”En mä aluks kuvitellukaan et siit tulee mitään, niin se on mun mielest niinko paras työ mitä oon tehny tähän asti” Yrittäjämäinen pedagogiikka käsityön opetuksessa: 7. luokkalaisten riskin ottaminen käsityöprosessin aikana.

Käsityöoppiaine sekä sille ominainen toiminnallinen ja vuorovaikutteinen oppimisympäristö tarjoavat erinomaiset puitteet yrittäjyyskasvatuksen toteuttamiseen. Käsityöoppiaineen sekä yrittäjyyskasvatuksen tavoitteet, sisällöt ja toimintatavat ovat yhteneviä. Yrittäjyydessä sekä elämässä ylipäätään yksi olennainen ominaisuus on kyky ottaa riskejä. Käsitöiden tekeminen on luovaa toimintaa, johon riskin ottaminen liittyy olennaisesti. Tässä tutkielmassa tarkastellaan oppilaiden kokonaiseen käsityöprosessiin liittyviä riskinottotilanteita Ristimäen (2004) yrittäjyyteen liittyvien taloudellisen, psykologisen ja sosiaalisen riskin luokittelun näkökulmasta. Tutkielmassa luodaan omat määritelmät edellä mainituille riskeille käsityöprosessissa, koska aiempia määritelmiä ei ole saatavilla. Tutkielmassa selvitetään, millaisia riskinottotilanteita oppilaat kohtaavat käsityöprosessinsa eri vaiheissa, ja voidaanko riskinottotilanteista erottaa taloudelliset, psykologiset ja sosiaaliset riskit. Lisäksi tutkitaan eroaako oppilaiden riskin ottaminen ryhmässä, jossa käsityön opetus on järjestetty yrittäjämäistä pedagogiikkaa noudattaen verrattuna ryhmään, jonka opetuksessa yrittäjämäistä pedagogiikkaa ei ole huomioitu, ja mikäli eroja on, niin millaisia ne ovat. Tutkimusaineistona on kaksi eri aineistoa: ensimmäinen on Turun yliopiston opettajankoulutuslaitoksen Rauman yksikön tutkimushankkeessa vuonna 2013 kerätty haastatteluaineisto 7. luokkalaisten oppilaiden tekstiilityön ja teknisen työn opetusjaksoilta. Toinen aineisto on kerätty keväällä 2016 7. luokkalaisten oppilaiden oppimispäiväkirjateksteinä tekstiilityön opetusjaksolta, jossa opetus toteutettiin yrittäjämäistä pedagogiikkaa noudattaen. Tutkimusaineistot analysoidaan käyttäen teoriasidonnaista sisällönanalyysia. Edellä mainitut Ristimäen (2004) riskin luokittelut ohjaavat analyysia. Tulokset osoittavat, että oppilaat kohtaavat kaikissa käsityöprosessin vaiheissa riskinottotilanteita, joissa he joko ottavat riskejä tai välttävät niiden ottamista. Suunnitteluvaiheessa riskinottotilanteet liittyivät päätöksentekoon valmistettavasta tuotteesta sekä valmistustekniikasta. Valmistusvaiheessa riskinottotilanteina korostuivat työn aloittaminen sekä työvaiheissa eteneminen. Arviointivaiheessa riskinottotilanteet liittyivät sosiaalisen ympäristön reaktioihin omaa valmista tuotetta kohtaan. Osoittautui, että näistä riskinottotilanteista on mahdollista erottaa taloudellisia, psykologisia ja sosiaalisia riskejä. Tämän tutkielman tulosten perusteella yrittäjämäisellä pedagogiikalla ei ole merkittävästi yhteyttä siihen, miten oppilaat riskin ottamiseen suhtautuvat. Erot, joita oppilaiden riskin ottamisessa ilmeni, liittyivät opetuksen toteuttamisen menetelmää enemmän oppilaiden valmistamaan tuotteeseen ja sen valmistustekniikkaan.

