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(2021)Forests have become one of the key themes when it comes to sustainability and have been subject to many demands and pressures from various stakeholders in recent years. In particular, the responsible use of forests has surfaced in discussions during the past 20 years. As the world and overall atmosphere is becoming more favorable to sustainability, companies are required to adapt to a changed society. The forest industry has a significant impact on Finland’s sustainable development and economy. In the forest industry corporate responsibility work has developed as demands from society have grown and the work has become more holistic over time as knowledge on the matter has increased. Pressures on companies' environmental work have come from outside companies from society, as well as from internal actors within companies. One well established forest industry company in Finland is Metsä Group, which has long been profiled as an environmentally responsible company. Therefore, corporate responsibility work and its development in Metsä Group has been selected as my research topic. In this thesis, I examine how environmental responsibility work has changed from the point of view of sustainability professionals in Metsä Group during the years 2000-2020. As research material, I use interviews with Metsä Group's sustainability professionals, as well as responsibility and annual reports from 2000-2020. For the interviews, I selected former and current Metsä Group employees who have worked in the company, either the whole duration or partly, during the years 2000-2020. I have analyzed the materials using content analysis as a method. The interviews revealed that, in general, employees felt that, compared to the early 2000s, in 2020 Metsä Group took more and more holistic action for its environmental responsibility. In general, it was seen that the most significant factors influencing corporate responsibility were megatrends such as climate change and biodiversity loss, as well as a general change in societal perceptions related to corporate responsibility. Leaders’ commitment and attitude towards environmental responsibility, as well as customer requirements, were also perceived as significant factors in promoting responsibility work. It was generally felt that if customers or management saw the matter as important it was invested in. All in all, the environmental corporate responsibility of Metsä Group had steadily improved during 2000-2020. Overall, the interviewees felt that environmental responsibility has been incorporated into Metsä Group's strategy, and that corporate responsibility work has become more holistic and professionalized.
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(2021)The way environmental issues are discursively constructed matters for how they are understood and what possibilities there are to solve them. This makes it relevant to investigate discourses around environmental issues and their proposed solutions. One such solution is ecological compensation, which has been widely implemented as a way to avoid environmental degradation and achieve no net loss of biodiversity. Compensation is also a contested mechanism, however, which has been shaped by the interplay of various discourses with diverging understandings of nature conservation. In this study, I investigate how ecological compensation is constructed by experts in Finland. Using the concept of storylines (Hajer 1995) I analyse 9 interviews conducted with experts involved in the discussion around ecological compensation, which is currently being implemented into Finnish legislation. Three storylines are identified which construct ecological compensation either as 1) a way to enable private actors to take environmental responsibility, 2) additional legislation to fill a “gap” in current conservation practices, or 3) a possibility to modify the relationship with nature by fostering local deliberations. This shows diverging understandings of ecological compensation among the experts and contestation over the way it should be implemented. What is at stake in the discussion are questions of how nature conservation should be understood, which makes ecological compensation pivotal for reconfiguring the field of nature conservation by shifting understandings of the roles and responsibilities involved. At the same time, consensus exists about the need to implement ecological compensation, which the experts all agree is the only possibility to stop biodiversity loss in Finland. This is traced to the interpretative flexibility of the ecological compensation concept which accommodates conflicting understandings and enables the experts to agree about the need for ecological compensation while contesting the “details” of how the implementation should take place. In that way, the implementation is supported despite contestation, and the discussion is focused on how – rather than whether – ecological compensation should be implemented.
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(2021)Microbial diversity can be found everywhere around us. The diversity is however declining globally and the diversity loss is most visible in highly urbanized areas. The lack of microbial biodiversity has been linked to increased risk of certain im-mune mediated diseases most prevalent within urban population. Understanding how diversity differs between urban and rural areas can help us to figure out mechanisms behind biodiversity loss and higher frequency of immune-mediated dis-eases and develop prevention methods for the latter. The aim of the thesis is to study how bacterial communities differ between urban and rural areas using indicator species as proxy. The aim is also to find out if the results support the biodiversity hypothesis. The results of the thesis found out significant differences in diversity indexes between bacterial communities in urban and rural areas, which supports the biodiversity hypothesis. The study also found differences in Proteobacteria diversity index-es, which have been linked to some immune mediated diseases in previous studies.
