Regulation of B Cell Gene Expression and Function by Ikaros, Helios and Bcl6

B lymphocytes constitute a key branch of adaptive immunity by providing specificity to recognize a vast variety of antigens by B cell antigen receptors (BCR) and secreted antibodies. Antigen recognition activates the cells and can produce antibody secreting plasma cells via germinal center reaction that leads to the maturation of antigen recognition affinity and switching of antibody effector class. The specificity of antigen recognition is achieved through a multistep developmental pathway that is organized by interplay of transcription factors and signals through BCR. Lymphoid malignancies arise from different stages of development in abnormal function of transcriptional regulation. To understand the B cell development and the function of B cells, a thorough understanding of the regulation of gene expression is important. The transcription factors of the Ikaros family and Bcl6 are frequently associated with lymphoma generation. The aim of this study was to reveal the targets of Ikaros, Helios and Bcl6 mediated gene regulation and to find out the function of Ikaros and Helios in B cells. This study uses gene targeted DT40 B cell lines and establishes a role for Ikaros family factors Ikaros and Helios in the regulation of BCR signaling that is important at developmental checkpoints, for cell survival and in activation. Ikaros and Helios had opposing roles in the regulation of BCR signals. Ikaros was found to directly repress the SHIP gene that encodes a signaling lipid-metabolizing enzyme, whereas Helios had activating effect on SHIP expression. The findings demonstrate a balancing function for these two Ikaros family transcription factors in the regulation of BCR signaling as well as in the regulation of gene expression. Bcl6 was found to repress plasma cell gene expression program while maintaining gene expression profile of B cells. Analysis of direct Bcl6 target genes suggested novel mechanisms for Bcl6-mediated suppression of plasma cell differentiation and promoting germinal center phenotype.

Towards increased productivity and quality in software development using agile, lean and collaborative approaches

