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Policy makers must make decisions regarding budget allocation between policies and research. Only actions improve the state of the system, but knowledge increases the probability of effective action. The outcome of environmental policies is usually uncertain, and the question remains: should we invest more in research or use resources for additional policies? Under uncertain decision making, it is clear, that doing and knowing go hand in hand. Still, there is a lack of scientific analyses about the relationship between these features. This paper analyses how uncertainty in policy implementation outcome (i.e., Value- of- control, VoC) effects the need for additional knowledge, which is measured using value- of- information (VoI) analysis. Additionally, the paper analyses how the obtained results can help in the allocation of resources between policy and research. To answer these questions, I use a published Bayesian decision model by Helle et al. (2015) as a source for further analysis. It is an influence diagram model, consisting of two decision variables, 40 random variables and 13 utility variables which are described in euros, allowing the monetary summarisation of utility. I introduce levels of implementation uncertainty to the other decision by placing an additional random variable to influence the successor variables of the decision. I define the levels of uncertainty using two co-variation methods, first one being the proportional co-variation method, and the second one the order- preserving uniform method. By this way, I analyse and compare the effect of distinct levels of decreased controllability (VoC) to VoI analysis results. These two methods describe two alternative ways of modelling the uncertainty in implementation, which is always uncertain when we consider future actions that have yet to been implemented. I conduct the analysis separately for both co-variation approaches, and 10 alternative levels of implementation uncertainty, to enable systematic comparability between the chosen methods, and to learn alternative ways to consider the relationship of controllability and knowledge. First, I preform the VoI analysis only for the policy that is subjected to implementation uncertainty. Secondly, the analysis is done for both decision variables and the Single Policy Updating (SPU) algorithm of Hugin software is used for detecting optimal policies for different implementation uncertainty levels. In other words, I show how the various levels of controllability of the system impact the needs to carry out research. I argue, that this is a fundamental question for many environmental policy questions, such as climate change, eutrophication, loss of biodiversity and in my example, risks of oil spills. The results show a consistent, but interesting effect of decreased controllability to the VoI analysis results. Increase in implementation uncertainty raises the overall VoI and increases the number of variables presented with VoI, i.e., once the estimated uncertainty of controllability increases, the chance to achieve desired results increases only by knowing more. When only one decision variable is included, VoI increases to the point of no control, indicating that VoI is zero when controllability of the system is zero, indicating that there is no point of carrying out research, if the knowledge cannot be used to improve the effects of actions. When both decision variables are included in this case, VoI increases to a certain point and decreases after that. This study highlights the need for such analyses in decision problems, where uncertainty in policy implementation is often overlooked. This is the case with most deterministic, point estimate models. I argue, that this type of analysis would lead to more effective solving of environmental problems.

Phage lysins are enzymes that degrade bacterial cell wall. A wild-type Lactococcus lactis strain LAC460 secretes three phage lysins, LysL, LysP, and LysT, encoded by three different prophages. Unlike common phage lysins, these enzymes do not break down the host's cell wall. Therefore, these lysins can attack other L. lactis strains and behave like bacteriocins, antimicrobial proteins. The binding of a phage lysin to bacterial cell wall requires a specific cell wall binding domain (CBD) in the lysin. However, nothing about the CBDs of LysL, LysP and LysT is known. This study aimed to determine the CBDs of these three lysins and the target specificity of the lysins. Putative CBD regions of the lysins were fused with green fluorescent protein (GFPuv). GFPuv-CBD-LysL and GFPuv-CBD-LysT were ligated into the pASG-IBA4 vector and cloned in Escherichia coli DH5α. After all, only the construction of the GFPuv-CBD- LysL was successful resulting in fluorescent transformants. To analyse the binding of GFPuv- CBD-LysL to cells of different L. lactis strains, the fusion proteins were mixed with the LysL sensitive L. lactis MG1614, LysL resistant L. lactis LM0230, and the LysL producing LAC460 cells. With fluorescence microscope it could be seen that the GFPuv-CBD-LysL decorated the cell surface of L. lactis MG1614 with green fluorescence, but LM0230 and LAC460 cells remained non-fluorescent. The fluorescence of the cells was also measured with a fluorometer, showing strong fluorescence from MG1614, but nothing from the other two strains. This showed that the fusion protein specifically bound to the MG1614 cell surface, but it did not bind to the LysL resistant strain LM0230 or the LysL producer LAC460 cell. In conclusion, the results demonstrate that the C-terminus of LysL contains a specific cell wall binding domain. In addition, the results provide an explanation for how LAC460 can secrete LysL without autolysis, as phage lysins not able to bind onto peptidoglycan are unable to lyse cells.