”Näytä pukumieheltä. Puhu kuin pukumies. Keksi itsellesi elämä.” Raha, uusi työ ja kuluttajasubjekti 2010-luvun kotimaisessa kaunokirjallisuudessa

Tutkimuksen tavoitteena on tarkastella miten uusliberalistisia kuluttajasubjekteja rakennetaan kaunokirjallisessa kerronnassa. Lähestyn aihetta tarkastelemalla aineistossani esiintyvää talousdiskurssia: rahan merkitystä toiminnan mahdollistavana välineenä, uuden työn rakenteita, jotka määrittelevät kuluttajasubjektin toiminta-aluetta sekä kulutusvalintoja, joilla subjekti haluaa ilmaista kuuluvansa johonkin sosiaaliseen ryhmään tai erottautuvansa siitä. Aineistona on kolme romaania, jotka kritisoivat ja parodioivat ironian keinoin kapitalistisen yhteiskunnan arvoja ja toimintamalleja. Miika Nousiaisen Metsäjätti (2011) käsittelee palkkatyöläisyyden ja yhteisökulttuurin muutosta jälkiteollisen työn globaaleille työmarkkinoille, joilla pärjäävät ne, jotka onnistuvat haalimaan eniten resursseja: koulutusta, verkostoja, rahaa. Tuomas Kyrön Kerjäläisessä ja jäniksessä (2011) romanikerjäläinen matkustaa ihmissalakuljettajan avulla Suomeen tavoittelemaan parempaa tulevaisuutta ja pääsyä länsimaiseen kulutuskulttuuriin. Venla Hiidensalon Mediahuora (2012) kertoo pakkopienyrittäjyydestä, jossa tekstityöläisellä, vapaalla toimittajalla, ei ole oikeutta tai mahdollisuutta sosiaaliturvaan tai moraaliin, mikäli haluaa pysyä työn syrjässä kiinni ja ruokkia jälkikasvunsa. Tutkimusta ohjaavina käsitteinä käytetään kuluttajasubjektia, talousdiskurssia, yhteiskuntaluokkaa ja identiteettiä. Teoriani rakentuu yhdistämällä kirjallisuudentutkimusta, kulttuurintutkimusta sekä sosiologista tutkimusta. Käytän Pierre Bourdieun pääoma-mallia, Beverley Skeggsin yhteiskuntaluokkatutkimusta, Jussi Ojajärven tutkimuksia kaunokirjallisuuden uusliberalistisesta käänteestä, Stuart Hallin identiteetti- ja subjektiteoria sekä Raija Julkusen uuden työn tutkimusta. Tutkimukseni osoittaa, että huumorin keinoja käyttävä kaunokirjallisuus toimii kritiikkinä luonnollistunutta valtaideologiaa sekä puhetapoja kohtaan ja tekee uusliberalismin ongelmia näkyväksi, samalla kerrontatapa uusintaa uusliberalistista menestystarinaa, kun se näyttää, että kuluttajasubjektit päätyvät sattumanvaraisesti haluamiinsa yhteiskuntaluokkiin ja kuluttajaryhmiin.

Oikeasuhtaisen seuraamuksen valinta moitittavan sananvapauden käytössä työ- ja virkasuhteissa

Tässä tutkielmassa tarkastellaan TSL:n, KvhL:n ja VirkamL:n sääntelemää oikeasuhtaisen seuraamuksen valintaa moitittavan sananvapauden käytössä oikeuskäytännössä muotoutuneiden kriteerien valossa. Tarkasteltavana on, millaisilla arviointiperusteilla huomautuksen ja varoituksen antamiskynnys sekä palvelussuhteen päättämiskynnys täyttyvät moitittavan sananvapauden käytössä työ- ja virkasuhteissa. Menetelmänä käytetään lainoppia tarkastelemalla TSL:n, KvhL:n ja VirkamL:n sanamuodosta tehtäviä tulkintoja hyödyntäen lain esitöitä ja oikeuskirjallisuutta sekä oikeuskäytäntöä. Valtion virkamiehen, kunnallisen viranhaltijan sekä työntekijän sananvapaus on turvattu PL 12 §:ssä ja EIS 10 art.:ssa. Sananvapaus ei oikeuta loukkaamaan muiden oikeuksia, ja työnantajalla on oikeus reagoida sananvapauden käyttöön, mikäli sillä esimerkiksi loukataan yrityksen tai viranomaisen oikeuksia taikka muiden työntekijöiden tai ulkopuolisten asiakkaiden oikeuksia. Sananvapauden rajoittamisen oikeusperustana on työsuhteessa TSL 3:1 mukainen lojaliteettivelvoite ja virkasuhteissa asiallisuusvaatimus (KvhL 17 §, VirkamL 14 §). Keskeistä seuraamusharkinnassa on työntekijän, viranhaltijan, valtion virkamiehen sananvapauden käytön moitittavuuden aste. Seuraamuskynnystä nostaa ja madaltaa oikeuskäytännössä muotonsa saaneet vertailuperusteet, joita tutkielmassa tarkastellaan. Mitä moitittavampaa sananvapauden käyttö on, sen todennäköisemmin seuraamuskynnys ylittyy. Arviointi tehdään kokonaisarviona in casu. Kaikkiin tapauksiin soveltuvaa yleissääntöä ei ole mahdollista asettaa lakien joustavien sananmuotojen vuoksi.