Dagens programvaruindustri står inför alltmer komplicerade utmaningar i en värld där programvara är nästan allstädes närvarande i våra dagliga liv. Konsumenten vill ha produkter som är pålitliga, innovativa och rika i funktionalitet, men samtidigt också förmånliga. Utmaningen för oss inom IT-industrin är att skapa mer komplexa, innovativa lösningar till en lägre kostnad. Detta är en av orsakerna till att processförbättring som forskningsområde inte har minskat i betydelse. IT-proffs ställer sig frågan: “Hur håller vi våra löften till våra kunder, samtidigt som vi minimerar vår risk och ökar vår kvalitet och produktivitet?” Inom processförbättringsområdet finns det olika tillvägagångssätt. Traditionella processförbättringsmetoder för programvara som CMMI och SPICE fokuserar på kvalitets- och riskaspekten hos förbättringsprocessen. Mer lättviktiga metoder som t.ex. lättrörliga metoder (agile methods) och Lean-metoder fokuserar på att hålla löften och förbättra produktiviteten genom att minimera slöseri inom utvecklingsprocessen. Forskningen som presenteras i denna avhandling utfördes med ett specifikt mål framför ögonen: att förbättra kostnadseffektiviteten i arbetsmetoderna utan att kompromissa med kvaliteten. Den utmaningen attackerades från tre olika vinklar. För det första förbättras arbetsmetoderna genom att man introducerar lättrörliga metoder. För det andra bibehålls kvaliteten genom att man använder mätmetoder på produktnivå. För det tredje förbättras kunskapsspridningen inom stora företag genom metoder som sätter samarbete i centrum. Rörelsen bakom lättrörliga arbetsmetoder växte fram under 90-talet som en reaktion på de orealistiska krav som den tidigare förhärskande vattenfallsmetoden ställde på IT-branschen. Programutveckling är en kreativ process och skiljer sig från annan industri i det att den största delen av det dagliga arbetet går ut på att skapa något nytt som inte har funnits tidigare. Varje programutvecklare måste vara expert på sitt område och använder en stor del av sin arbetsdag till att skapa lösningar på problem som hon aldrig tidigare har löst. Trots att detta har varit ett välkänt faktum redan i många decennier, styrs ändå många programvaruprojekt som om de vore produktionslinjer i fabriker. Ett av målen för rörelsen bakom lättrörliga metoder är att lyfta fram just denna diskrepans mellan programutvecklingens innersta natur och sättet på vilket programvaruprojekt styrs. Lättrörliga arbetsmetoder har visat sig fungera väl i de sammanhang de skapades för, dvs. små, samlokaliserade team som jobbar i nära samarbete med en engagerad kund. I andra sammanhang, och speciellt i stora, geografiskt utspridda företag, är det mera utmanande att införa lättrörliga metoder. Vi har nalkats utmaningen genom att införa lättrörliga metoder med hjälp av pilotprojekt. Detta har två klara fördelar. För det första kan man inkrementellt samla kunskap om metoderna och deras samverkan med sammanhanget i fråga. På så sätt kan man lättare utveckla och anpassa metoderna till de specifika krav som sammanhanget ställer. För det andra kan man lättare överbrygga motstånd mot förändring genom att introducera kulturella förändringar varsamt och genom att målgruppen får direkt förstahandskontakt med de nya metoderna. Relevanta mätmetoder för produkter kan hjälpa programvaruutvecklingsteam att förbättra sina arbetsmetoder. När det gäller team som jobbar med lättrörliga och Lean-metoder kan en bra uppsättning mätmetoder vara avgörande för beslutsfattandet när man prioriterar listan över uppgifter som ska göras. Vårt fokus har legat på att stöda lättrörliga och Lean-team med interna produktmätmetoder för beslutsstöd gällande så kallad omfaktorering, dvs. kontinuerlig kvalitetsförbättring av programmets kod och design. Det kan vara svårt att ta ett beslut att omfaktorera, speciellt för lättrörliga och Lean-team, eftersom de förväntas kunna rättfärdiga sina prioriteter i termer av affärsvärde. Vi föreslår ett sätt att mäta designkvaliteten hos system som har utvecklats med hjälp av det så kallade modelldrivna paradigmet. Vi konstruerar även ett sätt att integrera denna mätmetod i lättrörliga och Lean-arbetsmetoder. En viktig del av alla processförbättringsinitiativ är att sprida kunskap om den nya programvaruprocessen. Detta gäller oavsett hurdan process man försöker introducera – vare sig processen är plandriven eller lättrörlig. Vi föreslår att metoder som baserar sig på samarbete när processen skapas och vidareutvecklas är ett bra sätt att stöda kunskapsspridning på. Vi ger en översikt över författarverktyg för processer på marknaden med det förslaget i åtanke.

Regulation of cell growth, death, and polarization by ERBB4

Regulation of cell growth, death, and polarization by ERBB4 ErbB4 is a member of the epidermal growth factor receptor (EGFR, ErbB) family. The other members are EGFR, ErbB2 and ErbB3. ErbB receptors are important regulators for example in cardiovascular, neural and breast development but control key cellular functions also in many adult tissues. Abnormal ErbB signaling has been shown to be involved in various illnesses such as cancers and heart diseases. Among the ErbBs, ErbB4 has been shown to have unique signaling characteristics. ErbB4 exists in four alternatively spliced isoforms that are expressed in a tissue-specific manner. Two of the isoforms can be cleaved by membrane proteases, resulting in release of soluble intracellular domains (ICD). Once released into the cytosol, the ICD is capable of translocating into the nucleus and participating in regulation of transcription. The functional differences and the tissue-specific expression patterns suggest isoformspecific roles for ErbB4 isoforms. However, the abilities of ErbB4 isoforms to differently regulate cellular functions were discovered only recently and are not well understood. This study aimed to determine the expression patterns of ErbB4 in normal and diseased tissue, and to define whether the cleavable and non-cleavable isoforms could regulate different target genes and therefore, cellular functions. In this study, a comprehensive ErbB4 expression pattern in several normal tissues, various cancers and non-neoplastic diseases was determined. In addition, the data demonstrated that the cleavable and non-cleavable ErbB4 isoforms could regulate different cellular functions and target genes. Finally, this study defined the cellular responses regulated by ErbB4 during kidney development.

