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Semliki Forest virus (SFV) is an enveloped virus with positive-sense single-stranded RNA genome that encodes nine proteins, of which four non-structural proteins, nsP1-4, form the replication/transcription complex (RTC) along with several host proteins, which play an important role in the replication of the virus. To establish the interactome of SFV RTC, a promiscuous biotin ligase capable of biotinylating proximal endogenous proteins in the presence of exogenous biotin was genetically fused to nsP3. After establishing the stability, kinetics and functionality of this virus, BHK-21 cells were infected with this mutant SFV at multiplicity of infection of 50 plaque forming units per cell. At an early time point of 2.5 hours post infection, 50 μM biotin was added to medium for 15 minutes. Cells were lysed, and biotinylated proteins were enriched with streptavidin beads, and analyzed through tandem mass-spectrometry. We were able to identify several key host protein interactions, some of which were already established before, but also a several new ones. Many of the host proteins detected were involved with the formation of stress granules, including G3BP’s, or contained a SH3-binding domain (SRC homology 3) like CD2AP, SH3KBP1 and BIN1, and some of them also had RNA binding motifs. In future, we wish to study the role of these identified host proteins in the replication of SFV through gene silencing as well as their co-localization with the RTC and nsP3 with the help of Immunofluorescence.

The literature review dealt with the chemical and physical properties of chitin polymer and its degradation product glucosamine. Particular emphasis was given to studies on the analytical methods of determination of chitin from various products including gravimetrical, spectrophotometric and chromatographic methods. The aim of this study was to optimize an analytical method to determine insect chitin and to apply the method to quantify chitin in whole insects and their soluble protein fractions. Two species of insects were selected: mealworm larvae (Tenebrio molitor) and cricket (Acheta domesticus). Whole insects and their protein fractions were treated with alkaline to remove protein. Chitin in de-proteined insect material was then hydrolysed into glucosamine using 6M HCl. Glucosamine from the hydrolysate was determined using chromatographic and spectrophotometric methods, where chromatographic method was optimized and validated. UPLC-FLR method was specific for glucosamine and the UPLC system could separate two target peaks (glucosamine isomers: α and β). Glucosamine content was calculated using a calibration curve which showed excellent linearity in range 0.0033~24.0 ng/inj. with the determination coefficient more than 0.999 during the study period. The instrumental limit of detection and limit of quantification were 0.00095 ng/inj. (2 µL) and 0.0033 ng/inj. (7 µL), indicating a satisfied sensitivity. Recovery of glucosamine spiked to sample matrix (de-proteined cricket flour) following chitin hydrolysis was not satisfactory (~75%) using HPLC-FLR method, which indicated that spectrophotometric method gave chitin amount closer to the true value due to a higher recovery (>90%). Major findings on chitin amount was ~5% on dry matter basis in both mealworm and cricket. A small percentage of chitin was found in insect protein fractions extracted by 0.1M NaCl.

As the impacts of climate change have become more tangible and severe, it has become clear that changing the approach to resource consumption is crucial. The housing and construction sector has a major role to play in the fight against climate change, as it accounts for around 50% of all material consumption, and the construction sector is also responsible for over 35% of the EU's waste generation. The circular economy (CE) is commonly viewed as a substitute for the traditional linear economic model and therefore it is widely seen as one of the possible solutions for decarbonizing the housing and construction sector and, at the same time, as one of the solutions for tackling climate change. This thesis focuses on exploring the sustainability and circular economy activities of stakeholders in the Finnish housing and construction sector. A particular focus is on the circular economy actions, which will be analyzed with the ReSOLVE framework. Also, the suitability of the framework for mapping circular economy actions in the housing and construction sector will be explored. The thesis thus aims to answer the following research questions: how sustainability and the circularity are visible in the activities of stakeholders in the Finnish housing and buildings sector, and how suitable the ReSOLVE framework is for assessing circularity in this sector. The qualitative research data consists of fourteen stakeholder interviews, which were also complemented by background documents provided by the stakeholders. By utilizing content analysis and the ReSOLVE framework, it was found that the self-designated sustainability activities of the stakeholders were mostly either knowledge generation, dissemination or influencing activities, but the ReSOLVE framework also revealed a wide range of circularity activities already present in the stakeholders’ actions. The framework also worked as a tool to bring forward different aspects of circularity that would not necessarily have been discovered with other more traditional CE frameworks. The qualitative results suggest that within the stakeholder group there are different ways of implementing the circular economy across the different CE dimensions, and that the circular economy is therefore not a one-dimensional activity that is either implemented by an actor or not. The results also support previous literature that a holistic and systemic approach is needed to achieve sustainability transition in the construction and housing sector.

