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Gas chromatography (GC) is one of the most widely used method for analysis of lipids and it is commonly combined to a flame ionization detection (FID), since GC-FID is an excellent combination for quantitative analysis of wide range of lipids. Before GC analysis can be performed, lipids must be extracted from the sample and derivatized. Some common extraction methods include Soxhlet and Soxtec extraction, Folch extraction and extraction with the help of acid hydrolysis. To be able to analyse the extracted fatty acids by GC, they must be in a volatile form. This can be achieved by forming fatty acid derivatives by, for example, methylation. The commissioner wishes to have a way to analyse the fatty acid composition of pork by GC in their laboratory. The aim of the study was to take in use an easy, safe-to-use and simple method for determination of fatty acid composition of pork by gas chromatography, using the instruments and equipment already found from the laboratory. The research had three different parts. First, heptane-isopropanol extraction, Bligh & Dyer extraction and Caviezel extraction were compared. Then the chosen extraction method was validated. Finally, two different pork samples were compared to test the method’s ability to differentiate them. Caviezel extraction was chosen as the best extraction method, and it was validated by parameters repeatability and robustness. The method was able to differentiate samples that had similar, but slightly different fatty acid compositions. However, method still requires some fine tuning since the GC column seemed to be highly sensitive to any disturbances.

I denna avhandling analyseras svenskarnas förhållande till sitt hem och skandinavisk designs be-tydelse för dem genom att analysera de möbler som informanterna nämner som sina favoriter, och hur de överhuvudtaget pratar om inredningen. För att göra detta har 13 unga vuxna intervju-ats i Göteborgs region i Sverige. Därtill har tidigare intervjumaterial från Finland och Tyskland använts som sekundärdata. Analysen, som består av tematiseringen, lyfter fram två olika sidor av det svenska hemmet. Å ena sidan är det viktigt att ha ett hem där man kan vila i. I ett sådant hem består inredningen av möbler som personen känner väl och som har någon historia eller koppling till personen. I den här skyddshamnstankegången är funktionalitet ett centralt värde i inredningen. Å andra sidan verkar svenskarna vara mycket entusiastiska inredare. Den här andra tankegången - det representativa hemmet - betonar design och att hemmet ser bra ut också ur gästens synvin-kel. Förutom status är det möjligt att sådan inredningsstil fungerar väl i Sverige eftersom den är ett lätt och trevligt diskussionsämne. Den skandinaviska designen passar Sverige utmärkt ef-tersom den kombinerar stiligheten och funktionaliteten.

This master thesis aimed to develop a calibration model of the whipping cream for FTIR spectrophotometer MilkoScanTM FT3 and identify the effects of disruptive factors on whipping cream measurement. In the previous calibration model of whipping cream, there could be improvements in the level of the results, especially for lactose. The experimental part of this research included the development of a calibration model, calibration of lactose, total solids assay, and calculation of measurement uncertainty. The calibration model was developed utilizing the statistical methods PCA and PLS in the FossCalibrator program. Lactose calibration was carried out with the LactoSens method, utilizing whipping cream and α-D-lactose, and the results from the reference laboratory were used as reference. The accuracy and functionality of the calibration of total solids wanted to verify, which was why the experimental part included the total solid assay. Total solids were determined by a gravimetric method, and the results were compared with those of the reference laboratory. The measurement uncertainty of the calibration model was calculated with the results of the whipping cream sample reproducibility, repeatability, and accuracy. The new calibration model of whipping cream was verified utilizing validation samples, and the check results were at the expected level. The result of lactose calibration was considerably more accurate than the previous model. The total solids result from the gravimetric method varied slightly with the FT3 results, which, however, did not affect practically. The measurement uncertainty was relatively good, but the calculation will be improved as the number of reference results increases. The main subject of this research was achieved, i.e., the development of the calibration model was successful. The research on the disruptive factors was less than initially intended.