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(2021)This thesis explores the drivers and barrier to greening in the Finnish music festival scene. The term greening originates in literature on sustainable business and refers to the process of a company achieving greater sustainability through investments, managerial and organisational decisions and implementing policies and processes. The concept of greening is applied to music festivals, a type of event, that cause significant negative environmental impacts. Sustainable event management attempts to mitigate negative environmental impacts and to cause positive impacts for instance through environmental education and inspiring behaviour change. As a theoretical framework, this thesis uses drivers and barriers impacting the greening of music festivals identified in literature. The drivers (organisational and personal values, competitive advantage, consumer demand and the desire to educate) and barriers (lack of finance or support from stakeholders, lack of control over venues, lack of control over patron behaviour, lack of time, lack of sustainable supplies and suppliers), networking in the festival scene and the use of external environmental audits form the basis for a qualitative content analysis. The findings based on interview data from five Finnish music festival managers and a representative from an environmental auditing service suggests that the most significant drivers are value-based and the motivation for greening stems from personal pro-environmental values and the desire to inform and educate audiences. The barriers to overcome are external, related to stakeholders, infrastructure and the supply chain as well as unsustainable attendee behaviour. Additionally, there is among festival managers some uncertainty regarding best practices. A need for increased support from stakeholders and for accessible information highlights the importance of networking. External auditing services should offer flexible and contextual tools and a more inspirational tone. In conclusion, sustainability is seen by the Finnish music festival scene as a key issue and one of increasing priority in the future.
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(2020)Iron-manganese (FeMn) concretions are found on soft sediment bottoms both in the deep sea and coastal sea areas, formed as a result of a combination of biogeochemical and microbial processes. It has been estimated that concretions occur at least in 11 % of the Finnish marine areas. Concretions form hard substrates on predominantly soft seafloors, and they are therefore suggested to increase geodiversity and habitat complexity of the seafloor. This has been found to correlate with biodiversity of the benthic fauna. Despite their widespread occurrence in the northern Baltic Sea, the ecological importance of FeMn concretions has been left unaddressed. In the recent assessment of threatened habitat types in Finland, concretion fields were classified as a data deficient habitat type. The aim of this study is to examine the role of FeMn concretions as habitats in the Baltic Sea ecosystem. Benthic biodiversity was investigated utilizing two approaches; the abundance of mobile fauna and sessile macrofauna were studied with point-dives. The data was compared to pre-existing data from similar soft bottoms where there are no observations of concretions, collected in the Finnish Inventory Programme for the Underwater Marine Environment (VELMU). Samples for sediment in-fauna were taken with a Van Veen Grab Sampler, and additional data was gathered also from Environmental Information System HERTTA (administered by Environmental Administration). The shape and quantity of concretions appear to affect the abundance of sediment in-fauna. Similarities to the invertebrate composition of soft sediment habitats depends of the soft sediment availability in the habitat, which is dependent on concretion shape and quantity. Crusts seem to affect the faunal composition more than spheroidal and discoidal concretions, as they offer the most complex habitats, significantly different from bare seafloors. Based on this study, the concretion fields should not necessarily be considered as just one habitat type, since the faunal composition appears to differ according to the shape of the concretions.
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(2013)Lepidurus arcticus (Pallas, 1793) is a keystone species in High Arctic ponds, which are exposed to a wide range of environmental stressors. This thesis provides information on the ecology of this little studied species by paying particular focus on the sensitivity of L. arcticus to acidification and climate change. Respiration, reproduction, olfaction, morphology, salinity and pH tolerance of the species were studied in the laboratory and several environmental parameters were measured in its natural habitats in Arctic ponds. Current global circulation models predict 2–2.4 °C increase in summer temperatures on Spitsbergen, Svalbard, Norway. The L. arcticus respiration activity was tested at different temperatures (3.5, 10, 16.5, 20, 25 and 30 °C). The results show that L. arcticus is clearly adapted to live in cold water and have a temperature optimum at +10 °C. This species should be considered as stenothermal, because it seems to be able to live only within a narrow temperature range. L. arcticus populations seem to have the capacity to respond to the ongoing climate change on Spitsbergen. Changes can be seen in the species' reproductive capacity and in the individuals' body size when comparing results with previous studies on Spitsbergen and in other Arctic areas. Effective reproduction capacity was a unique feature of the L. arcticus populations on Spitsbergen. L. arcticus females reached sexual maturity at a smaller body size and sexual dimorphism appeared in smaller animals on Spitsbergen than anywhere else in the subarctic or Arctic regions. L. arcticus females were able to carry more eggs (up to 12 eggs per female) than has been observed in previous studies. Another interesting feature of L. arcticus on Spitsbergen was their potential to grow large, up to 39.4 mm in total length. Also cannibalistic behaviour seemed to be common on Spitsbergen L. arcticus populations. The existence of different colour morphs and the population-level differences in morphology of L. arcticus were unknown, but fascinating characteristic of this species. Spitsbergen populations consisted of two major (i.e. monochrome and marbled) and several combined colour morphs. Third interesting finding was a new disease for science which activated when the water temperature rose. I named this disease to Red Carapace Disease (RCD). This High Arctic crustacean lives in ponds between the Arctic Ocean and glaciers, where the marine environment has a strong impact on the terrestrial and freshwater ecosystems. The tolerance of L. arcticius to increased water salinity was determined by a LC50 -test. No mortality occurred during the 23 day exposure at low 1–2 ‰ water salinity. A slight increase in water salinity (to 1 ‰) speeded up the L. arcticus shell replacement. The observations from natural populations supported the hypothesis that the size of the animals increases considerably in low 1.5 ‰ salt concentrations. Thus, a small increase in water salinity seems to have a positive impact on the growth of this short-lived species. Acidification has been a big problem for many crustaceans, invertebrates and fishes for several decades. L. arcricus does not make an exception. Strong acid stress in pH 4 caused a high mortality of mature L. arcticus females. The critical lower limit of pH was 6.1 for the survival of this acid sensitive species. Thus, L. arcticus populations are probably in danger of extinction due to acidification of three ponds on Spitsbergen. A slight drop (0.1–1.0) in pH values can wipe out these L. arcticus populations. The survival of L. arcticus was strongly related to: (1) the water pH, (2) total organic carbon (TOC) and pH interaction, (3) the water temperature and (4) the water salinity. Water pH and TOC values should be monitored in these ponds and the input of acidifying substances in ponds should be prevented.