Effects of Country of Origin on Consumer Product Evaluations: Finnish clothing products in the minds of Russian tourists

Companies operating in today’s highly internationalized markets consider product differentiation the key priority in pursue to attain a constant competitive advantage in challenging global environment (Baker and Ballington 2002, 158). The main driver affecting companies’ differentiation actions was described as early as 1912 by one of the marketing pioneers A. W. Shaw (1912, 710) as meeting human wants more accurate than the competition, and thus increasing customers’ perceived value and satisfaction. Dickson and Ginter (1987, 2) point out in their study based on earlier research by Chamberlin (1965) and Porter (1976) that differentiation can be based on either tangible characteristics of a product such as design or intangible characteristics such as a brand name and country of origin (hereafter referred to as COO). The concept of COO and its impact on consumers’ evaluation of a product as an extrinsic product cue has been one of the most noteworthy topics in international marketing, having been voluminously examined by over 780 authors in more than 750 academic publications in the past 40 years (Papadopoulos and Heslop 2002, 294). Many of these studies accentuate the significant effect the COO has on consumers’ product attribute evaluations. People routinely associate country images with products and services in order to judge and categorize them based on perceived quality and risk levels; thereby COO can influence the likelihood of a purchase (Peterson and Jolibert 1995, 883-884; Verlegh and Steenkamp 1999, 523). Based on the vast research related to COO in the field of international business, it is widely recognized that the country associated with a product can act in a similar way as the name of a brand and even become a part of product’s total image. Thereby depending on customer’s values and perceptions, the product-country image can either increase or decrease perceived value.

DEVELOPING A CROWDSOURCED VIDEOGRAPHIC RESEARCH METHOD FOR CONSUMER CULTURE RESEARCH

The purpose of this study is to develop a crowdsourced videographic research method for consumer culture research. Videography provides opportunities for expressing contextual and culturally embedded relations. Thus, developing new ways to conduct videographic research is meaningful. This study develops the crowdsourced videographic method based on a literature review and evaluation of a focal study. The literature review follows a qualitative systematic review process. Through the literature review, based on different methodological, crowdsourcing and consumer research related literature, this study defines the method, its application process and evaluation criteria. Furthermore, the evaluation of the focal study, where the method was applied, completes the study. This study applies professional review with self-evaluation as a form of evaluation, drawing from secondary data including research task description, screenshots of the mobile application used in the focal study, videos collected from the participants, and self-evaluation by the author. The focal study is analyzed according to its suitability to consumer culture research, research process and quality. Definitions and descriptions of the research method, its process and quality criteria form the theoretical contribution of this study. Evaluating the focal study using these definitions underlines some best practices of this type of research, generating the practical contribution of this study. Finally, this study provides ideas for future research. First, defining the boundaries of the use of crowdsourcing in various parts of conducting research. Second, improving the method by applying it to new research contexts. Third, testing how changes in one dimension of the crowdsourcing models interact with other dimension. Fourth, comparing the quality criteria applied in this study to various other quality criteria to improve the method’s usefulness. Overall, this study represents a starting point for further development of the crowdsourced videographic research method.