The increasing recognition of eco-friendly packaging in the packaging industry makes paperboard one of the most promising green packaging materials. The global demand for paperboard has increased rapidly in the past two decades. The purposes of the study are: firstly, to provide an overview of the consumption, production, imports and exports of paperboard products in different regions worldwide between 1992 and 2010. Secondly, as the Finnish paperboard industry has been highly export-oriented, the study also aims to provide quantitative analysis of the main determinants influencing the demand for Finnish paperboard exports in its main destinations-Germany and Russia by applying economic models from the first quarter of 1995 to the third quarter of 2011. Finally, trends projection for the future development of Finnish paperboard is presented based on results of analysis. The research is both descriptive and explanatory based on secondary data collected from various databases. The regional analysis of paperboard market from 1992 to 2010 revealed that North America and Western Europe’s paperboard production exceeded the consumption, where Asia, Eastern Europe and Latin America’s consumption exceeded the production. The consumption and production growth was extremely fast in Asia and it has been the largest consumer and producer of paperboard products since 2000s. Western Europe has been the largest exporter and importer throughout the years. The empirical modeling of Finnish paperboard export demand showed that the export demand for Finnish paperboard in Germany was highly affected by the total paperboard imports of Germany and its GDP. It also has high sensitivity to the income changes of Germany in long term, but the price effect was not significant. The export demand for Finnish paperboard in Russia has low income elasticity and the price effect was statistically significant. Oil price has also significant influence on Finnish paperboard export demand to Russia. According to the linear trends projection for Finnish paperboard market to 2020, the consumption and imports will grow very slowly or even stagnate, but the production and exports will continuously increase at a faster pace.

Salicylic acid (SA) is a well-known phytohormone involved in pathogen defense, development and controlling the cellular redox balance. In response to stresses, Arabidopsis thaliana synthesizes SA in the chloroplasts mainly via the isochorismate (IC) pathway: IC synthase (ICS) uses chorismate to produce IC, which is in turn converted to SA. ICS1 is the rate limiting enzyme in SA biosynthesis. The ICS1 protein localizes in the chloroplasts and contains a chloroplast transit peptide sequence, which is the first 45 amino acids in the N-terminus. Under different light conditions, plants employ different stress defense strategies. In a previous study, wild type (WT) Arabidopsis thaliana ecotype Columbia-0 grown under different light conditions [short day (SD; 8h light/16h dark), long day (LD; 16h light/8h dark) and 12h light/12h dark] was exposed to ozone for 1 hour (350 ppb). In a phosphoproteomic approach to study signaling mechanisms, ICS1 has been found as an in vivo phosphoprotein in ozone-treated plants grown under SD condition by mass spectrometry (MS). To analyze the phosphorylation of ICS1 in vitro, WT and phospho-negative ICS1 proteins were produced recombinantly as GST-fusion proteins in E. coli and used as the substrates for targeted kinase assays. Phospho-negative ICS1 protein was generated by mutating the phospho-sites to alanines by site-directed mutagenesis. ICS1 could be phosphorylated either in the cytosol or in the chloroplasts. Therefore, different cytosolic kinases, which are involved in various stress signaling events, and a chloroplast protein kinase that functions as a dominant regulator of chloroplast processes, were selected as the kinases to be tested. In this study, GST-ICS1 could be successfully phosphorylated in vitro. To a similar extent, GST-phospho-negative ICS1 was phosphorylated, indicating that ICS1 was phosphorylated in vitro at sites that were different from the ones found by MS from plant material. In order to study the identified phospho-sites in vivo, transgenic Arabidopsis carrying WT, phospho-positive, and phospho-negative ICS1 proteins were generated. Transgenic plants were confirmed by genotyping. Western blotting was carried out to evaluate ECFP-WT ICS1 protein expression. However, none of the transgenic plants could be confirmed to carry ECFP-WT ICS1. Additionally, an antibody against AtICS1 produced by the company Agrisera was tested for its specificity against the endogenous ICS1 protein. While the antibody could detect recombinant GST-ICS1 protein, ICS1 could not be detected from a total protein extract.

