Browsing by Title

Diabetic ovarian cancer patients who take metformin as part of their anti-diabetic medication generally respond better to DNA-damaging cancer treatment. The molecular mechanisms of the anti-cancer effects of metformin are currently being investigated, but they remain poorly elucidated. Not much is understood about the metformin effect on DNA damage in ovarian cancer cells, where it is of particular importance. When chemotherapy-induced double-stranded DNA breaks are unrepaired, cells reach a point when they cannot tolerate the accumulated DNA damage and die. However, some ovarian cancer cells efficiently employ DNA repair mechanisms, the most prominent being homologous recombination (HR), to overcome DNA damage. Efficient HR causes chemoresistance. An important question is whether metformin has the ability to induce the HR-deficient state in cancer cells, thereby sensitizing them to treatment. This study did not examine HR directly, but it assessed HR indirectly by observing the effect of metformin on recovery from DNA damage in two ovarian cancer cell lines: OVCAR4 (HR-proficient) and Kuramochi (HR-deficient). Additionally, this study evaluated the metformin effect on cell proliferation and apoptosis. OVCAR4 and Kuramochi cells were exposed to varying metformin concentrations (0,5 mM, 5 mM, 10 mM, 15 mM, 20 mM and 25 mM) and for varying durations (24 hours and 48 hours). This study also tested how metformin pretreatment affected the cells’ ability to repair externally (ionizing irradiation) induced DNA damage. The cells were imaged with a high-content imaging system, and percentages of nuclei that were positive for markers for different cellular processes (i.e., DNA damage, proliferation, and apoptosis) were calculated. The study found that only high metformin concentrations, such as 20 mM were able to increase DNA damage and reduce cell proliferation in HR-proficient OVCAR4 cells, both non-irradiated and irradiated. The HR-deficient Kuramochi cell line was generally more sensitive to metformin, particularly with regards to DNA damage, which increased using metformin concentrations < 20 mM. However, 20 mM concentration resulted in the most significant effects. Similarly, only high metformin concentration (25 mM) increased apoptosis, although data were obtained only for a limited number of Kuramochi cells. More experiments on apoptosis would be beneficial. Also, more extensive experiments for the irradiation part are needed to validate these preliminary findings, as well as examining whether high metformin concentrations (> 20 mM) affect specifically the HR-mediated DNA repair pathway.

Enhancement of soil carbon sink has large potential to mitigate climate change. Earlier studies have suggested that improved management practices could promote climate change mitigation and improve soil fertility. To find out if the carbon sink of a clay soil under improved grassland management in Southern Finland can be enhanced by increasing mowing height at harvest, an experiment was set up with two different mowing heights (6 and 15 cm). Net ecosystem carbon exchange, based on total ecosystem respiration and photosynthetic capacity were monitored with chamber methods during three growing seasons from 2019 to 2021. Also, plant biomass, leaf area index, soil temperature, soil pH, soil water retention capacity, and soil grain size distribution were studied at both mowing height treatments. In this study, negative value is the CO2 flux from the atmosphere to the ecosystem and positive value is the CO2 flux from the ecosystem to the atmosphere. Negative NEE means that the ecosystem gains C when the absolute value of GPP is greater than TER and vice versa. The higher mowing height increased CO2 uptake by plants and caused more negative NEE for the higher mowing height after the grass was harvested. These results indicate that higher mowing height might be better for mitigating climate change. However, mowing height did not have a significant effect on biomass, LAI, TER or soil properties in the experiment. Short lasting and non-existent differences between mowing heights are probably explained by more pronounced compensation growth reaction at the lower mowing height as growth conditions were otherwise similar except for mowing height treatment at both treatments. More frequent measurements, especially after the harvest, could better reveal the dynamics of grass height differences and its effects on GHGs. Better detection of the effect of mowing height on the carbon balance would require even more regular and continuous measurements after harvesting and fertilization in different soil types with experimental setups such as applied in this study.