Kim jestem? Kim się staję? W dialogicznym napięciu JA-TY. Źródła teoretyczne

W niniejszym artykule przedstawiono komponenty wchodzące w skład źródeł teoretycznych, będących podstawą powstania konspektu warsztatów. Treść artykułu nie wychodzi poza obszar metodologii teologicznej. Badania ukazujące przesłanki procesów zaaranżowanych w czasie warsztatów oraz podane wnioski końcowe wynikają z badań przeprowadzonych w polu analiz teologii fundamentalnej. Ta dziedzina teologiczna zakłada interdyscyplinarne łączenie metod i osiągnięć teologii i nauk antropologicznych. W kolejnych odsłonach ukazane zostały trzy aspekty odkrywania ludzkiego JA w dialogicznym napięciu z TY. Dzięki analizie transakcyjnej Erica Berne’a można ukazać, przy pomocy prostych technik edukacyjnych, że człowiek w poznaniu siebie potrzebuje drugiego w każdym aspekcie swego istnienia. Tym, co umożliwia mu bycie z sobą i innymi do zdolność wczucia. Tę kategorię badawczą analizowała Edyta Stein jako podstawę realnych relacji JA-TY, gdzie poprzez TY może zostać określony sam człowiek jako nie-ja, inny, drugi, czy ostatecznie osobowy Bóg. Możliwość zastosowania procesów opartych na wczuciu w przestrzeni relacji wiary doprowadza ku teologii duchowości. W jej ramach wypracowany został model antropologiczny, który widzi człowieka w ciągłym stawaniu się wobec innych i Boga. Przyjęta do badań praktyczno-biblijna antropologia Ignacego Loyoli pozwala na uwiarygodnienie kroków podjętych w czasie warsztatów, tak by nie były manipulacją a służyły wolnemu wyborowi uczestników w świadomym poszukiwaniu sensu i celu istnienia.

Varsinaissuomalaisten nuorten kouluhyvinvointi ja sen muutokset vuosina 2008 - 2012

