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Soil nitrogen content and isotope ratios in urban remnant forests: What have dogs got to do with it? (2024)Keeping dogs (Canis familiaris) affects the environment. One significant effect of dogs in the environment is the production and deposition of urine. Unlike other waste, dog urine cannot be easily collected by owners, leading to unfiltered introduction into the soil. Dog urine is high in urea, which hydrolyzes to ammonium and nitrate. These soil nitrogen forms have the potential to change other soil properties that may cause harm in plant or microbial health, and contributing to eutrophication. In this thesis I investigate the impacts of dogs on soil nitrogen properties and their connection with human population density and closeness to walking paths in urban remnant forests. To evaluate the level of impact of dogs and to gain knowledge on the current nitrogen content and δ15N in urban remnant forests, five urban remnant forests were examined. Within each forest, soil nitrogen forms, δ15N in the soil and foliage samples, as well as total carbon and nitrogen content of the soil were compared, besides basic soil properties between dog-affected and control areas. The study revealed elevated levels of soil electrical conductivity, ammonium, and nitrate, as well as soil and foliage δ15N in dog-affected areas. Unexpectedly, soil acidity was lower in samples taken next to paths where dogs can potentially urinate. Organic matter content and total nitrogen and carbon content of the soils were higher where dogs are not frequent visitors, meanwhile the carbon-to-nitrogen ratio was also higher. These findings show that the impacts of dog urine on the soil should be considered in urban planning and further research is needed to understand the significance of it.
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(2021)Underwater light climate in mountain lakes is controlled by dissolved organic carbon concentrations and by lake ice regimes. Both are affected by local, regional and global variables linked to anthropogenic disturbances such as climate change and atmospheric pollution. Aim of this work was to investigate changes in underwater light climate over the past ~200 years in two oligotrophic mountain lakes and how it reflects on diatom (Bacillariophyceae) guild distribution. For these aims, diatom communities and ecological guilds were analyzed from sediment core and contemporary habitat samples along a depth gradient. In addition, sediment inferred chlorophyll a (CHLa) and lake water total organic carbon (TOC) were analyzed to detect development of primary production and lake water carbon content. Results showed that acidification of the lakes together with climate induced changes have been important drivers of the ecology of the lakes. Lake water TOC showed a decline and subsequent increase in line with the acidification and subsequent recovery of the lakes, likely affecting underwater light climate in the lakes. However, this did not reflect unambiguously into changes in diatom functionality. Warming has likely contributed to diversification of the diatom community over the study period while no distinct increases were observed in whole lake primary production. Overall, if the present study could not distinguish the exact role of underwater light in driving changes in diatom communities and functional traits, the result show that human pressures have left distinct imprints in the development of biotic communities in these remote mountain lakes.
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(2017)Chitinolytic enzymes belong to a group of pathogenesis-related proteins, which are induced in plants by a pathogenic attack. They have also been shown to function in abiotic stresses and related to signalling. Chitinases catalyze the degradation of β-1,4-N-acetyl glucosamine units of chitin. Chitin is not found in plants, but for example the shells of invertebrates, cell walls of certain fungi, algae and bacteria contain it abundantly. Lately chitinases have gathered attention because of their potential utilization possibilities in medicine, agriculture, food industry and biofuels. Strong chitinase gene expression and also chitinolytic activity has been found in a spruce (Picea abies L. Karst.) cell culture, which produces apoplastic lignin into the medium. Plant chitinases are known to be induced also in developmental processes. Chitinase-like genes have been shown to be essential in cellulose synthesis and in development of cell walls and tracheary elements and also affected the localisation of lignin in tissues. Therefore, the secretion of chitinases into the culture medium has not necessarily been caused by stress and chitinases might use also some endogenic substrate in addition to chitin. Because chitin is a structural component of plant pathogens but not present in plants and the growing circumstances of the studied aceptic spruce cell culture were stable, the research hypothesis was, that the secreted chitinases in the culture medium were not a stress reaction but related to development and besides chitin of the pathogens they utilize some own polysaccharide substrate of spruce. Additional hypothesis was connection of the strong expression of chitinases and chitinase-like genes in the culture medium and the production of apoplastic lignin by the spruce cell line. The aim of the study was to characterize the chitinases and chitinase-like enzymes of the spruce cell suspension culture medium and developing xylem by isolating, purifying and cloning them and by producing them heterologously. The aim was also to identify the putative endogenous substrates for the chitinases and chitinase-like proteins to reach better understanding about the meaning of the chitinases in the development of the spruce tissues. Two chitinases of the spruce cell suspension culture were isolated and purified by chromatographic methods. One chitinase gene in spruce cell culture and one chitinase-like gene in spruce xylem were cloned, after which the chitinases and chitinase-like proteins were produced in Pichia pastoris yeast. The total proteins of the culture medium, the purified chitinases isolated from the medium and the chitinases and chitinase-like proteins produced in P. pastoris were used to examine their substrate alternatives with the carbohydrate microarray method. The connection between apoplastic lignin and the chitinolytic enzymes in culture medium was not found and obvious candidates for the endogenous substrates were not detected. However, the mannan degradation in the presence of the chitinases was strong enough to give reason to further analyses.