The literature survey reviewed principles of oxidation of edible oils, adverse effects of lipid oxidation and analysis of volatile oxidation products by SPME-GC-MS. The main aim of the experimental research was to study the influence of relative humidity (RH) on the release of volatile oxidation products from spray-dried emulsions with natural and cross-linked casein as emulsifier. The release of volatiles was determined by SPME-GC-MS. The sub aims were to study the effects of stabilization time at specific RHs, of temperature and agitation speed during the SPME extraction. The spray-dried emulsions were oxidised at 40oC in order to reach a certain level of oxidation. Next, the powders were stabilised under five RHs (0%, 11%, 33%, 54% and 75%) for one or two weeks in order to observe the effect of the RH and the stabilisation time on the release of volatiles. After adjusting the RHs, volatile compounds were analysed by SPME-GC-MS. The following SPME extraction conditions were tested: C1: temperature 40oC, agitation speed 250 rpm, C2: 50oC, 250 rpm, C3: 40oC, 500 rpm and C4: 50oC, 500 rpm. Identification of the compounds was carried out by matching their MS spectra with the NIST database. Altogether 45 volatiles released from the powders could be identified, and 18 of them were found in most samples. RH had an important effect on the release of volatiles from the encapsulated samples. The highest release was always observed at 11% and 33% RH, whereas the lowest release was found at 0% and/or 75% RH, depending on the SPME extraction conditions. The stabilisation time did not have a significant effect on the release of volatiles in most RHs. During the SPME extraction step, elevation of the temperature from 40oC to 50oC, as well as the agitation speed from 250 rpm to 500 rpm, facilitated higher release. However, the effect of temperature was greater than that of agitation speed. Although it was suspected that cross-linking of sodium-caseinate would enhance retention of volatiles, our experiment showed greater peak areas of most volatiles from the cross-linked samples than from the natural ones. By controlling the SPME parameters, it was possible to obtain repeatable volatile compound results. The SPME-GC-MS method applied in this study can be reliably used to analyse volatile oxidation products from spray-dried emulsions. Only at very low or high RH the release of volatiles may differ from samples stored at 11% -54% RH.

3-Chloro-1,2-propanediol (3-MCPD), 2-chloro-1,3-propanediol (2-MCPD) and 2,3-epoxy-1-propanol (glycidol) and their fatty acid esters are contaminants formed during processing fat containing foodstuffs at high temperatures. Mainly MCPD and glycidyl esters have been found to be formed in the deodorization process of oils, and in vegetable oils, such as palm oil, they have been measured at high concentrations. In accordance with the restrictions imposed by the European Commission, the levels of glycidyl esters must be especially monitored, as they have been identified as potentially carcinogenic compounds. The aim of the study was to introduce and validate a gas chromatographic analysis method for glycidyl esters and MCPD esters for the Customs Laboratory. The method was validated for two matrices: first for oils and then for powdered infant formulas. In addition, the success of the validation was examined by analyzing various oil samples previously received by the Customs Laboratory. The Customs Laboratory is also involved in the activities of the European Union Reference Laboratory, for which it was intended to participate in the reference measurement organized by the EU Reference Laboratory. The method for the determination of 3-MCPD, 2-MCPD and glycidyl esters in oils and infant formulas was carried out according to the guidelines of the European Union Reference Laboratory for Contaminants (EURL-PC). Determination of MCPD and glycidyl ester concentrations in oils and infant formulas included the following steps: fat extraction by liquid-liquid extraction (for infant formulas), addition of standards, solid-phase extraction, conversion of glycidyl esters to 3-MBPD esters, transesterification, neutralization, salting out, derivatization and analysis with gas chromatography-mass spectrometry system. Concentrations were determined using internal standard method. The method was validated for the following parameters: specificity, selectivity, limit of detection and quantitation, reproducibility, repeatability, trueness, linearity and working range, stability and measurement uncertainty. The analytical method developed for the determination of MCPD and glycidyl esters was successfully validated for oils and powdered infant formulas. The developed method proved to be specific and selective. The limit of determination was found to be 6.3 µg/kg, 1.3 µg/kg and 0.8 µg/kg for the oil matrix 3-MCPD, 2-MCPD and glycidyl esters. The limits of determination for the infant formula were 5.4 µg/kg, 3.0 µg/kg and 1.6 µg/kg for 3-MCPD, 2-MCPD and glycidyl esters. Recoveries for MCPD and glycidyl esters in the oil and powdered infant formulas were 83-105%. R2 for calibration lines were greater than 0.99, and the lines were linear over the entire measurement range of 2-1000 µg/kg. The relative standard deviation of repeatability and reproducibility was less than 20% for both matrices. The expanded measurement uncertainty for the MCPD and glycidyl esters of the oil and powdered infant formula was less than 50%. For all parameters, the requirements set by the Customs Laboratory and the performance requirements of Regulation (EU) 1881/2006 were met. A method validated for two matrices can then be accredited. The customs laboratory may use the developed method in the future to control 3-MCPD, 2-MCPD and glycidyl esters levels of oils and powdered infant formulas. In the future, the method could also be validated for new matrices, such as liquid infant formulas.

