Browsing by Title

Työn tavoitteena oli muodostaa kokonaiskuva sähköjärjestelmän sisäisistä ja ulkoisista vuorovaikutussuhteista. Sähköntuotantoa tarkasteltiin ympäristöönsä kytkeytyvänä systeeminä tieteidenvälisestä näkökulmasta. Tätä tarkoitusta varten rakennettiin menetelmä laajan kokonaisuuden hahmottamiseksi ja sovellettiin sitä sähköntuotannon alalle. Menetelmäperustuu grounded theoryyn ja systeemiajatteluun: Aineisto kerättiin ja luokiteltiin grounded theorya soveltaen. Systeemiajattelua käytettiin aineiston analyysissä ja aihealuetta kuvaavien mallien luomisessa. Aineisto kerättiin haastattelemalla sähköntuotannon ja -kulutuksen asiantuntijoita. Sen pohjalta rakennettiin useita alamalleja kuvaamaan sähköntuotantokompleksin osajärjestelmiä sekä näitä yhdistävä päämalli. Alamallien aihealueiksi muodostuivat tekninen sähköjärjestelmä, sähköjärjestelmän talous, sähkönsäästö, yhteiskunnallinen vaikuttaminen, ympäristö sekä tuotantorakenteen uusiminen. Lisäksi työssä esitettiin aineistosta nousseita kriteerejä hyvälle sähköjärjestelmälle ja sähköjärjestelmään liittyviä toimijoita. Mallien yhteydessä käsiteltiin myöseri tuotantomuotojen ominaisuuksia. Työn tuloksia tarkasteltiin kriittisesti kirjallisuuden valossa ja arvioitiin luotujen mallien sovellettavuutta. Malleja on mahdollista hyödyntää tieteenalarajoja ylittävän energiatutkimuksen jäsentäjänä sekä arvioitaessa kestävämmän energiapolitiikan edellytyksiä järjestelmätasolla.

Tiivistelmä–Referat–Abstract Background: Cancer is one of the leading causes of death around the world and in Finland. Ambitious research projects have been carried out for decades investigating cancer and how it spreads. Over 35 years ago, the systems that regulate vascular formation were discovered; the vascular endothelial growth factor (VEGF)-VEGF receptor VEGFR system and the more recent Angiopoietin-TIE system. These are the main endothelial growth factor receptor pathways involved in regulation of vessel quiescence and angiogenesis. The VEGF-VEGFR system is the first discovered endothelial cell (EC) specific receptor tyrosine kinase (RTK) signaling system. VEGF is a major growth factor involved in proangiogenic activity and vascular permeability when bound to its receptor, VEGFR2. Tumor cells take advantage of the VEGF-VEGFR2 system by secreting VEGF to stimulate angiogenesis in surrounding tissue to create new blood vessels allowing for greater access to nutrients and oxygen for tumor growth. The Angiopoietin-TIE system is the second EC specific RTK signaling system that was discovered. Angiopoietin-1 (ANG1) is the ligand for the TIE2 RTK. ANG1 is an obligatory TIE2 agonist and its effects on intracellular signaling, cell cytoskeleton, and junction-related molecules allows ANG1 to restrict the amount and size of gaps that are formed at EC junctions in inflamed vessels, increasing barrier function and decreasing vascular permeability. Angiopoietin-2 (ANG2) is an autocrine context-dependent TIE2 agonist/antagonist which is implicated in stimulating pathological angiogenesis, inflammation and vascular permeability. Integrins are important cell surface receptors that all cells use to communicate with their environment. Recently, it has been discovered that ANG2 is capable of inducing pathological angiogenesis, and can destabilize ECs when bound to integrin, specifically β1-integrin, via ANG2 N-terminal region. Objectives: The general aim of this study was to discover which part of angiopoiten-2’s N-terminus region was responsible for integrin activation. Materials and Methods: Fibronectin fragment containing type III 7-10 domains was produced and fluorescently labeled with Alexa 647. Integrin activation was measured using the fluorescently labeled Fibronectin III 7-10 and angiopoietins. Fluorescence activated cell sorting (FACS) was used to collect the results, which were analyzed using excel. Chimeric angiopoietins were produced using retroviral vectors and used for FACS experiments. A cell internalization assay was performed in Hela cells using CellTracker™ Orange CMRA and angiopoietin proteins, stained with secondary antibody anti-human Alexa 488 and Texas Red Phallodin. Results: Optimization of the FACS assay defined the minimum number of cells required to reliably measure integrin activation and showed that BD Accuri FACS machine was better suited than Guarva FACS machine for the assay and that the amount of integrin varied between cell passages used for the assay. In addition, it was essential to ensure a homogenous mix of cells and recombinant proteins during the assay and the quality of the produced FN III 7-10 was critical for the success of the assay. Results from the FACS assay confirmed that ANG2 is capable of activating integrin. In addition, chimeric angiopoietins that were expressed and secreted from CHO cells, were capable of activating integrins to a variable degree. The results confirmed the importance of ANG2 N-terminus in integrin activation. Cell internalization assay visually demonstrated angiopoietin binding to Hela cells. ANG2 was internalized by the cell and resistant to the acid wash, while the majority of ANG1 bound to the cell surface was washed away by acid wash. Conclusions: In this thesis work, integrin activation assays were optimized and carried out, along with cell internalization assays, to determine which specific part of ANG2 is responsible for inducing integrin activation. The findings from this work confirmed that ANG2 is capable of activating integrin. Several chimeric constructs were successfully expressed in CHO cells, and the cell supernatants were used to activate integrins. However, more studies are needed to determine which specific region of ANG2 is responsible for integrin activation. Investigating angiopoietin induced integrin activation would allow for a better understanding of the angiopoietin signaling pathway with potential translational significance.