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(2020)Extracellular vesicles (EVs) are lipid bilayer-enclosed nano-sized particles that are found in all body fluids. EVs are part of normal cell functions and they can carry for example proteins, RNA and lipids between cells. This makes them potential candidates as drug delivery vehicles. When nanoparticles are introduced to blood plasma, a plasma protein structure is formed on their surface, called the protein corona. The formation of a protein corona is a dynamic process, and the proteins are binding to the corona depending on their affinity to the nanoparticle surface. When administrating nanoparticles to cells, protein corona has a big impact on the half-life and cellular uptake of the particles. The aim of this work was to study the plasma protein corona of PC-3 derived EVs, and to investigate methods for protein corona isolation. First, EVs and lipoprotein particles were removed from fresh frozen plasma by membrane filtration. Using this filtrated plasma, we then tested plasma-EV incubation and removing of free plasma proteins by simple ultrafiltration. Removing of free plasma proteins was also examined by size-exclusion chromatography (SEC). The resulting EV-protein corona complex was visualized with SDS-PAGE. EVs were characterized with nanoparticle tracking analysis and western blotting. Of these two methods used, SEC appeared to be more convenient and efficient way to remove the free plasma proteins from the EV-protein corona complex. The method still requires further development and testing in order to perform optimally.
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(2023)The thesis explores possibilities for mental health promotion in urban planning trough a case study of the community space in Lapinlahti, Helsinki. Lapinlahti is a former psychiatric hospital, and the area has in the 2010’s been developed through community efforts into a center for mental wellbeing and culture in Helsinki, a “Lapinlahti for all”. The future use of the area has been contested for the past years, with the City of Helsinki and the organizations at Lapinlahti having different views on how the area and it’s cultural and historical mental health legacy should be preserved and developed. In the discussions, statements and visions, the area’s mental health value has also been translated into urban planning, taking many forms. Mental health is a value that has traditionally not been prioritized or regarded as a value of its own in urban planning, and although the relationship between urban form and wellbeing is well known, urban (mental) health is seldom tied to cities physical- and land-use planning (Kim et al, 2022; Corburn 2004). The thesis approaches this topic by analyzing mental health as a planning objective, discussing how different understandings and approaches to mental health have affected the planning visions surrounding Lapinlahti. The thesis uses a theoretical framework of urban governance and critical discourse analysis (CDA) to seek an understanding to the different approaches to the area’s development, and identify the different understandings, connections and visions that surround Lapinlahti. The study identifies three discursive constructions of Lapinlahti: Lapinlahti as a site for cultural heritage, Lapinlahti as a place for mental health, and Lapinlahti as a site for development. The results show that all actors in their development schemes have considered the area’s mental health value from a cultural-historical perspective, viewing the area as a site for mental health history. This history is however understood and manifested in different ways for the actors, influencing different visions for how the area should be developed and what interests and/or values that should guide the development. The thesis continues with presenting three approaches for mental health promotion in urban planning, concluding that planning should be attentive of how the space is governed, the relationship between mental health and the space, and the activities and meanings embedded in space.
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(2023)Temporal lobe epilepsy (TLE), a condition defined by unprovoked and recurrent seizures originating from the temporal lobe, is among the most ubiquitous of the various forms of epilepsy. Despite being chronic and highly prevalent, the available treatment options concerning the same remains a critical issue. Since the current therapeutic condition of epilepsy requires more development, renewed focus studying its molecular mechanisms and therapies is imminent. One of the longstanding theories trying to decode the molecular perturbations in TLE has been deficits in GABAergic inhibition resulting in abnormal neuronal activation. K+ - Cl- co-transporter (KCC2) activity is vital for maintaining a hyperpolarizing GABA response. The past decades have intimately and causally linked the prognosis of the seizures observed in TLE with deficits in KCC2 functioning. However, the precise mechanisms relevant to the disruption of KCC2 activity are still blurry. Here we show how KCC2 de-stabilization/localization in the neuronal bilayer is a characteristic of epileptic animal tissue. With the help of co-immunoprecipitation assays, western blot, and mass spectrometry, we found that in normal healthy brain tissue, GM1 ganglioside present in the membrane has specific and direct interactions with the KCC2 cotransporter. However, in the pilocarpine model of TLE, the interaction of this complex was significantly disturbed, primarily in the hippocampus and to some extent in the cortex. Our results act as an extension to previous research which stated that the structural association of the KCC2 clusters with neuronal lipid rafts is crucial for the functionality of the KCC2 cotransporter. Having learned about the unique nature of the pathophysiology of TLE, it is imminent to note that additional research in the direction of studying its biochemical pathways is required. The findings of this experimental study support the claim that KCC2 and GM1 as a complex are closely associated in the epileptic conditions and hence, this research paves the way to further explore the role of KCC2 and GM1 as a consequential complex in the pathophysiology of TLE.
