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Increased intestinal permeability and its role in autoimmune, metabolic and chronic intestinal diseases is under extensive research as the “leaky gut” is considered as a potential target for preventive and therapeutic strategies in wide range of diseases. Zonulin, an eukaryotic analogue of Vibrio cholerae derived Zonula occludens toxin, which induces tight junction disassembly, has recently become a popular serum-based biomarker of intestinal permeability in biomedical research, even though the link between serum zonulin levels and functional measures of intestinal permeability has never been validated properly in humans. In addition, surprisingly little is known about the location and regulation of zonulin expression in the humans despite the protein was discovered almost two decades ago. Zonulin, also known as pre-haptoglobin-2, is an uncleaved precursor form of haptoglobin that is abundantly expressed in the liver. Zonulin, in turn, based on studies on rats, rabbits and monkeys, is expressed in the small intestine and stimulated by exposure to bacteria and gliadin, but no other stimulators have been described so far. It is also unclear, if different bacteria can induce different responses in zonulin secretion as only the effect of gram-negative enterobacteria has been documented so far. The aim of this study was to evaluate the effect of selected intestinal bacteria and of two known upregulators of haptoglobin, interleukin-6 (IL-6) and bacterial lipopolysaccharide (LPS), on zonulin secretion in vitro. The impact of two gram-positive probiotic bacteria (Lactobacillus rhamnosus GG & Bifidobacterium bifidum) and of two commensal gram-negative bacterial strains (Escherichia coli DH5α & Escherichia coli RY13) were tested for zonulin secretion in HT-29 intestinal epithelial cells, in addition to IL-6. Two separate lineages of immortalized human hepatocytes were tested for zonulin secretion by stimulation of LPS and IL-6. In addition, different immunological methods were assessed for quantification of zonulin, as the potential cross-reactivity of our primary analysis method, a commercial zonulin ELISA kit from Immundiagnostik AG that is also the main method used in the published zonulin studies, became more evident at the beginning of this thesis project. The main findings of this study were that the widely used commercial zonulin ELISA from Immundiagnostik AG is not specific for zonulin, but instead cross-reacts at least with complement C3, in line with the results published by other group during this work. Our further experiments comparing the signals of the above-mentioned zonulin ELISA and complement C3 ELISA for serum samples showed that there was only weak correlation between the obtained signals, suggesting that the zonulin antibody does not directly bind to complement C3. By using dot blot, western blot and immunoprecipitation, we found that the cross-reaction only occurred in native conditions. Based on zonulin ELISA measurements of the cell culture media from the in vitro experiments, very low signal was obtained for both intestinal and hepatic cells. Among the tested bacteria, only exposure to Lactobacillus rhamnosus GG led to a significant increase in the release of target protein. In hepatic cells, LPS had no effect, while IL-6 led to a significant increase of zonulin ELISA signal in one of the tested hepatic lines. However, it is currently difficult to differentiate if the low detected “zonulin” levels in this study are due to low level of secretion, or rather due to the lack of a proper method to detect zonulin. Taken together, these observations suggest that the most commonly used zonulin ELISA and other related, commercially available antibody-based methods for zonulin detection should be utilized with caution, as these antibodies cross-react with other protein(s). Hence, the serum “zonulin” cannot be considered as a biomarker of intestinal permeability until the captured protein(s) are identified, and similarly the anticipated effects of intestinal bacteria on zonulin expression cannot be reliably investigated with the currently available antibodies.

