Interactions of natural products with β-lactoglobulins, members of the lipocalin family

Natural products constitute an important source of new drugs. The bioavailability of the drugs depends on their absorption, distribution, metabolism and elimination. To achieve good bioavailability, the drug must be soluble in water, stable in the gastrointestinal tract and palatable. Binding proteins may improve the solubility of drug compounds, masking unwanted properties, such as bad taste, bitterness or toxicity, transporting or protecting these compounds during processing and storage. The focus of this thesis was to study the interactions, including ligand binding and the effect of pH and temperature, of bovine and reindeer β-lactoglobulin (βLG) with such compounds as retinoids, phenolic compounds as well as with compounds from plant extracts, and to investigate the transport properties of the βLG-ligand complex. To examine the binding interactions of different ligands to βLG, new methods were developed. The fluorescence binding method for the evaluation of ligand binding to βLG was miniaturized from a quartz cell to a 96-well plate. A method of ultrafiltration sampling combined with high-performance liquid chromatography was developed to assess the binding of compounds from extracts. The interactions of phenolic compounds or retinoids and βLG were investigated using the 96-well plate method. The majority of flavones, flavonols, flavanones and isoflavones and all of the retinoids included were shown to bind to bovine and reindeer βLG. Phenolic compounds, contrary to retinol, were not released at acidic pH. Those results suggest that βLG may have more binding sites, probably also on the surface of βLG. An extract from Camellia sinensis (L.) O. Kunze (black tea), Urtica dioica L. (nettle) and Piper nigrum (black pepper) were used to evaluate whether βLG could bind compounds from plant extracts. Piperine from P. nigrum was found to bind tightly and rutin from U. dioica weakly to βLG. No components from C. sinensis bound to βLG in our experiment. The uptake and membrane permeation of bovine and reindeer βLG, free and bound with retinol, palmitic acid and cholesterol, were investigated using Caco-2 cell monolayers. Both bovine and reindeer βLG were able to cross the Caco-2 cell membrane. Free and βLG-bound retinol and palmitic acid were transported equally, whereas cholesterol could not cross the Caco-2 cell monolayer free or bound to βLG. Our results showed that βLG can bind different natural product compounds, but cannot enhance transport of retinol, palmitic acid or cholesterol through Caco-2 cells. Despite this, βLG, as a water-soluble binding protein, may improve the solubility of natural compounds, possibly protecting them from early degradation and transporting some of them through the stomach. Furthermore, it may decrease their bad or bitter taste during oral administration of drugs or in food preparations. βLG can also enhance or decrease the health benefits of herbal teas and food preparations by binding compounds from extracts.

Rhetoric and Representation : Exploring the Cultural Meaning of the Natural Sciences in Contemporary Popular Science Writing and Literature

