Studies on chemoenzymatic synthesis: Lipase-catalyzed acylation in multistep organic synthesis

In this thesis, biocatalysis is defined as the science of using enzymes as catalysts in organic synthesis. Environmental aspects and the continuously expanding repertoire of available enzymes have firmly established biocatalysis as a prominent means of chemo-, regio- and stereoselective synthesis. Yet, no single methodology can solve all the challenges faced by a synthetic chemist. Therefore, the knowledge and the skills to combine different synthetic methods are relevant. Lipases are highly useful enzymes in organic synthesis. In this thesis, an effort is being made to form a coherent picture of when and how can lipases be incorporated into nonenzymatic synthesis. This is attempted both in the literature review and in the discussion of the results presented in the original publications contained in the thesis. In addition to lipases, oxynitrilases were also used in the work. The experimental part of the thesis comprises of the results reported in four peer-reviewed publications and one manuscript. Selected amines, amino acids and sugar-derived cyanohydrins or their acylated derivatives were each prepared in enantio- or diastereomerically enriched form. Where applicable, attempts were made to combine the enzymatic reactions to other synthetic steps either by the application of completely separate sequential reactions with isolated intermediates (kinetic and functional kinetic resolution of amines), simultaneously occurring reactions without intermediate isolation (dynamic kinetic resolution of amino acid esters) or sequential reactions but without isolating the intermediates (hydrocyanation of sugar aldehydes with subsequent diastereoresolution). In all cases, lipase-catalyzed acylation was the key step by which stereoselectivity was achieved. Lipase from Burkholderia cepacia was a highly selective enzyme with each substrate category, but careful selection of the acyl donor and the solvent was important as well.

Making the impact on research and society : a case study: crowdsourcing solutions developed for the linguistic research and citizen science

Can crowdsourcing solutions serve many masters? Can they be beneficial for both, for the layman or native speakers of minority languages on the one hand and serious linguistic research on the other? How did an infrastructure that was designed to support linguistics turn out to be a solution for raising awareness of native languages? Since 2012 the National Library of Finland has been developing the Digitisation Project for Kindred Languages, in which the key objective is to support a culture of openness and interaction in linguistic research, but also to promote crowdsourcing as a tool for participation of the language community in research. In the course of the project, over 1,200 monographs and nearly 111,000 pages of newspapers in Finno-Ugric languages will be digitised and made available in the Fenno-Ugrica digital collection. This material was published in the Soviet Union in the 1920s and 1930s, and users have had only sporadic access to the material. The publication of open-access and searchable materials from this period is a goldmine for researchers. Historians, social scientists and laymen with an interest in specific local publications can now find text materials pertinent to their studies. The linguistically-oriented population can also find writings to delight them: (1) lexical items specific to a given publication, and (2) orthographically-documented specifics of phonetics. In addition to the open access collection, we developed an open source code OCR editor that enables the editing of machine-encoded text for the benefit of linguistic research. This tool was necessary since these rare and peripheral prints often include already archaic characters, which are neglected by modern OCR software developers but belong to the historical context of kindred languages, and are thus an essential part of the linguistic heritage. When modelling the OCR editor, it was essential to consider both the needs of researchers and the capabilities of lay citizens, and to have them participate in the planning and execution of the project from the very beginning. By implementing the feedback iteratively from both groups, it was possible to transform the requested changes as tools for research that not only supported the work of linguistics but also encouraged the citizen scientists to face the challenge and work with the crowdsourcing tools for the benefit of research. This presentation will not only deal with the technical aspects, developments and achievements of the infrastructure but will highlight the way in which user groups, researchers and lay citizens were engaged in a process as an active and communicative group of users and how their contributions were made to mutual benefit.

