Characterisation and processing of amorphous binary mixtures with low glass transition temperature

The number of drug substances in formulation development in the pharmaceutical industry is increasing. Some of these are amorphous drugs and have glass transition below ambient temperature, and thus they are usually difficult to formulate and handle. One reason for this is the reduced viscosity, related to the stickiness of the drug, that makes them complicated to handle in unit operations. Thus, the aim in this thesis was to develop a new processing method for a sticky amorphous model material. Furthermore, model materials were characterised before and after formulation, using several characterisation methods, to understand more precisely the prerequisites for physical stability of amorphous state against crystallisation. The model materials used were monoclinic paracetamol and citric acid anhydrate. Amorphous materials were prepared by melt quenching or by ethanol evaporation methods. The melt blends were found to have slightly higher viscosity than the ethanol evaporated materials. However, melt produced materials crystallised more easily upon consecutive shearing than ethanol evaporated materials. The only material that did not crystallise during shearing was a 50/50 (w/w, %) blend regardless of the preparation method and it was physically stable at least two years in dry conditions. Shearing at varying temperatures was established to measure the physical stability of amorphous materials in processing and storage conditions. The actual physical stability of the blends was better than the pure amorphous materials at ambient temperature. Molecular mobility was not related to the physical stability of the amorphous blends, observed as crystallisation. Molecular mobility of the 50/50 blend derived from a spectral linewidth as a function of temperature using solid state NMR correlated better with the molecular mobility derived from a rheometer than that of differential scanning calorimetry data. Based on the results obtained, the effect of molecular interactions, thermodynamic driving force and miscibility of the blends are discussed as the key factors to stabilise the blends. The stickiness was found to be affected glass transition and viscosity. Ultrasound extrusion and cutting were successfully tested to increase the processability of sticky material. Furthermore, it was found to be possible to process the physically stable 50/50 blend in a supercooled liquid state instead of a glassy state. The method was not found to accelerate the crystallisation. This may open up new possibilities to process amorphous materials that are otherwise impossible to manufacture into solid dosage forms.

Understanding solid-state transformations during dehydration: new insights using vibrational spectroscopy and multivariate modelling

Many active pharmaceutical ingredients (APIs) have both anhydrate and hydrate forms. Due to the different physicochemical properties of solid forms, the changes in solid-state may result in therapeutic, pharmaceutical, legal and commercial problems. In order to obtain good solid dosage form quality and performance, there is a constant need to understand and control these phase transitions during manufacturing and storage. Thus it is important to detect and also quantify the possible transitions between the different forms. In recent years, vibrational spectroscopy has become an increasingly popular tool to characterise the solid-state forms and their phase transitions. It offers several advantages over other characterisation techniques including an ability to obtain molecular level information, minimal sample preparation, and the possibility of monitoring changes non-destructively in-line. Dehydration is the phase transition of hydrates which is frequently encountered during the dosage form production and storage. The aim of the present thesis was to investigate the dehydration behaviour of diverse pharmaceutical hydrates by near infrared (NIR), Raman and terahertz pulsed spectroscopic (TPS) monitoring together with multivariate data analysis. The goal was to reveal new perspectives for investigation of the dehydration at the molecular level. Solid-state transformations were monitored during dehydration of diverse hydrates on hot-stage. The results obtained from qualitative experiments were used to develop a method and perform the quantification of the solid-state forms during process induced dehydration in a fluidised bed dryer. Both in situ and in-line process monitoring and quantification was performed. This thesis demonstrated the utility of vibrational spectroscopy techniques and multivariate modelling to monitor and investigate dehydration behaviour in situ and during fluidised bed drying. All three spectroscopic methods proved complementary in the study of dehydration. NIR spectroscopy models could quantify the solid-state forms in the binary system, but were unable to quantify all the forms in the quaternary system. Raman spectroscopy models on the other hand could quantify all four solid-state forms that appeared upon isothermal dehydration. The speed of spectroscopic methods makes them applicable for monitoring dehydration and the quantification of multiple forms was performed during phase transition. Thus the solid-state structure information at the molecular level was directly obtained. TPS detected the intermolecular phonon modes and Raman spectroscopy detected mostly the changes in intramolecular vibrations. Both techniques revealed information about the crystal structure changes. NIR spectroscopy, on the other hand was more sensitive to water content and hydrogen bonding environment of water molecules. This study provides a basis for real time process monitoring using vibrational spectroscopy during pharmaceutical manufacturing.