Tässä tutkimuksessa tarkastellaan Varsinais-Suomalaisten 9.luokkalaisten nuorten kouluhyvinvointia vuosina 2008, 2010 ja 2012. Tutkimuksen aineisto koostuu Turun lapsi- ja nuorisotutkimuskeskus Cyrin, Nuoret luupin alla -tutkimusaineistoihin edellä mainituilta vuosilta. Tämän tutkimuksen aineisto koostui yhteensä 4946 nuoren vastauksesta. Tutkimukseen otettiin mukaan kaikki Varsinais-suomen alueella asuvat nuoret, jotka olivat vastanneet Nuoret luupin alla –tutkimukseen vuosina 2008, 2010 ja 2012. Kouluhyvinvointia tutkitaan tässä tutkimuksessa Konun (2002) kouluhyvinvointimallia apuna käyttäen. Kouluhyvinvointimallista käytetään niitä osatekijöitä, jotka ovat rinnastettavissa Allardtin (1976) hyvinvointiteorian osatekijöihin (having, loving, being), joihin myös Konun kouluhyvinvointimalli perustuu. Kouluhyvinvoinnilla tarkoitetaan tässä tutkimuksessa Konun (2002) luoman kouluhyvinvointimallin kolmea osatekijää; koulun olosuhteita, sosiaalisia suhteita koulussa sekä itsensä toteuttamisen mahdollisuuksia koulussa. Tutkimuksen tulosten perusteella voidaan todeta, että nuorten kouluhyvinvoinnissa ei ole tapahtunut radikaaleja muutoksia tutkimusvuosina. Nuorten varsinaissuomalaisten oppilaiden kokemusten suunta kouluhyvinvoinnista on ollut parempaan päin. Tilastollisesti merkitseviä eroja sukupuolten sekä asuinpaikan mukaan löytyy eri kouluhyvinvoinnin osatekijöissä eri tutkimusvuosina. Maaseudulla asuvat oppilaat kokivat koulunolosuhteet paremmiksi kahtena ensimmäisenä tutkimusvuotena, mutta erot ovat tasaantuneet viimeiseen tutkimusvuoteen tultaessa. Asuinpaikka ei vaikuttanut oppilaiden kokemuksiin itsensä toteuttamisen mahdollisuuksista koulussa eikä kokemuksiin opettajien ja oppilaiden välisistä sosiaalisista suhteista. Asuinpaikka vaikutti oppilaiden kokemuksiin oppilaiden keskinäisistä sosiaalisista suhteista, vain keskimmäisenä tutkimusvuotena siten, että maaseudulla asuvat oppilaat kokivat suhteet paremmiksi. Sukupuolten välillä oli eroja koetuissa koulun olosuhteissa kahtena viimeisenä tutkimusvuotena, siten että ensin pojat kokivat koulun olosuhteet paremmiksi ja viimeisenä tutkimusvuotena tytöt kokivat olosuhteet paremmiksi. Pojat kokivat myös itsensä toteuttamisen mahdollisuudet koulussa paremmiksi kahtena viimeisenä tutkimusvuotena. Sukupuolten kokemuksissa oppilaiden keskinäisistä sosiaalisista suhteista ei ollut tilastollisesti merkitsevää eroa, mutta opettajien ja oppilaiden sosiaaliset suhteet pojat kokivat tilastollisesti merkitsevästi paremmiksi kaikkina tutkimusvuosina.

Urban agriculture : a growing field of research workshop at INTERACT 2013

Growing food presents diverse challenges and opportunities within the urban environment. As cities develop, population density rises, land prices rise, and the opportunity to use land for traditional farming and gardening diminishes. Counter to this trend there are a growing number of both community gardens, city farms, guerrilla gardening, rooftop and vertical gardens, pot plants, windowsill herbs, and other balcony or backyard gardens cropping up in different cities, all with a purpose to produce food. This workshop brings to-gether practitioners and researchers in the field of urban agriculture and Hu-man-Computer Interaction to explore and opportunities for technology design to support the different forms of growing practice and foster local food production in cities. This 1-day workshop will serve as an active forum for researchers and practi-tioners across various fields including, but not limited to, agriculture and gar-dening, education, urban planning, human-computer interaction, and communi-ty engagement. This workshop has three distinct points of focus: i) Individual and small-scale gardening and food production, and how to connect like minded people who are involved in these practices to share their knowledge ii) Com-munities involved in urban agriculture, either through community gardens, city farms, or grassroots movements, often dependant on volunteer participation, providing the challenge of managing limited resources iii) Environmental and sociocultural sustainability through urban agriculture. The participants will have an opportunity to present their own work. This will be followed by a visit to a nearby city farm, which will provide a local context for a group design exercise. Finally the workshop will conclude with panel dis-cussions to review opportunities for further research and collaborations beyond the conference. For more information, please visit the workshop website, at http://www.urbaninformatics.net/resources/interact2013cfp/

One-stop shopping : some notes on the concept and some Australian initiatives

This paper aims to review the notion of the one-stop shop, particularly as it has developed in Australia. It has three main components. The first focuses on a significant initiative taken by the Coombs Royal Commission on Australian Government Administration as a form of action research in the mid-1970s. The second highlights the associated work on access theory by the late Bernard Schaffer and his associates and explains why this work was of fundamental importance for the development of the idea of the one-stop shop. The third deals with subsequent Australian understandings and a pp lications of the idea.