Antibiotic resistance is an increasing, terrible threat to human health, leading to a growing need for alternative therapies. Phage therapy, using bacterial viruses to fight infections, is a promising alternative to antibiotic therapy. However, several obstacles need to be overcome. Regrettably, phage therapy remains inaccessible to many laboratories worldwide due to the need for expensive machinery to establish sensitivity of bacteria to phage. Moreover, shipping phages between laboratories remains challenging. In the current study a device-free bacteriophage typing PhagoGramAssay was developed. In the assay bacteria suspended in soft agar were poured onto a 60-well Terasaki plate containing phages suspended in fibrillated nanocellulose separated from the bacteria by a seal. Phages were released into the bacterial agar layer by puncturing the seal to test for sensitivity observable with the naked eye. Contrast between lysis zone and bacterial lawn was enhanced using 2,3,5-triphenyltetrazolium chloride. Optimized parameters included the amount of bacteria and phage added, volume of phage suspension, agar percentage and thickness and puncturing tool size. In addition, a prototype of such a puncturing tool was developed. The optimized PhagoGramAssay was tested using several bacteria-phage combinations. Moreover, infectivity and stability of phages stored on Terasaki plates was followed over the course of 4 weeks. The optimal bacterial amount added was found to be a 1:300 dilution in soft agar taken from a OD600 = 1 culture. Phage suspensions used in the assay were found to need to have a titer of at least 108 PFU/ml in the original lysate, with 8 µl of 1:10 dilution in fibrillated nanocellulose present in the wells. Optimal agar conditions were found to be 0.4% – 0.5% (w/v) with a thickness of 2 mm – 3 mm. The optimal puncturing tool shape was found to be a slit with a thickness of 0.5 mm. When using these conditions sensitivity could be established for a vast number of bacteria-phage combinations. All phages remained stable and infective over the course of 4 weeks . The newly developed PhagoGramAssay can be further developed into a kit-like phage typing assay that would enable laboratories to test for sensitivity on site whenever a multi-drug resistant bacterial strain is isolated from a patient sample, effectively making phage therapy accessible to laboratories that cannot afford expensive machinery. Additionally, the use of fibrillated nanocellulose should enable laboratories to exchange phages. The final form of such a kit, however, is dependent on manufacturers and investors and may need to be adjusted accordingly.

The conundrum between market entry and business development activities within innovation companies is generally regarded as a challenge. The energy industry as a whole is in a flux and the sustainable future requires drastic actions to be taken to reduce the effects of the global warming and in adaptation of a circular economy model. Using the industrial innovation company St1 Biofuels Oy as a case, this thesis will identify the decision-making components of an opportunity based target market analysis in a company, which operates in an industry with notable resource scarcity, policy regulations and variable business models. In response to this hypothesis, this study suggests an opportunity based target market analysis model that illustrates a new framework to study target markets in a systematic and analytical manner. For the purpose of the case with St1 Biofuels Oy, a market intelligence tool was created to store and process the market data and illustrate the most essential components of the theoretical model. The case study demonstrates the utilization of the opportunity model presenting the internationalization criteria and justification to a potential new R&D concept investment decision. The implications of this thesis contribute to the decision-making of the case and aid in demonstrating analytical justification for internationalization on a strategic decision-making level. This thesis introduces relevant literature to the topic and reflects the existing theories to the new model concept design. Structure and empiric base in this study were drawn together from two-stage data collection, including extensive market research and investment calculations along with semi-structured interviews with the specialists of the case company. The results of this thesis present a theoretical model and the functioning of the model is then piloted with the case study variables of St1 Biofuels Oy. Based on the discussion in this thesis, further research is suggested reflecting the model as a theoretical framework in strategic marketing planning and value-based selling studies.

This thesis studies the current level of environmental management in the Ship Power business division within Wärtsilä corporation and aims to identify the related development needs. Hitherto environmental management has been mainly coordinated at the corporate level and implemented in local companies distributed geographically. Due to the recent organizational changes, however, the significance of division level environmental management in Ship Power has increased. The research goal is approached by examining the central elements of corporate environmental management and the challenges that the organizational structure places for it. Based on the findings, suggestions for further actions are given in order to develop the environmental management in Ship Power. Empirical data was collected through 35 qualitative interviews with Wärtsilä employees from different functions, business lines, and local companies in order to get a comprehensive view of environmental management in the Ship Power related activities. The interviews were semi-structured in order to provide answers for certain areas of concern, but also to enable the disclosure of topics not defined by the interviewer. The data is categorized into themes according to the theoretical background, and its analysis is based on inductive reasoning. Based on the findings, the environmental management in Ship Power is divided into two dimensions. The product-related environmental questions are handled in the business lines and the operational issues in the local companies. This fragmentation of the environmental knowledge causes inconsistent environmental focus at different organizational levels, and creates challenges to information sharing across the organization. The lack of corporate instructions on the environmental management system implementation has led to diverse practices in the local companies, and the lack of standardized documentation makes the internal comparison between them difficult. Therefore the experience gained from the local management systems does not support organizational learning throughout the corporation. While the product-related environmental aspects are the core of the business strategy, more attention could be paid to operational environmental management in Ship Power. For instance, the sharing of environmental knowledge could be strengthened in order to enhance employee awareness of the corporate practices and to facilitate the discussion of the best practices between the local units. A standardized documentation system would facilitate internal benchmarking and provide a means for centralized environmental performance follow-up. Because the local management systems are not sufficient to cover the global business processes, it would also be important to identify the environmental aspects in the Ship Power division. Furthermore, visible communication of the common environmental targets would help to create consistent environmental focus in Ship Power.

