The antitumoural cyanoguanidine CHS 828 has shown promising activity in a number of preclinical and clinical studies. However, the mechanisms underlying the cell death induced by CHS 828 has not been clarified. This thesis describes in vitro studies of the cellular pharmacology of CHS 828. CHS 828 induced cell death with necrosis like features in the lymphoma cell line U-937 GTB. Addition of 3-aminobenzamide, an inhibitor of ADP-ribosylation, resulted in a decreased sensitivity to CHS 828 and a shift in the mode of cell death towards apoptosis. Mouse fibroblasts lacking the enzyme PARP-1 were more sensitive to CHS 828 compared to normal fibroblasts. CHS 828 was able to induce p53 in normal fibroblasts but this effect does not seem to be necessary to induce cell death. Characterization of two CHS 828 resistant cell lines indicated that they were selectively resistant to cyanoguanidines. Known mechanisms of anticancer drug resistance did not seem to account for the cyanoguanidine resistance. One possible resistance mediating protein, which was upregulated in the resistant cells, was epidermal fatty acid binding protein. A novel high content screening assay was also developed. The assay was shown to be suitable both for screening of potential novel antitumoural substances as well for mechanistic studies. In the assay, CHS 828 induced caspase-3 activity and reduction in mitochondrial membrane potential, both signs of apoptosis, in U-937 GTB cells. However, nuclei in exposed cells did not show nuclear fragmentation, one of the hallmarks of apoptosis. CHS 828 was also shown to indirectly inhibit the proteasome activity in U-937 GTB cells. In conclusion, the results presented provide new insights into the metabolic and molecular events involved in cell death induced by CHS 828.

The CNS monoaminergic systems are considered as the head engine regulating neuropsychiatric functions and personality. Transcription factor AP-2 is known to be essential for the development of the brainstem including the monoaminergic nuclei, and has the ability to regulate many genes in the monoaminergic systems. The ability of transcription factors to regulate specific gene expression, has lately made them hot candidates as drug targets. In this thesis, results indicating a role of AP-2 in the molecular effects of the antidepressant drugs citalopram and phenelzine, are presented. A polymorphism in the second intron of the gene encoding AP-2ß has previously been associated with anxiety-related personality traits as estimated by the Karolinska Scales of Personality (KSP). In this thesis, results confirming this association, gained by using a larger material and several different personality scales, are presented. Furthermore, data is presented showing an association between the activity of platelet monoamine oxidase, a trait-dependent marker for personality, and the genotype of the AP-2ß intron 2 polymorphism. The functional importance of the AP-2ß intron 2 polymorphism has not yet been elucidated. Included in this thesis are results showing that the AP-2ß intron 2 polymorphism is not in linkage disequilibrium with the only other described polymorphism in the AP-2ß gene, i.e. in the AP-2ß promoter (-67 G/A). Introns have in several studies been shown to include binding sites for regulatory proteins, and thus, to be important in transcriptional regulation. Results are presented demonstrating that one human brain nuclear protein binds only to the long variant of the AP-2ß intron 2 polymorphism. If this protein is involved in the regulation of the AP-2ß gene, it would affect the expression levels of the AP-2ß protein. In general, this thesis further establishes the role of transcription factor AP-2 as a regulatory factor of importance for personality and monoaminergic functions.

Semicarbazide-sensitive amine oxidase (SSAO, EC 1.4.3.6) belongs to a family of copper-containing amine oxidases. SSAO exists as a membrane bound protein in endothelial-, smooth muscle-, and adipose cells as well as soluble in plasma. SSAO catalyses oxidative deamination of primary monoamines, which results in the production of corresponding aldehydes, hydrogen peroxide and ammonia. These compounds are very reactive and potentially cytotoxic, and are able to induce vascular damage if produced in high levels. Patients with diabetes mellitus, and with diabetic complications in particular, have a higher SSAO activity in plasma compared to healthy controls. It has therefore been speculated that high SSAO activity is involved in the development of vascular complications associated with diabetes. The aim of this thesis is to investigate the importance of SSAO in the development of disorders of a vascular origin. We have studied the transcriptional regulation of the SSAO gene, by inducing diabetes in NMRI and in transgenic mice, overexpressing the human form of SSAO in smooth muscle cells. We found that the increase in SSAO activity in diabetes is accompanied by reduced mRNA levels of the endogenous mouse gene, suggesting a negative feedback on the transcription of the SSAO gene. In addition, the transgenic mice exhibited an abnormal phenotype in the elastic tissue of aorta and renal artery. These mice have a lower mean artery pressure and an elevated pulse pressure. These results indicate that high SSAO activity in smooth muscle cells is associated with a change in the morphology of large arteries. This is likely contributing to the development of vascular complications in diabetes.