The objective of this thesis is to assess the recreational value (access value) provided by Yuelu Mountain Park in China applying travel cost method (TCM) which is commonly used to estimate non-market benefits. Also, a fee that would maximize the entrance fee income is tentatively calculated. The potential trips to be lost next year are estimated based on local respondents visiting intentions among different age groups. The travel cost demand function is estimated by using basic count data travel cost model-Poisson regression, and survey data collected on-site. Average access values per trip were estimated to be € 0.75 for local and € 64.52 for non-local individuals producing aggregate annual access value of € 20.43 million. Based on the travel cost demand function, an entrance fee of € 5.43 would maximize the revenue collected from the visitors. This would mean more than doubling of the present entrance fee. The result could potentially be utilized when deciding on the entrance fees. It is also suggested that the park management could further study visitors' intentions and reasons either to visit or not to visit the park in the future. Estimated consumer surpluses as well as suggested entrance fee must however, be considered with caution because truncation of the on-site survey data is not accounted for in the Poisson model estimations of this study.

Background: ​In adults, vegetarian and vegan diets have been associated with decreased cardiovascular disease (CVD) risk. At the same time, lower plasma LDL-concentrations are reported in vegans than in omnivores. Classification of a vegetarian diet can be challenging, as the diets vary in the degree and categories of the avoided food products. Recently, plant-based diet indices (PDI), such as the provegetarian food pattern, have been introduced as​ a priori ​method to study vegetarianism as a continuum. Plant-based dietary indices weigh positively foods derived from plant sources, whereas animal foods are weighed negatively. Greater adherence to PDI has been associated with lower CVD mortality and lower incidence of coronary heart disease (CHD). Meanwhile, little is known about the associations of plant-based eating and the risk of CVD in children. Aims:​ The aim of this thesis is to study plant-based eating as a continuous phenomenon in a group of Finnish preschoolers. Furthermore, the focus is on the animal source energy and its associations with dietary intakes, and cholesterol and bile acid metabolism. Materials and methods: ​In this thesis, the data from the cross-sectional MIRA Helsinki Study is used. In 2017, City of Helsinki started a pilot project, in which a vegan food option was offered in municipal daycare. At the same year, MIRA Helsinki Study was started to examine the nutritional status of children following a vegan diet. Altogether 42 children, aged 1–7-years, participated. At daycare, 18 of the children consumed a vegan diet, whereas 24 were omnivores. However, these diet categories did not apply to the child’s diet outside daycare. Children’s diet was assessed with 4-day food records and with food frequency questionnaires (FFQs). Anthropometric measurements were taken and fasting blood samples collected. Blood samples were analysed with standard laboratory tests. Gas-liquid chromatography (GLC) and liquid chromatography with tandem mass spectrometry (HPLC-MS/MS) methods were used to examine serum cholesterol metabolism biomarkers and bile acids, respectively. For each food item (n=719) in the food record data, the weight proportion of animal content was estimated. The animal-origin energy content of each food item was calculated by multiplying the known energy content of the food item with the aforementioned score. The proportion of animal source energy (ASE) per subject across the 4-day food record was calculated by dividing the energy derived from animal sources with total energy intake. The correlations of ASE proportion with dietary intakes and surrogate markers of cholesterol and bile acid metabolism were investigated with the Pearson correlation analysis. A multiple linear regression model was used to test whether the ASE proportion explained the plasma LDL-cholesterol (LDL-C) concentration when controlling for the child’s sex and maternal education. In addition, the associations between the ASE proportion and serum concentrations of 20 bile acid variables were examined with the Pearson correlation coefficient. The Benjamini-Hochberg procedure was used to correct for multiple analyses. Results: ​Among all of the participants, ASE proportions ranged from 0.00 to 0.52, with a median value of 0.275 (that is, 27.5% of total energy derived from animal sources).​ ​I​ntake of animal source energy correlated positively with SAFA (r=0.815, p<0.000), cholesterol (r=0.707, p<0.000) and protein (r=0.493, p<0.001) intakes. There was a strong negative correlation between the ASE proportion and fibre intake (r=–0.836, p<0.000). In a linear regression model, adjusted for child’s sex and mother’s educational level, the model explained 44% (p<.000) of the variance in the plasma LDL-C concentration. Every 0.1 increase in the ASE proportion (10E% increase in energy derived from animal sources) was associated with a 0.32 mmol/L higher plasma LDL-C concentration. Associations of ASE proportion and cholesterol surrogate markers were present: a higher intake of ASE correlated with lower concentrations of cholesterol absorption markers. A similar negative association with ASE proportion was present for two out of four cholesterol synthesis biomarkers. In addition, the bile acid synthesis biomarker (​7α-hydroxy-4-cholesten-3-one) correlated negatively with the ASE proportion. From the 20 serum bile acid variables studied only the ratio of tauro- to glyco-conjugated bile acids was significantly associated with the ASE proportion after correcting for multiple analyses. Conclusion:​ Among the study participants, animal source energy ratio was a powerful predictor of plasma LDL-C concentrations. Dietary patterns characterised by high intake of animal source energy were less likely to meet the dietary recommendations of fatty acid (SAFA and PUFA) intake. Our results on cholesterol metabolism need to be confirmed and replicated in future studies, which should include data on absolute measurements of cholesterol synthesis and absorption.