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(2019)GRACILE (Growth Retardation, Aminoaciduria, Cholestasis, Iron Overload, Lactic Acidosis, and Early death) syndrome (Fellman disease, MIM603358) is a mitochondrial disorder that belongs to the Finnish disease heritage and follows an autosomal recessive inheritance pattern. It is a lethal neonatal disease and the affected infants usually survive only a couple of days, but in some cases up to four months. The disease is very rare affecting about 1 in 47,000 infants in Finland. GRACILE syndrome is caused by a homozygous Finnish founder mutation (c.A232G) in the BCS1L gene, which encodes an assembly factor for the mitochondrial respiratory chain complex III. The missense mutation (c.A232G) causes an amino acid change (p.S78G) in the BCS1L protein. Alternative oxidase (AOX) is a terminal oxidase that is not naturally present in mammals. It enables the respiratory chain electron flow to bypass complexes III and IV. The AOX pathway works parallel with the respiratory chain and gets activated under stress conditions in plants and lower animals. In mitochondrial diseases with a complex III deficiency, such as GRACILE syndrome, AOX expression could alleviate the symptoms caused by the complex III dysfunction. The aim of this study was to investigate the effects of AOX expression on early-onset manifestations of the disease in the Bcs1lc.A232G mouse model of GRACILE syndrome. The mice used in this study have a genetic background with a short survival to P35-40. The respiratory chain function in freshly isolated mitochondria from the liver and kidney was studied, as well as complex III activity, mitochondrial mass and liver and kidney histology. The findings of this study suggest that AOX has a strong beneficial effect on both liver and kidney histology and it is able to restore the glycogen stores to some extent, thus alleviating the glycogen depletion seen in the mice. The Bcs1lc.A232G mice also had an improved body weight in the presence of AOX, suggesting a less severe energy deficiency due to the activity of the AOX transgene. CI- and CII-linked respiration was also robustly improved in the liver. Overall, the condition of the Bcs1lc.A232G mice was improved by the presence of the AOX transgene compared to the Bcs1lc.A232G mice not expressing AOX. These results are highly encouraging for further studies on the rescue effects that AOX seems to have on this disease model.
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(2022)Light pollution, or artificial light at night, is a globally increasing environmental problem that threatens especially nocturnal organisms dependent on darkness. Modern lighting technology offers opportunities for mitigation of the ecological impacts of light pollution, but effective implementation requires better understanding of how different artificial light qualities, such as light spectrum, influence its effects on wildlife. The common glow-worm, Lampyris noctiluca, is an example of a species believed to be suffering from light pollution. Artificial light has been found to interfere with glow-worm reproduction by decreasing the success of females in attracting males with their glow. In this study, I investigated how the color (spectrum) of artificial light affects the attraction of male glow-worms towards a female mimicking stimulus, in order to find out whether certain colors of artificial light are less detrimental to glow-worm reproduction than others. I used dummy female traps to capture male glow-worms in the field and compared the catch success of traps in different treatments: illuminated from above with blue, white, yellow or red artificial light, or left unilluminated as a control. I also conducted a laboratory experiment where male glow-worms were given two choices. One of the choices was an unilluminated dummy female, and the other was either a dummy female illuminated with yellow or red light, or a red light illuminated area with no dummy female. Traps illuminated with short wavelength artificial light (blue and white) caught significantly fewer males than unilluminated traps or traps illuminated with long wavelength artificial light (yellow and red). There was no significant difference in the number of males caught between unilluminated traps and traps illuminated with long wavelength artificial light. In the laboratory, males significantly preferred an unilluminated dummy female over a dummy female illuminated with yellow light. However, the males chose a red light illuminated dummy female or area more often than an unilluminated dummy female, although this difference in preference was not significant. The results show that mate attraction in the glow-worm is influenced by artificial light color, with short wavelength artificial light decreasing the mate attraction success of female glow-worms more than long wavelength artificial light. This could point to yellow-tinted artificial lighting presenting an ecologically friendly alternative to cool white lighting. However, the specifics of how long wavelength artificial light affects male glow-worm perception of female attractiveness are still unclear. Furthermore, male glow-worms show signs of attraction towards long wavelength artificial light, which could form an evolutionary trap for them. The impacts of artificial light spectrum on organisms are thus not straightforward, but can vary depending on both species and situation.