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(2024)Prostate cancer is one of the most common cancers diagnosed in men. Somatic copy number alterations (SCNA), such as the deletion of PTEN or NKX3.1 and the amplification of MYC, have been associated with prostate cancer progression and could serve as potential biomarkers during diagnosis. One approach to utilize this information would involve screening a large number of prostate tissue sections for SCNAs and subsequently validating the findings using a secondary method. This process could enable more personalized treatment options for cancer patients. This thesis aimed to create a robust and reproducible workflow for SCNA identification. This was achieved by optimizing a chromogenic immunohistochemistry (IHC) protocol using immunostaining chambers and open-source 3D-printed laboratory hardware. The optimized protocol was then transferred to an automated liquid handling robot, and a panel of three antibodies for PTEN, NKX3.1, and MYC was developed for SCNA screening with IHC. Additionally, a chromogenic in situ hybridization (CISH) protocol was optimized to validate the results of the IHC. The immunostaining chambers required a lower antibody dilution to perform comparably to the manual IHC stainings. The automated protocol using the liquid handling robot produced high-quality stains with optimized dilutions. The optimized CISH protocol successfully identified the presence of the target gene, but unclear signals and merging of the signals obstructed detailed analysis. While complete deletion of PTEN was detectable, determining the number of gene copies per cell proved challenging due to signal variability and sample-dependent problems. Further optimization of the CISH protocol or development of an automated analysis workflow tailored to address these challenges is needed for more accurate analysis.
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(2021)The Glanville fritillary (Melitaea cinxia) butterfly inhabiting the fragmented meadows and pastures in the Åland Islands, Finland, has a classic metapopulation structure: its long-term persistence depends on frequent re-colonization events counter-balancing the extinctions of local populations. The spatial structure and the temporal dynamics of the metapopulation are likely to influence genetic variation within and among local population networks. With high population turnover, population declines accelerate genetic drift, potentially leading to a reduction in neutral genetic diversity. This loss is likely to be counteracted with immigration bringing in new alleles especially in well-connected populations. Dispersal has indeed been shown to be a key mechanism in maintaining genetic variation and adaptive potential in fragmented landscapes. In my MSc-thesis, I am utilizing long-term monitoring and genetic data collected from semi-independent networks from the main Åland region (Saltivik) and from two isolated island networks Föglö and Sottunga. Specifically, I investigate how genetic variation varies in time and space, in relation to demographic change and whether the responses vary among well- and poorly-connected networks and/or between island and mainland networks. My results showed that the total number of nests fluctuated in all the networks across time. Heterozygosity appeared to track the changes in population abundance but this tracking varied among the networks. Although connectivity did not explain the change in heterozygosity during the decline years, allele frequencies shifted over time and the speed of change in allele frequencies differed among networks.
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(2023)Grasslands occur on every continent except Antarctica and are a key part of agricultural and urban landscapes in many parts of the world. In Finland, many of the grassland species are threatened nowadays. Fragmentation, worsening quality and shrinking area of habitats are the biggest threats to grassland species. Many grassland plants, however, can be found in urban habitats such as road verges, different meadows and rocky outcrops. Suitable urban habitats could be important for these species if the degradation of their habitats continues elsewhere. Indicator species could provide information about the ecological quality of urban habitats and thus the suitability for grassland specialists. My aim for this thesis was to determine how indicator species can be used to assess the ecological quality of urban grasslands by answering the following research questions: 1. Does habitat type affect the abundance and occurrence of known quality indicator species of semi-natural grasslands? 2. Which species are characteristic to different urban grassland habitat types? 3. How different sites relate to each other in terms of vegetation? 4. How do the measures used to answer the other three questions compare together? I collected vegetation data from 28 study sites in the cities of Espoo, Helsinki and Vantaa, southern Finland. In addition, a dataset consisting of 116 sites from these cities, collected by another person, was included here. The study sites were divided into eight habitat types: dry and mesic landscape grasslands, dry and mesic road verges, dry and mesic valuable grasslands, rocky outcrops and valuable rocky outcrops. From all study sites, I collected a list of plant species observed within a standardized time frame by doing an on-site vegetation inventory. In addition, I observed the frequencies of plant species within 1 x 1 m vegetation quadrats. I used one-way ANOVA and Tukey-Kramer test as well as Welch’s ANOVA and Games-Howell test to study the effect of habitat type on abundance and occurrence of semi-natural grassland indicator species. I used IndVal to identify which species are indicators for the habitat types and their combinations. I used NMDS to study the grouping of different habitat types and their species. Habitat type affects the abundances and occurrences of semi-natural grassland indicator species in urban settings and these species can be used in evaluation of the ecological quality of urban grasslands. Indicator species produced from observed data seem to have a limited potential in evaluating the ecological quality of urban grasslands, but they provide a good method for classifying urban grassland habitats, especially valuable rocky outcrops and valuable grasslands. Study sites do form groups according to their vegetation, but these groups do not entirely follow the habitat type division used in the study. With right standardization for the use, indicator species can be a valuable tool in assessing the ecological quality of urban grasslands.