The novel cyanoguanidine CHS 828 has shown promising antitumor activity in many in vitro and in vivo studies. The long-term 14 days in vitro hollow fiber cultures, where tumor cells from different tumor cell lines were cultured inside semipermeable fibers, were more resistant to CHS 828 and other cytotoxic drugs than the shorter-term 3 days cultures. CHS 828 was generally more effective against haematological than solid tumor cells from both cell lines and patients samples. In vivo, the hollow fibers were implanted into immunocompetent rats and the pharmacokinetics, tumor response and/or toxicity (pharmacodynamics) of CHS 828 were successfully assayed. CHS 828 showed higher activity in this model when a more protracted schedule was used. The quantitative relationships between dose, plasma concentration and response (PK/PD model) developed for CHS 828 explained this phenomenon partly by dose-dependent fraction absorbed and partly by a schedule-dependent pharmacodynamic effect. Modelling of the in vitro CHS 828 and standard cytotoxic drugs concentration-time effect data in different tumor cell types and characterization of pattern of change of the potency and the slope of the concentration-time effect curves were performed. The results suggest two different mechanisms of action for CHS 828 and that CHS 828 cytotoxicity may depend on the schedule used. The NF-kB pathway that regulates the transcription of anti-apoptotic genes proved to be inhibited by CHS 828 in different tumor cell lines and the inhibition was correlated to the cell death induced by this agent. CHS 828 did not seem to induce the NF-kB inhibition by affecting the proteasome activity. The in vitro and in vivo hollow fiber methods were also used successfully to evaluate the new paclitaxel formulation, Pacliex. Pacliex had a similar activity to that of the clinically used formulation Taxol®.

Protein folding and export to the membrane are crucial events in the cell. Both processes may be initiated already at the ribosome, assisted by factors that bind to the polypeptide as it emerges from the ribosome. The signal recognition particle (SRP) scans the ribosome for nascent peptides destined for membrane insertion and targets these ribosomes to the site for translocation in the membrane. Trigger factor (TF) is a folding chaperone that interacts with nascent chains to promote their correct folding, prevent misfolding and aggregation. In this thesis, we first investigated membrane targeting and insertion of two heterologous membrane proteins in E. coli by using in vitro translation, membrane targeting and cross-linking. We found that these proteins are dependent on SRP for targeting and that they initially interact with translocon components in the same way as native nascent membrane proteins. Moreover we have characterised the SRP and TF interactions with the ribosome both with cross-linking experiments and with quantitative binding experiments. Both SRP and TF bind to ribosomal L23 close to the nascent peptide exit site where they are strategically placed for binding to the nascent polypeptide. Quantitative analysis of TF and SRP binding determined their respective KD values for binding to non translating ribosomes and reveals that they bind simultaneously to the ribosome, thus having separate binding sites on L23. Finally, binding studies on ribosome nascent chain adds clues as to how TF functions as a chaperone.

Spontaneous reporting systems (SRSs) for adverse drug reactions (ADRs) have been developed as a result of the thalidomide disaster, whereby thousands of children world-wide were born with birth defects. The Swedish Adverse Drug Reactions Advisory Committee was established in 1965. Since 1975, reporting has been compulsory for all suspected serious or new ADRs. International collaboration started in 1968 with countries contributing their ADR reports to an international database set up by the World Health Organization. ADRs represent the negative side of the benefit-to-risk balance that in theory needs to be counteracted by perceived or established positive drug effects. All drugs are subject to preclinical and clinical testing prior to marketing authorization. However, these studies are insufficient to detect rare ADRs, ADRs that occur after long-term administration or with latency, ADRs that occur in special patient groups such as children, the elderly, patients with renal or hepatic insufficiency or patients on concomitant drug treatment, and ADRs that represent a modest increase in the risk of diseases (including mortality) that are prevalent in the study population. Postmarketing surveillance of drugs is therefore essential, and regulatory action may be needed on the basis of new ADR information. SRSs are important sources of ADR information as exemplified here by the evaluation of peripheral sensory disturbances with fluoroquinolones, hyponatremia with antidepressants, blood dyscrasias with dipyrone, glucose intolerance with atypical antipsychotics, pulmonary embolism with combined oral contraceptives and extrapyramidal symptoms with selective serotonin reuptake inhibitors. SRSs can be used to study clinical manifestations of ADRs (that can give insights into potential ADR mechanisms), risk factors for the ADR or for specific outcomes of the ADR, and ADR reporting incidences when combined with sales data. Signals from SRSs may need to be studied further e.g., by use of large-scale epidemiologic studies based on record linkage between drug prescription databases and health databases. Owing to the rapid availability of information, however, SRSs are likely to remain of major importance for the post-marketing surveillance of drugs.