Functional magnetic resonance imaging (fMRI) has become an important tool in measuring the connectivity of the brain. Based on the connectivity maps acquired from scanning, research is being done for example on how to improve fMRI-guided targeting of transcranial magnetic stimulation (TMS). One of the means is to improve the quality of the imaging. There are some challenges in producing high quality images due to technical limitations and due to patient behavior, most important of which are movement and state of alertness. The aim of this thesis was to find out, if natural viewing during scanning would reduce movement, increase alertness and increase comfortableness of the patients compared to resting state scanning. We had 19 (N=19) patients suffering from treatment resistant MDD (major depressive disorder) scanned to receive fMRIguided TMS -treatment. We divided the fMRI into four sessions: two movie and two resting state sessions. After each session we asked the patients to rate their level of alertness and comfortableness on a visual analogue scale from 0 to 100. We also computed two different movement measures (mean FD [= framewise displacement] Jenkinson and number of FD Power >0.5) of the scanning. We could see a significant difference (p=0.014) in mean FD Jenkinson between movie vs rest indicating that natural viewing reduced movement during scanning. We also could see a significant difference in alertness (p=0.002) between sessions indicating that longer duration of scanning diminishes the state of alertness of the patients. In other parameters we did not see a significant difference between movie and resting state. Our research supports and adds to previous findings that showing a movie during fMRI could reduce movement and that state of alertness could decrease during longer scanning times.

To study the feasibility of replacing dairy proteins with plant proteins to stabilise emulsions, 9 emulsions were prepared at pH= 3 with sunflower oil and 1% protein solutions containing WPI, SC, PPI individual proteins and their blends (3:1, 1:1, 1:3). Changes in droplet size and zeta-potential were measured to investigate the emulsion stability. The interfacial composition, surface loads, and the protein adsorption behaviour were studied to analyse the interface. Furthermore, the dilatational rheology of the interface was studied as the interfacial properties play a critical role in emulsion stability. All proteins showed good emulsifying and stabilising abilities at pH=3, as they contributed to form emulsions with relatively small droplets (0.355-0.411µm), and prevented destabilisation for 2 weeks. Such results were related to the strong electrostatic repulsion between the droplets and the viscoelastic film formed by proteins. No synergistic or antagonistic effect was observed by using dairy and pea protein blends, indicating pea proteins could be used to partially replace the dairy proteins. A preferential adsorption of dairy proteins over pea proteins was found, with SC being the most dominating type at the interface due to the higher diffusion rate and structural flexibility. However, globular WPI and PPI formed stronger interfacial films than SC and its blend owing to the extensive intermolecular interactions.

Current diets with high meat consumption have several adverse effects on the environment and human health. Different alternative foods such as whole plant foods (WPFs), plant-based meat substitutes, and novel/future foods (NFFs) are potential options to reduce the environmental pressure caused by meat. The aim of this thesis was to study the effect of replacing meat with alternative foods on the climate impact, energy use, and nutrient intakes in Finnish diets. The study was carried out by formulating scenarios for the current diet using the National FinDiet2017 survey, in which all meat or half of meat was substituted with WPFs, PBMSs, and NFFs. The daily climate impact and energy use were the highest for the current diet while substituting meat with alternative foods reduced the climate impact by 13%−37% and energy use by 15%−40% from the current diet. The largest decrease in the environmental impacts was found when all meat was replaced, especially with WPFs. Moreover, a similar reduction of the climate impact and energy use was found for all alternative scenarios when the environmental impacts were integrated with nutritional properties by using the NRD index. Regarding nutrient intakes in Finnish diets, substituting meat resulted in several beneficial changes such as increased levels of folate and fiber or decreased levels of saturated fatty acid. On the other hand, full replacement of meat with plant-based alternatives led to decreased intake of some nutrients, such as vitamin B12. Even though some NFFs are still under development, this thesis showed that they may provide nutritious options in future diets. To conclude, replacing even half of the meat may be beneficial for improving nutrient intakes and reducing the environmental pressure in terms of lower climate impact and energy use.

