The mechanism of HAMLET-induced cell death - cellular signalling, oncogenes and clinical perspectives

Despite recent advances in cancer treatment, truly innovative approaches are required to move beyond the modest benefits achieved to date. HAMLET is a human protein-lipid complex originally discovered in breast milk able to kill a wide range of tumour cells while leaving healthy, differentiated cells unaffected. The aim of this thesis was to identify mechanisms dictating HAMLET sensitivity and to define the events that lead to cell death in response to HAMLET. In Paper I we show that the properties sensitising cells to HAMLET coincide with the ‘’Hallmarks of cancer’’. Using a combination of shRNA screens, proteomic and metabolomic technology, we identified the c-Myc oncogene and hexokinase as essential determinants of HAMLET sensitivity. HAMLET sensitivity was modified by the glycolytic state of tumor cells and HAMLET induced a metabolic paralysis in tumour cells. In paper II the initiating events for tumour cell death are elucidated. By rapid activation of a single ion channel HAMLET disturbs the flux of cations in tumour cells and subsequent cellular responses are shown to rely on this mechanism for their initiation. Ion fluxes activated p38 MAPK signalling, which was found to be crucially involved in tumour cell death. Paper III identifies oleate as the functional ligand in HAMLET formation and shows that oleate actively contributes to the induction of the cellular response to HAMLET. Both protein and lipid are needed for HAMLET’s tumoricidal effect, however. Finally, in paper IV we identify HAMLET as a new therapeutic agent in colon cancer. HAMLET caused a significant reduction in tumour numbers as well as mortality in mice carrying a human APC mutation. In tumours surviving HAMLET challenge a reduction in onco-protein expression was detected as well as an increase in expression of glycolic enzymes. By long-term prophylaxis, prevention of tumour development was achieved

Examen philosophicum – between the idea of universitas and the schoolification of the university

Examen philosophicum hviler på en idé om at alle som forsker, underviser eller studerer ved universitetet inngår i et fellesskap. I artikkelen blir dette fellesskapet omtalt som ideen om universitas og består av to elementer: Det ene er en spesifikk kunnskapsform der man søker helhetlig kunnskap for sin egen del, mens det andre er en spesifikk symmetrisk og jevnbyrdig omgangsform som blir nødvendig nettopp for å søke denne kunnskapen. Denne ideen om universitas er imidlertid truet av vår tids massifisering, byråkratisering og instrumentalisme, med den konsekvens at universitetet minner mer og mer om skolen. Implementeringen av Kvalitetsreformen i 2003 kan sies å være et tydelig uttrykk for denne skolegjøringen. Ved Universitetet i Bergen var imidlertid undervisningen ved examen philosophicum preget av skolegjøring før Kvalitetsreformen ble innført. Ved å skissere opp et filosofisk og historisk bakteppe for ideen om universitas og å sette skolegjøringen av universitetet i et bredere perspektiv, ønsker artikkelen å sette fokus på en noe paradoksal rolle examen philosophicum har fått: Examen philosophicum er både en siste skanse for ideen om universitas og en pådriver for tendenser som vil underminere denne ideen på samme tid.publishedVersio

Du kan, du skal – skolevegring, digitalisering og selvets tretthet i den elevsentrerte skolen

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Metabolite Profiling of LADA Challenges the View of a Metabolically Distinct Subtype

Latent autoimmune diabetes in adults (LADA) usually refers to GAD65 autoantibodies (GADAb)-positive diabetes with onset after 35 years of age and no insulin treatment within the first 6 months after diagnosis. However, it is not always easy to distinguish LADA fromtype 1 or type 2 diabetes. In this study, we examined whether metabolite profiling could help to distinguish LADA (n = 50) from type 1 diabetes (n = 50) and type 2 diabetes (n = 50). Of 123 identified metabolites, 99 differed between the diabetes types. However, no unique metabolite profile could be identified for any of the types. Instead, the metabolome varied along a C-peptide-driven continuum from type 1 diabetes via LADA to type 2 diabetes. LADA was more similar to type 2 diabetes than to type 1 diabetes. In a principal component analysis, LADA patients overlapping with type 1 diabetes progressed faster to insulin therapy than those overlapping with type 2 diabetes. In conclusion, we could not find any unique metabolite profile distinguishing LADA from type 1 and type 2 diabetes. Rather, LADA was metabolically an intermediate of type 1 and type 2 diabetes, with those patients closer to the former showing a faster progression to insulin therapy than those closer to the latter.Peer reviewe

Single-Cell Profiling of Coding and Noncoding Genes in Human Dopamine Neuron Differentiation

Dopaminergic (DA) neurons derived from human pluripotent stem cells (hPSCs) represent a renewable and available source of cells useful for understanding development, developing disease models, and stem-cell therapies for Parkinson's disease (PD). To assess the utility of stem cell cultures as an in vitro model system of human DA neurogenesis, we performed high-throughput transcriptional profiling of ~20,000 ventral midbrain (VM)-patterned stem cells at different stages of maturation using droplet-based single-cell RNA sequencing (scRNAseq). Using this dataset, we defined the cellular composition of human VM cultures at different timepoints and found high purity DA progenitor formation at an early stage of differentiation. DA neurons sharing similar molecular identities to those found in authentic DA neurons derived from human fetal VM were the major cell type after two months in culture. We also developed a bioinformatic pipeline that provided a comprehensive long noncoding RNA landscape based on temporal and cell-type specificity, which may contribute to unraveling the intricate regulatory network of coding and noncoding genes in DA neuron differentiation. Our findings serve as a valuable resource to elucidate the molecular steps of development, maturation, and function of human DA neurons, and to identify novel candidate coding and noncoding genes driving specification of progenitors into functionally mature DA neurons