Driven by the growing world population and increased food demand, the conversion of natural habitats into agricultural lands is occurring at an unprecedented rate. This is in turn the leading driver of the current biodiversity crisis, particularly in tropical forest areas characterised by prominent species diversity. Due to an accelerating cash-crop expansion, the once forest-savannah landscape of Northern Guinea-Bissau is now notably occupied by cashew orchards, in addition to scattered forest patches and floodable rice paddies. So far, it remains unknown how these kinds of human-modified landscapes can sustain mammal species over contrasting seasons, and how the habitat use varies across feeding guilds. To address this gap, this study aimed to understand how medium-sized mammals make use of mosaic-like landscapes in Northern Guinea-Bissau by considering the interactive effects of (1) habitat type: forest patch, cashew orchard and rice paddy and (2) season: before (June-July 2022) and after (October-November 2022) the peak of the rainy season. To do so, in each of the two seasons we conducted a 30-day consecutive camera-trap survey across seven landscapes, each of which comprised the three habitat types, amounting to 21 sampling sites. I evaluated mammal species richness, abundance (number of records) and composition across habitat types and seasons at the assemblage-level and discussed the species-specific habitat dependency. I further analysed species abundance across four feeding guilds: carnivores, insectivores, herbivores and omnivores. Based on a sampling effort of 1200 camera-trap days and 940 records, I identified 21 mammal species from 10 families and five orders. At the assemblage-level, species richness and abundance were similar between habitat types, but higher after the peak of the rainy season, except for rice paddies. Forest and cashew habitats shared similar species composition, which differed from rice paddies. Habitat-dependent species were found in each of the habitat types. At the feeding guild-level, in both seasonally periods carnivores and insectivores were less abundant in cashew orchards, while omnivores were more abundant. The results demonstrate that habitat conversion is likely to disrupt the functional structure of mammal assemblages. Maintaining heterogeneous landscapes, including both forested and open-area habitats, is crucial to maximize the integrity of mammal assemblages in Northern Guinea-Bissau. These findings can be used as baseline information in effective conservation measures in Guinea-Bissau and other tropical regions undergoing rapid conversion for cashew cultivation.

Climatic cycles lead to changes in habitat suitability, which in turn can lead to allopatry, i.e. isolation, between populations. Lack of gene flow between allopatric populations causes them to diverge through accumulation of genetic differences that can create incompatibilities between lineages upon secondary contact in the form of lowered survivability or reproduction rate in hybrids. Incompatible genes act as reproductive barriers and keep lineages isolated by selection against hybrids, while gene flow and recombination work as a counterforce to selection promoting admixture. Reproductive barriers like these are most often found inside hybrid zones and are well demonstrated in nature, but the isolating effect of individual genetic incompatibilities on genome-wide gene flow is still an open question. Here we test if selection counteracting gene flow maintains a narrow hybrid zone between two subspecies of the meadow grasshopper Pseudochorthippus parallelus. We targeted 0,01 % of the 13 GB genome, recovering a 29,1 mean coverage per locus per individual in targeted regions, when mapping against a transcriptome. We find that, for the nuclear markers, the hybrid zone is narrower than expected under a neutral scenario of no selection, suggesting that it is maintained by selection against hybrids. We also find significant isolation by distance, suggesting gene flow across the hybrid zone despite selection against hybrids. Different parts of the genome show significant excess or deficit of heterozygotes, suggesting that selection and gene flow are heterogeneous throughout the genome. Combined, our results show that reproductive isolation between recently diverged lineages can evolve quickly despite gene flow in neutral and positively selected sites.

