climate change is predicted to increase flooding in several places globally in the future, and secondly, because even a small risk could be realised in the right conditions. The methods are a case study of flood adaptation in Helsinki, qualitative content analysis, interviews, and a scenario. Land use planning is chosen as the context of the case study, because densification challenges flood preparedness. The material consists of the zoning plan of Helsinki, its flood risk management related appendixes and interviews with city experts. The qualitative content analysis aims to answer the first research question: How does land use planning consider extreme floods in Helsinki? The scenario, in turn, aims to answer the second research question: In what ways might an extreme flood challenge the current land use planning in Helsinki? The interviews are mainly used to support the other methods. The results lead to one main argument, for which I present several justifications. The argument is that the flood risk management and land use planning in Helsinki, the urban structure of which is densifying, do not sufficiently consider the risk related to extreme floods, even though climate change is increasing the likelihood of such. In the end, I present some policy recommendations to change this.
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(2022)A common understanding of partnership goals is widely acknowledged to be one of the most crucial factors of a partnership project’s success. This thesis examines a partnership between the city of Espoo and five company partners and looks for processes and conditions that support a common understanding of the project goals. The aim is to support future sustainability-partnerships by giving managers concrete tools for facilitating goal-alignment by answering the research question “how did the partners accept and adapt to a common sustainability goal?” The data consists of interviews conducted with the project employees from both the city and the private entities, the project contract and web communications published by the partners. The data was analyzed using theme categories derived from the literature, seeking to first find the answer of whether the official partnership goal was adopted an accepted by the partners, and then comparing those observations to the experiences that partners had had working in the collaboration. It seems that the project goal was adopted and accepted in this case, and based on the findings five key mechanisms for how that was achieved and what future managers can therefore consider were recognized: 1) the organizational goals of the partner organizations were sufficiently compatible with the partnership goal, 2) the partnership goal was broad enough to leave room for later adjustments, 3) the partners were further divided into smaller sub-tasks with supporting sub-goals, 4) there was a lot of mandatory group-work and 5) all of the above-mentioned features were taken into account already in the design-phase of the partnership, and programmed into the day-to-day activities of the collaboration.
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(2020)Early life conditions have long-term effects on the fitness and survival of individuals. Foetal development is an especially crucial period and even small changes may have large impacts on the development of individuals. Mammal foetuses may be exposed to additional testosterone either from their male littermates or their mother. This additional prenatal androgen exposure leads to masculinization of female features and behavior. In males the effects of additional prenatal androgen exposure are less drastic due to their own testosterone production. The anogenital distance, defined as the distance between the anus and genitalia, has been used to determine the sex of young mammals since males have longer anogenital distances than females. An elongated anogenital distance is an indicator of additional prenatal androgen exposure in females, and in some species also in males. It correlates with for example increased aggressiveness in both females and males. In females a longer anogenital distance has also been connected to delayed puberty and decreased fertility. I studied the effects of additional prenatal androgen exposure on weight and important life-history traits in banded mongooses (Mungos mungo) with data from a long-term study. Banded mongooses are small co-operatively breeding mammals living in family groups of 10-30 individuals across sub-Saharan Africa. Breeding is extremely synchronized within groups and in most cases all pregnant females give birth on the same day. The resulting communal litter is cared for by most adults in the group regardless of relatedness. Adults escort the pups until three months of age, providing the pup with food, grooming and protection. This early life care has long-term fitness benefits for the pups. Pregnant females may change the phenotype of their offspring via maternal effects. When the competition faced by breeding females is more intense, they compensate by investing more resources to their foetuses, making them bigger. Using the anogenital distance as a proxy for additional prenatal androgen exposure, I measured its effects on weight at early life and maturity, the amount of care received as pups and whether the individual reproduced in its lifetime or not. I hypothesized that a longer anogenital distance may be an indicator of increased competitiveness in the banded mongoose. It could lead to a cumulative advantage since more aggressive individuals may be able to access more food and care, which leads to higher maturity weight and lifetime reproductive success. I also measured the effects of resource abundance and intensity of competition during gestation on the anogenital distance of the pups. I hypothesized that mothers may prepare their offspring for future competitive environment by exposing them to androgens during gestation. In males a longer anogenital distance predicted higher weight both at early life and maturity. Higher weight at the beginning of the escorting increased the amount of care received, which in turn increased weight at maturity. A longer anogenital distance therefore has both direct and indirect fitness benefits in male banded mongooses. In females, a longer anogenital distance predicted lighter weight at maturity, suggesting that it may have negative effects on female growth and development. This study offers evidence that additional prenatal androgen exposure has long-term fitness consequences on banded mongooses and that these consequences are sex specific. Future research should focus on confirming the connection between additional prenatal androgen exposure and longer anogenital distance in this species, as well as assessing the effects of prenatal androgen exposure on survival, puberty and growth of especially female individuals.