Responsible reconstruction : The architect's role

This paper investigates the role of the architect in post-disaster reconstruction and questions their ability to facilitate per- manent building solutions. There is an ever-increasing population of refugees and internally displaced persons due to disasters and conflicts who have a basic need for shelter. To date, housing solutions for such people has tended to focus on short-term, temporary shelter solutions that have been largely unsuccessful. This increasing demand for shelter has led to an emerging group of architects skilled in post-disaster reconstruction. These architects acknowledge that shelter is critical to survival, but believe architects should focus on rebuilding in a manner that is quick, durable but permanent. They believe that an architect skilled in post-disaster reconstruction can produce solutions that meet the requirement of the emergency phase, through to semi-permanent and even permanent homes, without wasting time and money on interim shelters. Case Study Research was used to examine and evaluate the assistance provided by Emergency Architects Australia (EAA) to the Kei Gold community in the Solomon Islands after the 2007 earthquake and tsunami. The results indicate that an architect’s response to a disaster must go beyond providing temporary shelter; they must create permanent building solutions that respond to the site and the culture while servicing the needs of the communi- ty. The vernacular reconstruction methods implemented by EAA in Kei Gold Village have been successful in develop- ing permanent housing solutions. Further research and development is required to gain a broader understanding of the role of the architect in disasters of varying scales and typologies.

Roles of forced nicotine exposure and Comt gene disruption in the development of addiction-related behavioural and neurochemical changes in mice

Activation of midbrain dopamine systems is thought to be critically involved in the addictive properties of abused substances. Drugs of abuse increase dopamine release in the nucleus accumbens and dorsal striatum, which are the target areas of mesolimbic and nigrostriatal dopamine pathways, respectively. Dopamine release in the nucleus accumbens is thought to mediate the attribution of incentive salience to rewards, and dorsal striatal dopamine release is involved in habit formation. In addition, changes in the function of prefrontal cortex (PFC), the target area of mesocortical dopamine pathway, may skew information processing and memory formation such that the addict pays an abnormal amount of attention to drug-related cues. In this study, we wanted to explore how long-term forced oral nicotine exposure or the lack of catechol-O-methyltransferase (COMT), one of the dopamine metabolizing enzymes, would affect the functioning of these pathways. We also wanted to find out how the forced nicotine exposure or the lack of COMT would affect the consumption of nicotine, alcohol, or cocaine. First, we studied the effect of forced chronic nicotine exposure on the sensitivity of dopamine D2-like autoreceptors in microdialysis and locomotor activity experiments. We found that the sensitivity of these receptors was unchanged after forced oral nicotine exposure, although an increase in the sensitivity was observed in mice treated with intermittent nicotine injections twice daily for 10 days. Thus, the effect of nicotine treatment on dopamine autoreceptor sensitivity depends on the route, frequency, and time course of drug administration. Second, we investigated whether the forced oral nicotine exposure would affect the reinforcing properties of nicotine injections. The chronic nicotine exposure did not significantly affect the development of conditioned place preference to nicotine. In the intravenous self-administration paradigm, however, the nicotine-exposed animals self-administered nicotine at a lower unit dose than the control animals, indicating that their sensitivity to the reinforcing effects of nicotine was enhanced. Next, we wanted to study whether the Comt gene knock-out animals would be a suitable model to study alcohol and cocaine consumption or addiction. Although previous work had shown male Comt knock-out mice to be less sensitive to the locomotor-activating effects of cocaine, the present study found that the lack of COMT did not affect the consumption of cocaine solutions or the development of cocaine-induced place preference. However, the present work did find that male Comt knock-out mice, but not female knock-out mice, consumed ethanol more avidly than their wild-type littermates. This finding suggests that COMT may be one of the factors, albeit not a primary one, contributing to the risk of alcoholism. Last, we explored the effect of COMT deficiency on dorsal striatal, accumbal, and prefrontal cortical dopamine metabolism under no-net-flux conditions and under levodopa load in freely-moving mice. The lack of COMT did not affect the extracellular dopamine concentrations under baseline conditions in any of the brain areas studied. In the prefrontal cortex, the dopamine levels remained high for a prolonged time after levodopa treatment in male, but not female, Comt knock-out mice. COMT deficiency induced accumulation of 3,4-dihydroxyphenylacetic acid, which increased further under levodopa load. Homovanillic acid was not detectable in Comt knock-out animals either under baseline conditions or after levodopa treatment. Taken together, the present results show that although forced chronic oral nicotine exposure affects the reinforcing properties of self-administered nicotine, it is not an addiction model itself. COMT seems to play a minor role in dopamine metabolism and in the development of addiction under baseline conditions, indicating that dopamine function in the brain is well-protected from perturbation. However, the role of COMT becomes more important when the dopaminergic system is challenged, such as by pharmacological manipulation.