Investigating physical properties of solid dosage forms during pharmaceutical processing : Process analytical applications of vibrational spectroscopy

In order to improve and continuously develop the quality of pharmaceutical products, the process analytical technology (PAT) framework has been adopted by the US Food and Drug Administration. One of the aims of PAT is to identify critical process parameters and their effect on the quality of the final product. Real time analysis of the process data enables better control of the processes to obtain a high quality product. The main purpose of this work was to monitor crucial pharmaceutical unit operations (from blending to coating) and to examine the effect of processing on solid-state transformations and physical properties. The tools used were near-infrared (NIR) and Raman spectroscopy combined with multivariate data analysis, as well as X-ray powder diffraction (XRPD) and terahertz pulsed imaging (TPI). To detect process-induced transformations in active pharmaceutical ingredients (APIs), samples were taken after blending, granulation, extrusion, spheronisation, and drying. These samples were monitored by XRPD, Raman, and NIR spectroscopy showing hydrate formation in the case of theophylline and nitrofurantoin. For erythromycin dihydrate formation of the isomorphic dehydrate was critical. Thus, the main focus was on the drying process. NIR spectroscopy was applied in-line during a fluid-bed drying process. Multivariate data analysis (principal component analysis) enabled detection of the dehydrate formation at temperatures above 45°C. Furthermore, a small-scale rotating plate device was tested to provide an insight into film coating. The process was monitored using NIR spectroscopy. A calibration model, using partial least squares regression, was set up and applied to data obtained by in-line NIR measurements of a coating drum process. The predicted coating thickness agreed with the measured coating thickness. For investigating the quality of film coatings TPI was used to create a 3-D image of a coated tablet. With this technique it was possible to determine coating layer thickness, distribution, reproducibility, and uniformity. In addition, it was possible to localise defects of either the coating or the tablet. It can be concluded from this work that the applied techniques increased the understanding of physico-chemical properties of drugs and drug products during and after processing. They additionally provided useful information to improve and verify the quality of pharmaceutical dosage forms

Mapinduzi Daima – Revolution Forever : Using the 1964 Revolution in Nationalistic Political Discourses in Zanzibar

This Master's thesis examines two opposite nationalistic discourses on the revolution of Zanzibar. Chama cha Mapinduzi (CCM), the party in power since the 1964 revolution defends its revolutionary and "African" heritage in the current multi-party system. New nationalists, including among others the main opposition party Civic United Front (CUF), question both the 1964 revolution and the post-revolution period and blame CCM for empty promises, corruption and ethnic discrimination. This study analyzes the role of a significant historical event in the creation of nationalistic ideology and national identity. The 1964 revolution forms the nucleus of various debates related to the history of Zanzibar: slavery, colonialism, racial discrimination and political violence. Representations of these Social constructivist principles form the basis of this study, and central concepts in the theoretical framework are nationalism, national identity, ethnicity and race. I use critical discourse analysis as my research method, lean on the work by Teun A. van Dijk and Norman Fairclough as the most significant researchers in this field. I examine particularly the ways in which linguistic methods, such as stereotypes and metaphors are used to form in- and out-groups ("us" vs. "others"). My material, both in Swahili and English, was collected mainly in Tanzania in the fall of 2007 and from online sources in the spring of 2009. It includes publications by the Zanzibari government between the years of 1964-2000 (12), official speeches for the Revolution Day or the Union Day (12), articles from Tanzanian newspapers from the 1990s until the year of 2009 (15), memoirs and political pamphlets (10), blog posts and opinion pieces from four different websites (8), and interviews or personal communication in Zanzibar, Dar es Salaam and Uppsala (8). Nationalistic rhetoric often creates enemy images by using binary good-bad oppositions. Both discourses in this study build identities on the basis of "otherness" and exclusion, with the intent of emphasizing the particularity of the own group and excluding "evilness" outside the own reference group. These opposite views on the 1964 revolution as the main axis of the history of Zanzibar build different portraits of the nation and Zanzibari-ness (Uzanzibari). CCM still relies on the pre-revolutionary enemy images of Arabs as selfish rulers and cruel slave traders. For CCM, Zanzibar is primarily an "African" nation and a part of Tanzania which is threatened by "Arabs", the outsiders. In contrast, the new nationalists stress the long history of Zanzibar as multi-racial, cosmopolitan and formerly independent country which has its own, separate culture and identity from mainland Tanzanians. Heshima, honour/respect, one of the basic values of Swahili culture, occupies a central role in both discourses: the main party emphasizes that the revolution returned "heshima" to the Zanzibari Africans after centuries of humiliation, whereas the new nationalists claim that ever since the revolution all "non-Africans" have been humiliated and lost their "heshima". According to the new nationalists, true Zanzibari values which include tolerance and harmony between different "races" were lost when the "foreign" revolutionaries arrived from the mainland. Consequently, they see the 1964 revolution as Tanganyikan colonialism which began with the help of Western countries, and maintain that this "colonialism" still continues in the violent multi-party elections.