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(2021)Adeno-Associated Viruses (AAVs) are quickly becoming one of the most applied vectors for gene therapy applications. In the recent years three new AAV-based gene therapies have been approved by U.S. Food and Drug Administration (FDA) and European Medicines Agency (EMA). The regulatory bodies require accurate and reliable characterisation of the clinical grade viral vectors during and after production. Analytic methods measuring the purity, potency and safety of the product support the up-stream and down-stream processes during the production and are used for final-drug substance characterisation. Median Tissue Culture Infectious Dose (TCID50) is a well-established method for measuring the infectious titer of a virus. Here, an assay for determining the infectious titer of AAVs, which has previously been used to characterise the existing AAV2 Reference Standard Material (AAV2RSM) was set up and optimised for research use at Kuopio Center for Gene and Cell Therapy (KCT). The assay utilizes the HeRC32-cell line, a HeLa clone, that stably expresses AAV Rep and Cap -proteins and in presence of adenovirus, enables the replication of recombinant AAV-vectors. The cells were grown in 96-well plates and infected with 10-fold dilution series of AAV vectors (AAV2 and AAV6) using human adenovirus type 5 as the co-infector. 72 hours post infection the vector genome replication of AAV was detected with quantitative PCR (qPCR). Thresholds for qPCR determined copy number and cycle threshold (Ct) were set and used for the determination of infection-positive wells. The 50-percent endpoint was observed and used to calculate the infectious titer according to the Spearman-Kärber method. The assay was set up and optimised with the AAV2 Reference standard material (AAV2RSM) using two different primer-probe sets (targeted sequences were; SV40 polyadenylation signal and AAV inverted terminal repeats (ITRs)). Plates infected with AAV2RSM were analysed separately with both primers resulting in mean infectious titers of 8.07 ± 3.13 x 108 TCID50 Infectious Units (IU) / mL (n = 9) and 1.27 ± 0.464 x 109 TCID50 IU/mL (n = 9) for SV40 and ITR, respectively. After the assay was set up with the AAV2RSM, an in-house AAV6 product was analysed with the ITR primers yielding 6.09 ± 3.94 x 109 TCID50 (IU) / mL (n = 5). The assay protocol was successfully set up for research use at the KCT laboratory. Improvements were added to the original protocol to increase assay robustness, accuracy and precision as well as to minimize the possibility of over-estimation of the infectious titer. The assay can be further optimised for a particular therapeutic AAV product in the research laboratory or technology transferred to a production facility for optimisation and validation for the analytics needs of a production pipeline.
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(2023)Diets of wild owls have been studied by pellets, when the nutritional value of owl prey remains unclear. Fatty acids are an essential part of animal nutrition, acting as energy reserve, building blocks of membranes and precursors for signaling molecules. I studied whether Finnish owl species have species-specific characteristics in the fatty acid profiles of adipose tissue due to different diets and feeding habits. I also compared fatty acid profiles between wild and captive owls to elucidate if the diet of captive owls resembles the diet in the wild. Fatty acids from visceral adipose tissue and a liver were transesterified to fatty acid methyl esters, which were identified and quantified by gas-chromatographs linked to either a mass spectrometry or flame ionisation detector. The fatty acid compositions were compared with Principal Component Analysis and statistical significance of the separations between species, origins and tissues were studied by Soft Independent Modelling of Class Analogies. Differences in the relative concentrations of individual fatty acids and their structural category sums were evaluated by the Kruskal-Wallis test followed by the Wilcoxon rank-sum test. Compared to wild owls, captive owl (snowy owl Bubo scandiacus and Eurasian eagle-owl Bubo bubo) adipose tissues had lower relative concentrations of many long-chain polyunsaturated fatty acids (PUFAs), such as arachidonic acid, eicosapentaenoic acid, and docosahexaenoic acid. The captive owls also had lower n-3 PUFA/n-6 PUFA ratio than the wild owls. I suggest that these differences arise from anthropogenic food given to the prey of the captive owls. In the future, the feeding and breeding of captive owls could be improved and potential health hazards prevented by giving the owls a diet rich in long-chain PUFAs, either by giving the prey more versatile diet or adding supplements to the owl’s diet. The adipose tissues of wild Eurasian eagle-owls contained higher total relative concentrations of monounsaturated fatty acids than the species belonging to the Strix genus. On the other hand, long-chain PUFAs were more prominent in the Strix owls. I suggest that these differences in the fatty acid composition are due to the disparities in feeding behaviour: the Eurasian eagle-owls eat carrions and large prey that cannot be swallowed whole, which may lead to consuming more the body outer parts of the prey. The fatty acid composition of the eagle-owls can also be affected by urban prey that have consumed anthropogenic food. In the future, the fatty acid composition of wild prey should be examined to be able to evaluate the prey composition of owls and its effect on the owl fatty acid profile with physiological consequences, the knowledge that could be used in the future decision-making that supports the conservation of owls.