Tämä lisensiaatin tutkimus koostuu kolmesta osasta: kirjallisuuskatsauksesta, kokeellisesta osasta ja liitteistä. Hevosten mahahaavat ovat yleinen sairaus. Maha-suolikanavan mukoosa on vahingoittunut ja haavauma voi yltää lamina propriaan asti. Sairauden diagnosointi tapahtuu tyypillisesti kliinisten oireiden ja hoitovasteen perusteella. Ainoa varma diagnosointimenetelmä on mahan tähystys. Tähystys on kallista ja vaatii erityistaitoa, jonka vuoksi olisi tarve löytää vaihtoehtoinen diagnosointimenetelmä. Terve suoliston mukoosa ei läpäise sakkaroosia ja se hydrolysoidaan ohutsuolessa monosakkarideiksi, joka imeytyy ohutsuolen mukoosan läpi. Jos hevosella on mahahaava, mukoosa on vahingoittunut ja sakkaroosi pääsee imeytymään sen läpi. Imeytynyt sakkaroosi voidaan todeta veressä. Tämän perusteella seerumin sakkaroosi pitoisuuden mittaaminen voisi olla hyvä vaihtoehto mahahaavan diagnosointiin. Sakkaroosin pitoisuus mahdollisesti myös korreloi mahahaavan vakavuuden kanssa. Tämän hypoteesin selvittämiseksi on suunnitteilla tutkimus, jossa suuri määrä hevosia tähystetään, jonka jälkeen niille annostellaan sakkaroosia ja otetaan verinäyte. Verikokeista määritellään sakkaroosin määrä ja verrataan tulosta mahahaavan vakavuuteen. Tähystystä varten hevoset on rauhoitettava. Yleisimmin hevosen rauhoituksessa käytetään detomidiniä ja butorphanolia. Useiden tutkimusten mukaan nämä aineet vaikuttavat mahalaukun tyhjentymiseen hidastavasti. Jos mahalaukun tyhjentyminen hidastuu, sakkaroosi pysyy mahdollisesti pidempään mahalaukussa, mikä voi vaikuttaa sakkaroosin pitoisuuteen seerumissa. Sakkaroosin pitoisuuteen seerumissa voisi siis vaikuttaa se, onko hevonen rauhoitettu vai ei. Tämän tutkimuksen tavoite onkin selvittää rauhoituksen vaikutusta sakkaroosin imeytymiseen. Materiaalimme koostui kymmenestä terveestä hevosesta. Kaikille hevosille suoritettiin kliininen tutkimus ja niiltä otettiin verinäytteet maha-suolikanavan sairauden poissulkemiseksi. Ensimmäisessä kokeessa hevosille annosteltiin 16 tunnin paaston jälkeen 500 grammaa sakkaroosia 10 % -liuoksena nenämahaletkulla suoraan mahaan ja verinäytteet otettiin 0,45, 90, 135, 180 ja 225 minuuttia annostelun jälkeen. Seuraavassa kokeessa hevoset rauhoitettiin 16 tunnin paaston jälkeen ja niille suoritettiin mahalaukun tähystys. Tämän jälkeen niille annosteltiin sakkaroosi ja otettiin verikokeet ensimmäisen kokeen mukaisesti. Sakkaroosin pitoisuus veressä määritettiin GC-FID metodilla. Sakkaroosin pitoisuudesta tietyillä ajanhetkillä hevosen ollessa rauhoitettuna ja ei-rauhoitettuna muodostettiin kuvaajat. Sakkaroosi voitiin havaita seerumissa 45 minuutin kuluttua annostelusta ja sitä oli vielä havaittavissa 225 minuutin kohdalla. Ei-rauhoitetuilla hevosilla sakkaroosin seerumipitoisuudet olivat suuremmat kaikkina mittaus ajankohtina. Kuvaajan malli oli sama kummallakin ryhmällä. Ei-rauhoitetuilla hevosilla huippu tuli jo 45 minuutissa, kun se rauhoitetuilla hevosilla tuli 90 minuutissa. Rauhoitetuilla hevosilla sakkaroosin imeytyminen oli hitaampaa. Tähän selityksenä voi olla tähystys ja siihen liittyvä mahalaukun täyttäminen ilmalla, joka suoritettiin ainoastaan rauhoitetuilla hevosilla. Kuvaajien muotojen yhteneväisyyden perusteella sakkaroosin seerumipitoisuutta voidaan verrata rauhoitetuilla ja ei-rauhoitetuilla hevosilla, mutta viiterajojen pitää olla kummallakin ryhmällä erilaiset.