The roughness and imaging characterisation of different pharmaceutical surfaces

The surface properties of solid state pharmaceutics are of critical importance. Processing modifies the surfaces and effects surface roughness, which influences the performance of the final dosage form in many different levels. Surface roughness has an effect on, e.g., the properties of powders, tablet compression and tablet coating. The overall goal of this research was to understand the surface structures of pharmaceutical surfaces. In this context the specific purpose was to compare four different analysing techniques (optical microscopy, scanning electron microscopy, laser profilometry and atomic force microscopy) in various pharmaceutical applications where the surfaces have quite different roughness scale. This was done by comparing the image and roughness analysing techniques using powder compacts, coated tablets and crystal surfaces as model surfaces. It was found that optical microscopy was still a very efficient technique, as it yielded information that SEM and AFM imaging are not able to provide. Roughness measurements complemented the image data and gave quantitative information about height differences. AFM roughness data represents the roughness of only a small part of the surface and therefore needs other methods like laser profilometer are needed to provide a larger scale description of the surface. The new developed roughness analysing method visualised surface roughness by giving detailed roughness maps, which showed local variations in surface roughness values. The method was able to provide a picture of the surface heterogeneity and the scale of the roughness. In the coating study, the laser profilometer results showed that the increase in surface roughness was largest during the first 30 minutes of coating when the surface was not yet fully covered with coating. The SEM images and the dispersive X-ray analysis results showed that the surface was fully covered with coating within 15 to 30 minutes. The combination of the different measurement techniques made it possible to follow the change of surface roughness and development of polymer coating. The optical imaging techniques gave a good overview of processes affecting the whole crystal surface, but they lacked the resolution to see small nanometer scale processes. AFM was used to visualize the nanoscale effects of cleaving and reveal the full surface heterogeneity, which underlies the optical imaging. Ethanol washing changed small (nanoscale) structure to some extent, but the effect of ethanol washing on the larger scale was small. Water washing caused total reformation of the surface structure at all levels.

Crystallization as a Tool for Controlling and Designing Properties of Pharmaceutical Solids

The ability to deliver the drug to the patient in a safe, efficacious and cost-effective manner depends largely on the physicochemical properties of the active pharmaceutical ingredient (API) in the solid state. In this context, crystallization is of critical importance in pharmaceutical industry, as it defines physical and powder properties of crystalline APIs. An improved knowledge of the various aspects of crystallization process is therefore needed. The overall goal of this thesis was to gain better understanding of the relationships between crystallization, solid-state form and properties of pharmaceutical solids with a focus on a crystal engineering approach to design technological properties of APIs. Specifically, solid-state properties of the crystalline forms of the model APIs, erythromycin A and baclofen, and the influence of solvent on their crystallization behavior were investigated. In addition, the physical phenomena associated with wet granulation and hot-melting processing of the model APIs were examined at the molecular level. Finally, the effect of crystal habit modification of a model API on its tabletting properties was evaluated. The thesis enabled the understanding of the relationship between the crystalline forms of the model APIs, which is of practical importance for solid-state control during processing and storage. Moreover, a new crystalline form, baclofen monohydrate, was discovered and characterized. Upon polymorph screening, erythromycin A demonstrated high solvate-forming propensity thus emphasizing the need for careful control of the solvent effects during formulation. The solvent compositions that yield the desirable crystalline form of erythromycin A were defined. Furthermore, new examples on solvent-mediated phase transformations taking place during wet granulation of baclofen and hot-melt processing of erythromycin A dihydrate with PEG 6000 are reported. Since solvent-mediated phase transformations involve the crystallization of a stable phase and hence affect the dissolution kinetics and possibly absorption of the API these transformations must be well documented. Finally, a controlled-crystallization method utilizing HPMC as a crystal habit modifier was developed for erythromycin A dihydrate. The crystals with modified habit were shown to posses improved compaction properties as compared with those of unmodified crystals. This result supports the idea of morphological crystal engineering as a tool for designing technological properties of APIs and is of utmost practical interest.