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(2020)Bipotential gonads are precursor structures for testes and ovaries. Steroidogenic factor-1 (SF1) is one of the most important transcription factors in an embryo needed for the development and maintenance of bipotential gonads. If SF1 is not expressed, bipotential gonads fail to develop, and genitalia and kidneys are not formed. Later, SF1 expression persists high in testes, where it supports Sertoli and Leydig cell formation and development. If SF1 is not expressed enough in males, the bipotential gonads differentiate into ovaries. The factors activating and regulating SF1 are not currently fully known. By getting more knowledge of how SF1 is controlled, regulatory mechanisms behind normal fetal development of gonads and disorders of sex development (DSD) can be understood better. The aims of this thesis were to study whether growth factors, that naturally regulate differentiation of developing gonads, promote differentiation of human induced pluripotent stem cells (hiPSC) into Sertoli-like cells (SLCs) and whether SF1 expression is induced by the addition of these growth factors. For conducting the study, we used hiPSCs, which have an SF1 activation domain cassette previously introduced to the cells by clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9) (CRISPR/Cas9) method. SF1 could be activated by adding doxycycline (DOX) and trimethoprim (TMP). These hiPSCs were differentiated into intermediate mesoderm (IM) on the first four days according to the protocol published earlier by the group. After this, the differentiation to SLCs was guided by adding growth factors to the culture medium. Basic fibroblast growth factor (bFGF), fibroblast growth factor 9 (FGF9) and prostaglandin-2 (PGD2) were tested separately and in a combined cocktail also including follicle stimulating (FSH) and glial cell-derived neurotrophic factor (GDNF). In a control condition, cells were differentiated without additional growth factors. In all tested conditions, cells first differentiated into IM were further differentiated either in the presence or absence of DOX and TMP for 8 days. The differentiation medias were changed to the cells every day and lysis samples for quantitative PCR (qRT-PCR) were taken every other day. The relative gene expression levels of bipotential gonad, testis and steroidogenic gene markers from each condition were monitored with qRT-PCR and compared to the levels of the undifferentiated hiPSCs. Immunocytochemistry was performed to see the changes in protein production. Against our hypothesis and the previous studies by others, none of the tested growth factors induced the cells to differentiate into SLCs. However, SF1 expression was triggered by chemical induction with DOX and TMP. Also, the expression levels of bipotential gonadal and testicular gene markers increased in control conditions with/without chemical induction. PGD2 conditions were the only ones to resemble the gene expression and morphology of control conditions while the others differed. These results indicated that the addition of bFGF, FGF9, FSH and GDNF did not improve the differentiation of iPSCs into SLCs and in fact, bFGF and FGF9 hindered their differentiation into SLCs. As a future perspective the optimal concentrations for each growth factor and the duration of growth factor supplementation ought to be tested to refine the protocol.
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(2021)Lake ecosystems are shaped by water chemistry processes that affect the lake environment and the species communities within. Changes in the water chemistry thus have far-reaching consequences. Water colour is one variable that affects water chemistry and stems from humic substances in the water. Dark water reduces light availability and also affects nutrient and oxygen availability. A trend of brownification of freshwater systems has been observed in recent years and it is expected to influence species community’s diversity and composition. The aim of this thesis was to study whether brownification is an ongoing issue in the study lakes and whether it has had a negative effect on phytoplankton diversity and resulted in shifts in the phytoplankton composition. A data set including about a 100 lakes in Finland with measurements from 1965 up until now served as the study system which was analysed with statistical methods. The results indicated a brownification trend in the past decades. The brownification so far had a positive impact on species richness but a negative impact on beta diversity. Brownification also affected species composition. Flagellates and autotrophic species increased in darker waters but mixotrophic species that are known to dominate in dark water colour, did not show a clear increase with water colour. Other hydrological variables than water colour could have had a bigger impact on the phytoplankton community than water colour but future monitoring of the phytoplankton community is recommended to see if water colour will have a negative impact on species diversity in the future.