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(2021)The focus of this thesis is on environmental agency in two different video games, The Sims 4 Eco Lifestyle and Final Fantasy VII Remake. The research questions aim to answer how the player can act as an environmental agent in these two games and what are the key differences between these games. The study is executed in a form of qualitative two-case case study with the help of close reading. With close reading eight different types of agencies that form the typology of this study, will be analysed in order to answer the research questions. The data for this thesis was collected by playing both games and taking notes by following close reading. The notes were then analysed with the different types of agencies. The results showed clear overlapping of the types of agencies, and it could be said that environmental agency can be used better in the game when the overlapping is happening. The agencies complemented each other and made the possible learning process in the game more fulfilling. The main difference between the game seems to be that The Sims 4 Eco Lifestyle as a life simulation game offers more diverse possibilities for environmental player agency. The studies about environmental games are mainly focused on serious games and not so much on commercial games. Video games hold great potential to engage people in environmental things especially with the help of player agency. It offers the player the ability to make meaningful choices and if they are structure well, the player can see the consequences of their agency which serves as an effective feedback which could lead to positive learning. In this case, the environmental agency in the game could be transformed into real-life environmental agency. As video games have become more immersive and their environments more realistic, it could be worth considering that separating virtual environment from the real-life one might not be necessary anymore. Therefore, games like The Sims 4 Eco Lifestyle and Final Fantasy VII Remake could serve as an example of how environmental agency within them could be harnessed into wider use.
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(2024)Although scientific knowledge is centered around academic institutions of the Global North, publications claiming to be “global” have surged recently. This thesis used a rigorous systematic literature review to identify articles with “global” in the title or author keywords, then analyzed the exact locations from where their dataset originates. Furthermore, it examined circulation metrics of each assessed article, as determined by their journal’s impact factor, to investigate if there are any discrepancies among an article’s impact factor and its global representativeness. Finally, it compared the impact factor of these “global” papers to similar non-global articles to get an idea if “global” papers are published in more reputable journals than their non-global counterparts. The results of this thesis show there is an overrepresentation of “global” environmental science data from Eastern Asia, Northern America, Western Europe, Southern Europe and Northern Europe, and an underrepresentation of environmental science data from Southern America, Eastern Europe and Northern Asia, Eastern Africa, Western Africa, Northern Africa, Middle Africa, and Central Asia. Moreover, all journals, regardless of their prestige, publish articles with these globally unrepresentative results. Yet, articles with “global” in the title or keywords circulate in journals with higher impact factors than non-global papers. My results signal that data from only a few regions dilute “global” environmental studies, while marginalizing many parts of the world. Therefore, “global” environmental science research needs a stricter threshold of globality.