From Polymorph Screening to Dissolution Testing - Solid Phase Analysis during Pharmaceutical Development and Manufacturing

Solid materials can exist in different physical structures without a change in chemical composition. This phenomenon, known as polymorphism, has several implications on pharmaceutical development and manufacturing. Various solid forms of a drug can possess different physical and chemical properties, which may affect processing characteristics and stability, as well as the performance of a drug in the human body. Therefore, knowledge and control of the solid forms is fundamental to maintain safety and high quality of pharmaceuticals. During manufacture, harsh conditions can give rise to unexpected solid phase transformations and therefore change the behavior of the drug. Traditionally, pharmaceutical production has relied on time-consuming off-line analysis of production batches and finished products. This has led to poor understanding of processes and drug products. Therefore, new powerful methods that enable real time monitoring of pharmaceuticals during manufacturing processes are greatly needed. The aim of this thesis was to apply spectroscopic techniques to solid phase analysis within different stages of drug development and manufacturing, and thus, provide a molecular level insight into the behavior of active pharmaceutical ingredients (APIs) during processing. Applications to polymorph screening and different unit operations were developed and studied. A new approach to dissolution testing, which involves simultaneous measurement of drug concentration in the dissolution medium and in-situ solid phase analysis of the dissolving sample, was introduced and studied. Solid phase analysis was successfully performed during different stages, enabling a molecular level insight into the occurring phenomena. Near-infrared (NIR) spectroscopy was utilized in screening of polymorphs and processing-induced transformations (PITs). Polymorph screening was also studied with NIR and Raman spectroscopy in tandem. Quantitative solid phase analysis during fluidized bed drying was performed with in-line NIR and Raman spectroscopy and partial least squares (PLS) regression, and different dehydration mechanisms were studied using in-situ spectroscopy and partial least squares discriminant analysis (PLS-DA). In-situ solid phase analysis with Raman spectroscopy during dissolution testing enabled analysis of dissolution as a whole, and provided a scientific explanation for changes in the dissolution rate. It was concluded that the methods applied and studied provide better process understanding and knowledge of the drug products, and therefore, a way to achieve better quality.