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(2019)Understanding the link between the gut microbiota, diet and the enteric nervous system is of significant importance in the prevention of gastrointestinal disorders. The aim of the study was to answer two questions: Firstly, is butyrate able to stimulate the luminal release of serotonin? Secondly, in which parts of the gastrointestinal tract does this possibly occur? These questions are of interest, due to the importance of the serotonergic signalling in the enteric nervous system. We created a luminal perfusion system to investigate the effect of butyrate in the gastrointestinal tract of male Wistar rats (500-550g). We isolated the stomach and 4 cm long segments of the duodenum, jejunum and colon. To our knowledge this form of physiological ex vivo studies investigating the entire gastrointestinal tract have not been done previously. The isolated stomach and the isolated intestinal segments were luminally perfused with 100 mM butyrate for 10 min respectively 45 min. The tissues were homogenized after the luminal perfusion. Serotonin and its main metabolite 5-hydroxyindoleacetic acid (5-HIAA) were assayed using commercial ELISA kits. Our results showed that butyrate significantly stimulates the release of 5-HIAA in the stomach, duodenum, jejunum and colon. Butyrate seems also to have a positive trend-effect on the release of serotonin itself in the stomach, duodenum, jejunum and colon. Although, there is a future potential for preventing gastrointestinal disorders with the help of diet and gut microbiota, the possible clinical significance of our results should be considered carefully.
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(2019)Epithelial cells form a barrier between the tissue and the external environment. Epithelial morphogenesis refers to the shaping of epithelial layers and is a key step in the development of organisms. The actin cytoskeleton provides the cell its form and during epithelial morphogenesis, produces force to shape the cells. To achieve this, the actin cytoskeleton is organized into protrusive and contractile networks. In a living cell, these actin networks are dynamic, as the filaments are constantly undergoing assembly and disassembly. Actin-binding proteins regulate the turnover of actin filaments, but in epithelial morphogenesis, the regulatory role of most of these proteins is still relatively unknown. In all multicellular organisms, actin disassembly is controlled by ADF/cofilin. ADF/cofilin activity is furthermore enhanced by other actin-binding proteins, one of which is cyclase-associated protein (CAP). CAP promotes actin turnover by accelerating ADF/cofilin mediated actin disassembly and in recycling actin monomers to sites of actin polymerization. Unlike ADF/cofilin that regulates actin disassembly throughout the whole cell, CAP could be subject to more specific spatial regulation, as loss of CAP leads to F-actin accumulation on the apical side of epithelial cells. However, the role of CAP in morphogenetic cell rearrangements remains poorly known. In addition, the in vivo role of the biochemical functions of CAP has not been elucidated. The aim of this master’s thesis is to describe the role of CAP in regulating the actin cytoskeleton in the follicular epithelium of the fruit fly Drosophila melanogaster. For this purpose, chimeric mutant flies with homozygous CAP loss of function mutation were generated. Subsequently, the effect of the CAP loss of function was observed in follicle cell populations undergoing morphogenetic changes. In addition, CAP loss of function was rescued with different transgenes producing mutant CAP proteins to identify the protein domains of CAP with in vivo significance. In addition, a Drosophila CAP specific antibody was purified to be used in immunostaining. The ovaries were imaged using confocal microscopy. In this thesis, it is shown that CAP loss of function caused accumulation of filamentous actin in all observed follicular cell populations. Surprisingly, the actin turnover was rescued by all of the used CAP rescue transgenes, but the mutant transgenes exhibited phenotypes resembling the CAP loss of function in other epithelial tissues. Moreover, CAP loss of function caused defects in the follicle cell movement and cell spreading. The loss of function also caused expression changes in other actin-binding proteins. The findings of these thesis support the current knowledge of CAP importance for functional actin turnover in the follicle cells, even though the protein domain necessary for in vivo function could not be deciphered. Moreover, this project provides indication that CAP has an indispensable role in dynamic morphogenetic processes in the epithelium. Together with other actin-binding proteins, CAP could regulate epithelial actin turnover in spatially directed manner, providing force for epithelial cell adhesions or protrusions.