An Open Source, DDI-based Curation System for Social Science Data

Poster at Open Repositories 2014, Helsinki, Finland, June 9-13, 2014

The neural processes generating visual phenomenal consciousness: ERP and neuronavigated brain stimulation studies

One of the greatest conundrums to the contemporary science is the relation between consciousness and brain activity, and one of the specifi c questions is how neural activity can generate vivid subjective experiences. Studies focusing on visual consciousness have become essential in solving the empirical questions of consciousness. Th e main aim of this thesis is to clarify the relation between visual consciousness and the neural and electrophysiological processes of the brain. By applying electroencephalography and functional magnetic resonance image-guided transcranial magnetic stimulation (TMS), we investigated the links between conscious perception and attention, the temporal evolution of visual consciousness during stimulus processing, the causal roles of primary visual cortex (V1), visual area 2 (V2) and lateral occipital cortex (LO) in the generation of visual consciousness and also the methodological issues concerning the accuracy of targeting TMS to V1. Th e results showed that the fi rst eff ects of visual consciousness on electrophysiological responses (about 140 ms aft er the stimulus-onset) appeared earlier than the eff ects of selective attention, and also in the unattended condition, suggesting that visual consciousness and selective attention are two independent phenomena which have distinct underlying neural mechanisms. In addition, while it is well known that V1 is necessary for visual awareness, the results of the present thesis suggest that also the abutting visual area V2 is a prerequisite for conscious perception. In our studies, the activation in V2 was necessary for the conscious perception of change in contrast for a shorter period of time than in the case of more detailed conscious perception. We also found that TMS in LO suppressed the conscious perception of object shape when TMS was delivered in two distinct time windows, the latter corresponding with the timing of the ERPs related to the conscious perception of coherent object shape. Th e result supports the view that LO is crucial in conscious perception of object coherency and is likely to be directly involved in the generation of visual consciousness. Furthermore, we found that visual sensations, or phosphenes, elicited by the TMS of V1 were brighter than identically induced phosphenes arising from V2. Th ese fi ndings demonstrate that V1 contributes more to the generation of the sensation of brightness than does V2. Th e results also suggest that top-down activation from V2 to V1 is probably associated with phosphene generation. The results of the methodological study imply that when a commonly used landmark (2 cm above the inion) is used in targeting TMS to V1, the TMS-induced electric fi eld is likely to be highest in dorsal V2. When V1 was targeted according to the individual retinotopic data, the electric fi eld was highest in V1 only in half of the participants. Th is result suggests that if the objective is to study the role of V1 with TMS methodology, at least functional maps of V1 and V2 should be applied with computational model of the TMS-induced electric fi eld in V1 and V2. Finally, the results of this thesis imply that diff erent features of attention contribute diff erently to visual consciousness, and thus, the theoretical model which is built up of the relationship between visual consciousness and attention should acknowledge these diff erences. Future studies should also explore the possibility that visual consciousness consists of several processing stages, each of which have their distinct underlying neural mechanisms.

Membrane proteins: functional and structural studies using reconstituted proteoliposomes and 2-D crystals

Reconstitution of membrane proteins into lipid bilayers is a powerful tool to analyze functional as well as structural areas of membrane protein research. First, the proper incorporation of a purified membrane protein into closed lipid vesicles, to produce proteoliposomes, allows the investigation of transport and/or catalytic properties of any membrane protein without interference by other membrane components. Second, the incorporation of a large amount of membrane proteins into lipid bilayers to grow crystals confined to two dimensions has recently opened a new way to solve their structure at high resolution using electron crystallography. However, reconstitution of membrane proteins into functional proteoliposomes or 2-D crystallization has been an empirical domain, which has been viewed for a long time more like "black magic" than science. Nevertheless, in the last ten years, important progress has been made in acquiring knowledge of lipid-protein-detergent interactions and has permitted to build upon a set of basic principles that has limited the empirical approach of reconstitution experiments. Reconstitution strategies have been improved and new strategies have been developed, facilitating the success rate of proteoliposome formation and 2-D crystallization. This review deals with the various strategies available to obtain proteoliposomes and 2-D crystals from detergent-solubilized proteins. It gives an overview of the methods that have been applied, which may be of help for reconstituting more proteins into lipid bilayers in a form suitable for functional studies at the molecular level and for high-resolution structural analysis.