During recent years obesity has been under extra scrutiny due to its globally rising prevalence, multifaceted effects on the human body and common occurrence of comorbidities. It is estimated that one third of the Finnish population over age 40 will be obese by 2028 (THL, 2022). Consequently, development of mitigation strategies has become a high priority in today’s societies leading to a rising need for new treatments. Several studies have shown how pathological adipogenesis has deleterious effect on brain functionality. The neuropathology of obesity could be explained by increased blood-brain barrier (BBB) leakage, oxidative stress, neuroinflammation and glial activation. Pathologically activated astrocytes (astrogliosis) exhibit phenotypical and functional differences compared to healthy astrocytes, typically exhibiting enlarged cell bodies and swollen cytoskeleton. Astrogliosis has been mainly studied in the context of CNS diseases. Recent studies also shed light on the role of astrocytes in the progression of peripheral diseases including cancer metastasis or inflammation (Ma et al., 2023). However, the active astrocytic profile in obesity is relatively underexplored. In this study we report astroglial phenotypic shifts induced by high-fat diet (HFD) feeding and weight loss (WL). No significant change in GFAP expression was seen between mice that were given an HFD for different durations and their corresponding controls. However, we noted a non-significant trend for increased GFAP expression in response to shorter timepoints (5 days of diet change). This suggested an early astrocytic response to diet, which later normalizes over time. We reported healthy morphologies in astrocytes from chow group exhibiting typical simple thin cytoskeleton with long cell protrusions. Astrocytes in HFD-conditions exhibited reactive phenotypes evidenced by swollen cytoskeletal structures and high GFAP immunofluorescence, extensive lipid droplet (LD) accumulation and upregulated metabolic activity. These observations indicated stressful conditions caused by the diet. Astrocytes in WLconditions exhibited varying phenotypes displaying both reactive and healthy characteristics, slight increase of metabolic activity and lipid accumulation. In addition, we reported different immunofluorescence profiles between glial differentiation promoting marker Meteorin and ER stress regulated cytoprotective marker MANF between the experimental groups. These results show that HFD-induced obesity and consecutive weight loss induce a reactive-like phenotypic shift on astrocytes involving both morphological and functional changes.

Luontopohjaiset ratkaisut (NBS, Nature-based solutions) ovat tietoisesti suunniteltuja ratkaisuja komplekseihin haasteisiin, ja niiden ytimessä ovat ”luonnon” elementit, kuten valikoidut mikrobit ja kasvillisuus. Nämä elementit ovat luontopohjaisen ratkaisun ytimessä, ja näin tekevät NBS:stä usein ekologisesti kestävämmän vaihtoehdon perinteisille teknisille maisemaratkaisuille. Suuri osa olemassaolevasta kirjallisuudesta on siten keskittynyt NBS:n ekologiseen ulottuvuuteen huolimatta niiden sisäsyntyisestä moniulotteisuudesta. Sosiaaliset elementit ovat kuitenkin paitsi olennainen osa NBS:n suunnittelua ja toimintaa, myös niiden pitkän tähtäimen onnistumisen edellytys. Aiempi tutkimus on osoittanut, että luontopohjaisten ratkaisujen menestys riippuu ennen kaikkea niiden paikallisen yhteisön saavuttamasta yhteydestä ja omistajuudesta. Näiden suhteiden muodostuminen tapahtuu usealla tasolla, mukaan lukien psykologisella ja asuinaluetasolla. Siksi tämä tutkimus tarkastelee luontopohjaisiin ratkaisuihin yhdistettäviä merkityksiä ja näkemyksiä paikallisen, karttapohjaisen, audio-visuaalisen kyselyn kautta. Vastauksista nousee esiin kaksi primääristä huomiota: Ensiksi, aineistosta on mahdollista erottaa NBS- ja äänimaailmamieltymysten, sekä avointen paikan merkitysten pohdintojen kautta vähintään kaksi paikallista arkkityyppiä, jotka lähestyvät luontopohjaisia ratkaisuja uniikeilla tavoilla. Toiseksi, erot näiden arkkityyppien reaktioissa paikallisiin luontopohjaisiin ratkaisuihinsa voivat selittyä osin luontosuhteen ja paikallishistorian kautta. Nämä löydöt korostavat yhtäältä olemassaolevan henkilökohtaiseen psykologiseen ulottuvuuteen keskittyvän NBS-tutkimuksen merkitystä, ja toisaalta paikallisen ja yhteisötason ymmärrystä kokonaisen