Upregulation and Functionality of Neuronal Nicotinic Acetylcholine Receptors

Cigarette smoking is, in developed countries, the leading cause of premature death. In tobacco smoke, the main addictive compound is nicotine, which in the brain binds to neuronal nicotinic acetylcholine receptors (neuronal nAChRs). These have been implicated in addiction, but also in several neurological disorders including Alzheimer's and Parkinson's diseases, Tourette's syndrome, attention-deficit hyperactivity disorder (ADHD), schizophrenia, pain, depression, and autosomal-dominant noctural frontal lobe epilepsy; all of which makes nAChRs an intriguing target of study. Chronic treatment with nicotine leads to an increase in the number of nAChRs (upregulation) in the brain and changes their functionality. Changes in the properties of nAChRs are likely to occur in smokers as well, since they are exposed to nicotine for long periods of time. Several nAChR subtypes likely play a role in the formation of nicotine addiction by participating in the release of dopamine in the striatum. The aim of this study was to clarify at cellular level the changes in nAChR characteristics resulting from chronic nicotine treatment. SH-SY5Y cells, endogenously several nAChR-expressing, and SH-EP1-h-alfa7 cells, transfected with the alfa 7 nAChR subunit gene were treated chronically with nicotine. The localisation of alfa 7 and beta2 subunits was studied with confocal and electron microscopy. Functionality of nAChRs was studied with calcium fluorometry. Effects of long-term treatment with opioid compounds on nAChRs were studied by means of ligand binding. Confocal microscopy showed that in SH-SY5Y cells, alfa7 and beta2 subunits formed clusters, unlike the case in SH-EP1-h alfa7 cells, where alfa7 nAChRs were distributed more diffusely. The majority of nAChR subunits localised on endoplasmic reticulum (ER). The isomers of methadone acted as agonists at alfa7 nAChRs. Acute morphine challenge also stimulated nAChRs. Chronic treatment with methadone or morphine led to an increased number of nAChRs. In animal studies, mice received nicotine for 7 weeks. Electron microscopical analysis of the localisation of nAChRs showed in the striatum that alfa7 and beta2 nAChR subunits localised synaptically, extrasynaptically, and intracellularly, with the majority localising extrasynaptically. Chronic nicotine treatment caused an increase in the number of nAChR subunits at all studied locations. These results suggest that the alfa7 nAChR and beta2 subunit-containing nAChRs respond to chronic nicotine treatment differently. This may indicate that the functional balance of various nAChR subtypes in control of the release of dopamine is altered as a result of chronic nicotine treatment. Compounds binding both to opioid and nACh receptors may be of clinical importance.

The Effects of Nicotine on the Regulation of Neuronal Alpha7 Nicotinic Acetylcholine Receptors and Intracellular Signalling Pathways

Nicotine, the addictive compound of tobacco products, exerts its effects in the brain by binding to neuronal acetylcholine nicotinic receptors (nAChRs). The aim of the present study was to increase the knowledge of nicotine s complex effects, the focus being on homomeric alpha7-nAChRs that are widely expressed in the brain. Nicotinic regulation of differential signalling molecules including transcriptional regulators was also studied. We found that the number of alpha7-nAChRs is increased in specific brain regions in mice, in a time-dependent manner after chronic oral nicotine administration. Our results suggest that in addition to alpha4beta2-nAChRs, the other major nAChR subtype expressed in the brain, the number of alpha7-nAChRs is affected by chronic presence of nicotine. We suggest that when studying the long-term effects of nicotine, the duration on administration is of great importance. Next, we observed that nicotine exposure induces accumulation of cAMP in cell cultures expressing nAChRs. Furthermore, nicotine-induced alpha7-nAChR upregulation was potentiated by treatments enhancing cAMP-signalling, suggesting a role for cAMP in the upregulation process. Protein kinase C (PKC) was found essential for the basal regulation of alpha7-nAChR number. The nicotine-evoked alpha7-nAChR upregulation could be further increased by PKC overexpression. Thirdly, the effects of nicotine on dopamine and cAMP regulated phosphoprotein (DARPP-32) were characterised in rat brain. The results show that DARPP-32 is regulated by both acute and long-term nicotine treatment in the striatal subdivisions. The effect of acute nicotine is dose-dependent and the three striatal regions display differential sensitivities to nicotine. Chronic nicotine is also able to regulate DARPP-32 signalling with prominent effect seen in the nucleus accumbens (NAc), suggesting a role for DARPP-32 in the mediation of long-term effects of nicotine. Finally, the regulation of transcription factors Elk-1 and FosB/deltaFosB by nicotine was investigated. We found that Elk-1 is activated by acute nicotine selectively in the NAc core and hippocampal area CA1, whereas acute nicotine does not affect FosB/deltaFosB. Long-term intermittent or continuous nicotine increases the level of total Elk-1 in the same brain regions as acute nicotine. FosB/deltaFosB is also affected by chronic nicotine. Thus, similarly to other drugs of abuse, nicotine regulates transcriptional regulators Elk-1 and FosB/deltaFosB. These results bring further support for a common mechanism underlying the development of addiction. Nicotine s positive effects on learning and memory might involve the transcription factor Elk-1 based on the changes seen in the hippocampus, the key area in cognitive functions.