Solid organ transplantation is today an established form of treatment for end-stage organ disease. Monitoring of graft function and pharmacological therapy constitutes a maze of clinical observations and histological evaluations of biopsy specimens; with the biopsy results playing a decisive role. The aims of this doctoral research were to investigate the feasibility of detecting acute rejection of transplanted organs and monitoring the effect of anti-rejection treatment, with the use of ultrasmall superparamagnetic iron oxide particles (USPIO) and magnetic resonance (MR) imaging with a clinical MR scanner. Allogeneic and syngeneic heterotopic heart transplantations were performed in rats. Three different-sized USPIO were given to one allogeneic and one syngeneic group. The change in MR signal intensity (SI) over time was measured. An increase in SI was interpreted as damage to micro vessels due to the pronounced inflammatory reaction caused by acute rejection, which led to leakage of USPIO into the tissue. A decrease in SI was interpreted as normal vascular structure, since USPIO normally remains in the intravascular space. The same method, using one of the previously tested USPIO, was used in a treatment study in which acute rejection in transplanted rats was induced and subsequently treated. An attempt was also made to detect presence of macrophages in an acutely rejecting graft, since this cell type plays an important role in the acute rejection process; this was done by testing the ability of macrophages to phagocytose the UPSIO compound. In permeability studies with MR imaging all three USPIO tested discriminated between rejecting and non-rejecting grafts without any overlap of the groups. Factors that contributed to the ability to distinguish between grafts were the size and half-life of the particle. We were also able to monitor effects of anti-rejection treatment by studying the vascular permeability of USPIO and MR imaging. We did not succeed in detecting macrophages in the rejecting grafts with USPIO and MR imaging. This thesis presents a novel approach to detection of acute rejection of transplanted organs and to monitoring the effects of anti-rejection treatment using different USPIO contrast agents and MR imaging in a clinical MR scanner.

I inledningen ställs frågan om vilka beröringspunkter som finns mellan skådespelar- och undervisningskonst. Är beröringspunkterna desamma oavsett lärar- och undervisningsstil?Utifrån tidigare forskning belyses falska motsättningar likaväl som nyanserade och fördjupade kopplingar mellan skådespelar- och dramapedagogroll. Användning av olika teatrala och pedagogiska konventioner problematiseras och fördjupas teoretiskt. För att vidga synen på dramapedagogisk praktik fördjupas teoretiska perspektiv på relationen mellan konventioner, kompetenser och inre förhållningssätt.Studien syftar till att undersöka hur man kan analysera komplexiteten i den dramapedagogiska praktiken med estetiskt baserade referensramar ur ett lärarperspektiv. Frågor som ställs är: Med vilka kompetenser och förmågor hanterar och använder dramapedagogen komplexiteten i den interaktiva, teatralt gestaltande och meningsskapande formen av undervisning? Kan detta analyseras och synliggöras med hjälp av teatersemiotisk analys och de teoretiska perspektiven?Undersökningen är kvalitativ och har en hermeneutisk ansats, den vill också pröva teatersemiotik som en estetiskt baserad analysmetod. Datainsamlingen sker genom deltagande observation och ett löpande protokoll. Det som observerats är undervisningslektioner i processdrama för dramapedagogikstudenter. Två sekvenser, en med en instruerande lärarroll och en med "lärare- i-roll", analyseras och jämförs. Resultatet visar exempel på att teatersemiotisk analys kan utifrån sina styrkor och begränsningar vara en hjälpmetod för att synliggöra de konventioner, förmågor och kompetenser som både använder och hanterar komplexitet i den dramapedagogiska praktiken. De teatrala formerna används också i sig som pedagogiska arenor. Skådespelarteori tydliggör att det finns ett avgörande samband mellan kvalitén i det inre förhållningssättet, kompetensutveckling och användningen av yttre konventioner. Det är rimligt att konstatera att det finns anledning att belysa fler möjliga paralleller mellan skådespelar- och undervisningskonst, för att skapa en djupare förståelse för dramapedagogikens praktik.