The Art of Sympathy : Forms of Moral and Emotional Persuasion in Fiction

"The Art of Sympathy: Forms of Moral and Emotional Persuasion" in Fiction is an interdisciplinary study that looks closely at the ways that stories evoke sympathy, and the significance of this emotion for the development of moral attitudes and awareness. By linking readers' emotional responses to fiction with the potential impact of such responses on "the moral imagination," the study builds on empirical research conducted by literary scholars and psychologists into the emotional effects of reading fiction, as well as social psychological research into the connections between empathy/sympathy and moral development. I first investigate the dynamics of readers beliefs regarding characters in fictional narratives, and the nature of the emotions that they may experience as a result of those beliefs. The analysis demonstrates that there are important similarities between real emotions and emotions generated by fiction. Recognizing these similarities, I claim, can help us to conceptualize the nature of sympathetic responses to fictional characters. Building on these assertions, I then draw on research from social psychology and philosophy to develop a comprehensive definition of sympathy and to clarify the ways in which sympathy operates, both in people s daily lives and in readers sympathetic responses to fictional characters. Having established this definition and delineated its practical implications, I then examine how particular stories, through a variety of narrative techniques, persuade readers to feel sympathy for characters who are unsympathetic in certain ways. In order to verify my claims about the impact of these stories on readers emotions, I also review the results of tests that I conducted with nearly 200 adolescent readers. Through these tests, which were constructed and scored according to methods prevalent in social psychological research, it was determined that a majority of readers felt sympathy for the protagonists in two of the stories included in the study. These results were combined with data from an additional test, a standard measure of empathy and sympathy in the field of social psychology. The cross-tabulation of these results suggests that there was not a strong connection between readers responses and their general tendencies to feel sympathy for others. This finding would appear to support my hypotheses regarding the sympathetic persuasiveness of the stories in question. In light of these results, finally, I consider the potential contribution that fiction can make to adolescent emotional and moral development and the implications of that potential for future language arts curricula in the schools. In particular, I suggest the pedagogical importance of providing adolescents with opportunities to engage with the lives of fictional characters, and especially to experience feelings of sympathy for individuals towards whom they ordinarily might feel aversion.