Agroforestry is a collective name for land-use systems and technologies where woody perennials are deliberately used on the same land-management units as agricultural crops and/or animals. Silvopasture is a type of agroforestry that integrates trees, livestock, and forage crops on the same piece of land. This topic is specifically interesting because silvopasture is already present in Finland, mostly in the form of traditional biotopes, despite these being endangered habitats. Agroforestry is also recognized by the Kyoto Protocol as an afforestation practice that has several advantages. These include environmental benefits, increased productivity, and animal welfare. However, there has been discussion on detrimental effects of animal presence to the trees as well. This research aims to explore the effects of silvopasture on trees, cattle, and the environment in South-western Finland traditional biotopes. Several individual criteria are assessed in three categories: woodland, animal performance and biodiversity. Data was collected on three farms on four pastures through field surveys and farmer interviews. The data is analyzed through multi criterium decision analysis (MCDA) with normalized values from agriculture and forestry to assess performance of silvopastures on three separate categories. Results from four pastures follow a somewhat similar pattern but variation occurs. High scores of forest indicators are reached in woodland category whereas biodiversity category results are lower. Silvopasture negatively affects forest regeneration but otherwise does not have adverse effects. Cattle performance is positive outcome whereas biodiversity category has room for improvement. To get more accurate and comprehensive results the research should be conducted during growing season and with several farms around Finland. However, silvopasture is a meaningful measure to diversify habitats, contribute to animal wellbeing and strengthen ecosystem services.

Due to climate change, the period with snow cover is shortening and the amount of snowfall is decreasing in the boreal region. In Finland, even the probability of snowless winters is increasing in southern and coastal areas of the country. Bilberry (Vaccinium myrtillus) is a keystone species of boreal forests and adapted to overwinter under insulative and protective snow cover. The effects of changing snow cover on bilberry are studied more in Artic and alpine ecosystems but are not well known in forest environments. In this master’s thesis, the effect of snow cover on growth and performance of bilberry was studied in field conditions using snowless treatment area and control area with natural snow cover in a forest environment in Southern Finland. The experiment was carried out during two consecutive winters between the years 2020 and 2022. In the snowless treatment area, the snow shelters prevented snow reaching the ground and shrubs overwintered without protective snow cover, which exposed them to lower air and soil temperatures. Shrubs at the control area overwintered under natural snow cover. Bilberry samples were collected, and field measurements were done in late August 2022. At the field, the number of dead ramets per square meter and the proportion of dead shoots per ramet were counted. In the laboratory, the elongation growth, leaf area and mass, and specific leaf area (SLA) were measured for growing season 2022, and the stem diameter growth was measured for growing seasons 2021 and 2022. After two consecutive snowless winters, the shoot and ramet mortality increased in snowless treatment area compared to control with natural snow cover. The length of an average shoot increased and the number of living shoots per ramet decreased in the treatment area compared to control. The treated ramets also had greater leaf area, leaf mass, and SLA. This indicates changes in bilberry’s resource allocation, and ability to compensatory growth after winter damages. However, increased mortality could lead to decrease in bilberry coverage in forest understory in a long-term. Changes in bilberry growth and performance can have unpredictable and complex effects on forest ecosystems, and there is a need for future research to show how longer snow-free periods and increased cold stress affect this keystone species in boreal forests.