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(2021)Natural scientists study a wide variety of species, but whether they have identified all studied samples correctly to species is rarely evaluated. Species misidentification in empirical research can cause significant losses of money, information, and time, and contribute to false results. Thus, I study the abundance of species misidentification and ecologists’ perceptions of such mistakes through a web survey targeting researchers from scientific institutes around the globe (including universities, research societies and museums) who completed their doctoral degree in any ecology-related field of science. I received 117 responses with either work or educational background from 30 countries. I found that species misidentification widely existed in respondents’ research: almost 70% of the respondents noticed species misidentification in their own research, while the estimated proportion of existing studies with species misidentification was 34% (95% CI: 28% - 40%). Although misidentification was mainly found during specimen collection, specimen handling and data analysis, misidentifications in reporting stages (writing, revision and after publishing) could persist until publication. Moreover, according to respondents, reviewers seldom comment about species identification methods or their accuracy, which may affect respondents’ (both leading and not leading a research team) low reporting frequency about the possibility of misidentification. Expert checking, training students, and DNA barcoding are the most prevalent approaches to ensure identification accuracy among respondents. My results imply that species misidentification might be widespread in existing ecological research. Although the problem of species misidentification is widely recognized, such an issue seldom be appropriately handled by respondents. To increase the accuracy of species identification and maintain academic integrity, I suggest that researchers need to focus more on the study species (e.g., sampling process, identification method, and accuracy) when writing and reviewing papers. Furthermore, I appeal for guidelines about reporting species identification methods and their accuracy in papers, as well as research on education about identification skills in universities, as these two topics may constrain the precision of species identification.
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(2020)The antibiotic resistance of pathogenic bacteria is becoming a major problem in treating bacterial infections and development of new antibiotics is very challenging. In traditional phage therapy the bacteriophages, viruses that infect bacteria, are being used as an optional treatment to eliminate infectious agents. Methicillin resistant Staphylococcus aureus (MRSA) is resistant to several currently used antibiotics and is one of the most common antibiotic resistant bacteria causing infections. Therefore, it is a potential target for the phage therapy. Some of the Staphylococcus aureus strains produce several different enzymes and toxins which can be harmful to patients. Products developed for phage therapy purposes need be free from the material originated from host bacteria. In this study, three different methods were tested for the purification of bacteriophages infecting S. aureus. The main goal was to produce phage lysates with purity and phage concentration suitable for therapeutic purposes using a fast and aseptic procedure upgradable for large volumes. The tested methods were ultrafiltration with filter tubes from two different manufacturers (Sartorius Vivaspin 6 ja Merck Millipore Amicon Ultra 4), polyethylene glycol (PEG) precipitation and ion exchange chromatography. Three different bacteriophage strains were used. One was isolated from a commercial Russian phage therapy product (vB_SauM_fRuSau02) and the other two from feces of pigs (vB_SauS_fPf-Sau02 and vB_SauS_fPfSau03). Host bacteria strains for the first bacteriophage were S. aureus strains TB4 and 13KP originally isolated from human infections. Two host strains for the latter two phages were MRSA strains isolated from healthy pigs. Purification of the phage lysates was evaluated by measurement of enterotoxins produced by S. aureus bacteria, measurement of free double stranded DNA (dsDNA), and by cytotoxicity test in cell cultures. All evaluation methods were commercially available tests. To determine how much of the bacteriophages were lost in the process, the phage concentrations of the lysates were determined before and after the purification and recovery rates were calculated for the viruses. After two separate ultrafiltrations, the recovery rates of the bacteriophages were mainly good, but there was a lot of variation in the results. The lowest recovery rate calculated was 5%, the highest 57%, and the mean of all the rates 24%. In this study the ion exchange chromatography was combined with ultrafiltration which was used in pre-cleaning of the lysates and changing the phages in a buffer suitable for the chromatography. The recovery rates from the ion exchange chromatography varied between 14-26% but the results may be affected by the ultrafiltration steps performed before and after, since a lot of variation was seen in ultrafiltration processes. PEG precipitation was performed for one phage lysate only in order to compare the laboriousness of the method and the rates of the recovery to the other methods used. The rate of recovery from the PEG precipitation was 9,5% which was fairly low. The purity of this lysate was not evaluated since the method was estimated to be too laborious compared to the other methods. Ultrafiltration turned out to be an efficient method in the removal of small protein molecules, such as enterotoxins from bacteriophage lysates. With two sequential ultrafiltrations 96-99% of the enterotoxins in the lysates were removed. The removal of the free dsDNA was also successful but there was variation between the phage lysates. Approximately 67-93% of the free dsDNA was removed but it is possible that some of the measured DNA originated from lysed bacteriophages as their genome also consists of dsDNA. Ion exchange chromatography produced extremely well purified phage products. The fractions had no enterotoxins left or the amount was below the detection limit of the test (<0,5-1 ng/g). Ion exchange chromatography was able to remove 96-99% of the free dsDNA of the lysates. It is possible that some of the DNA left in the lysates originated from the bacteriophages lysed during the process or in storage after that. When comparing how simple and fast the methods were, the ultrafiltration turned out to be superior. It can be used in fast production of bacteriophage products for the treatment of S. aureus infections. The purification achieved with the ultrafiltration should be adequate for a topical use of the product. When higher purity products are required, e.g. for administrating the product intravenously, ion exchange chromatography might be a safer option.