Catcher-protein and Tag-peptide originate from split CnaB domains of Gram-positive bacteria surface proteins, which are stabilized by spontaneous intramolecular isopeptide bonds formed between lysine and asparagine residues. However, there is a limited number of non-cross-reacting Catcher and Tag pairs available where the reaction occurs close to the diffusion limit, and which can be used in multiple fragment ligation to construct recombinant fusion proteins. Therefore, a new Catcher/Tag system – LplCatcher/LplTag – was developed in our group from CnaB domain of Lactobacillus plantarum. However, the ligation efficiency of this pair needs to be improved to expand the application possibilities. Therefore, there is a need for efficient library screening method, which allows to detect improved protein-peptide pairs where the covalent interaction takes place rapidly. In this study a new high-throughput in-vivo screening system was developed for visualizing the ligation of Catcher/Tag fusion proteins using splitFAST fluorogenic reporter system for detecting the phenotype, and Fluorescence-activated cell sorting (FACS) for separating the variants at single cell level based on fluorescence intensity. splitFAST is a system engineered by splitting a fluorescent protein named Fluorescence-Activating and absorption-Shifting Tag (FAST) into CFAST and NFAST. The system can be utilized in visualizing the protein interactions because once NFAST and CFAST associate, in the presence of a fluorogen, they form the active and highly fluorescent FAST protein. Herein, Catcher-protein was fused with CFAST and Tag-peptide with NFAST, which allowed detecting the Catcher-Tag ligation ratio based on fluorescence with splitFAST system. Next, a screening system was developed for detecting Catcher variants with improved ligation efficiency. The developed high-throughput screening system showed high potential since visualizing the protein ligation was possible, and hence the system could help in expanding the Catcher/Tag toolbox by allowing large mutant library analyzes.

The current multidisciplinary interests on human intestinal microbiomes have stimulated large scale research initiatives, involving collection and processing of up to thousands of fecal samples within a single study. Hence, there is a need for high throughput protocols that are cost-efficient and validated for their performance to ensure that the relative abundance of different bacteria, the main outcome of microbiota studies, is not biased due to technical artefacts originating from sample processing. Infant’s microbiota colonization is one of the central research areas in human microbiome research because of the long-lasting and profound health implications of the pioneering microbes. This experimental study aimed to develop and validate a high throughput fecal sample collection and processing system for microbial DNA extraction operating in 96 well format. This newly developed method was used to extract DNA from 647 fecal samples collected from mother-infant pairs within a clinical study that will study the effect of antenatal antibiotic prophylaxis on infant’s gut microbiota development. A subset of 28 mother-infant pair samples (14 from each antibiotic and non-antibiotic groups) were selected to study the prevalence of a probiotic bacterium, Lactobacillus rhamnosus GG (LGG), among infants and their mothers longitudinally from birth to 3 months by using species-specific PCR amplification method targeting sortase C gene. In addition, the prevalence of L. rhamnosus GG in 3-month-old infants was compared between the above samples and those (n=30) collected in another clinical trial conducted ~10 years earlier. From extensive testing and validation, an efficient high throughput system for fecal sample collection and processing for extraction of microbial DNA in 96 well format was established. Tests were performed to validate the performance of a) fecal sample collection system b) commercial, readymade bead beating tubes for bacterial cell lysis and c) selfmade wash buffers as part of the automatic DNA purification system. Performance was evaluated based on the quality and quantity of the resultant DNA. We show that this new fecal processing system can yield high quality microbial DNA from 96 fecal samples within ~6 hrs. Based on the ratios of dominant gram-positive and gram-negative bacteria evaluated using PCRs and next generation sequencing, the new DNA extraction method resulted into similar microbiota composition than the previously validated manual DNA extraction method. However, the DNA yield per sample was markedly lower due to the lower input volume of the sample. Based on the sortase C gene PCR tests, the prevalence of LGG was similar (~60%) among 3-month-old children in both clinical studies conducted ~10 years apart, although false negatives among the recent samples due to the low amount of DNA cannot be excluded. Following the temporal pattern of colonization, we observed no evidence for the transfer of LGG at the time of birth from the mother to her child, instead the infants became positive for LGG typically between 1-3 weeks after birth. The carriage of LGG seemed to be dependent on their diet. During this project, we found out that the PCR method employed for detection of LGG was not fully specific for this strain, and hence a more specific qPCR assay was developed.