User involvement in adolescents’ mental healthcare: a systematic review

More than one out of ten adolescents suffer from mental illness at any given time. Still, there is limited knowledge about their involvement in mental healthcare. Adolescents have the right to be involved in decisions affecting their healthcare, but limited research focuses on their engagement and decision-making. Therefore, this systematic review aims to explore the existing experiences with, the effectiveness of, and safety issues associated with user involvement for adolescents’ mental healthcare at the individual and organizational level. A systematic literature review on user involvement in adolescents’ mental healthcare was carried out. A protocol pre-determined the eligibility criteria and search strategies, and established guidelines were used for data extraction, critical appraisal, and reporting of results. Quantitative studies were analysed individually due to heterogeneity of the studies, while qualitative studies were analysed using thematic synthesis. A total of 31 studies were included in the review. The experiences with user involvement were reported in 24 studies with three themes at the individual level: unilateral clinician control versus collaborative relationship, capacity and support for active involvement, the right to be involved; and two themes at the organizational level: involvement outcomes relevant to adolescents’ needs, conditions for optimal involvement. The effectiveness of user involvement was reported in seven studies documenting fragmented evidence related to different support structures to facilitate adolescents’ involvement. The safety associated with user involvement was not reported in any studies, yet a few examples related to potential risks associated with involvement of adolescents in decision-making and as consultants were mentioned.publishedVersio

Teacher educators reflecting on case-based teaching – a collective self-study

The current study is a collective self-study on how we as 15 teacher educators at a university in Norway tried to improve our teaching through working with cases with the aim of better supporting student teachers in making links between theory and practice. We wanted to address the common criticism in teacher education concerning a perceived gap between practice and theory. Our presupposition was that one way to prepare student teachers for work and bring together theoretical and practical knowledge would be through case-based teaching. We agreed that we wanted to try different ways of working with cases and to follow our own actions with research and conduct a self-study. Throughout the project, each teacher educator experienced to learn about case-based teaching, but our joint learning was limited due to practical issues and lack of time. With teacher education as a shared responsibility, our conclusion is that teacher educators need time to develop as a team, not only as individual teachers.publishedVersio

Bacterial control of host gene expression through RNA polymerase II

The normal flora furnishes the host with ecological barriers that prevent pathogen attack while maintaining tissue homeostasis. Urinary tract infections (UTIs) constitute a highly relevant model of microbial adaptation in which some patients infected with Escherichia coil develop acute pyelonephritis, while other patients with bacteriuria exhibit an asymptomatic carrier state similar to bacterial commensalism. It remains unclear if the lack of destructive inflammation merely reflects low virulence or if carrier strains actively inhibit disease-associated responses in the host. Here, we identify a new mechanism of bacterial adaptation through broad suppression of RNA polymerase II-dependent (Pol II-dependent) host gene expression. Over 60% of all genes were suppressed 24 hours after human inoculation with the prototype asymptomatic bacteriuria (ABU) strain E. coil 83972, and inhibition was verified by infection of human cells. Specific repressors and activators of Pol II-dependent transcription were modified, Pol II phosphorylation was inhibited, and pathogen-specific signaling was suppressed in cell lines and inoculated patients. An increased frequency of strains inhibiting Pol II was epidemiologically verified in ABU and fecal strains compared with acute pyelonephritis, and a Pol II antagonist suppressed the disease-associated host response. These results suggest that by manipulating host gene expression, ABU strains promote tissue integrity while inhibiting pathology. Such bacterial modulation of host gene expression may be essential to sustain asymptomatic bacterial carriage by ensuring that potentially destructive immune activation will not occur

Novel subgroups of adult-onset diabetes and their association with outcomes : a data-driven cluster analysis of six variables

Background Diabetes is presently classified into two main forms, type 1 and type 2 diabetes, but type 2 diabetes in particular is highly heterogeneous. A refined classification could provide a powerful tool to individualise treatment regimens and identify individuals with increased risk of complications at diagnosis. Methods We did data-driven cluster analysis (k-means and hierarchical clustering) in patients with newly diagnosed diabetes (n=8980) from the Swedish All New Diabetics in Scania cohort. Clusters were based on six variables (glutamate decarboxylase antibodies, age at diagnosis, BMI, HbA(1c), and homoeostatic model assessment 2 estimates of beta-cell function and insulin resistance), and were related to prospective data from patient records on development of complications and prescription of medication. Replication was done in three independent cohorts: the Scania Diabetes Registry (n=1466), All New Diabetics in Uppsala (n=844), and Diabetes Registry Vaasa (n=3485). Cox regression and logistic regression were used to compare time to medication, time to reaching the treatment goal, and risk of diabetic complications and genetic associations. Findings We identified five replicable clusters of patients with diabetes, which had significantly different patient characteristics and risk of diabetic complications. In particular, individuals in cluster 3 (most resistant to insulin) had significantly higher risk of diabetic kidney disease than individuals in clusters 4 and 5, but had been prescribed similar diabetes treatment. Cluster 2 (insulin deficient) had the highest risk of retinopathy. In support of the clustering, genetic associations in the clusters differed from those seen in traditional type 2 diabetes. Interpretation We stratified patients into five subgroups with differing disease progression and risk of diabetic complications. This new substratification might eventually help to tailor and target early treatment to patients who would benefit most, thereby representing a first step towards precision medicine in diabetes.Peer reviewe