För att hjälpa barn och ungdomar som lider av ätstörningar krävs specialistkompetens inom området. Specialistsjukvården måste kunna möta såväl nyinsjuknade som långvarigt sjuka barn och ungdomar. Syftet med denna studie var att belysa sjuksköterskors upplevelser av att vårda barn och ungdomar med ätstörningar. En intervjustudie har genomförts med sju sjuksköterskor som arbetar på två olika specialistenheter där barn och ungdomar vårdas för ätstörningar. Intervjuerna har analyserats med kvalitativ innehållsanalys. Utifrån analysen skapades tre kategorier: ”att finnas till för patient och anhöriga”, ”sjuksköterskans behov av stöd” och ”utmaningar i yrket” samt sex subkategorier: ”att möta och stödja patienten på ett professionellt sätt”, ”sjuksköterskans relation till anhöriga”, ”balans mellan arbete och fritid”, ”kollegial anda och ledarskap”, ”sjuksköterskans upplevelse av svårigheter” och ”känslor”. Huvudfynden visade att sjusköterskor som arbetar på specialistenheter för ätstörningar där barn och ungdomar vårdas kände stor meningsfullhet med arbetet.  Med professionalism och lugn kände sjuksköterskorna att de kunde hjälpa sina patienter trots att arbetet stundom kunde upplevas både utmanande och svårt. En balans mellan arbete och fritid, god kollegial anda, ett gott samarbete med anhöriga, ett gott ledarskap samt en tro på sig själv och sin förmåga som sjuksköterska upplevdes som väsentligt för god omvårdnad av dessa barn och ungdomar. / To help children and adolescents who suffer from eating disorders specialist skills are needed. Specialist Health Services need not only respond to children and adolescents with recent onset but also those who have been long-term ill. The purpose of this study was to illustrate nurses’ experiences of caring for children and adolescents with eating disorders. An interview study was carried out with seven nurses working at two specialist units treating children and adolescents with eating disorders. The interviews were analyzed with qualitative content analysis. Based on the analysis three categories were created: “to be there for the patient and their relatives”, “the nurse’s need of support” and “challenges of the profession”. Six sub-categories were created out of these three main categories: “to meet and support patients in a professional manner”, “the nurse’s relationship with the patient’s relatives”, “balance between work and leisure”, “collegial spirit and leadership”, “the nurse’s experience of difficulties” and “feelings”. Major findings showed that nurses working in specialized units for eating disorders felt great meaningfulness with their work. With professionalism and tranquility nurses felt they could help their patients even though the work sometimes was both challenging and difficult. A balance between work and leisure, good collegial spirit, a good way to interact with the patients relatives, good leadership and a belief in herself and her nursing ability were perceived essential for good care of children and adolescents suffering from eating disorders.

The aim of this thesis is to analyze the academic discipline translation studies; the analysis centers upon bibliometric aspects, and is carried out chiefly by means of quantitative methods, enabled by the databases Web of Science and Scopus as well as by the corpus-linguistic software Wordsmith; however, this quantitative focus is complemented by auxiliary qualitative means of investigation, i.e. a discourse-analytically oriented study of a corpus of domain-endemic reviews. A number of core journals are analyzed in order to show what and whom are cited in translation studies, and, furthermore, what adjacent domains hold sway over it; attention is also given to influential authors, regions and languages operating within the domain. It is shown that the domain under scrutiny is quite differentiated, even fragmentary; the term pluricentric is used to describe this tendency towards sprawling. No superior authors (in terms of publication frequency) emerge, which arouses the suspicion that the domain may be conditioned by a hierarchical divide between journals and monographs. Furthermore, it is clear that translation studies is a eurocentric domain: European languages dominate. This is, however, a contested state-of-affair; debates rage within the domain, evidencing a vibrant yet also discordant characteristic. The thesis also contributes to a meta-theoretical advancement within bibliometrics; it does so by showing the benefits and limitations of using quantitative tools to examine a domain located squarely within the humanities; the main contribution, however, lies in suggesting that qualitative methods are relevant to bibliometric endeavors, and in illustrating how such methods can be brought to bear on a somewhat overlooked genre, namely the scholarly review. Also worthy of note within this thesis is the cultural-studiesque critique of reified analytical categories within bibliometrics; such humanities-fueled critique is to be encouraged, and will hopefully flourish in future scholarly projects. This is a two years master’s thesis in the field of library and information science.