Examining life-course influences on chronic disease: the importance of birth cohort studies from low- and middle- income countries. An overview

The objectives of this overview are to describe the past and potential contributions of birth cohorts to understanding chronic disease aetiology; advance a justification for the maintenance of birth cohorts from low- and middle-income countries (LMIC); provide an audit of birth cohorts from LMIC; and, finally, offer possible future directions for this sphere of research. While the contribution of birth cohorts from affluent societies to understanding disease aetiology has been considerable, we describe several reasons to anticipate why the results from such studies might not be directly applied to LMIC. More than any other developing country, Brazil has a tradition of establishing, maintaining and exploiting birth cohort studies. The clear need for a broader geographical representation may be precipitated by a greater collaboration worldwide in the sharing of ideas, fieldwork experience, and cross-country cohort data comparisons in order to carry out the best science in the most efficient manner. This requires the involvement of a central overseeing body - such as the World Health Organization - that has the respect of all countries and the capacity to develop strategic plans for `global' life-course epidemiology while addressing such issues as data-sharing. For rapid progress to be made, however, there must be minimal bureaucratic entanglements.

Studies on some precursors involved in meat flavour formation

The effect of some precursors on the formation of meat flavour during heating has been investigated. A comparison of the influence of three different precursors, inosine-5'-monophosphate (5'-IMP), cysteine and thiamine, added to the meat systems, showed that formation of certain heterocyclic compounds, like sulfur-containing furans, dithiolanones and thiophenes, was significantly affected by changes in the concentration of precursors. However, aliphatic compounds, such as hydrocarbons, alcohols and ketones were not changed by these additions. Inosine-5'-monophosphate was established to be more effective than cysteine or thiamine in the formation of some "meaty" volatiles, i.e. the furanthiols, when its concentration was increased 10 times in raw meat.

Fiber-reinforced composite fixed dental prostheses: Studies of the materials used as pontics

Fiber-reinforced composite fixed dental prostheses – Studies of the materials used as pontics University of Turku, Faculty of Medicine, Institute of Dentistry, Department of Biomaterials Science, Finnish Doctoral Program in Oral Sciences – FINDOS, Annales Universitatis Turkuensis, Turku, Finland 2015 Fiber-reinforced composites (FRC), a non-metallic biomaterial, represent a suitable alternative in prosthetic dentistry when used as a component of fixed dental prostheses (FDPs). Some drawbacks have been identified in the clinical performance of FRC restorations, such as delamination of the veneering material and fracture of the pontic. Therefore, the current series of studies were performed to investigate the possibilities of enhancing the mechanical and physical properties of FRC FDPs by improving the materials used as pontics, to then heighten their longevity. Four experiments showed the importance of the pontic design and surface treatment in the performance of FRC FDPs. In the first, the load-bearing capacities of inlay-retained FRC FDPs with pontics of various materials and thicknesses were evaluated. Three different pontic materials were assessed with different FRC framework vertical positioning. Thicker pontics showed increased load-bearing capacities, especially ceramic pontics. A second study was completed investigating the influence of the chemical conditioning of the ridge-lap surface of acrylic resin denture teeth on their bonding to a composite resin. Increased shear bond strength demonstrated the positive influence of the pretreatment of the acrylic surfaces, indicating dissolution of the denture surfaces, and suggesting potential penetration of the monomer systems into the surface of denture teeth. A third study analyzed the penetration depth of different monomer systems on the acrylic resin denture teeth surfaces. The possibility of establishing a durable bond between acrylic pontics and FRC frameworks was demonstrated by the ability of monomers to penetrate the surface of acrylic resin denture teeth, measured by a confocal scanning type microscope. A fourth study was designed to evaluate the load-bearing capacities of FRC FDPs using the findings of the previous three studies. In this case, the performance of pre-shaped acrylic resin denture teeth used as pontics with different composite resins as filling materials was evaluated. The filling material influenced the load-bearing capacities, providing more durable FRC FDPs. It can be concluded that the mechanical and physical properties of FRC FDPs can be improved as has been shown in the development of this thesis. The improvements reported then might provide long lasting prosthetic solutions of this kind, positioning them as potentially permanent rehabilitation treatments. Key words: fiber-reinforced composite, fixed dental prostheses, inlay-retained bridges, adhesion, acrylic resin denture teeth, dental material.