Abstract Strategic development in the forest sector has been slow and not until the 21st century stake-holder focus came as a part of the strategic development. New innovations are now having important role while renewing forest sector into bioeconomy. Corporate Social Responsibility (CSR)has traditionally been as a target for development in forest sector because of the high usage of natural resources, but from the 1990s it has gained more attention inspired by the global megatrends and now CSR can be seen as the base of the consept of bioeconomy. The study explored the citizen’s perception towards forest sector innovation efforts from the year 2000 and within the next 20 years. The results were drawn from an analysis of two data sets. Data one consisted of multivariate survey data based on 218 valid responses where current state of forest industry innovative-ness was disclosed by using 13 bioeconomy products and services. Data two consisted of qualitative managerial interviews where forest industry professionals’ opinions of forest in-dustry contributions to sustainable innovations through the lens of three-dimensional innova-tion model/pyramid. Additionally, the aim was to get some reflections from survey results. Concluding the results of this study, the public opinion of forest sector innovations seemed to focus on constructions related innovation efforts and biofuels. In addition, results showed that perceptions of material substitution with wood and reducing environmental impacts of industry were improved since year 2000. The image of future innovation targets of forest sector was quite wide and most heavily it was pointed towards wood building systems, con-struction materials, brand development and material substitution with wood. Statistically sig-nificant differences found between the “past” and “future” innovativeness inquire strengthen-ing of both industry R & D and functioning of innovation systems.

Agriculture’s contribution to climate change is controversial as it is a significant source of greenhouse gases but also a sink of carbon. Hence its economic and technological potential to mitigate climate change have been argued to be noteworthy. However, social profitability of emission mitigation is a result from factors among emission reductions such as surface water quality impact or profit from production. Consequently, to value comprehensive results of agricultural climate emission mitigation practices, these co-effects to environment and economics should be taken into account. The objective of this thesis was to develop an integrated economic and ecological model to analyse the social welfare of crop cultivation in Finland on distinctive cultivation technologies, conventional tillage and conservation tillage (no-till). Further, we ask whether it would be privately or socially profitable to allocate some of barley cultivation for alternative land use, such as green set-aside or afforestation, when production costs, GHG’s and water quality impacts are taken into account. In the theoretical framework we depict the optimal input use and land allocation choices in terms of environmental impacts and profit from production and derive the optimal tax and payment policies for climate and water quality friendly land allocation. The empirical application of the model uses Finnish data about production cost and profit structure and environmental impacts. According to our results, given emission mitigation practices are not self-evidently beneficial for farmers or society. On the contrary, in some cases alternative land allocation could even reduce social welfare, profiting conventional crop cultivation. This is the case regarding mineral soils such as clay and silt soils. On organic agricultural soils, climate mitigation practices, in this case afforestation and green fallow give more promising results, decreasing climate emissions and nutrient runoff to water systems. No-till technology does not seem to profit climate mitigation although it does decrease other environmental impacts. Nevertheless, the data behind climate emission mitigation practices impact to production and climate is limited and partly contradictory. More specific experiment studies on interaction of emission mitigation practices and environment would be needed. Further study would be important. Particularly area specific production and environmental factors and also food security and safety and socio-economic impacts should be taken into account.

Discussion around climate crisis and companies’ role in its mitigation has been accelerating especially in the past few years. Companies are in a crucial role if the targets set in the Paris Agreement are wished to be fulfilled. Companies have also noted the importance of the topic. Corporate environmental responsibility and sustainability themes have gotten a firm foothold in corporate world and companies can control them by utilizing different corporate governance mechanisms. This thesis aims to examine the importance of corporate governance and sustainability management in companies. Purpose is to study whether there is a link between the level of companies’ climate maturity and different corporate governance mechanisms that are used to manage companies’ sustainability. In addition, this thesis examines if there are differences in the results depending on size, industry, or country where companies are headquartered. Scope of this thesis is corporate environmental responsibility and climate sustainability in the context of greenhouse gas emissions. Companies are divided into two groups based on their climate maturity which is determined by whether they have set science-based emission reduction targets validated by the Science Based Targets initiative. Analysis is conducted with statistical analysis, logistic regression and is carried out with Stata. Data is originally from a corporate study and consists of 46 medium and large-sized Nordic companies from various industries. Sustainability criteria in management’s incentive plans and in companies’ investment decisions had a positive and significant link to companies’ climate maturity. Chief Sustainability Officer and board-level sustainability committee were insignificant in the model. Larger companies were more connected to climate maturity most likely because they have more resources to develop their sustainability and corporate environmental responsibility. In addition, larger companies are often obligated to disclose their sustainability performance and face pressure from the public to decrease their negative effects which can encourage them to set more advanced targets. It seems that the most effective measures are mechanisms with concrete criteria, compared to the more symbolic measures with no direct effect. Companies should focus on creating actions with impactful measures that create change in their organizations whereas policy makers should aim to create regulation directing companies towards these measures. Scientific research can help by providing knowledge of the most impactful corporate governance mechanisms. Sample size was relatively small, which prevents from making highly generalized conclusions. With a larger dataset, companies’ maturity could have been determined on a wider scale, different analysis methods could have been used and sustainability could have been considered in a more comprehensive perspective.