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(2019)Climate change is one of the most major environmental challenges of our time. The upcoming decades will be crucial for delimiting the global average temperature rise to 1.5 - 2 degrees, as pointed out by the IPCC. Tackling climate change requires system-wide changes, emission reductions from all sectors and a shift towards low-carbon society. The global food system is responsible for circa 20-30 percent of global greenhouse gas emissions. Therefore, achieving the goal of carbon neutrality necessitates profound changes in the food system, i.e. in the ways our food is being produced and consumed today. This, in turn, necessitates a transition towards more plant-based diets. There is a scientific consensus that plant-based diets are - in addition to human health - also beneficial for the climate, as they produce less greenhouse gas emissions. Carbon-intensive lifestyles are especially characteristic for people in industrialized countries. Food covers roughly one fifth of the annual carbon footprint of a Finn. Finnish diets produce relatively large emissions mainly due to high consumption of meat and dairy. As food choices contribute to climate change, it is crucial to seek solutions in order to steer sustainable food consumption. In addition to the efforts of the public sector, proactive companies are also capable of steering or “nudging” sustainable food consumption also by other than conventional means, such as information provision and financial incentives. These means and their acceptability are addressed in this thesis. This thesis explores how Finnish consumers allocate responsibilities between the government, companies and consumers for mitigating climate change. In addition, this thesis elucidates how Finnish consumers perceive food manufacturers’ role and contribution (means) in steering food consumption towards more climate-friendly choices. The study conducted for this thesis was a consumer survey (n=504) and the target population was Finnish consumers aged between 18-74 years. The material was collected during August 2019, and statistical methods were mostly applied to analyze the results. One of the key findings of this study is that half of the survey respondents perceive a shared responsibility between the government, companies and consumers in mitigating climate change. While least responsibility is assigned for consumers, their role is not, however, underestimated. It also seems like food manufacturers play an important role in steering climate-friendly food consumption in the respondents’ opinion. In addition, the respondents seem to be relatively accepting towards the fact of being steered by food companies in order to make more climate-friendly food choices. Also the means to steer consumer behaviour, which were addressed in this study, received a promising level of support from the respondents. The findings of the study shed light on the food industry’s opportunities to influence climate-friendly food consumption and can help to develop policies that aim to mitigate climate change and promote more sustainable culinary culture.
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(2020)Jäkälät ovat sieniosakkaan ja yhteyttävän levän muodostamia symbiooseja, joista suurin osa elää maaekosysteemeissä ympäri maapalloa. Jäkälät toimivat ekosysteemeissä tärkeinä indikaattorilajeina, sillä ne ovat usein hyvin herkkiä elinympäristössä tapahtuville muutoksille, esimerkiksi ilmanlaadun huononemiselle. Sticta-suvun syanojäkäliä on tähän mennessä kuvattu noin 200 lajia erityisesti subtrooppisilta ja trooppisilta alueilta. Todellisuudessa lajimäärän arvioidaan olevan paljon suurempi. Afrikan jäkälistä tiedetään vielä suhteellisen vähän, ja tietomme Itä-Afrikan Sticta-lajeista pohjautuvat pääasiassa 1980-luvulla tehtyyn tutkimukseen. Tässä tutkimuksessa tavoitteenani oli tuottaa DNA- analyysiin perustuva selvitys Sticta-suvusta Kenian Taitavuorilla. Tutkimuksessa selvitin myös Sticta-lajien symbioottisten syanobakteerigenotyyppien levinneisyyttä Taitavuorten eri metsäsaarekkeissa ja eri Sticta-lajeissa. Tutkimukseni aineistona oli 176 jäkälänäytettä, jotka ovat kerätty Taitavuorten alueelta sekä Elgonvuoren rinteeltä Länsi-Keniasta Jouko Rikkisen ja Ulla Kaasalaisen toimesta vuosina 2009- 2017. Alustava lajinmääritys morfologian perusteella tapahtui mikroskopoimalla. Neljänkymmenenkahdeksan näytteen tarkempi lajinmääritys tehtiin DNA-menetelmin, selvittämällä jokaisesta näytteestä sienen sekä syanobakteerin genotyyppi. DNA-eristyksen, PCR:n sekä elektroforeesin tein Helsingin luonnontieteellisen museon laboratoriossa kesällä 2018. DNA- sekvensointi tehtiin Sanger-menetelmällä Saksassa GATC Biotech-yrityksen toimesta. Laboratoriotöiden tuloksena sain sieniosakkaan DNA-sekvenssin kaikkiaan 48 jäkälänäytteistä. Sekvenssien perustella 46 näytettä voitiin määrittää ennestään tunnetuiksi Sticta-lajeiksi, ja kaksi näytettä osoittautui Sticta pseudosylvatica -lajille sukua olevaksi, toistaiseksi kuvaamattomaksi lajiksi. Näytteistä löytyi kaikkiaan 21 erilaista syanobakteerigenotyyppiä. Eri metsäsaarekkeista kerättyjen näytteiden syanobakteerikoostumus ei näyttänyt poikkeavan toisistaan, mutta Sticta- lajien syanobakteerikannat poikkesivat toisistaan.