Kirjallisuuden- ja kulttuurintutkimus on viimeisten kolmen vuosikymmenen aikana tullut yhä enenevässä määrin tietoiseksi tieteen ja taiteen suhteen monimutkaisesta luonteesta. Nykyään näiden kahden kulttuurin tutkimus muodostaa oman kenttänsä, jolla niiden suhdetta tarkastellaan ennen kaikkea dynaamisena vuorovaikutuksena, joka heijastaa kulttuurimme kieltä, arvoja ja ideologisia sisältöjä. Toisin kuin aiemmat näkemykset, jotka pitävät tiedettä ja taidetta toisilleen enemmän tai vähemmän vastakkaisina pyrkimyksinä, nykytutkimus lähtee oletuksesta, jonka mukaan ne ovat kulttuurillisesti rakentuneita diskursseja, jotka kohtaavat usein samankaltaisia todellisuuden mallintamiseen liittyviä ongelmia, vaikka niiden käyttämät metodit eroavatkin toisistaan. Väitöskirjani keskittyy yllä mainitun suhteen osa-alueista popularisoidun tietokirjallisuuden (muun muassa Paul Davies, James Gleick ja Richard Dawkins) käyttämän kielen ja luonnontieteistä ideoita ammentavan kaunokirjallisuuden (muun muassa Jeanette Winterson, Tom Stoppard ja Richard Powers) hyödyntämien keinojen tarkasteluun nojautuen yli 30 teoksen kattavaa aineistoa koskevaan tyylin ja teemojen tekstianalyysiin. Populaarin tietokirjallisuuden osalta tarkoituksenani on osoittaa, että sen käyttämä kieli rakentuu huomattavassa määrin sellaisille rakenteille, jotka tarjoavat mahdollisuuden esittää todellisuutta koskevia argumentteja mahdollisimman vakuuttavalla tavalla. Tässä tehtävässä monilla klassisen retoriikan määrittelemillä kuvioilla on tärkeä rooli, koska ne auttavat liittämään sanotun sisällön ja muodon tiukasti toisiinsa: retoristen kuvioiden käyttö ei näin ollen edusta pelkkää tyylikeinoa, vaan se myös usein kiteyttää argumenttien taustalla olevat tieteenfilosofiset olettamukset ja auttaa vakiinnuttamaan argumentoinnin logiikan. Koska monet aikaisemmin ilmestyneistä tutkimuksista ovat keskittyneet pelkästään metaforan rooliin tieteellisissä argumenteissa, tämä väitöskirja pyrkii laajentamaan tutkimuskenttää analysoimalla myös toisenlaisten kuvioiden käyttöä. Osoitan myös, että retoristen kuvioiden käyttö muodostaa yhtymäkohdan tieteellisiä ideoita hyödyntävään kaunokirjallisuuteen. Siinä missä popularisoitu tiede käyttää retoriikkaa vahvistaakseen sekä argumentatiivisia että kaunokirjallisia ominaisuuksiaan, kuvaa tällainen sanataide tiedettä tavoilla, jotka usein heijastelevat tietokirjallisuuden kielellisiä rakenteita. Toisaalta on myös mahdollista nähdä, miten kaunokirjallisuuden keinot heijastuvat popularisoidun tieteen kerrontatapoihin ja kieleen todistaen kahden kulttuurin dynaamisesta vuorovaikutuksesta. Nykyaikaisen populaaritieteen retoristen elementtien ja kaunokirjallisuuden keinojen vertailu näyttää lisäksi, kuinka tiede ja taide osallistuvat keskusteluun kulttuurimme tiettyjen peruskäsitteiden kuten identiteetin, tiedon ja ajan merkityksestä. Tällä tavoin on mahdollista nähdä, että molemmat ovat perustavanlaatuisia osia merkityksenantoprosessissa, jonka kautta niin tieteelliset ideat kuin ihmiselämän suuret kysymyksetkin saavat kulttuurillisesti rakentuneen merkityksensä.

Datorstödda studier i hälso- och socialvård : MuPSiNet-projektet

The aim of the study was to explore why the MuPSiNet project - a computer and network supported learning environment for the field of health care and social work - did not develop as expected. To grasp the problem some hypotheses were formulated. The hypotheses regarded the teachers' skills in and attitudes towards computing and their attitudes towards constructivist study methods. An online survey containing 48 items was performed. The survey targeted all the teachers within the field of health care and social work in the country, and it produced 461 responses that were analysed against the hypotheses. The reliability of the variables was tested using the Cronbach alpha coefficient and t-tests. Poor basic computing skills among the teachers combined with a vulnerable technical solution, and inadequate project management combined with lack of administrative models for transforming economic resources into manpower were the factors that turned out to play a decisive role in the project. Other important findings were that the teachers had rather poor skills and knowledge in computing, computer safety and computer supported instruction, and that these skills were significantly poorer among female teachers who were in majority in the sample. The fraction of teachers who were familiar with software for electronic patient records (EPR) was low. The attitudes towards constructivist teaching methods were positive, and further education seemed to utterly increase the teachers' readiness to use alternative teaching methods. The most important conclusions were the following: In order to integrate EPR software as a natural tool in teaching planning and documenting health care, it is crucial that the teachers have sufficient basic skills in computing and that more teachers have personal experience of using EPR software. In order for computer supported teaching to become accepted it is necessary to arrange with extensive further education for the teachers presently working, and for that further education to succeed it should be backed up locally among other things by sufficient support in matters concerning computer supported teaching. The attitudes towards computing showed significant gender differences. Based on the findings it is suggested that basic skills in computing should also include an awareness of data safety in relation to work in different kinds of computer networks, and that projects of this kind should be built up around a proper project organisation with sufficient resources. Suggestions concerning curricular development and further education are also presented. Conclusions concerning the research method were that reminders have a better effect, and that respondents tend to answer open-ended questions more verbosely in electronically distributed online surveys compared to traditional surveys. A method of utilising randomized passwords to guarantee respondent anonymity while maintaining sample control is presented. Keywords: computer-assisted learning, computer-assisted instruction, health care, social work, vocational education, computerized patient record, online survey