Climate change is challenging farmers around the world, undermining food security and livelihoods. Smallholder farmers are encountering changing environmental conditions and extreme weather events as a significant negative impact on their livelihoods and food security when the resources needed to adapt are not sufficiently available. Previous studies show that the impacts of climate change are currently most noticeable in the developing countries of the Global South, where local agriculture and smallholder farming still remain an important part of people's livelihoods and local food security. This study focused on the experiences and outcomes of the Building Resilience -tool I (BR-I) for smallholder farmers in 5 different African countries between 2021 and 2022. Through the tool, farmers assessed the impacts of climate change on their livelihoods and their own capacities and needs to adapt to the challenges caused by climate change. The data used in this study consisted of reports filled in when using the tool collecting farmers' responses, and separate feedback on their experiences of using the tool. The results showed that climate change reduced the predictability of environmental conditions and increased the irregularity of the growing seasons. Drought, floods, and storms were the main threats to crops, often leading to yield losses or poor crop quality. Other losses included livelihood-related losses of material assets, for example during floods and storms. Indirect negative impacts of climate change included loss of farmers' livelihoods, increased poverty and reduced local food security and weaker nutritional quality of food. To adapt to climate change impacts, farmers often used more sustainable and higher quality crop varieties and, for example, built more efficient irrigation systems. However, the results showed that to adapt to climate change, farmers need more material and financial resources and research-based knowledge to improve their livelihood activities.

Imbalance of intestinal microbiota is called dysbiosis. Signs of dysbiosis are altered abundance of different bacterial species and reduced diversity together with altered interactions between bacterial species and microbiota and the host. Dysbiosis of intestinal microbiota is connected to many intestinal diseases and today many studies are focused to find so called “next generation” probiotics to be used for the alleviation of dysbiosis instead of traditional antibiotic treatments. The study was made in the Human Microbiome Research Program, Faculty of Medicine, University of Helsinki. Aim of the study was to isolate spore-forming bacterial species for the treatment of intestinal inflammation and infections with bacterial therapy. For this purpose, feces from a healthy adult who had acted as a donor for fecal microbiota transplantation was used to isolate spore-forming commensal bacteria. The isolated bacteria were identified and their ability to adhere into intestinal epithelium and strengthen it was investigated. Also anti-inflammatory potential of these isolated bacterial strains was investigated. For isolating bacteria three different heat treatments and ethanol and methanol treatments were used as a pre-treatment step. Pre-treated samples were cultivated on YCFA-media and isolates were picked from plates at different growth points for further cultivation. Selected isolates were purified, their DNA was isolated and they were identified by partial 16S rRNA -gene sequencing. From these identified isolates four isolates were chosen for further investigation and their full length 16S rRNA -gene was sequenced. These isolates were studied also by using API and aerotolerance tests. Potential anti-inflammatory and adhesion properties of the isolates were investigated by attenuation, adhesion and TER-experiments. In the isolation, the effect of different pre-treatments on the recovery of isolates was clear and based on sequencing isolates that were spore-forming anaerobic bacteria were selected for further investigation. Three of the isolates were Clostridium butyricum and one Blautia wexlerae species. Anti- and pro-inflammatory properties of these isolates were very different depending on isolate and one of them was potentially anti-inflammatory. Isolates also adhered differentially and two of them possibly strengthened gut epithelial barrier so they are promising for further research and in the future investigation with these isolates continues. Experience and results with different cultivation methods can be used to for further development of cultivation for anaerobic intestinal bacteria.