The aim of this research is to examine the sphere of legitimate State measures. Member States intervene in many ways in the operation of the internal market and in the activities of private undertakings. When public authorities exercising public powers take measures, which can affect the competition between private entities, the pursuit may lead to the distortion of competition within the internal market. Similarly, fundamental freedoms can be restricted, if access to the market is prohibited or rendered less attractive. The main legal instrument of this study is Article 106(1) TFEU, which prohibits Member States from circumventing Treaty rules by granting tasks of public authority to public or private undertakings. The provision will be assessed as regards to competition law (Article 102 TFEU) and fundamental freedoms (Article 49 TFEU regarding freedom of establishment and Article 56 TFEU regarding freedom to provide services). In addition, Article 106(2) TFEU, which provides a derogation from Treaty rules for the Member States in certain situations, will be addressed. State measures which violate the above-mentioned Articles can in some cases be justified in order to pursue the Union’s objective of social market economy. They can be regarded legitimate when they are correcting market failures on areas where certain important activities or services would not be provided merely by the market mechanism. In addition, justifications based on overriding reasons of general/public interest, may materialize when restricting fundamental freedoms. In summary, the first research question is whether State measures create obstacles to the internal market. Additionally, it will be examined, when State measures can be justified. Often the CJEU carries out a balancing act between the restrictive measure and a possible justification. In the end, the fulfilment of fundamental freedoms will be assessed in cases related to State measures, by applying the market access test.

Lactase is a digestive enzyme, and its principal function is to break down lactose, a disaccharide found in milk. The main site for lactase expression is the intestines, however, it is also expressed in other tissues, including the brain. Because the primary substrate, lactose, is not present in the central nervous system, it can be assumed that lactase serves another function besides lactose breakdown outside the digestive system. In C57BL/6NCrl mice, lactase expression is higher in the ventral hippocampus after chronic social defeat stress in comparison to controls. This suggests that lactase expression is to some extent affected by stress. Although lactose metabolism is only necessary for mammals, some other animals – including the zebrafish (Danio rerio) – possess a gene that codes for lactase. Research on the zebrafish lactase gene is scarce, and the expression pattern of its two transcripts, the primary lct-201 and the secondary lct-202, is not known. This study focused on measuring lactase expression in adult wild type zebrafish – both on the gene and on the protein level as enzymatic activity. The effect of stress on lactase expression was also examined by applying two different stress models: netting handling stress as a form of physiological stress, and chronic social defeat as a model for psychosocial stress. Real-time polymerase chain reaction (q-RT-PCR) showed lct-201 expression in all five tissues investigated in this study – the forebrain, the mid-hindbrain, higher intestines, lower intestines, and skeletal muscle, whereas lct-202 was only expressed in the higher and lower intestines. The expression level of lct-201 in the muscle was only fifth of that in the lower intestines. Lactase activity assay on the whole brain and whole intestines displayed enzymatic activity in both tissues, with the activity in the intestines being more than seven-fold compared to the brain. q-RT-PCR on both stressed and control fish whole brain and intestines revealed higher lactase expression in the stressed fish intestines, however, the effect was only seen with a primer pair targeting both transcripts simultaneously, and not for either of them separately. Lactase expression was on average approximately 40 % higher in physiologically and 55 % higher in psychosocially stressed fish in comparison to their respective controls. Neither physiological nor psychosocial stress affected lactase expression in the brain. These findings suggest that the two zebrafish lactase transcripts have distinct expression patterns, which might imply different functional roles for lct-201 and lct-202. Furthermore, these results indicate that lactase is expressed in the zebrafish brain, suggesting that it has a specific function in the central nervous system. Based on the findings in this study, lactase gene expression might be connected to experienced stress – both physiological and psychosocial.