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(2013)The bioluminescent and toxin-producing dinoflagellate Alexandrium ostenfeldii forms dense blooms in coastal areas of the Baltic Sea. Bioluminescence, light production by an organism through a chemical reaction, is a nocturnal, rhythmic phenomenon in surface algae. In this study, the bioluminescence pattern and rhythm of A. ostenfeldii was under investigation. Procedures for continuous bioluminescence measurements, to support dedicated environmental monitoring of toxic dinoflagellate blooms, were developed. The study consisted mainly of laboratory experiments. Semi-continuous field measurements were included for comparison. In the laboratory, the light production of monocultures of A. ostenfeldii was measured with a spectroluminometer or bathyphotometer, continuously during the night, or for several consecutive days. The method to stimulate bioluminescence was varied, as well as the recovery period of the cells after stimulation. Light regimes during growth and pre-measurement adaptation were also taken into account. The experiments confirm that bioluminescence in A. ostenfeldii follows a circadian pattern and can be stimulated with the chosen methods. Bioluminescence could also be stimulated after culturing in continuous light. Measurement parameters for rhythm experiments (stimulation frequency and recovery period), were optimised. Multi-day experiments in complete darkness suggested that sufficient energy was available to maintain bioluminescent response during one night, although an endogenous rhythm remained present. These experiments gave insight to the phenomenon of bioluminescence regulation in A. ostenfeldii, but also gave rise to new questions. Some repeated measurements resulted in very low bioluminescence intensity, without an obvious reason. The light regime is not the only factor controlling bioluminescence. The interplay between bioluminescence and the growth and condition of the cultures is of interest.
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(2014)Havsekosystemen runt om i världen lider av antropogen inverkan såsom näringsutsläpp och utfiske. Speciellt utfiske av högre trofiska nivåer dvs. stora rovfiskar, anses ha ändrat dynamiken i flera näringsvävar. Ändringarna har lett till kraftiga trofiska kaskader som ökat eller minskat primärproduktionen speciellt vid kustliga vattenområden. Speciellt mesopredatorer har ökat i antal, och i Östersjön är storspiggen (Gasterosteus aculeatus) en art som börjat dominera samhället. Storspiggen konsumerar växtavbetare av trådalger, och kan därmed ha en effekt på trådalgstillväxten i Östersjön. Undersökningens syfte är att granska vilken roll storspiggens predation har i trådalgsbältet, och hur ändringarna syns i näringsväven. Undersökningen bestod av ett mesokosmexperiment och en fältundersökning vid Tvärminne zoologiska station. Mesokosmexperimentet bestod av två behandlingar: 10 mesokosm med två trofinivåer (utan storspigg), och 10 mesokosmer med tre trofinivåer (med storspigg). Trofinivåerna bestod av trådalger (Cladophora sp.), märlkräftor (Gammarus sp.) och storspigg. Experimentet pågick 4 veckor, vartefter mesokosmerna tömdes och innehållet analyserades. Med en veckas mellanrum gjordes fältbesök till den närbelägna ön Långskär, för trådalgsprovtagning, observation av storspiggstäthet och magsäcksanalysering av fångade storspiggar. Trådalgsproverna analyserades under mikroskop. Fältundersökningens resultat visade att mängden storspiggshanar och fångade storspiggar minskade längs med säsongen, medan trådalgsbiomassan ökade. Antalet vuxna storspiggar korrelerade negativt med märlkräftor. Mesokosmexperimentets resultat visade att mesokosmen med storspigg hade i medeltal högre trådalgs-biomassa (24%), högre epifytmängder (72%), färre märlkräftor (79%) och lägre färgintensitet än mesokosmerna utan storspigg. I Mesokosmerna utan storspigg hade trådalgsbiomassan inte ökat nämnvärt, men trådalgen var tydligt grönare i färg och saknade epifyter. Av resultaten framgår att storspiggen påverkar primärproduktionen i ekosystemet, genom en trofisk kaskad, då växtavbetarna regleras kraftigt. Primärproduktionen ökar mest i form av epifyter och mikroalger, vilket leder till att trådalgen kvävs och börjar föråldras. Trådalgen och växtavbetarna har därmed ett mutualistiskt förhållande. För att minska mängden skadliga algblomningar i Östersjön, borde näringsvävarnas stabilitet och mångfald tas i beaktande vid fiskeriförvaltningen för att effektivera Östersjöns rehabilitering.
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(2022)Sustainable consumption has become a widely debated topic in academic discussions, politics, the media, and consumer culture. The objective of this thesis was to identify the stories that are told about overconsumption and how responsibilities for the issue are distributed to different actors in these stories. Stories, in which events logically unfold and actors are positioned into different roles have a key role in the articulation of understandings of phenomena. 13 online news articles and two editorials published in three Finnish news media and the related news discussion forums were chosen as an empirical example of public discussion that shapes and reflects common perceptions on the issue. The news articles and editorials include references to the Earth Overshoot Day, limiting the context of the thesis to overconsumption that is defined based on the limits of the planet’s biocapacity. With the help of a qualitative narrative analysis, four storylines were constructed. These storylines are overconsumption by overpopulation, greed and impossible endless growth; one world, shared challenge, and technology as the problem and the solution. The most frequently mentioned actors across the storylines are citizens, developing and wealthy nations, decision-makers and politicians, businesses, and wealthy elite. Besides reflecting both academic discussions and previous literature on narratives about sustainable consumption, the presence of population dynamics and the conflict between socio-economic development and environmental well-being was notable in the analyzed discussions. The incomplete narrative structures, missing roles, and diverse elements included in the storylines reflect the complexity of the issue and the struggles related to articulating a coherent story with a sufficient solution and actors capable of solving the issue. To clarify the discussion on the topic, it is suggested that the distribution of the responsibilities and opportunities to improve the situation amongst actors should be addressed in the communication on sustainable consumption and the Earth Overshoot Day.