Master's thesis project includes the backbone assignment of the human activity-regulated cytoskeleton-associated protein C-lobe (hArc, Uniprot ID: Q7LC44), 7-fluoroindole-based tryptophan-labeling method, and comparing that with the 100% double-labeled and 20%(13C) fractionally labeled samples. The project focuses on the effects of 7-fluoroindole-based fluorotryptophan-labeling. hArc C-lobe has only one tryptophan, which makes the analysis easier. Typically fluorotryptophan-labeling is a costly method – fluorotryptophan itself is very expensive and attaching the fluorine to the tryptophan while expressing is expensive and complicated. Fluoroindolebased labeling circles around the problem, as indole and serine are used in procaryotic systems for tryptophan biosynthesis – meaning that fluoroindole, which is cheap, could be used as an alternative for previous methods. Fluoro-labeled tryptophan is used in protein NMR; for example, in binding studies – fluorine-probes are sensitive, and binding of ligand or protein would move these peaks, indicating binding. This project aims to get an insight into the application of this labeling method. The goal is to see if one could utilize one sample with both (1H, 15N, 13C) labeling and 7-fluorotryptophan labeling for binding and structural studies. However, fluorine is very electronegative, affecting surrounding structures and possibly sequentially nearby amino acids. This possible effect will be observed and determined by comparing the 1H15N-chemical shifts between well-established labeling methods and fluoroindolebased labeling. To determine what amino acids in the protein are affected, if they are affected, will be determined by using the backbone assignment results and the results from the sample comparisons.

Ecosystem accounting is a new framework for integrating the value of nature into decision-making. Previous measures of natural capital accounting have not been able to achieve policy relevancy and have only limited use cases. It is important to look into the usability of ecosystem accounting, to ensure that it is implemented in a way that supports decision-making. The goal of my research is to provide a categorization of barriers and enablers of use, as well as provide some insight into how to make ecosystem accounting usable for decision-making. There is only limited literature on ecosystem accounting, so I conducted a scoping literature review on barriers and enablers of use of natural capital accounting to look into the issues and opportunities in making ecosystem accounting usable. I complemented this literature review with interviews of ecosystem accounting experts from Norway and Germany. Both countries have started the implementation journey and thus provide a view into the active development phase. My main results are the categorization of barriers and enablers of use of ecosystem accounting, as well as main messages for creating usable accounts. I categorized the barriers and enablers into 12 categories under three themes of political, structural and relevancy related barriers and enablers. The categories can be used when planning implementation to identify potential issues and to plan accounts to be as usable as possible. I have three main messages for compiling usable ecosystem accounts. First, sufficient resources are essential in gathering comprehensive accounts. Second, compiling the accounts rely on cooperation of multiple institutions, thus there is a need to agree on forms of coordination. Third, the categories of barriers and enablers are interconnected, and thus there are positive feedback loops that can support making ecosystem accounts usable for policymaking.

Oil spills in aquatic environments are devastating disasters with both biological and economic impacts. Fish populations are among the many subjects of these impacts. In literature, there are numerous assessments of oil spill impacts on fish populations. From all applied research methods, the focus of this thesis is on Bayesian methods. In prior research, several Bayesian models have been developed for assessing oil spill impacts on fish populations. These models, however, have focused on the assessment of impacts from past spills. They have not been used for predicting impacts of possible future oil spills. Furthermore, the models have not utilized data from laboratory studies. Some examples can be found of models assessing economic impacts of oil spills on fish populations however, none of them assess the economic impacts that follow from decreases in biomass. The aim of this thesis is to develop a Bayesian bioeconomic prediction model, which would be able to predict oil spill impacts on Baltic Sea main basin herring population, and the consequential economic impacts on fishermen. The idea is to predict the impacts of several hypothetical oil spill scenarios. As a result of this thesis, a bioeconomic prediction model was developed, which can predict both biological and economic impacts of oil spills on Baltic Sea main basin herring through additional oil induced mortality of herring eggs. The model can be applied to other fish populations in other regions as well. The model utilizes laboratory studies for assessing population level impacts. The model can be used for both assessing risks of the impacts of possible future oil spills, and for decision analysis after a spill has already occurred. Furthermore, the model can be used for assessing unknown aspects of past oil spills. The economic predictions can be used, for example, to estimate the compensations that could possibly be paid to fishermen. In the future, the prediction model should be developed further, especially regarding its stock-recruitment relationship assumptions. In addition, the model’s assumptions regarding the calculation of oil induced additional mortality and the economic impacts, should be expanded.