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(2022)The general question of this research is how beta oscillations are implicated in stopping an ongoing movement. Previous studies regarding movement cancellation have found a significant increase in beta activity in sensorimotor areas, especially in the form of transient increases in beta oscillations, called beta bursts. However, the functional role of beta band activity in stopping is still unclear, mainly because the behavioural tasks used cannot measure the exact timing when the subjects start the stopping process and therefore it is only possible to infer the stopping time. To resolve this, we used head-fixed rats running on a treadmill while performing a Go/NoGo task. In some NoGo trials, the rat starts to run, realizes the mistake and stops before crossing a fixed distance threshold. These are the events being analyzed, called near-mistake events (N=39,366). We found a single beta burst occurring prior to stopping in all five brain regions analyzed (from 44.2±20.1 ms to 55.8±16.0 ms) and positive correlations of beta burst number and power increase with movement speed before stopping. We also found a single alpha burst prior to and during stopping in all brain regions (from 45.9±20.1 ms to 57.1±19.3 ms), supporting previous studies’ findings of alpha band involvement in inhibitory motor actions. Our findings on beta bursts suggest a causality role in stopping an ongoing movement while our results of alpha bursts need to be further analyzed to understand its functional role.
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(2022)The climate crisis is such major environmental threat that it has negative impacts on both nature and people. Recently, the effects of climate change on people’s mental health have also been noticed. Children and adolescents are especially vulnerable group in terms of climate anxiety. The amount of climate anxiety among youth has been found to be increasing, yet the studies on it are still scarce. It is important to find out how much climate anxiety there is among youth and does it influence their behavior. The objective of this master’s thesis is to find out how much climate anxiety there is and is climate anxiety interconnected with the pro-environmental behavior among youth. Also, its aim is to find out whether attending the climate course affects the amount of climate anxiety. In addition, the study focuses on whether gender, home country and parental education level are interconnected with the amount of climate anxiety. This study was conducted as part of the D.Game project which includes a climate course for youth aged 12–18 years. Data for this research was collected through a survey during the climate course. The schools and students participating in the D.Game climate course and the climate anxiety survey were selected through an open recruitment during autumn of 2021. The data consists of 48 students who responded to the pre survey and of 13 students who responded to the post survey. The participants were 15–17 years old. On the contrary to other research, there is not much climate anxiety among youth. There is also a strong positive correlation between climate anxiety and pro-environmental behavior, which differs from other research. On the other hand, there were no significant differences in climate anxiety between the groups divided by divided by gender, home country and parental education level, which was expected result. Based on the data, the climate course did not have statistically significant effect on the amount of climate anxiety among youth. Also, there is need for further research. Larger sample could provide more valid results. The causality of the correlation between climate anxiety and pro-environmental behavior would be interesting to resolve. Whether attending to a climate course affects the amount of climate anxiety could be found out by development of climate courses and comparing different courses.
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(2024)Background: ProSAAS is a neuroendocrine peptide precursor implicated in various physiological pathways and several disorders. Despite its potential significance, there is a notable lack of studies exploring the roles of proSAAS and its derived peptides. Objectives: This hybrid systematic review aims to provide an overview of the neuroprotective role of proSAAS in brain-related disorders and its potential as a biomarker. The hypothesis is that the loss of proSAAS, known for its many neuroprotective properties, would affect dopaminergic and serotonergic neurons in zebrafish brains. Methods: Following PRISMA 2020 guidelines, this review includes studies on proSAAS in brain-related disorders and its biomarker potential, excluding non-brain-related physiological aspects. The focus is on dopaminergic and serotonergic systems in zebrafish. Searches were conducted on PubMed using keywords like "proSAAS," "aminergic system in zebrafish," "dopaminergic neurons in zebrafish," and "serotonergic neurons in zebrafish" on 04.05.2024, 09.05.2024, 11.05.2024, and 12.05.2024. Risk of bias was evaluated using the Cochrane Collaboration’s and AMSTAR 2 tools. For the experimental part, immunohistochemical analysis was conducted on zebrafish aged 4, 5, and 6 days post-fertilization. Results: A total of 103 studies were included in the systematic literature review. Six studies highlighted the neuroprotective role of proSAAS in neurodegenerative diseases. Two studies linked proSAAS to homeostatic upscaling, and five identified it as a potential biomarker for neurological conditions. Furthermore, nine studies investigated the role of proSAAS-derived peptides. Experimental results from immunohistochemical analysis showed no significant changes in dopaminergic and serotonergic systems between wild type, heterozygote, and knockout zebrafish. Discussion: Limitations include potential bias from included studies, small sample sizes and limited repetitions of the experiments. The review suggests proSAAS is critical for brain function and neurological conditions, though experimental findings did not show significant effects on dopaminergic and serotonergic neurons in zebrafish.