Microbial cellulases, e.g. cellobiohydrolases, are able to degrade cellulose and lignocellulosic biomass to smaller glucose-containing monomers and oligomers. Cellulases are often multi-domain enzymes comprised of different protein domains (i.e. modules), which have different functions. The main two components, which often appear in cellulases, are the cellulose-binding module (CBM) and the catalytic domain. The CBMs bind to cellulose, bringing the catalytic domains close to their substrate and increasing the amount of enzymes on the substrate surface. The catalytic domain performs the cleavage of the substrate, e.g. in the case of cellobiohydrolases hydrolyses or “cuts” the crystalline cellulose chain into smaller soluble saccharides, mainly cellobiose. Unlike aerobic fungi, which utilize free extracellular enzymes to break down cellulose, anaerobic microbes often use a different kind of strategy. Their cellulases are organized and bound to the cell surface in a macromolecular protein complex, the cellulosome. The core of the cellulosome is formed of a scaffolding protein (the scaffoldin) consisting mainly of multiple consecutive cohesin domains, into which the catalytic subunits of enzymes attach via a dockerin domain. This creates a protein complex with multiple different catalytic domains and activities arranged in close proximity to each other. Dockerins and cohesins are known to bind each other with one of the strongest receptor-ligand -pair forces known to nature. Dockerin containing fusion proteins have also been successfully combined in vitro with proteins containing their natural counterparts, cohesins, to create functional multiprotein complexes. In this Master’s thesis work the goal was to 1) produce fusion proteins in which different CBMs were connected to dockerin domains, 2) combine these fusions with cohesin-catalytic domain fusion proteins to create stable CBM and catalytic domain containing enzyme complexes, 3) to characterize these enzyme complexes in respect of their thermostability and cellulose hydrolysis capacity and 4) to ultimately create a robust and fast domain shuffling method for multi-domain cellobiohydrolases (CBH) to facilitate their faster screening. The hypothesis of the experiments was that different CBMs fused with a dockerin domain and the cellobiohydrolase catalytic domain fused with a cohesin domain could be produced separately and then be combined to produce a functional two-domain enzyme with a dockerin-cohesin “linker” in between. In this way time and work could be saved because not every different CBM- catalytic domain -pair would have to be cloned and produced separately. Several CBM-dockerin fusion proteins (in which the CBM were of fungal or bacterial origin) were tested for expression in heterologous hosts, either in Saccharomyces cerevisiae or Escherichia coli. The purified proteins were combined with a fungal glycoside hydrolase family 7 (GH7) cellobiohydrolase-cohesin fusion protein produced in S. cerevisiae. The characterization of the catalytic domain-CBM -complexes formed through cohesin-dockerin interaction included thermostability measurements using circular dichroism and activity assays using soluble and insoluble cellulosic substrate. The results were compared to enzyme controls comprising of the same CBM and catalytic domain connected by a simple peptide linker. The results showed that the cohesin-dockerin –linked cellobiohydrolase complex performed in the cellulose hydrolysis studies in a similar manner as the directly linked enzyme controls at temperature of 50˚C and 60 ˚C. At temperatures of 70 ˚C the complex did not perform as well as the control enzymes, apparently due to the instability of the dockerin-cohesin interaction. The thermostability measurements of the enzymes, together with the previously published data supported the hydrolysis results and this hypothesis. The future work should be aimed at enhancing the thermostability of the cohesin-dockerin interaction as well as on verifying the results on different cellulase fusion complexes.

The textile industry is in a need of various solutions to address its sustainability and responsibility issues and goals the industry players have set. Sustainable textile material sourcing plays a remarkable role as a solution for such issues. In addition to sourcing cotton, polyester or other conventional textile materials, next generation textile fibres are a part solution for the industry. However, market diffusion of such innovative products is a complex process. Purpose of the study is to study the innovation commercialisation and innovation selling from the point of view of a start-up or an innovator company that develops such technologies for the market. The literature part of the study defines the key determinants for a successful innovation commercialisation, which are reflected to results of semi-structured interviews with innovators from the industry. By collecting views and perceptions by textile brand owners and consumers, the study forms a practical pathway for innovators to make their commercialisation process succeed, ultimately, in support to tackle the sustainability issues of the industry. Results of the study elaborate the importance of flowing exchange of information throughout the textile value chain. Information flow is based on the value propositions of the innovation, which function as core values of innovators’ team building and communication with the value chain and the consumers, to build grounds for innovation acceptance and market diffusion. Reciprocally, collecting feedback from both is crucial to adapt to expectations flexibly. Lack of consumer awareness and, therefore, currently slow implementation pace of next generation textile fibres by the brand owners limit the resources of innovators and slow down market diffusion. The study finds that the role of brand owners in shifting consumer perceptions of prevailing issues in the industry is centric. Generally, consumers find the sustainability topics the brand owners promote and state to be addressing, as the most concerning. Therefore, as the brands are sparsely involving or offering next generation textile fibres as solutions for the issues, consumers do not see them as sufficient solutions. The study concludes that commercialisation of next generation textile fibres requires better facilitated educative information flow from innovators with all stakeholders of the process to build consumer demand and to activate the value chain. The most evident measure for it would be the better utilisation of various key opinion leaders. Well facilitated information flow and early start of educative sharing of information are centric for the successful market diffusion.