The biodegradation of wood by fungi in natural forest environment is of vital importance for carbon and nutrient replenishment. Additionally, the ecological functions, maintenance of biodiversity and fungal succession in forest ecosystems is partly dependent on the availability of utilizable organic substances. The fungal group of basidiomycetes are known to play important functional roles in these processes. There is however scarcity of information on how co-cultures of multiple fungi affect wood biomass decomposition as well as the consequent effect of interspecific interaction on niche colonization and substrate utilization. This study investigated and evaluated the role and efficiency of interspecific fungal interaction on biodegradation of Scots pine (Pinus sylvestris L), Norway spruce (Picea abies L. (H) Karst) and birch (Betula pubescens Ehrh) wood. Five fungal isolates were selected for the test in co-cultivation assay: Antrodia sinuosa P. Karst, Bjerkandera adusta (Willd.) P. Karst, Gloeophyllum sepiarium (Wulfen) P. Karst, Heterobasidion annosum (Fr.) Bref, and Phlebiopsis gigantea Jülich (Basidiomycota). The results revealed that antagonistic and competitive fungal interaction does not largely appear to speed up wood decay process. However, the rare combination of Phlebiopsis, Antrodia and Gloeophyllum had significant positive effect on degradation of spruce wood whereas Heterobasidion, Phlebiopsis, and Gloeophyllum had synergistic effect on pine wood decay. It is therefore likely that in nature, wood decay by fungi is mostly facilitated by parallel niche colonization and by fungal succession.

The spread of antibiotic resistance is a global health threat. Hospitals are a potential source of antibiotic-resistant bacteria and antibiotic resistance genes (ARGs), which may disseminate into the environment via wastewater. Hospital water environments, such as sink traps and shower drains, are known to harbor antibiotic-resistant bacteria, which might spread from the drains to the patients causing nosocomial infections that are hard to treat because of the limited number of treatments available. However, the current understanding of antibiotic resistance in the drains of residences, and how it relates to the situation in hospitals is limited. The aim of this study was to compare the microbial communities and ARGs in the water environments of homes and hospitals. The sink traps and shower drains of three hospital rooms and eighteen homes were sampled for metagenomic sequencing, and bioinformatic tools were used to detect the microbial taxa and ARGs in the metagenomes. The resistomes of hospital environments were distinct from those of homes and exhibited a higher diversity of ARGs. On the other hand, the microbial communities of homes and hospital rooms could not be clearly distinguished, although there were some differences in the abundances of certain taxa. The abundance of ARGs was higher in the hospital shower drains than in the corresponding samples in homes, but there was no statistical difference in the abundance of ARGs between the sink traps of homes and the hospital. Although the study had limitations, such as the low number of hospital samples, it indicates that the water environments of hospitals have a resistome that is distinct from that of homes and highlights the role of hospital sink traps and shower drains as potential hotspots of antibiotic resistance.