Urbanisation threatens species and biodiversity globally. Consequent habitat loss and habitat fragmentation force species upon one another, inevitably also increasing human-wildlife conflicts. Despite the situation growing dire for many organisms, studies also show species from a variety of taxa being able to adapt to urban environments. Most studies of primates’ ability to adapt to urban environments have been done on diurnal species. To my knowledge, the African lesser bushbaby (Galago moholi) is the only nocturnal primate in which this has been studied. To assess urban adaptability in another nocturnal primate, I present transect and recording data on the abundance of the white-tailed small-eared greater galago (Otolemur garnettii lasiotis) in an urban and rural environment in the biodiversity hotspot Taita Hills, Kenya. Sampling was done in Wundanyi town and Ngangao forest and the two locations were then compared by fitting a negative binomial as well as a Poisson model for recording and transect count data. Additionally, preliminary observations are made regarding behavioural and spectral acoustic adaptation, increased sociality, and colouration coupled traits. My results indicate O. g. lasiotis to be significantly more abundant in the town than in the forest. In the town, I observed a two-fold increase in total number of vocalizations and mean vocalization rate, and a seven-fold increase in total number of sightings and mean encounter rate, as indicated by recordings and transects respectively. This discrepancy in estimated abundance differences in location from transect and recording data (two-fold versus seven-fold) suggest that urban individuals vocalize less than rural individuals. The spectral profile of three vocalization types were studied and urban individuals exhibited significantly higher high frequencies in the clustered squawk vocalization. I found town dwelling individuals to be a part of a larger group or pair considerably more often than forest dwelling individuals. A dark morph, as opposed to a light morph, was significantly more frequently encountered in the urban location, whereas both morphs were encountered almost equally often in the rural location. In summary, O. g. lasiotis seems to be capable of adapting to an urban environment, as indicated by its’ greater abundance in the town than in the forest. My data suggest decreased vocalization rate, increased sociality, and colouration coupled traits being possible adaptations affecting O. g. lasiotis’ capability to inhabit an urban environment. Further research is required to draw conclusions on what factors and adaptations might allow for the high abundance of O. g. lasiotis in the town.

Innovation in food packaging interlinks many sustainability challenges ranging from food loss and waste through the value chains, to resource extraction and growing amounts of plastic waste globally. Food packaging innovations arising from regulation often focus on material waste and ignore other facets of sustainability such as food loss and waste. Simultaneously, conventional notions of innovations are focused on firm growth and competitiveness. This study investigates the perceptions of sustainability in food packaging among expert actors in Finland. Moreover, it examines how notions of Responsible Research and Innovation (RRI) are reflected in the research and development processes in the field. Here, RRI is understood as a framework for examining the role of socio-ethical considerations in research and development. The study aimed to find out which packaging attributes are considered sustainable, what motivations actors in the field have, what type of obstacles exist to innovation in the field, and which actor groups are perceived to be responsible for accelerating the food packaging transition towards sustainability. Semi-structured expert interviews were conducted with 14 participants, and the interview data were analyzed using qualitative content analysis (QCA). The results show that perceptions of sustainability in food packaging vary across the field. However, reducing food waste and loss was considered the most important facet of sustainability in food packaging. Actors in the field are motivated by personal reasons and the anticipated profitability of sustainable innovations. However, innovations in the field are slowed down because of regulatory issues, food safety requirements, unpredictable future changes, and technological lock-ins. Finally, the results of this study indicate that actors in the sector believe the Finnish government and brand owners in the food and beverage industries should be responsible for driving innovation towards improved sustainability. However, the qualitative approach taken here limits the generalizability of the results. The results suggest an ongoing narrative shift in innovation towards greater inclusion of social and ethical considerations in the research and development process.

The need to develop and expand urban areas is increasing in most countries, but urbanization also increases the threat for global biodiversity. Some cities have acknowledged this challenge and formed strategies and action plans for biodiversity preservation. How can we ensure that such strategies are realized in city planning? Negotiations are a crucial part of urban planning, and therefore can be a leverage point of intervention to effectively implement strategies to pro-tect biodiversity. However, little is known about the dynamics of the actual negotiation process in city planning. I applied a game theoretic approach to study how information availability influences the suc-cess and efficiency of negotiations. A role-playing game was used to simulate a negotiation on specific measures to preserve biodiversity in a residential building project. Eleven urban devel-opment specialists played the game with different sets of information. In addition to the direct outcomes of the negotiation, I analysed the post hoc discussion and arguments used to gain in-sights into perceptions of biodiversity-related negotiations in urban planning. Results indicate that information availability can increase the efficiency of negotiations. Partici-pants favour principled and integrative negotiation, but incomplete information seems to push them to take a more positional stance. The post hoc discussion also reveals some issues rele-vant to the design of urban planning process for biodiversity. The overall results suggest that a simple game-theoretic framework, implemented in (a) game-like simulation with quasi-experimental control and (b) qualitative analysis of discussions, holds potential for both under-standing (i) how decision makers frame and resolve the negotiation with conflicting interests and (ii) how to design efficient administrative processes taking into account not only the partic-ipants’ preferences but also wider public interests, such as biodiversity preservation.