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(2024)As global temperatures rise due to anthropogenic greenhouse gas emissions, it is becoming increasingly important to mitigate climate change. Agriculture, one of the major contributors to greenhouse gas emissions, has the potential for mitigation by implementing carbon sequestration practices. Biochars, carbon-rich materials derived from biomass pyrolysis, have drawn attention for their potential to sequester carbon and enhance crop growth. However, limited yield effects have been reported in boreal agricultural soils after applying pure biochars to soils. Therefore, application of activated and nutrient enriched biochar has been proposed as an amendment for getting most use of the practice due to slow release of nutrients and carbon sequestration. However, the effects of these new activated biochars on soil biota remain unknown. The study investigates the impact of acid and slurry activated biochar on earthworm community in agricultural soil in southern Finland, focusing on their implications for soil physical properties and ecosystem health. Through a split-plot designed field experiment, with biochar (applied at rate 0 t ha-1 and 20 t ha-1) being the main factor and fertilizer level the sub-plot factor, the interaction between nutrient enriched biochar and earthworm population was examined by assessing earthworms' abundance, biomass, and mean weight. There was no significant effect of the treatments on earthworm abundance, biomass, or mean weight. However, the mean weight showed a notable response to the interaction effect of biochar x fertiliser. Biomass and mean weight were negatively correlated by temperature. Among the other variables, soil water content had positive correlation with temperature. These findings highlight a possible association influenced by soil physicochemical parameters, microbial dynamics, and environmental conditions, even though biochar had no apparent impacts on earthworm density, biomass, or mean weight. In this first-ever field experiment with a high rate of acid and slurry-activated biochar applied to agricultural soil, biochar had no adverse effect on earthworm populations. Thus, the present findings confirm that the application of large amounts of biochar treated with acid and cattle slurry in Nordic agricultural soil is a safe C sequestration method for earthworms at least for a short time period. However, the long-term effect is still a knowledge gap. Addressing this knowledge gap is crucial to apply sustainable agriculture methods that enhance soil properties and health.
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(2022)Colorectal cancer is one of the most common cancers in the world, and in 2020 it was the cause of nearly 1 million deaths. A major reason for the high death rate is treatment resistance; eventually, almost all colorectal cancer patients with metastatic disease stop responding to chemotherapy. The problem of treatment resistance is not specific to this type of cancer, but it is a widespread issue for all cancer treatments. Chemotherapy resistance is the sum of several cellular and non-cellular factors that together enable sustained cell growth despite the treatment. The non-cellular factors are related to the tumor microenvironment, whereas the cellular factors are related to changes in gene expression, which facilitate e.g. the repair of drug-induced damage or lead to changes in drug metabolism. Lately, researchers have turned their interest to translational control in chemotherapy resistance. This is because translational control plays a major role in stress adaptation. During cellular stress, global translation rates are reduced and those messenger RNAs that are most important for cell survival are translated efficiently. Moreover, translation is fine-tuned by transfer RNA (tRNA) modifications. These modifications are chemical groups that are added to the ribose and the nucleobase of the tRNA molecule, and they affect all aspects of tRNA function, ranging from the structure and stability of the molecule to reading frame maintenance and rate of translation. tRNA modifications are dynamic and change in response to the cellular state, thus facilitating adaptation by translational control. Given the major role of translational control and tRNA modifications in cellular stress responses, their role in the chemotherapy response and adaptation should be thoroughly investigated. The aim of this thesis was to study how chemotherapy affects translation and tRNA modifications in a colon adenocarcinoma model. The cell lines SW480 (from a primary colorectal tumor) and SW620 (from a metastasis) were treated with 5-fluorouracil, oxaliplatin, and leucovorin (FOLFOX), a common combination of chemotherapeutics used in colorectal cancer treatment. The cells were subjected to long-term cyclic treatment as well as 24 h pulse treatment. Chemotherapy resistant cell lines were established by increasing the concentration of FOLFOX for each round of treatment. The effect on translation was studied by polysome profiling, which revealed that FOLFOX treatment causes immediate translational stress, as evidenced by the “shoulders” in the polysomal fractions in the profiles of the pulse treated cells. We hypothesized that these shoulders represent halfmers, polyribosomes without the large subunit. No difference was observed between the long-term treated cells and controls, possibly indicating that the cells had adapted to FOLFOX. The resistant cells exhibited slightly reduced translational activity, which might be due to altered function of ribosomes following the exposure to 5-fluorouracil. Changes in tRNA modification levels were quantified by liquid chromatography mass spectrometry. Several anticodon loop modifications exhibited altered levels after the pulse treatment. In addition, 5-FUrd, a metabolite of 5-fluorouracil, was incorporated into the tRNA. The long-term treated or resistant cells exhibited no differences in the modification levels. In conclusion, this study provided insights on the immediate effects of FOLFOX treatment on translation. This constitutes the first step towards understanding how RNA-based regulatory mechanisms may contribute to the effect and possible resistance to chemotherapy.