Target Cell Topography and Cytoskeletal Reorganization in Natural Killer Cell Activity

The immune system has to recognize and destroy abnormal or infected cells to maintain homeostasis. Natural killer (NK) cells directly recognize and kill transformed or virus-infected cells without prior sensitization. We have studied both virus-infected and tumor cells in order to identify the target structures involved in triggering NK activity. Mouse/human cell hybrids containing various human chromosomes were used as targets. The human chromosome responsible for activating NK cell killing was identified to chromosome number 6. The results suggest that activated NK cells recognize ligands that are encoded on human chromosome 6. We showed that the ligand on the target cell side was intercellular adhesion molecule 2 (ICAM-2). There was no difference in the level of expression of ICAM-2, however, but a drastic difference was seen in the distribution of the molecule: ICAM-2 was evenly distributed on the surface of the NK-resistant cells, but almost totally redistributed to the tip of uropods, bud-like extensions, which were absent from the parental cells. Interestingly, the gene coding for cytoskeletal linker protein ezrin has been localized to human chromosome 6, and there was a colocalization of ezrin and ICAM-2 in the uropods. Furthermore, the transfected human ezrin into NK cell-resistant cells induced uropod formation, ICAM-2 and ezrin redistribution to newly formed uropods, and sensitized target cells to NK cell killing. These data reveal a novel form of NK cell recognition: target structures are already present on normal cells; they become detectable only after abnormal redistribution into hot spots on the target cell membrane. NK cells are central players in the defence against virus infections. They inhibit the spread of infection, allowing time for specific immune responses to develop. The virus-proteins that directly activate human NK cell killing are largely unknown. We studied the sensitivity of virus-specific early proteins of Semliki Forest virus (SFV) to NK killing. The viral non-structural proteins (nsP1-4) translated early in the virus cycle were transfected in NK-resistant cells. Viral early gene nsP1 alone efficiently sensitized target cells to NK activity, and the tight membrane association of nsP1 seems to be critical in the triggering of NK killing. NsP1 protein colocalized with (redistributed) ezrin in filopodia-like structures to which the NK cells were bound. The results suggest that also in viral infections NK cells react to rapid changes in membrane topography. Based on the results of this thesis, a new model of target cell recognition of NK cells can be suggested: reorganization of the cytoskeleton induces alterations in cell surface topography, and this new pattern of surface molecules is recognized as "altered-self".

Community forestry and environmental literacy in northern Thailand : Towards collaborative natural resource management and conservation