Vikuroivia vilkaisuja : Ruumis, sukupuoli, seksuaalisuus ja visuaalisen kulttuurin tutkimus

The PhD dissertation "Bucking Glances: On Body, Gender, Sexuality and Visual Culture Research" consists of theoretical introduction and five articles published between 2002-2005. The articles analyze the position of visual representations in the processes of knowledge production on acceptable genders, bodies, and sexualities in contemporary Wes¬tern societies. The research material is heterogeneous, consisting of representations of contemporary art, advertisements, and fashion images. The ideological starting point of the PhD dissertation is the politics of the gaze and the methods used to expose this are the concepts of oppositional gaze, close reading, and resisting reading. The study situates visual representations in dialogue with the concepts of the grotesque and androgyny, as well as with queer-theory and theories of the gaze. The research challenges normative meanings of visual representations and opens up space for more non-conventional readings attached to femininity and masculinity. The visual material is read as troubling the prevailing heteronormative gender system. The dissertation also indicates how visual culture research utilizing the approach of queer theory can be fruitful in opposing and re-visioning changes in the repressive gender system. The article "A Heroic Male and A Beautiful Woman. Teemu Mäki, Orlan and the Ambivalence of the Grotesque Body" problematizes the concept of heroic masculinity through the analysis of the Finnish artist Teemu Mäki's masochistic performance The Good Friday (1989). It also analyzes cosmetic surgery, undertaken by the French artist Orlan, as a cultural tool in constructing and visualizing the contemporary, com¬mercial ideals of female beauty. The article "Boys Will Be Girls Will Be Boys Will Be Girls. The Ambivalence of Androgyny in Calvin Klein' Advertisements" is a close reading of the Calvin Klein perfume advertisement One (1998) in reference to the concept of androgyny. The critical point of the article is that androgynous male bodies allow the extension of the categorical boundaries of masculinity and homosexuality, whereas representations of androgynous women feed into the prevailing stereotypes of femininity, namely the fear of fat. The article "See-through Closet: Female Androgyny in the 1990s Fashion Images, New Woman and Lesbian Chic" analyzes the late 1990s fashion advertisements through the concept of female androgyny. The article argues that the figures of the masculine female androgynes in the late 1990s fashion magazines do not problematize the dichotomous gender binary. The women do not pass as men but produce a variation of heterosexual desirability. At the same time, the representations open up space for lesbian gazing and desiring. The article "Why are there no lesbian advertisements?" addresses the issue of femme gaze and desire in relation to heterosexual fashion advertisements from the British edition of the mainstream fashion magazine Vogue. The article considers possibilities for resistant femme visibility, identification, and desire. The article "Woman, Food, Home. Pirjetta Brander's and Heidi Romo's Works as Bucking Representations of Femininity" analyses the production and queering of heteronormative femininity and family through the analysis of art works. The article discusses how the term queer has been translated into Finnish. The article also introduces a new translation for the term queer: the noun vikuuri, i.e. faulty form and the verb vikuroida, i.e. to buck. In Finnish, the term vikuuri is the vernacular or broken form of the term figure, i.e. figuuri. Vikuuri represents all forms situated outside the norm and the normative.

Post-industrial Intervention : An Activity-Theoretical Expedition Tracing the Proximal Development of Forms of Conducting Interventions

Ei saatavilla

Valuation of reduced eutrophication in the Gulf of Finland : Choice experiment with attention to heterogeneous and discontinuous preferences and respondent uncertainty

Eutrophication of the Baltic Sea is a serious problem. This thesis estimates the benefit to Finns from reduced eutrophication in the Gulf of Finland, the most eutrophied part of the Baltic Sea, by applying the choice experiment method, which belongs to the family of stated preference methods. Because stated preference methods have been subject to criticism, e.g., due to their hypothetical survey context, this thesis contributes to the discussion by studying two anomalies that may lead to biased welfare estimates: respondent uncertainty and preference discontinuity. The former refers to the difficulty of stating one s preferences for an environmental good in a hypothetical context. The latter implies a departure from the continuity assumption of conventional consumer theory, which forms the basis for the method and the analysis. In the three essays of the thesis, discrete choice data are analyzed with the multinomial logit and mixed logit models. On average, Finns are willing to contribute to the water quality improvement. The probability for willingness increases with residential or recreational contact with the gulf, higher than average income, younger than average age, and the absence of dependent children in the household. On average, for Finns the relatively most important characteristic of water quality is water clarity followed by the desire for fewer occurrences of blue-green algae. For future nutrient reduction scenarios, the annual mean household willingness to pay estimates range from 271 to 448 and the aggregate welfare estimates for Finns range from 28 billion to 54 billion euros, depending on the model and the intensity of the reduction. Out of the respondents (N=726), 72.1% state in a follow-up question that they are either Certain or Quite certain about their answer when choosing the preferred alternative in the experiment. Based on the analysis of other follow-up questions and another sample (N=307), 10.4% of the respondents are identified as potentially having discontinuous preferences. In relation to both anomalies, the respondent- and questionnaire-specific variables are found among the underlying causes and a departure from standard analysis may improve the model fit and the efficiency of estimates, depending on the chosen modeling approach. The introduction of uncertainty about the future state of the Gulf increases the acceptance of the valuation scenario which may indicate an increased credibility of a proposed scenario. In conclusion, modeling preference heterogeneity is an essential part of the analysis of discrete choice data. The results regarding uncertainty in stating one s preferences and non-standard choice behavior are promising: accounting for these anomalies in the analysis may improve the precision of the estimates of benefit from reduced eutrophication in the Gulf of Finland.