Stability studies and shelf life estimation of a soy-based dessert

This work is aimed at evaluating the physicochemical, physical, chromatic, microbiological, and sensorial stability of a non-dairy dessert elaborated with soy, guava juice, and oligofructose for 60 days at refrigerated storage as well as to estimate its shelf life time. The titrable acidity, pH, instrumental color, water activity, ascorbic acid, and physical stability were measured. Panelists (n = 50) from the campus community used a hedonic scale to assess the acceptance, purchase intent, creaminess, flavor, taste, acidity, color, and overall appearance of the dessert during 60 days. The data showed that the parameters differed significantly (p < 0.05) from the initial time, and they could be fitted in mathematical equations with coefficient of determination above 71%, aiming to consider them suitable for prediction purposes. Creaminess and acceptance did not differ statistically in the 60-day period; taste, flavor, and acidity kept a suitable hedonic score during storage. Notwithstanding, the sample showed good physical stability against gravity and presented more than 15% of the Brazilian Daily Recommended Value of copper, iron, and ascorbic acid. The product shelf life estimation found was 79 days considering the overall acceptance, acceptance index and purchase intent.

Studies of chemical and enzymatic characteristics of Yacon (Smallanthus sonchifolius) and its flours

Due to the importance of studies on yacon related to health, its in natura pulp, in natura peel, pulp flour, and peel flour were chemically analyzed in terms of its centesimal composition, specific minerals, total dietary fiber and fractions, pH, total soluble solids, total titratable acidity, tannins, oxalic acid, and nitrate. The polyphenoloxidase and peroxidase enzymatic activities were evaluated for in natura pulp and peel only. Yacon pulp and peel flour presented average yield of 7.94% and 10.86%, respectively. The in natura pulp presented a higher moisture and carboydrate content and lower lipid, protein, total dietary fiber, and ash than those of the peel flour. The same pattern was observed for pulp flour when compared to peel flour. The highest tannin, nitrate, and oxalic acid levels were found in the peel flour, 15,304.5 mg.kg-1, 1,578.3 mg.kg-1, and 7,925.0 mg.kg-1 (wet weight), respectively. The polyphenoloxidase and peroxidase enzymes presented higher enzymatic activity in the yacon peel. Based on the results obtained, it can be said that the yacon and its derivatives are important dietary carbohydrate and mineral sources and contain antinutritional substance contents lower than those harmful to health.

Preliminary studies on nutritive and organoleptic properties in processed fish fillets obtained from Iran

The effects of boiling, frying, and roasting methods on nutrients and properties some of marine fishes were studies. AOAC official methods were used for the analyses of the samples. Frying reduced the protein content of Nemipterus japonicas, while roasting reduced the protein content of Carangoides malabaricus and Saurida undosquamis. Fresh and fried C. malabaricus had 71.2% and 57.05% protein content, respectively, while N. japonicas had 57.39% and 47.51%, respectively. The highest oil content in C.malabaricus was 16.80%, followed by N.japonicus 12.80%; while the lowest oil content in S.undosquamis was 5.88%. N.japonicusis protein content reduced with frying. Some cooking methods used in the processing can have an effect on of fish nutrient composition. The texture of fried and boiled fish fillets significantly differ from that of the roasted fillets (p<0.05).

In search of the interrelatedness of innovation and internationalisation processes in entrepreneurial life science companies