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(2021)Tropomyosins (Tpm) are coiled-coil proteins, which wind around actin filaments to form head-to-tail oligomers. Tpms control actin filament growth, movement and interactions with other actin-binding proteins like myosins and cofilin. Tpms play essential role in the construction and stabilization of complex three-dimensional actomyosin contractile structures called stress fibers, because Tpms regulate structural and functional attributes of actin filament populations. In mammals, there are 4 TPM genes encoding above 40 TPM variants, giving rise to many functional Tpm protein isoforms. They are responsible for several physiological mechanisms in cell such as morphogenesis, cytokinesis, vesicle transportation, metabolism, motility, organ development, and signaling. Even though several studies have been conducted to determine structures and functions of various Tpms, many questions are still to be answered about Tpm2.1 and its significance in cells. So far, Tpm2.1 isoform has been a difficult protein to study due to poor success rate at achieving its complete depletion from the cell. Its involvement in cytokinesis, cell movement, cancer progression, and association with mechanosensing ability of cells were recently reported, and this raised the interest of researchers to focus on unveiling its precise cell biological function. Conditional deletion, degradation or inactivation of a protein helps to determine its function in cells. A new revised Auxin Inducible Degron version-2 (AID2) approach employs the cell´s own ubiquitin mediated protein degradation process, ensuring efficient and rapid depletion of target protein by the help of expressing OsTIR1(F74G) auxin-receptor mutant in presence of 5-phenyl-indole-3 acetic acid (5-Ph-IAA) ligand. In this study, we established a pipeline to identify Tpm2.1’s functions using AID2 technology by integrating OsTIR1(F74G) mutant at AAVS1 locus of the homozygous knock-in U2OS clones, containing mAID-msGFP2-TPM2.1 fusion insert at their endogenous TPM2.1 locus. We aimed to deplete Tpm2.1 from cells using this approach by inducing with 5-Ph-IAA and observe the direct, immediate phenotypes during Tpm2.1 degradation. We succeeded in achieving almost complete Tpm2.1 depletion. By this approach, we revealed that Tpm2.1 controls actin reorganization, stress fiber stability, and maturation of focal adhesions in cultured cells. To our knowledge, Tpm2.1 is the first actin-binding protein to be studied using AID2 approach, and the promising outcome brings hope to study other complicated actin-regulating proteins with this approach.
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(2020)The production and consumption of foodstuffs has a strong impact on climate change, and vice versa. Agriculture and the food industry are responsible for over 25% of man-made greenhouse gas emissions. Thus, food choices are a significant way in which individuals can influence greenhouse gas emissions. By comprehensively changing one’s food consuming habits to align with the principles of sustainability, an individual can decrease the climate impact of their total consumption by approximately ten percent. Sustainable habits, such as food consumption, are part of wider social and cultural processes. Therefore, in research relating to food consumption, it is important to consider both the background of the research participants and the community and society in which they live. Through food choices, an individual expresses their identity, status, and belonging in the community. Eating is a social event that is affected by the values and attitudes of the surrounding community and society. These things strongly influence an individual’s food choices, but on the other hand, individuals can also reshape the attitudes and values of their community through their choices. In order to advance sustainable food decisions on a societal level, it is important to examine what factors influence people’s consuming and eating habits. There has been a considerable amount of research done on sustainable foods, but the research focus has not often been on aware consumers. Studying aware consumers provides information about which factors hinder the making of sustainable decisions when the obstacle is not a lack of awareness. As more is known about the reasons behind people’s food choices, it becomes possible to consider new methods for getting people to make more sustainable choices. The students of the Environmental Change and Global Sustainability program at the University of Helsinki are interesting subjects of research, because they presumably are aware of and interested in the impacts of their food choices. Therefore, in studying them it is possible to focus on other factors determining their food choices, rather than obstacles related to a lack of awareness. In addition, Helsinki as a study location offers good possibilities for making sustainable choices, because there is a diverse supply of sustainable foods in the urban centre. In this master’s thesis, I aim to answer the question: What kinds of perceptions of a sustainable diet do the students of the University of Helsinki program of Environmental Change and Global Sustainability have and what kinds of obstacles do they face when making sustainable food choices? My study is a qualitative case study. I gathered the research material by conducting semi-structured theme interviews with eight students. I analyzed the material by employing thematic analysis methods. My study indicates that the students found it most difficult to follow a totally plant-based diet, even though they consider it to be a sustainable choice. Choosing plant-based food was challenging for them especially in certain social situations. These included situations in which the students felt pressured into eating animal-based food, or situations in which they wanted to please another person by eating the animal-based food they were offering. In such situations, the students were inclined to make choices that differ from those they make in their everyday lives. In social situations that involve food and eating, people have a human need to indicate a sense of community and respect towards people important to them. The students I studied have a lot of knowledge and awareness of the sustainability impacts of their food choices. However, my study indicates that in some situations the need for social cohesion is more important than the need to make a sustainable food choice. The social meaning of eating is important to take into account when considering how to get people to make more sustainable food choices.