Efficient search for statistically significant dependency rules in binary data

Analyzing statistical dependencies is a fundamental problem in all empirical science. Dependencies help us understand causes and effects, create new scientific theories, and invent cures to problems. Nowadays, large amounts of data is available, but efficient computational tools for analyzing the data are missing. In this research, we develop efficient algorithms for a commonly occurring search problem - searching for the statistically most significant dependency rules in binary data. We consider dependency rules of the form X->A or X->not A, where X is a set of positive-valued attributes and A is a single attribute. Such rules describe which factors either increase or decrease the probability of the consequent A. A classical example are genetic and environmental factors, which can either cause or prevent a disease. The emphasis in this research is that the discovered dependencies should be genuine - i.e. they should also hold in future data. This is an important distinction from the traditional association rules, which - in spite of their name and a similar appearance to dependency rules - do not necessarily represent statistical dependencies at all or represent only spurious connections, which occur by chance. Therefore, the principal objective is to search for the rules with statistical significance measures. Another important objective is to search for only non-redundant rules, which express the real causes of dependence, without any occasional extra factors. The extra factors do not add any new information on the dependence, but can only blur it and make it less accurate in future data. The problem is computationally very demanding, because the number of all possible rules increases exponentially with the number of attributes. In addition, neither the statistical dependency nor the statistical significance are monotonic properties, which means that the traditional pruning techniques do not work. As a solution, we first derive the mathematical basis for pruning the search space with any well-behaving statistical significance measures. The mathematical theory is complemented by a new algorithmic invention, which enables an efficient search without any heuristic restrictions. The resulting algorithm can be used to search for both positive and negative dependencies with any commonly used statistical measures, like Fisher's exact test, the chi-squared measure, mutual information, and z scores. According to our experiments, the algorithm is well-scalable, especially with Fisher's exact test. It can easily handle even the densest data sets with 10000-20000 attributes. Still, the results are globally optimal, which is a remarkable improvement over the existing solutions. In practice, this means that the user does not have to worry whether the dependencies hold in future data or if the data still contains better, but undiscovered dependencies.

The high- and low-activity forms of human plasma phospholipid transfer protein (PLTP)