This study is motivated by the question how resource scarce innovative entrepreneurial companies seek and leverage global resources. This study takes a resource-seeking perspective a step forward and suggests that resources that enable the entrepreneurial internationalisation are largely accrued from the early stages of entrepreneurial life; that is from the innovation development. Consequently, this study seeks to explain how innovation and internationalisation processes are interrelated in the entrepreneurial internationalisation. This main objective is approached through three research questions, (1) What role do inter-organisational relationships in innovation have in the entrepreneurial internationalisation process? (2) What kind of inward–outward links do inter-organisational relationships create in the resource-seeking-based entrepreneurial internationalisation process? (3) What kind of capability to collaborate forms in the interaction of inter-organisational relationship deployment? The research design is a mixed methods design that consists of quantitative pilot study and qualitative multiple case study of five entrepreneurial life science companies from Finland and Austria. The findings show that innovation and internationalisation processes are tightly interwoven in pre-internationalisation state. The findings also reveal that the more experienced companies are able to take advantage of complexcross-border inter-organisational relationship structures better than the starting companies. However, very minor evidence was found on inward links translating into outward links in the entrepreneurial internationalisation process, despite the expectation to observe more of these links in the data. Combined intangible-tangible resource-seeking was the most preferred to build links between inward–outward internationalisation but also to develop competence to collaborate. By adopting a resource- instead of market-seeking approach, this study illustrated that internationalisation extends to early stages of innovative companies, and that in high-technology companies’ potentially significant cross-border relationships have started to form long before incorporation. Therefore, these observations justified the firmer inclusion of pre-company history in innovative entrepreneurship studies. The study offers a conceptualisation of entrepreneurial internationalisation that is perceived as a process. The main theoretical contributions are in the areas of international entrepreneurship and in the behavioural process studies of entrepreneurial internationalisation and resource-based internationalisation. The inclusion of the innovation-based discussion, namely the innovation process, in the internationalisation process theories has clearly contributed to the understanding of entrepreneurial internationalisation in the context of international entrepreneurship. Innovation development is a central act of entrepreneurial companies, and neglecting innovation process investigation from entrepreneurial internationalisation leaves potentially influential mechanisms unexplored.

Suomalaisten yliopistojen life science-alan spin-off-yritysten rahoitustilanne, -lähteet ja –riskit

Life science-ala on rahoituksellisesti erittäin haastava, koska tuotekehitysputket ovat 10-15 vuoden pituisia ja voivat vaatia suuria etupainotteisia investointeja. Monet life science-alan yritykset ovat niin Suomessa kuin kansainvälisestikin syntyneet suoraan yliopistosta niin sanottuina spin-off-yrityksinä. Yrityksen perustaminen tutkimustiedon pohjalle on yksi akateemisen yrittäjyyden muodoista. Tässä tutkimuksessa tutkitaan akateemista yrittäjyyttä life science-alalla Suomessa. Suomessa life science-alan osaaminen on kansainvälistä huippua ja alalle on syntynyt useita spin-off-yrityksiä viime vuosina. Aiemmat tutkimukset ovat keskittyneet erityisesti agentti-päämiesongelmiin akateemisessa yrittäjyydessä ja life science alaa puolestaan on käsitelty usein pelkästään yhtenä alana. Tutkimuksessa pyritään analysoimaan life science-alan eri painopiste-alueita tarkemmin rahoitusstrategisesta näkökulmasta, koska ala ei ole homogeeninen. Akateemisen yrittäjyyden agenttiongelmaa pyritään tarkoittamaan Suomen yliopistomaailmassa life science-alalta käsin. Tutkimus toteutetaan kvalitatiivisena haastattelututkimuksena ja sitä tukevana tilinpäätöstietojen analysointina. Tutkimuksen tulokset vahvistavat agentti-päämiesongelmien olemassaolon suomalaisessa yliopistomaailmassa. Ongelmien osapuolina ovat niin akateemikot, yliopiston innovaatiopalvelut kuin TEKES:kin. Yrittäjät kaipaavat kokonaisvaltaisempaa apua, koska kokevat alalla liiketoimintaosaamisen puuttuvan monin paikoin. Alan vaatimien runsaiden tuotekehityspanostusten vuoksi alan yritykset ovat pitkään raskaasti tappiollisia. Yritysten valitsemat rahoitusratkaisut vaihtelevat suuresti. Kaikki tutkitut yritykset pyrkivät kasvamaan nykyisten ydintuotteidensa mukana ja ovat jo jossain määrin hankkineet rahoitusta kasvustrategiaansa varten. Alan pääomaintensiivisyydestä johtuen ovat monet yritykset kuitenkin suunnittelemassa tai jo päätyneet tekemään yhteistyötä integroituen alalla joko vertikaalisesti tai horisontaalisesti. Tutkimuksen tulokset vahvistavat aiemman käsityksen, jonka mukaan life science-alan yrityksiä on vaikea arvioida ulkopuolelta pelkästään tilinpäätöstietojen pohjalta, koska alkuvaiheessa rahoituskierrokset ja tappiolliset vuoden seuraavat toisiaan ja toisaalta yrityksen arvo sitoutuu pitkiin tuotekehitysputkiin, jotka eivät välttämättä näy tilinpäätöksessä. Tulokset tuovat esille merkittäviä yritysten kokemia ongelmia rahoituksen saannissa life science-alalla Suomessa sekä akateemisen yrittäjyyden epäkohtia yritystoiminnan näkökulmasta. Näiden tekijöiden huomioonottaminen ja laajempi kansainvälinen vertailu voivat auttaa suomalaisia yrityksiä tällä vahvalla osaamisalalla eteenpäin.