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(2024)The mammalian circadian clock is critical to physiological homeostasis and oversees many important processes like sleep and nociception. Chronic pain, particularly, neuropathic pain (NP) is defined as pain caused by nerve injury and is commonly studied in rodents using the spared nerve injury (SNI) model. The SNI model strongly mimics NP pathology by causing hypersensitive pain responses. As pain and the circadian rhythm are actively linked (as proven by diurnal fluctuations in pain sensitivities in individuals with NP), these responses can disrupt overall well-being and increase an individual’s predisposition to diseases like cancer and depression. Therefore, avenues to discern the workings of the circadian clock have been explored, and some include the use of small-molecule modulators like TH301, that regulate specific core circadian genes. In this study, we explored the possible role of TH301 in normalizing disrupted circadian rhythms in a rodent model of NP, while also examining its potential as an analgesic agent.
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(2020)Northern peatlands have a major role in the global carbon cycle due to their carbon stocks and fluxes of carbon dioxide (CO2) and methane (CH4). Anthropogenic climate change may affect peatland carbon dynamics through changes in e.g. primary production, hydrology, and permafrost dynamics. It is uncertain whether these changes will lead to northern peatlands becoming significant sources of carbon to the atmosphere. Changes in moisture conditions especially can be an important factor in determining the carbon sink potential of northern peatlands. In this thesis I examine the palaeohydrology and peat accumulation over the past centuries in a permafrost peatland complex in Lovozero, Kola Peninsula, Russia. I used testate amoebae as a proxy of past changes in moisture conditions. Other study methods used here are detrended correspondence analysis (DCA) and 14C and 210Pb dating. The results were also supplemented with plant macrofossil and carbon accumulation data provided by other members of the research team. The results show varying responses of the peatland hydrology and peat accumulation to the past climatic shifts, suggesting that the changes have been driven more by autogenic factors rather than climate. However, all three sites indicated a drying trend and an increased peat accumulation for the last century. Yet, the last decade is charachterised by a wet shift. The wet shifts suggest that the peatlands may have crossed a threshold where increased evapotranspiration is exceeded by increased moisture due to thawing permfrost. The surface peat layers of all three sites were dominated by mixotrophic testate amoebae, which may have contributed to the high peat accumulation. The inconsistency of past successional pathways identified at Lovozero peatlands and the drying trend over the past century correspond to the previous studies from northern peatlands elsewhere. However, the most recent surface wetting during the last decade differs from what has been reported for the other northern sites. This suggests that the response mechanisms of peatlands to the anthropogenic climate change may not be uniform. Thus, further research is inevitably needed to increase our understanding of peatland-climate intercations.
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(2022)Microglia, the resident immune cells of the central nervous system, react to inflammatory stimuli in the brain in a variety of ways. These include migrating to the site of damage and releasing pro- and anti-inflammatory factors. Previous research indicates that these microglial functions require extensive intracellular calcium signaling. Microglial overactivation can exacerbate neuronal damage, especially in cases of chronic inflammation. The ability to modulate the microglial response to damage would therefore be of great clinical relevance. The endoplasmic reticulum (ER) acts as the cell’s main calcium store and regulates cellular calcium levels primarily through the activity of ryanodine receptors (RYR), inositol-triphosphate receptors (IP3R), and the sarco-endoplasmic reticulum calcium ATPase (SERCA) pump. Calcium depletion from the ER is associated with cellular stress and microglial reactivity and therefore the ER may be an important target for modulating the microglial reactive response. The aim of this study is to show whether ER calcium depletion in a microglial cell line causes changes in protein expression, cellular infiltration, and the release of key pro-inflammatory factors. Drugs that block the pumping of calcium from the cytosol via the SERCA pump, such as thapsigargin, effectively induce a state of calcium depletion in the ER. In the present study, treatment with the SERCA pump inhibitor thapsigargin was found to increase SERCA2 expression in BV2, but not SV40, microglial cell lines. Treatment of microglia with thapsigargin was associated with large increases in the release of pro-inflammatory factors IL-6 and TNF-alpha but had no effect on microglial migration.
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(2009)The Baltic Sea suffers from eutrophication caused by the increased use of nitrogen- and phosphorus based fertilizers in agriculture. When these nutrients end up in the water ecosystem, they increase the growth of filamentous algae causing turbidity at many locations. The three-spined stickleback (Gasterosteus aculeatus) breeds at the shallow coastal waters of the Baltic Sea, which are often eutrophied. In these locations turbidity of the water may interfere with the mating cues used by the three-spined stickleback, which in turn may lead to decreased fitness of the population. I attempted to find out how turbidity alters the use of visual and olfactory cues in the mate choice of the three-spined stickleback, as well as to see if these changes decrease the viability of the following generation. Female three-spined sticklebacks choose their mates based on visual and olfactory cues. During the reproductive season stickleback males turn bright red and attract females to their nests by a conspicuous courtship dance. Females use males' red colouration, size and courtship intensity as visual cues when choosing an appropriate mating partner. They also pay much attention to olfactory cues. Female sticklebacks are able to smell MHC-encoded peptides which are secreted to the males' skin. The allelic combination of MHC determines which pathogens the individual has resistance for, and this resistance may be inherited by the offspring. I empirically tested the use of olfactory and visual cues in the mate choice of the three-spined stickleback using turbid and clear water as treatments. In mate choice tests a female was made to choose from two males in circumstances where she was allowed to use only one of the cues (visual or olfactory) or both cues simultaneously. The redness and size of the males was measured. Artificial inseminations were performed to produce offspring, whose growth rate was measured to evaluate fitness. Based on the results of these experiments, turbidity alters the use of mating cues of the three-spined stickleback. Visual cues seem to be important in clear water, whereas in turbid water olfactory cues increase in importance in relation to visual cues. The sample size was limited to reliably test offspring fitness effects, but it seems that the alteration in the use of mate choice cues may influence population viability in the long term. However, additional research is needed to determine this.