Plasma phospholipid transfer protein (PLTP) plays a crucial role in high-density lipoprotein (HDL) metabolism and reverse cholesterol transport (RCT). It mediates the generation of pre-beta-HDL particles, enhances the cholesterol efflux from peripheral cells to pre-beta-HDL, and metabolically maintains the plasma HDL levels by facilitating the transfer of post-lipolytic surface remnants of triglyceride-rich lipoproteins to HDL. In addition to the antiatherogenic properties, recent findings indicate that PLTP has also proatherogenic characteristics, and that these opposite characteristics of PLTP are dependent on the site of PLTP expression and action. In human plasma, PLTP exists in a high-activity (HA-PLTP) and a low-activity form (LA-PLTP), which are associated with macromolecular complexes of different size and composition. The aims of this thesis were to isolate the two PLTP forms from human plasma, to characterize the molecular complexes in which the HA- and LA-PLTP reside, and to study the interactions of the PLTP forms with apolipoproteins (apo) and the ability of apolipoproteins to regulate PLTP activity. In addition, we aimed to study the distribution of the two PLTP forms in a Finnish population sample as well as to find possible regulatory factors for PLTP by investigating the influence of lipid and glucose metabolism on the balance between the HA- and LA-PLTP. For these purposes, an enzyme-linked immunosorbent assay (ELISA) capable of determining the serum total PLTP concentration and quantitating the two PLTP forms separately was developed. In this thesis, it was demonstrated that the HA-PLTP isolated from human plasma copurified with apoE, whereas the LA-PLTP formed a complex with apoA-I. The separation of these two PLTP forms was carried out by a dextran sulfate (DxSO4)-CaCl2 precipitation of plasma samples before the mass determination. A similar immunoreactivity of the two PLTP forms in the ELISA could be reached after a partial sample denaturation by SDS. Among normolipidemic Finnish individuals, the mean PLTP mass was 6.6 +/- 1.5 mg/l and the mean PLTP activity 6.6 +/- 1.7 umol/ml/h. Of the serum PLTP concentration, almost 50% represented HA-PLTP. The results indicate that plasma HDL levels could regulate PLTP concentration, while PLTP activity could be regulated by plasma triglyceride-rich very low-density lipoprotein (VLDL) concentration. Furthermore, new evidence is presented that PLTP could also play a role in glucose metabolism. Finally, both PLTP forms were found to interact with apoA-I, apoA-IV, and apoE. In addition, both apoE and apoA-IV, but not apoA-I, were capable of activating the LA-PLTP. These findings suggest that the distribution of the HA- and LA-PLTP in human plasma is subject to dynamic regulation by apolipoproteins.

Development of Novel Assays for Measuring Different Molecular Forms of Prostate Specific Antigen

Prostate cancer (PCa) is the most commonly diagnosed non-skin cancer and second leading cause of cancer-related death of men in developed countries. Measurement of prostate specific antigen (PSA) is a very sensitive method for diagnosing and monitoring of prostate cancer (PCa), but the specificity needs improvement. Measurements of different molecular forms of PSA have been shown to improve differentiation between PCa and benign prostatic diseases. However, accurate measurement of some isoforms has not been achieved in previous assays. The aim of the present study was to develop new assays that reliably measure enzymatically active PSA, PSA-α1-chymotryposin (PSA-ACT) and PSA-α1-protease inhibitor (PSA-API), and to evaluate their diagnostic value. Double-label immunofluorometric assays using a novel monoclonal antibody (MAb) and another antibody to either free PSA (fPSA) or total PSA (tPSA) were developed and used to measure PSA-ACT and fPSA or tPSA at the same time. These assays provide enough sensitivity for measurement of PSA-ACT in sera with low PSA levels. The results obtained confirmed that proportion of PSA-ACT to tPSA (%PSA-ACT) was as useful as proportion of fPSA to tPSA (%fPSA) for discrimination between PCa and benign prostatic hyperplasia (BPH). We developed an immunoassay for detection of PSA-API based on proximity ligation, which improved assay sensitivity 10-fold compared with conventional assays. Our results confirmed previous findings that the PSA-API level is somewhat lower in men with than without PCa, and the combination of %fPSA and proportion of PSA-API to tPSA (%PSA-API) provides diagnostic improvement compared with either method alone. Assays based on this principle should be applicable to other immunoassays in which the nonspecific background is a problem. An immunopeptidometric sandwich assay (IPMA) was developed to measure the enzymatically active PSA. This assay showed high specificity, but sensitivity was not good enough for measurement of PSA concentrations in the gray zone, 2-10 µg/L, in which tPSA does not efficiently differentiate between PCa and BPH. We further developed a solid-phase proximity ligation immunoassay, which provided a 10-fold improvement in sensitivity. This proof of concept study shows that peptides reacting with proteins are potentially useful for sensitive and specific measurement of protein variants for which specific MAbs cannot be obtained.