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(2006)The purpose of my thesis was to find out how Finnish dung beetles find their most important resource, dung. I tried to establish the most important sense for finding dung, to which part or parts of the scent dung beetles react to and how sensitive their ability to detect dung is. Dung beetles are an important part of the ecosystem and a long used model for population study. They are an interesting target for sensory research because their resource is both patchy and short-lived. Because of these aspects dung beetles have to locate dung very quickly and efficiently. I used dung beetles belonging to family Aphodiidae (Scarabaeoidea Laparosticti) that I collected from cow pastures in Viikki using pitfall traps. To track down how dung beetles approach dung I observed their approaching behaviour in the wild. To figure out the importance of sight and smell I eliminated one sense at a time and observed how it affected beetles' behaviour. I eliminated olfaction by cutting off antennas and sight by painting the beetles' eyes. To find out chemical attractants in question I first analyzed smelly, volatile organic compounds in cow dung and then offered them as bait alone and in different combinations in pitfall traps in the wild. I also tried to establish the sensitivity of olfaction by offering different amounts of cow dung in a long windless corridor to dung beetles. I noticed that dung beetles approach dung by flying low in headwind. Part of the beetles approached flying in a narrowing zigzag-pattern while others landed occasionally to orientate and approached in small spurts. Eliminating olfaction effectively prevented dung beetles from finding dung whilst blinded individuals performed as well as control beetles. From all the chemical components I used, only butane acid attracted some dung beetles. A combination of chemicals is what presumably attracts dung beetles best.
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(2023)Attention deficit hyperactivity disorder (ADHD) is a neuropsychiatric disorder characterized by inattention, hyperactivity and impulsivity. The symptoms appear in childhood and, if left untreated, can continue into adulthood affecting the quality of life. Currently, diagnosing a child's ADHD relies on subjective questionnaires filled out by a parent and an interview. The detection of changes in brain activity especially during everyday activities could bring important information that could help inform the diagnostics of ADHD. The changes in brain activity in persons diagnosed with ADHD during familiar, everyday events have been studied very little. However, ADHD brain imaging studies done during resting state and simple tasks have found changes in large-scale brain networks. These networks can be studied using functional connectivity approach, where the degree of synchronous activity in different brain regions is used to determine the connection strength between these regions. In this thesis, using functional magnetic resonance imaging, the differences in the functional connectivity of the whole brain between children diagnosed with ADHD (n=17) and controls (n=19) were investigated during a virtual reality task that simulates everyday life (EPELI), watching natural-like videos, and resting state. Connectivity matrices were generated with the NiLearn Toolbox program using Seitzman and colleagues' (2018) 300-area parcellation. The connectivity of the whole brain was examined using Network-Based Statistics. During movie watching the ADHD group showed increased connectivity compared to the control group in a network that included several areas of the motor cortex. This may indicate a role for these regions in the hyperactivity symptoms of ADHD. The same network also included the right superior temporal gyrus, which has previously been linked to impulsivity symptoms in individuals diagnosed with ADHD. During the virtual reality task and resting state, no differences in connectivity were observed between the groups. However, differences between the experimental situations were revealed in several networks when the connectivity was compared within the groups. Many of these networks were very extensive and included several subcortical and cerebellar structures in addition to cortical areas. Both the control and ADHD groups showed increased connectivity in the resting state compared to EPELI. This could possibly be due to the differences in the participants' actions during the task performance. To the best of the author's knowledge, this study is the first to examine the functional connectivity of brains diagnosed with ADHD during naturalistic stimuli. The clear differences between the controls and the ADHD group during movie watching are promising for future naturalistic brain studies. Based on the results, network models are effective in studying the functional connectivity of ADHD under different conditions. However, consideration for similarity of activities during the virtual task could have led to the detection of larger differences.
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