Conservation and sustainable management of tropical forests needs a holistic approach: in addition to ecological concerns, socio-economic issues including cultural aspects must be taken into consideration. An ability to adapt practices is a key to successful collaborative natural resource management. Achieving this requires local participation and understanding of local conceptions of the environment. This study examined these issues in the context of northern Thailand. Northern uplands are the home of much of the remaining natural forest in Thailand and several ethnic minority groups commonly referred to as hill tribes. The overall purpose of this study was to grasp a regional view of an ethnically diverse forested area and to elicit prospects to develop community forestry for conservation purposes and for securing people s livelihood. Conservation was a central goal of management as the forests in the area were largely designated as protected. The aim was to study local perceptions, objectives, values and practices of forest management, under the umbrella of the concept environmental literacy, as well as the effects of forest policy on community management goals and activities. Environmental literacy refers to holistic understanding of the environment. It was used as a tool to examine people s views, interests, knowledge and motivation associated to forests. The material for this study was gathered in six villages in Chiang Mai Province. Three minority groups were included in the study, the Karen, Hmong and Lawa, and also the Thai. Household and focus group interviews were conducted in the villages. In addition, officials at district, regional and national levels, workers of non-governmental organisations, and academics were interviewed, and some data were gathered from the students of a local school. The results showed that motivation for protecting the forests existed among each ethnic group studied. This was a result of culture and traditions evolved in the forest environment but also of a need to adapt to a changed situation and environment and to outside pressures. The consequences of deforestation were widely agreed on in the villages, and the impact of socio-economic changes on the forests and livelihood was also recognised. The forest was regarded as a source of livelihood providing land, products and services essential to the people inhabiting rural uplands. Traditions, fire control, cooperation, reforestation, separation of protected and utilisable areas, and rules were viewed as central for conservation. For the villagers, however, conservation meant sustainable use, whereas the government has tended to prefer strict restrictions on forest resource use. Thus, conflicts had arisen. Between communities, cooperation was more dominant than conflict. The results indicated that the heterogeneity of forest dwellers, although it has to be recognised, should not be overemphasised: ethnic diversity can be considered as no major obstacle for successful community forestry. Collaborative management is particularly important in protected areas in order to meet the conservation goals while providing opportunities for livelihood. Forest management needs more positive incentives and increased dialogue.

Natural regeneration and ecological succession in Pinus kesiya watershed plantations in northern Thailand : Implications on plantation management

Establishment of Pinus kesiya Roy. ex Gord. plantations in Thailand began in the 1960s by the Royal Forest Department. The aim was to reforest abandoned swidden areas and grasslands in order to reduce erosion and to produce timber and fuel wood. Today there are about 150, 000 ha of P. kesiya plantations in northern Thailand. Most of these plantations cannot be harvested due to a national logging ban. Previous studies have suggested that Pinus kesiya plantations posses a capability as a foster environment for native broadleaved tree species, but little is known about the extent of regeneration in these plantations. The general aim of the study was to clarify the extent of forest regeneration and interactions behind it in Pinus kesiya plantations of the Ping River basin, northern Thailand. Based on the results of this study and previous literature, forest management proposals were produced for the area studied. In four different pine plantation areas, a total of seven plantations were assessed using systematic data collection with clustered circular sample plots. Vegetation and environmental data were statistically analysed, so as to recognise the key factors affecting regeneration. Regeneration had occurred in all plantations studied. Regeneration of broadleaved trees was negatively affected by forest fire and canopy coverage. A high basal area of mature broadleaved trees affected the regeneration process positively. Forest fire disturbance had a strong effect also on plantation structure and species composition. Because of an unclear future forest management setting as regards forest laws in Thailand, a management system that enables various future utilisation possibilities and emphasises local participation is recommended for P. kesiya watershed platations of northern Thailand.

Reconstructions of past climates from documentary and natural sources in Finland since the 18th century