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(2020)Spinal cord injury (SCI) in human patients is the most expensive clinical condition worldwide, restricting individuals’ ability to manage with daily-life activities independently. With very limited available treatment possibilities, the understanding and validating of regenerative mechanisms and treatment options in animal models is crucial for their translation to clinical practice. The majority of SCIs in human patients are contusive in the cervical level of the spinal cord. However, thoracic injury rodent model is more commonly studied, with only recent studies working with cervical contusion injury model. Chondroitin sulphate proteoglycans (CSPGs), and especially their CS chains, are thought to be the major inhibitory structures for neurite regeneration after SCI. However, current research has led to a new idea that the inhibitory effect of CS chains can be reversed to regeneration enhancing by heparin-binding growth-associated molecule (HB-GAM). This endogenously secreted molecule is highly up-regulated in the central nervous system (CNS) during postnatal development, but in the adult CNS the expression is down-regulated. This suggests that postnatal-level concentrations might be needed for inducing neurite regeneration in adult CNS. In this study, HB-GAM treatment was tested on both cervical hemicontusion and hemisection injury models. Here we show that repeated intrathecal injections of HB-GAM were sufficient to increase grey matter myelin optical density in mouse hemicontusion injury model, and partly induce functional recovery in hemisection model. Obtained anatomical evidence suggests that enhanced myelination is potentially involved in the repair mechanism of HB-GAM. The connection between HB-GAM treatment and functional recovery, and also other mechanisms of HB-GAM-induced regeneration need further exploration. In broader perspective, the results are promising for translation of a novel treatment approach to clinical use.
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(2021)Birdsong is information, communication and a target of sexual selection. Song complexity is the variation in bird song structure. Within-song complexity is variation in song components, in number and order of syllables and phrases. Song complexity is an honest trait under sexual selection and competition for mates increases it. When species expands its range front to a new area, the population density is low in the beginning. Males in the new population do not need to compete as hard as before, because population density is decreased, which decreases song complexity. Many warblers have complex songs and they do not have a colourful plumage. It is well possible that song complexity is the main target of sexual selection in warblers. The purpose of this study was to investigate if there were differences in song complexity in Eurasian reed warblers (Acrocephalus scirpaceus) breeding at their range front in Finland. The population of Finland is 100 years old and the population density is still decreased in compared to Central Europe. My hypothesis was that reed warblers would sing less complex songs at lower densities as competition for mates is low. I used song recordings of 51 reed warbler males, which I recorded during the breeding season 2019 in song complexity analyses. Recordings were from South Finland from 14 locations in 6 municipalities. Most of the recording places were coastal reedbeds. A couple of recording places were reedbeds in inland ponds and reedbeds in a flooded forest. I counted the male densities within 50 m and 200 m radius around the recorded males using Google Maps. I took account the males which were at least 20 m away from each other and were there in the same week than the recorded male. I extracted four within-song complexity variables from the recordings: Total number of syllables, number of syllable types, number of syllable transitions and mean frequency bandwidth. I measured song complexity with spectral analyses using Koe, a bioacoustic software. I did next analyses with R to see if there is association between local density and complexity. I found that one of the song complexity variables, total number of syllables, was significantly positively associated with the density of singing males within 200 m radius. The other variables were not significantly associated with male density, but number of syllable types and number of syllable transitions showed a similar relationship with singing male density within 200 m. Previous studies support the hypothesis that higher local density can increase some aspects of song complexity, for example number of syllables or length of phrases. Some males did not have neighbours within 50 m. Density within 50 m had no significant relationship with song complexity variables. However, mean frequency bandwidth, total number of syllables and number of syllable types showed more variation when neighbours were present, which suggest that males increase their effort when they have competition. Finally, I found that the number of syllable transitions decreased as the breeding season progressed. This could be explained with younger birds coming to breed later or with paired males singing less.
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