Pohjoismaissa syntypaikkalajitellun biojätteen kompostointi suurissa kompostointilaitoksissa on yleistynyt merkittävästi 2000-luvun aikana. Viime vuosina biojätteiden ja muiden orgaanisten jätteiden käsittelyssä on alettu suosimaan yhä enemmän mädätysprosessia sekä mädätyksen ja kompostoinnin yhdistävää osavirtamädätysprosessia. Mädätysprosessin avulla orgaanisten jätteiden sisältämästä energiasta pystytään tuottamaan biokaasua ja siitä edelleen uusiutuvaa energiaa. Tämän tutkimuksen lähtökohtana oli tarkastella biohiilen lisäyksen vaikutuksia kompostointiprosessiin, jossa yhdistyvät mädätys ja kompostointi. Aiempien tutkimusten perusteella biohiilen lisäyksellä on todettu olevan positiivisia vaikutuksia erilaisten orgaanisten jätteiden kompostointiprosessiin. Aiemmat tutkimukset on suurimmaksi osaksi tehty pienemmässä mittakaavassa ja ne ovat keskittyneet enemmän maatalousjätteiden kompostointiin. Tämän tutkimuksen tarkoituksena oli selvittää, voidaanko verrattain pienellä biohiilen lisäyksellä nopeuttaa orgaanisen aineksen hajoamista sekä parantaa typen pidättymistä kompostimassassa erilliskerätyn biojätteen osavirtamädätysprosessin kompostointivaiheessa. Tutkimuksen kokeellinen osuus koostui laboratoriomittakaavan kompostointikokeesta sekä laitosmittakaavan kompostointikokeesta Ämmässuon jätteenkäsittelykeskuksessa. Laboratoriomittakaavan kokeiden tarkoituksena oli saada tietoa eri biohiilipitoisuuksien vaikutuksesta kompostointiprosessiin ja toistomäärien kautta saada tuloksille tieteellistä pätevyyttä. Laboratoriokokeissa saatuja tuloksia sovellettiin laitosmittakaavan kokeissa. Tuloksia pyritään tarkastelemaan mikrobiologisen prosessin lisäksi osana jätehuoltoa ja sen kehittämistä. Tulosten perusteella biohiilen lisäyksellä oli myönteinen vaikutus kompostointiprosessiin osavirtamädätysprosessin kompostointivaiheessa. Laboratoriokokeissa biohiilen lisäyksellä oli merkitsevä vaikutus orgaanisen aineksen hajoamiseen ja siten kompostimassan mikrobiaktiivisuuteen. Vaikka laitosmittakaavan kokeessa biohiilen lisäys ei vaikuttanut orgaanisen aineksen hajoamiseen tilastollisesti merkitsevästi, voidaan kokonaisuutena arvioida, että biohiililisäsi kompostien mikrobiaktiivisuutta. Biohiilellä havaittiin merkitsevä vaikutus kompostien stabiloitumisnopeuteen ja ammoniumtypen pidättymiseen kokeen lopputilanteessa. Verrattain pienet biohiilipitoisuudet eivät vaikuttaneet haihtuneen ammoniakin määrään eivätkä kokonaistyppihäviöön kompostoinnin alkuvaiheessa. Biohiilikompostien stabiloitumisnopeudella ja mikrobiaktiivisuuden lisääntymisellä on kompostoinnin ympäristövaikutusten vähentämisen lisäksi kompostointilaitoksille myös taloudellinen merkitys, koska niiden avulla pystytään mahdollisesti säästämään tuotantokustannuksissa sekä säästämään kompostien jälkikypsytykseen tarvittavaa tilaa tai vastaavasti lisäämään kompostointilaitosten kapasiteettia. Tulevaisuudessa biohiili on teoriassa mahdollista saada osaksi orgaanisten jätteiden käsittelykokonaisuutta. Jäteperäisten biohiilten käytöllä voi tulevaisuudessa olla mahdollista parantaa resurssien kestävää käyttöä ja alueellisia kierrätysmääriä.