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(2022)Faculty: Faculty of Biological and Environmental Sciences Degree programme: Master’s Programme in Neuroscience Study track: Neuroscience Author: Emma-Helka Erkkilä Title: The brain physiology of stress and the effects of burnout on executive functions Level: Master’s thesis Month and year: 08/2022 Number of pages: 35 Keywords: executive functions, emotion, cognition, stress, burnout Supervisor or supervisors: Docent Kaisa Hartikainen and Lic.Med. Mia Pihlaja Where deposited: Helsinki University Library Additional information: Abstract: BACKGROUND- Burnout as a result of prolonged and excessive stress may impair higher order cognitive functions of the brain such as executive functions and their efficiency. This Master's thesis examines the effects of chronic stress on the brain, more specifically the effects of burnout on executive functions. The aim of this study was to specifically research the effects of burnout on executive and emotional functions and their interaction. The research was conducted at the Behavioral Neurology Research Unit, Tampere University Hospital as part of Sustainable Brain Health project funded by the European Social Fund. MATERIAL AND METHODS- 54 voluntary examinees of whom 51 were analyzed. The examinees were divided into two groups based on BBI-15 survey (27 suffering from burnout and 24 control subjects without burnout). The examinees performed a computer-based Executive reaction time (RT) test, during which a 64-channel electroencephalogram (EEG) was recorded. In additions all examinees received alternating transcutaneous vagus nerve stimulation (tVNS) and placebo stimulation. From the Executive RT test, we obtained objective measures reflecting the efficiency of executive functions (RT and total errors) and specific executive functions such as working memory, inhibition and attention. Additionally, the emotional stimulus included in the test enabled the assessment of the emotional functions and the interaction between emotional and executive functions. The EEG and tVNS results were not in the scope of this master’s thesis, and they will be reported later on. RESULTS- The results of this thesis are preliminary. Distinct positive correlation was observed between burnout assessment based on the BBI-15 survey and the results of the BRIEF-A self-report which measures the subjective experience of challenges in executive functions in daily life. There was no statistically significant (p<0.05) difference between the groups in RTs or errors made in the Executive function RT test. Instead, the groups differed on how the threatening emotional stimulus affected the accuracy of responses. Subjects suffering from burnout made less errors with a threatening emotional stimulus compared to a neutral stimulus and vice versa the control subjects made more errors with the threatening emotional stimulus compared to neutral. This difference was statistically significant (p=0,025). DISCUSSION- Challenges experienced in everyday executive functions were linked with burnout. However, RTs and errors in the Executive reaction time test did not correlate with the severity of the burnout nor were the self-evaluated problems in executive functions depicted in the test performance. Instead, the subjects suffering from burnout differed from the control group in how the threatening stimulus affected the accuracy of responses in the test. It is possible that the subjects suffering from burnout benefit from the increase in arousal caused by the threatening emotional stimulus which was shown as improved accuracy of responses when there was a threatening stimulus, whereas the control group's accuracy of responses was disrupted by the threatening stimulus. We speculate that if the control group’s baseline level of arousal was optimal then the threatening emotional stimulus may have increased arousal to suboptimal level causing decrease in performance. Subjectively experienced challenges in executive functions and objective changes in the interaction between emotions and the executive functions were observed in the study. In conclusion, burnout causes changes in executive functions.
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(2023)Urbanisation is rapidly increasing, along with the use of artificial lighting, which poses a growing threat to wildlife, especially nocturnal fauna. Bats are nocturnal mammals that prey on light-sensitive insects, making them particularly vulnerable to the effects of artificial light sources. Responses to artificial lights vary among bat species, with some being strongly repelled while others appear to benefit from them. However, little attention has been given to the impact of light pollution on bats in northerly latitudes, where bright summer nights may influence the effects of artificial light on these animals. In my thesis I compared the presence of bats in artificially illuminated and unilluminated areas in urban parks in Southern Finland. To see the effect of bright nights in the early summer I collected acoustic data during two time periods: bright early summer (June), and dark late summer (August). My aim was to determine if different bat species respond differently to artificial light, if lamp type affects their response, and how natural light influences their activity patterns. I found that northern bats (E. nilssonii) were drawn to street lights in both early and late summer, and they seemed to prefer HPS (high-pressure sodium) lamps to LED (light emitting diode) lamps. Conversely, more light shy bats from the genus Myotis strictly avoided street lamps at all times. The strongest effect of natural light on bats was observed on their activity patterns: the timing of peak activity shifted considerably along with changes in natural light conditions. My results indicate possible changes in the responses of nocturnal wildlife to the changes in spectral composition of illumination along with increasing popularity of LEDs. The significant differences in the responses of different bat taxa to illumination highlight the importance of conservation of dark spaces and times.