Triple-negative breast cancer (TNBC) accounts for 10-15% of all breast cancer cases and has the worst clinical outcome. Characterizing features of TNBC are high recurrence and mortality rates, and the absence of three commonly targetable breast cancer biomarkers estrogen receptor, progesterone receptor, and HER2, limiting the number of targetable therapy options. Cytotoxic CD8 positive T cells play a crucial role in the anticancer immune response and act as a major component of successful cancer immunotherapies. However, cancer cells can evade T cell-mediated killing by overexpressing programmed death-ligand 1 (PD-L1) resulting in T cell exhaustion and limited immune response via the interaction with programmed death protein 1 (PD-1). Systemic anti-PD-L1/PD-1 therapies aim to prevent this immunosuppressive mechanism, but they are burdened with potentially life-threatening autoimmunity-type adverse effects. Therefore, cancer cell-specific targets to downregulate PD-L1 could offer efficacious and less harmful ways to overcome PD_L1/PD-1 mediated immunosuppression. Serine protease hepsin is commonly overexpressed in many solid tumors where it is responsible for the activation of HGF/MET signaling pathway as well as degradation of desmosomes and hemidesmosomes leading to the loss of epithelial integrity, invasion, and metastasis. Earlier studies have linked hyperactive HGF/MET pathway to the upregulation of immune checkpoint molecule PD-L1. In this thesis, I show how pharmacological inhibition of hepsin leads to decreased MET activity and downregulation of PD-L1 in a panel of TNBC cell lines. My results demonstrate the potential of hepsin-mediated regulation of PD-L1 in tumor immunosuppression, and hint at the potential of hepsin as a therapeutic avenue towards safe and efficacious immunotherapy in the future. These results are part of a larger study addressing the role of hepsin as a regulator of PD-L1 breast cancer.

Climate change is expected to increase the frequency and intensity of heatwaves in Finland. Heatwaves increase morbidity and mortality, especially among the elderly and chronically ill. Home care services are in an important role supporting the functioning ability and quality of life among populations vulnerable to heat. This study presents a qualitative empirical case study to understand heatwave adaptation in municipal home care services in Helsinki. The purpose of this study is to explore, describe and analyse the effects of the 2018 heatwave in the context of home care services, and the services’ capacity to adapt to heat. The data was collected by conducting semi-structured interviews with home care workers both on the operational and strategical levels. Theoretical frameworks regarding extreme heat vulnerability and local adaptive practices were used to support the analysis. The results suggest that heatwaves have caused challenges for both workers and customers. While the working ability of home care staff may be reduced due to thermal stress, the customers’ reduced functioning ability may simultaneously increase the need for care. Some workers had experienced fatigue, and customers were described having typical heat exhaustion symptoms. Workers adaptive practices were mostly reactive responses aimed at reducing the vulnerability of customers to heat. Interviewees described having taken preventive measures to limit customers’ exposure to heat and prevent adverse heat-related health outcomes by monitoring customers’ hydration more carefully. No long-term preparedness measures were identified on the strategical level. The city had supported workers by offering mineral drink bottles and providing instructions for coping in hot weather. In conclusion, workers have an active role in adaptation, but it might be limited by their own thermal stress, customers’ different health conditions and resources, as well as service-level stressors such as lack of staff. Because heatwaves are expected to increase in the future, home care services’ preparedness to heatwaves might need to be improved. It would be important to consider adaptation needs of both workers and customers as they can be interrelated in the sense, that care is dependent on workers ability to function. In a wider societal framework, the adaptive capacity of home care should be secured through providing sufficient resources for these services. The latter is even more important in the light of future trends regarding the aging of population, increase in need for home-based services and decrease in workforce availability. More research is required on the effects of heatwaves on the most vulnerable populations, what adaptation needs there might be as well as how this relates to service provision.