In this paper both documentary and natural proxy data have been used to improve the accuracy of palaeoclimatic knowledge in Finland since the 18th century. Early meteorological observations from Turku (1748-1800) were analyzed first as a potential source of climate variability. The reliability of the calculated mean temperatures was evaluated by comparing them with those of contemporary temperature records from Stockholm, St. Petersburg and Uppsala. The resulting monthly, seasonal and yearly mean temperatures from 1748 to 1800 were compared with the present day mean values (1961-1990): the comparison suggests that the winters of the period 1749-1800 were 0.8 ºC colder than today, while the summers were 0.4 ºC warmer. Over the same period, springs were 0.9 ºC and autumns 0.1 ºC colder than today. Despite their uncertainties when compared with modern meteorological data, early temperature measurements offer direct and daily information about the weather for all months of the year, in contrast with other proxies. Secondly, early meteorological observations from Tornio (1737-1749) and Ylitornio (1792-1838) were used to study the temporal behaviour of the climate-tree growth relationship during the past three centuries in northern Finland. Analyses showed that the correlations between ring widths and mid-summer (July) temperatures did not vary significantly as a function of time. Early (June) and late summer (August) mean temperatures were secondary to mid-summer temperatures in controlling the radial growth. According the dataset used, there was no clear signature of temporally reduced sensitivity of Scots pine ring widths to mid-summer temperatures over the periods of early and modern meteorological observations. Thirdly, plant phenological data with tree-rings from south-west Finland since 1750 were examined as a palaeoclimate indicator. The information from the fragmentary, partly overlapping, partly nonsystematically biased plant phenological records of 14 different phenomena were combined into one continuous time series of phenological indices. The indices were found to be reliable indicators of the February to June temperature variations. In contrast, there was no correlation between the phenological indices and the precipitation data. Moreover, the correlations between the studied tree-rings and spring temperatures varied as a function of time and hence, their use in palaeoclimate reconstruction is questionable. The use of present tree-ring datasets for palaeoclimate purposes may become possible after the application of more sophisticated calibration methods. Climate variability since the 18th century is perhaps best seen in the fourth paper study of the multiproxy spring temperature reconstruction of south-west Finland. With the help of transfer functions, an attempt has been made to utilize both documentary and natural proxies. The reconstruction was verified with statistics showing a high degree of validity between the reconstructed and observed temperatures. According to the proxies and modern meteorological observations from Turku, springs have become warmer and have featured a warming trend since around the 1850s. Over the period of 1750 to around 1850, springs featured larger multidecadal low-frequency variability, as well as a smaller range of annual temperature variations. The coldest springtimes occurred around the 1840s and 1850s and the first decade of the 19th century. Particularly warm periods occurred in the 1760s, 1790s, 1820s, 1930s, 1970s and from 1987 onwards, although in this period cold springs occurred, such as the springs of 1994 and 1996. On the basis of the available material, long-term temperature changes have been related to changes in the atmospheric circulation, such as the North Atlantic Oscillation (February-June).

Integration of natural hazards, risk and climate change into spatial planning practices

Most countries of Europe, as well as many countries in other parts of the world, are experiencing an increased impact of natural hazards. It is often speculated, but not yet proven, that climate change might influence the frequency and magnitude of certain hydro-meteorological natural hazards. What has certainly been observed is a sharp increase in financial losses caused by natural hazards worldwide. Eventhough Europe appears to be a space that is not affected by natural hazards to such catastrophic extents as other parts of the world are, the damages experienced here are certainly increasing too. Natural hazards, climate change and, in particular, risks have therefore recently been put high on the political agenda of the EU. In the search for appropriate instruments for mitigating impacts of natural hazards and climate change, as well as risks, the integration of these factors into spatial planning practices is constantly receiving higher attention. The focus of most approaches lies on single hazards and climate change mitigation strategies. The current paradigm shift of climate change mitigation to adaptation is used as a basis to draw conclusions and recommendations on what concepts could be further incorporated into spatial planning practices. Especially multi-hazard approaches are discussed as an important approach that should be developed further. One focal point is the definition and applicability of the terms natural hazard, vulnerability and risk in spatial planning practices. Especially vulnerability and risk concepts are so many-fold and complicated that their application in spatial planning has to be analysed most carefully. The PhD thesis is based on six published articles that describe the results of European research projects, which have elaborated strategies and tools for integrated communication and assessment practices on natural hazards and climate change impacts. The papers describe approaches on local, regional and European level, both from theoretical and practical perspectives. Based on these, passed, current and future potential spatial planning applications are reviewed and discussed. In conclusion it is recommended to shift from single hazard assessments to multi-hazard approaches, integrating potential climate change impacts. Vulnerability concepts should play a stronger role than present, and adaptation to natural hazards and climate change should be more emphasized in relation to mitigation. It is outlined that the integration of risk concepts in planning is rather complicated and would need very careful assessment to ensure applicability. Future spatial planning practices should also consider to be more interdisciplinary, i.e. to integrate as many stakeholders and experts as possible to ensure the sustainability of investments.