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(2022)COVID-19 pandemic, caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has thus far claimed over six million lives. Vaccines against SARS-CoV-2 have successfully mitigated severe disease and eased the burden on healthcare systems. Neutralizing antibodies play crucial roles both in vaccine derived immunity, and as drugs widely utilized in monoclonal antibody therapy. Neutralizing antibodies primarily target the spike protein, where most of the neutralizing epitopes are located in the receptor binding domain (RBD). Elucidating the sites of vulnerability to neutralization on SARS-CoV-2 can facilitate the development of therapeutics. 7A12 is a newly-discovered IgG antigen-binding fragment (Fab) isolated from a COVID-19 patient in Finland that can neutralize SARS-CoV-2 by targeting the spike protein. In this thesis, a complex of the Fab 7A12 with SARS-CoV-2 spike ectodomain trimer was studied using cryogenic electron microscopy (cryo-EM) single-particle analysis to elucidate the epitope of 7A12 and to gain insight into the neutralization mechanism of 7A12. Cryo-EM data of the complex revealed that Fab 7A12 can bind to both “open” and “closed” conformations of RBD. Rigid-body fitting of the spike trimer and Fab 7A12 models into the cryo-EM density indicates that 7A12 recognizes an epitope in the RBD which is mainly located outside the ACE2 binding site. Together, these results suggest that the 7A12 epitope belongs to class III of SARS-CoV-2 neutralizing epitopes located in the RBD, indicating that 7A12 could neutralize by sterically hindering ACE2 and by preventing the adjacent RBD from changing to ”up” conformation. Furthermore, our results revealed an overlap of 7A12 epitope with the putative binding site of heparan sulfate, a host factor of SARS-CoV-2 infection, which might contribute to neutralization. 7A12-RBD interface mapping delineated the residues comprising the epitope, which do not include mutants found in Beta, Gamma, and Delta variants, while four mutants were found in Omicron within the epitope indicating that 7A12 is likely to neutralize Beta, Gamma, and Delta variants of concern.
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(1998)Cells in tissues have only three serious options in life; they can grow and divide, remain static, or die by apoptosis. Upon growth factor stimulation a cell enters the so called cell cycle which will eventually lead to the division of the cell. Cell cycle can be divided into four phases; G1, S, G2 and M. The current model of the cell cycle control holds that the transitions between different cell cycle states are regulated by cyclin dependent kinases (CDK) with their activator subunits, the cyclins. CDK regulation can be separated into four distinct mechanisms, one of which being phosphorylation on the so called T-loop leading to complete activation. This phosphorylating activity is mediated by apparently a single enzymatic activity termed the CDK activating kinase, CAK. CAK activity was originally isolated as a biochemical purification extract and the enzyme was surprisingly noticed to be structurally related to CDKs. Since a novel cyclin was identified to be associated to it, the enzyme exercising CAK activity was named CDK7 and the cyclin was designated cyclin H. An entirely new perspective on CDK7 function was opened when CDK7 was identified as a subunit of transcription factor IIH (TFIIH) and shown to phosphorylate the carboxy-terminal domain (CTD) of RNA polymerase II (RNAPII). CDK7 has also been suggested to be involved in irradiation sensitivity pathways and nucleotide excision repair functions. To elucidate the intriguing in vivo role of CDK7, proteins interacting with CDK7 were screened for using the yeast two-hybrid method as part of previous studies of the laboratory. The results showed that 15 out of 144 (10,4%) positive clones were identified to encode a peptide sequence of a protein previously known as the inhibitor/interactor of protein kinase C (PKCI). These yeast colonies had an unexpected phenotype; contradictory to a dark blue color of the colonies, indicating strong interaction, the size of the PKCI colonies was small compared to others, indicating a possible growth inhibition effect. Several DNA open reading frames (ORF) coding for proteins related to human PKCI have been identified in a broad range of species representing mammalian, plant, fungal and bacterial kingdoms, all these forming a HIT (conserved triad of histidines) protein family. Another human member, part of this now super family, named FHIT (fragile triad of histidines) was identified with a dinucleoside 5’,5’’’-P1,P3-triphosphate hydrolase activity. These molecules; substrates of FHIT and related enzymes have been proposed to have various intracellular functions, including signalling stress responses. The aim of this study was to extend the investigation of the interaction between CDK7 and PKCI observed in yeast two-hybrid by means of several genetic and biochemical approaches to determine if this observed interaction and growth phenotype has any physiological significance. Investigations included performing yeast two-hybrid screening for PKCI, developing yeast three-hybrid system and carrying out growth rate assays for yeast liquid cultures. These studies also included performing biochemical purifications of over-expressed proteins, immunoprecipitations, western blot analysis and kinase activity assays. Protein extracts originated from transformed yeast cells, transfected mammalian cells or from in vitro transcription and translation reactions. On basis of growth rate assays it can be concluded that PKCI has an inhibitory growth effect in yeast. The preliminary finding of a specific PKCI-CDK7 interaction in yeast two-hybrid, however could not be conclusively verified by the other methods that were used in this study. Studies of PKCI characterisation also included examination of the subcellular localisation of PKCI in mammalian cells by immunofluorescence labelling of HA-PKCI. Results showed PKCI to localize both in the nucleus and in the cytoplasm. Also, studies to elucidate the function of PKCI were performed; whether it possesses enzymatic activity related to that of FHIT. By NMR spectroscopy using bacterially produced GST-PKCI, hydrolase activity towards ADP was indeed observed. Future studies will include elucidation of possible links between growth inhibition and hydrolase activity, in the form of stress signalling functions. The main focus of our future studies will be the generation of mice with targeted PKCI alleles offering powerful means to reveal the function of PKCI through observing phenotypes and through detailed analysis of these mice harbouring wild type, hypomorphic or null alleles.
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