Pluripotent stem cells (PSC) can exists in both primed and naïve states. The conventionally derived human PSCs represent the later primed state of pluripotency during embryo development, while the naïve state resembles the inner cell mass (ICM) of pre-implantation blastocyst. Primed human PSCs can be reverted chemically by transient histone deacetylase (HDAC) inhibition back to the naïve state in vitro. The reverted PSCs can then be characterized based on their morphology and expression of selected naïve markers using immunocytochemistry and RT-qPCR assays. Leucine twenty homeobox (LEUTX) is one of the genes expressed during the early stages of embryo development and is capable of activating the transcription of multiple genes, including pluripotency-associated genes, which are upregulated during the human embryonic genome activation (EGA). LEUTX expression could potentially improve the naïve reversion efficiency or the maintenance of naïve PSCs by driving the transcriptome of primed PSCs back towards the earlier cell stages of embryo development, potentially even to cell stages that precede the naïve state. The aim of this thesis was to setup the naïve reversion protocol and study the effects of LEUTX on the reversion by using the generated and tested H9 activator cell line for targeted activation of endogenous LEUTX expression. First, a conditionally stabilized CRISPRa activator cell line was generated for targeted activation of endogenous gene expression in H9 cells. Then sequence-specific guide RNAs (gRNA) targeting LEUTX for activation were introduced to the activator cell line. Using the generated activator cell line during the naïve reversions allows the targeted activation of specific genes, here LEUTX, and thus enables studying the effects of these genes on PSCs during the naïve reversion protocol. The induced activator cells expressing LEUTX managed to form four times as many naïve resembling colonies during the reversion compared to the controls, but most of these were lost after changing the medium conditions towards the end of the protocol. After the reversion was complete, the reverted PSCs were characterized as naïve PSCs based on their domed morphology and the high expression of naïve markers NANOG, KLF17, TFCP2L1 and DNMT3L when compared to the primed PSCs. The naïve reversion protocol was set up and optimized successfully and can now be used as a reliable way of obtaining human naïve PSCs for further experiments studying and modelling the earlier developmental stages during embryo development. Furthermore, the generated H9 activator cell line worked as intended and can be utilized for studying the effects of other targeted genes during the reversion or in the reverted naïve PSCs.

Binge eating disorder (BED) is a common eating disorder that includes eating a large amount of food in a short period of time and is often associated with obesity. Patients can suffer from stress, anxiety, and metabolic syndrome caused by the weight gain, but no effective medication for both psychological and physiological issues have been discovered. This thesis studies the potential of short-chain fatty acids (SCFA) in reducing binge-like eating behaviour in a mouse model that does not include food restriction. The model includes a 24 h bingeing period with high-fat food once a week. SCFA butyrate, propionate, and acetate were administered to mice via 1 g/kg i.p. injections or 200 mM drinking water for three days, and their effects on energy intake during the bingeing period were measured. The results show that SCFA can significantly reduce binge-like eating behaviour in mice in the short term, but long-term effects vary. I.p. butyrate, propionate, and acetate decreased energy intake by 68%, 57%, and 62% during the first hour, respectively. SCFA via drinking water did not decrease energy intake a lot, and the results were inconsistent between animals. These results suggest a potential for SCFA to attenuate bingeing episodes when administered acutely, but the mechanisms remain to be discovered.