Computing the Stochastic Complexity of Simple Probabilistic Graphical Models

Minimum Description Length (MDL) is an information-theoretic principle that can be used for model selection and other statistical inference tasks. There are various ways to use the principle in practice. One theoretically valid way is to use the normalized maximum likelihood (NML) criterion. Due to computational difficulties, this approach has not been used very often. This thesis presents efficient floating-point algorithms that make it possible to compute the NML for multinomial, Naive Bayes and Bayesian forest models. None of the presented algorithms rely on asymptotic analysis and with the first two model classes we also discuss how to compute exact rational number solutions.

A Probabilistic Approach to the Primary Visual Cortex

What can the statistical structure of natural images teach us about the human brain? Even though the visual cortex is one of the most studied parts of the brain, surprisingly little is known about how exactly images are processed to leave us with a coherent percept of the world around us, so we can recognize a friend or drive on a crowded street without any effort. By constructing probabilistic models of natural images, the goal of this thesis is to understand the structure of the stimulus that is the raison d etre for the visual system. Following the hypothesis that the optimal processing has to be matched to the structure of that stimulus, we attempt to derive computational principles, features that the visual system should compute, and properties that cells in the visual system should have. Starting from machine learning techniques such as principal component analysis and independent component analysis we construct a variety of sta- tistical models to discover structure in natural images that can be linked to receptive field properties of neurons in primary visual cortex such as simple and complex cells. We show that by representing images with phase invariant, complex cell-like units, a better statistical description of the vi- sual environment is obtained than with linear simple cell units, and that complex cell pooling can be learned by estimating both layers of a two-layer model of natural images. We investigate how a simplified model of the processing in the retina, where adaptation and contrast normalization take place, is connected to the nat- ural stimulus statistics. Analyzing the effect that retinal gain control has on later cortical processing, we propose a novel method to perform gain control in a data-driven way. Finally we show how models like those pre- sented here can be extended to capture whole visual scenes rather than just small image patches. By using a Markov random field approach we can model images of arbitrary size, while still being able to estimate the model parameters from the data.

Exploring privacy for ubiquitous computing: Tools, methods and experiments

Ubiquitous computing is about making computers and computerized artefacts a pervasive part of our everyday lifes, bringing more and more activities into the realm of information. The computationalization, informationalization of everyday activities increases not only our reach, efficiency and capabilities but also the amount and kinds of data gathered about us and our activities. In this thesis, I explore how information systems can be constructed so that they handle this personal data in a reasonable manner. The thesis provides two kinds of results: on one hand, tools and methods for both the construction as well as the evaluation of ubiquitous and mobile systems---on the other hand an evaluation of the privacy aspects of a ubiquitous social awareness system. The work emphasises real-world experiments as the most important way to study privacy. Additionally, the state of current information systems as regards data protection is studied. The tools and methods in this thesis consist of three distinct contributions. An algorithm for locationing in cellular networks is proposed that does not require the location information to be revealed beyond the user's terminal. A prototyping platform for the creation of context-aware ubiquitous applications called ContextPhone is described and released as open source. Finally, a set of methodological findings for the use of smartphones in social scientific field research is reported. A central contribution of this thesis are the pragmatic tools that allow other researchers to carry out experiments. The evaluation of the ubiquitous social awareness application ContextContacts covers both the usage of the system in general as well as an analysis of privacy implications. The usage of the system is analyzed in the light of how users make inferences of others based on real-time contextual cues mediated by the system, based on several long-term field studies. The analysis of privacy implications draws together the social psychological theory of self-presentation and research in privacy for ubiquitous computing, deriving a set of design guidelines for such systems. The main findings from these studies can be summarized as follows: The fact that ubiquitous computing systems gather more data about users can be used to not only study the use of such systems in an effort to create better systems but in general to study phenomena previously unstudied, such as the dynamic change of social networks. Systems that let people create new ways of presenting themselves to others can be fun for the users---but the self-presentation requires several thoughtful design decisions that allow the manipulation of the image mediated by the system. Finally, the growing amount of computational resources available to the users can be used to allow them to use the data themselves, rather than just being passive subjects of data gathering.