Problem solving and computer-assisted instruction in science education : analysis of research findings and the research process /

The quantitative component of this study examined the effect of computerassisted instruction (CAI) on science problem-solving performance, as well as the significance of logical reasoning ability to this relationship. I had the dual role of researcher and teacher, as I conducted the study with 84 grade seven students to whom I simultaneously taught science on a rotary-basis. A two-treatment research design using this sample of convenience allowed for a comparison between the problem-solving performance of a CAI treatment group (n = 46) versus a laboratory-based control group (n = 38). Science problem-solving performance was measured by a pretest and posttest that I developed for this study. The validity of these tests was addressed through critical discussions with faculty members, colleagues, as well as through feedback gained in a pilot study. High reliability was revealed between the pretest and the posttest; in this way, students who tended to score high on the pretest also tended to score high on the posttest. Interrater reliability was found to be high for 30 randomly-selected test responses which were scored independently by two raters (i.e., myself and my faculty advisor). Results indicated that the form of computer-assisted instruction (CAI) used in this study did not significantly improve students' problem-solving performance. Logical reasoning ability was measured by an abbreviated version of the Group Assessment of Lx)gical Thinking (GALT). Logical reasoning ability was found to be correlated to problem-solving performance in that, students with high logical reasoning ability tended to do better on the problem-solving tests and vice versa. However, no significant difference was observed in problem-solving improvement, in the laboratory-based instruction group versus the CAI group, for students varying in level of logical reasoning ability.Insignificant trends were noted in results obtained from students of high logical reasoning ability, but require further study. It was acknowledged that conclusions drawn from the quantitative component of this study were limited, as further modifications of the tests were recommended, as well as the use of a larger sample size. The purpose of the qualitative component of the study was to provide a detailed description ofmy thesis research process as a Brock University Master of Education student. My research journal notes served as the data base for open coding analysis. This analysis revealed six main themes which best described my research experience: research interests, practical considerations, research design, research analysis, development of the problem-solving tests, and scoring scheme development. These important areas ofmy thesis research experience were recounted in the form of a personal narrative. It was noted that the research process was a form of problem solving in itself, as I made use of several problem-solving strategies to achieve desired thesis outcomes.

Gender bias in the Trends in Mathematics and Science Study 2003 (TIMMS) for Canadian students /

This study is a secondary data analysis of the Trends in Mathematics and Science Study 2003 (TIMSS) to determine if there is a gender bias, unbalanced number of items suited to the cognitive skill of one gender, and to compare performance by location. Results of the Grade 8, math portion of the test were examined. Items were coded as verbal, spatial, verbal /spatial or neither and as conventional or unconventional. A Kruskal- Wallis was completed for each category, comparing performance of students from Ontario, Quebec, and Singapore. A Factor Analysis was completed to determine if there were item categories with similar characteristics. Gender differences favouring males were found in the verbal conventional category for Canadian students and in the spatial conventional category for students in Quebec. The greatest differences were by location, as students in Singapore outperformed students from Canada in all areas except for the spatial unconventional category. Finally, whether an item is conventional or unconventional is more important than whether the item is verbal or spatial. Results show the importance of fair assessment for the genders in both the classroom and on standardized tests.