Eliölajit ovat jatkuvassa vuorovaikutuksessa niin fysikaalisen ympäristönsä, kuin muiden eliölajienkin kanssa. Nämä vuorovaikutukset voivat olla osapuolille negatiivisia, neutraaleja tai positiivisia. Fasilitaatio tarkoittaa sitä, että joku laji edesauttaa toisen lajin selviytymistä. Äärimmäinen esimerkki fasilitaatiosta on perustajalaji. Perustajalaji on laji, joka pelkällä läsnäolollaan tarjoaa muille eliöille kriittisiä resursseja, joita ilman muut lajit eivät voi selviytyä. Nämä resurssit voivat olla esimerkiksi ravintoa, suojaa tai elintilaa. Alustaansa kiinnittyvät simpukat ovat perustajalajeja, joita tavataan kaikilta maailman merialueilta. Esiintyessään tiheinä kasvustoina kalliorannoilla ne tarjoavat lukuisille selkärangattomille eläimille suojaa aallokon voimalta ja pedoilta, ravintoa ja elintilaa lisääntyneen sedimentaation myötä, sekä runsaasti sekundaarista kiinnittymispintaa. Alustaansa kiinnittyvät simpukat muodostavat perustan habitaatille, joka lopulta muotoutuu simpukoiden ja muiden lajien yhteisvaikutuksella. Vähäsuolaisessa ja -lajisessa Itämeressä sinisimpukka (Mytilus trossulus x edulis) on alustaansa kiinnittyvänä simpukkana erittäin tärkeässä roolissa. Sinisimpukkaa esiintyy pinnan läheltä aina kymmenien metrien syvyyteen asti, kunhan kiinnittymiseen sopivaa alustaa vain on tarjolla. Simpukoiden esiintymiseen Itämeressä vaikuttavat monet tekijät, joista tärkeimpinä ovat aallokko joka aiheuttaa fysikaalista stressiä, ja alhainen, itää ja pohjoista kohti pienentyvä suolapitoisuus joka aiheuttaa fysiologista stressiä. Paikoin erittäin runsaina esiintyvät sinisimpukat edesauttavat sedimentin kertymistä muodostamansa kolmiulotteisen matriisin onkaloihin mikä mahdollistaa myös pehmeän pohjan lajien esiintymisen kalliorannoilla. Sinisimpukkayhteisössä onkin piirteitä sekä pehmeistä, että kovista pohjista, mikä tekee niistä edullisia elinympäristöjä runsaalle määrälle erilaisia selkärangattomia eläimiä. Tässä tutkimuksessa tutkittiin sisäsaaristosta ulkosaaristoon voimistuvan aallokon vaikutusta sinisimpukka-rakkohauru -yhteisön rakenteeseen kolmella lähellä toisiaan sijaitsevilla merialueilla läntisellä Suomenlahdella, missä suolapitoisuus laskee läntisimmän ja itäisimmän tutkimusalueen välillä ainoastaan muutamilla promillen kymmenyksillä. Näytteitä noudettiin sukeltamalla 36:sta eri pisteestä kolmen metrin syvyydeltä rakkohauruvyöhykkeeltä, neljältä eri avoimuusvyöhykkeeltä. Näytteistä havaitut eläintaksonit tunnistettiin ja eläinten yksilömäärät laskettiin, ja sinisimpukoiden sekä rakkohaurun (Fucus vesiculosus) kuivapaino määritettiin. Tilastollisilla menetelmillä selvitettiin eliöyhteisön rakenteen eroja näillä kolmella alueella, sekä kasvavan avoimuuden vaikutusta. Lisäksi selvitettiin korreloivatko tavattujen eläinlajien yksilömäärät sinisimpukan tai rakkohaurun biomassan kanssa. Sinisimpukka oli kaikilla näytepisteillä runsain tavattu laji, ja jäljessä yksilömääristä puhuttaessa sinisimpukkayksilöitä ei huomioida. Alueet eivät eronneet toisistaan taksonien tai eläinyksilöiden kokonaismäärien osalta, mutta yksittäisillä taksoneilla eroja kuitenkin havaittiin. Avoimuudella havaittiin olevan merkittävä vaikutus eliöyhteisön rakenteeseen. Avoimuuden kasvaessa havaittujen taksonien määrä laski ja runsaimmat taksonit vaihtuivat sedimenttiä suosivista nilviäisistä vapaasti uiviin äyriäisiin, sen sijaan eläinten kokonaisyksilömäärissä merkitseviä eroja ei havaittu. Merkittävä osuus tavatuista yksittäisistä taksoneista korreloi negatiivisesti kasvavan avoimuuden kanssa, avoimuuden kanssa positiivisesti korreloivia taksoneita oli vähemmän. Sinisimpukoiden biomassan kanssa havaittiin korrelaatiota usealla taksonilla, kaikki positiivisia, sen sijaan rakkohaurun biomassan kanssa korrelaatiota havaittiin ainoastaan yhdellä taksonilla. Tämä tutkimus osoittaa, että avoimuudella on selkeä vaikutus perustajalajina toimivan sinisimpukan ympärille muotoutuneeseen eliöyhteisöön ja sen rakenteeseen. Suolapitoisuuden vähäinen lasku tutkimusalueiden välillä sen sijaan ei vielä merkitsevästi vaikuta sinisimpukkayhteisön rakenteeseen, vaikka yksittäisillä lajeilla - merkittävimpänä juuri sinisimpukka - viitteitä vaikutuksista havaittiinkin. Sinisimpukan merkitys muuhun lajistoon on merkittävä, rakkohaurun merkitys on sen sijaan varsin vähäinen.