Coronavirus disease 2019 (COVID-19) is still considered a global pandemic with novel immunoevasive variants constituting a potential threat to life for many susceptible individuals. Despite successful vaccination programmes, which ensued in early 2020, spread of the virus is still an unresolved issue. To address this, innovative prophylactic approaches are being continuously investigated to target the causative agent of COVID-19, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Prevention of infection primarily focuses on the targeting of the receptor-binding domain (RBD) on the spike protein of SARS-CoV-2, which is used to infect host cells presenting angiotensin-converting enzyme 2 (ACE2) on their surface. In 2023, a novel antibody mimetic targeting scaffold, namely the sherpabody platform (SH3; src-homology 3; Recombinant Protein Affinity), was introduced. Accordingly, an intranasally administered, RBD targeting trimeric sherpabody, TriSb92, was demonstrated to prevent infection by SARS-CoV-2 and its recent variants of concern by targeting a conserved region within the spike RBD in vitro and in vivo. This study was performed to further investigate and develop the use of sherpabodies in SARS-CoV-2 prophylaxis. Various homo- and heteromultimeric constructs were assembled and the efficiency of their bacterial production was assessed. Additionally, their functionality, specificity and avidity was analysed. Specifically, the combination of different functionalities within a single molecule – receptor blocking and fusion prevention – was studied. Newly discovered RBD-targeting sherpabodies assembled into multimers were able to neutralize SARS-CoV-2 variants, including the latest Omicron subvariants BA.2.75.2 and XBB.1.5. These multimeric sherpabodies were shown to be easily manufacturable, highly target-specific and multifunctional when desired, making them excellent candidates for intranasally administered SARS-CoV-2 prophylaxis.

Introduction Kenya has recently acquired lower-middle income country status and is facing the triple burden of malnutrition. There is a shortage of data on food intake habits of children and adolescents especially in the rapidly changing urban environments. To be able to reliably measure food intake, one must be able to accurately estimate food portion sizes. Children’s ability to recall portion sizes consumed can vary widely. When a photographic food atlas designed for children with applicable portions is used, it can improve children’s estimation of food portions. Objectives The aim of this study was to develop a photographic food atlas to be used in assessing portion sizes among Kenyan adolescents aged 9-14 years living in urban areas, to support a quantitative 7-day food frequency questionnaire. The second aim was to assess the usability of the atlas amongst 9-14-year-olds and professionals working in the field of nutrition. Methodology A steering group of Finnish and Kenyan nutritionists was formed to oversee the development of the atlas. Literature and other official documents were reviewed to identify the most commonly consumed foods among 9-14-year-old Kenyans. To obtain weighed portion size data, participants were recruited in Nairobi sub-counties Embakasi Central and Langata to represent low- and middle-socioeconomic status respectively. Twenty-one participants aged 9-14 years participated in the weighing of portion sizes, food portions from street markets were also weighed. Three portion sizes (A, B, C) were calculated for most of the 88 food items in the photographic food atlas. Portion B was the average of all weighed portion sizes, portion A was half of B, and portion C was one and half times B. Cooking demonstrations were arranged with the families of participants and the food portions were weighed out and photographed. A photographic food atlas was compiled, and its usability was tested amongst eight adolescents and four nutrition professionals. The usability survey consisted of Likert scale and open-end questions to ascertain acceptability of the atlas. Verbal feedback and observations were also recorded. Results Based on the usability survey, the photographic food atlas received the Usability Score of “OK” and “Good” from adolescents and nutrition professionals respectively. All eight adolescents agreed that the atlas helped them recall portion sizes, but half disagreed and one was unsure whether they could use the atlas on their own. All four professionals agreed they would use the atlas in their work, but all found the quality of photographs poor. Two adolescents disagreed when asked if the portion sizes were small enough and one disagreed when asked if the portion sizes were large enough. However, all professionals agreed that portion sizes were reasonable for the age group. Professionals gave verbal suggestions on improvements, for example, which foods were missing, how to adjust layout as well as the shapes of portion sizes. Conclusion An atlas consisting of 88 most commonly consumed Kenyan foods was developed based on weighed portion sizes of 9-14-year-old Kenyans. The shapes of portion sizes as well as range of portion sizes were crucial for its usability. Poor picture quality hampered recognition of pictures. Clear instructions and explanation of the purpose of the atlas were crucial. A second version of the atlas was developed based on the feedback. The updated atlas, including 173 food items, was used in a cross-sectional study in Nairobi. Further research is recommended to validate the photographic food atlas in order to identify the possible bias it may introduce to portion size estimation.