Regulation of epidermal tight junctions by calcium ATPases and p38

The epidermis is the upper layer of the skin and keratinocytes are its most abundant cells. Tight junctions are cell junctions located in the granular layer of the epidermis. They maintain the polarity of the cells and regulate the movement of water-soluble molecules. Epidermal tight junctions may lose their integrity when there are defects in intercellular calcium regulation. Hailey-Hailey and Darier´s disease are dominantly inherited, blistering skin diseases. Hailey-Hailey disease is caused by mutations in the ATP2C1 gene encoding a calcium/manganese ATPase SPCA1 of the Golgi apparatus. Darier´s disease is caused by mutations in the ATP2A2 gene encoding a calcium ATPase SERCA2 of the endoplasmic reticulum. p38 regulates the differentiation of keratinocytes. The overall regulation of epidermal tight junctions is not well understood. The present study examined the regulation of tight junctions in the human epidermis with a focus on calcium ATPases and p38. Skin from Hailey-Hailey and Darier´s disease patients was studied by using immunofluorescence labeling which targeted intercellular junction proteins. Transepidermal water loss was also measured. ATP2C1 gene expression was silenced in cultured keratinocytes, by siRNA, which modeled Hailey-Hailey disease. Expression of intercellular junction proteins was studied at the mRNA and protein levels. Squamous cell carcinoma and normal human keratinocytes were used as a model for impaired and normal keratinocyte differentiation, and the role of p38 isoforms alpha and delta in the regulation of intercellular junction proteins was studied. Both p38 isoforms were silenced by adenovirus cell transduction, chemical inhibitors or siRNA and keratinocyte differentiation was assessed. The results of this thesis revealed that: i.) intercellular junction proteins are expressed normally in acantholytic skin areas of patients with Hailey-Hailey or Darier´s disease but the localization of ZO-1 expanded to the stratum spinosum; ii.) tight junction proteins, claudin-1 and -4, are regulated by ATP2C1 in non-differentiating keratinocytes; and iii.) p38 delta regulates the expression of tight junction protein ZO-1 in proliferating keratinocytes and in squamous cell carcinoma derived cells. ZO-1 silencing, however, did not affect the expression of other tight junction proteins, suggesting that they are differently regulated. This thesis introduces new mechanisms involved in the regulation of tight junctions revealing new interactions. It provides novel evidence linking intracellular calcium regulation and tight junctions.

Improvement of wet and dry web properties in papermaking by controlling water and fiber quality

Pappersindustrin står inför många utmaningar och bör uppfylla krav som ställs av t.ex. marknadstrender och slutanvändare. Snabbare och effektivare processer ställer även större krav på pappersbanans mekaniska egenskaper. Avbrott i produktionen sker inte enbart pga. låg styrka utan orsaken kan även vara en för låg spänning i pappersbanan. De flesta problemen sker vid våtpressen och/eller i början av torkpartiet då papprets torrhalt är 30-70 %. Spänningen hålls inte konstant efter töjning, utan sjunker pga. papprets relaxering, som till största delen sker då pappret flyttas från pressen till torkpartiet. Ur forskningssynvinkel öppnar en smidigt fungerande verksamhet med potentiella energibesparingar upp möjligheterna för förbättring av befintliga processer. I detta arbete undersöktes hur olika faktorer inverkar på fibernätverkets avvattning, initiala våtstyrka och relaxering samt också på slutproduktens mekaniska egenskaper och ytegenskaper. I första delen ändrades processvattnets egenskaper, såsom pH, konduktivitet och ytspänning. Ytspänningen varierades genom tillsatts av ett nonjoniskt ytaktivt ämne, s.k. surfaktant. Fiberegenskaperna modifierades också. Fibern maldes antingen genom en mild eller genom en hård process. Effekten av finmaterial undersöktes genom att tillsätta finmaterial till den ursprungliga massan eller genom att avlägsna finmaterialet från den malda massan. I den sista delen användes en hemicellulosa, som finns i stora mängder i gran, som tillsatsmedel. Förutom naturliga galaktoglukomannaner (GGM), tillverkades också en katjoniserad, en karboximetylerad samt en iminerad, amfifil GGM. Dessa användes i pappersmassa eller sprayades på ytan av ett nybildat ark. Resultaten, som erhölls i denna avhandling, bildar en värdefull kunskapsbas, som kan användas för kontroll och reglering av pappersmaskinens körbarhet och papperskvalitet.

Regulation of cell fate by c-FLIP phosphorylation

Programmed cell death is an important physiological cellular process that maintains homeostasis and protects multicellular organisms from diseases. Apoptosis is the principal mode of cell death, which eliminates unwanted cells and an enormous effort has been made to understand the molecular mechanisms of the signaling pathway and its regulatory systems. Irregular apoptosis often has life-threatening consequences to humans, including cancer, autoimmune diseases and degenerative diseases. In cancer for example, cell death is an attractive target to eradicate uncontrollably proliferating cells that have disregard pro-apoptotic signaling. Targeted therapeutic approaches are not as effective as once expected, since now we know that the cell death pathways are not sole entities in cells, but are highly associated with various cellular processes. Proteins that regulate apoptosis can also control non-apoptotic signaling pathways. For example, c-FLIP is a protein that can either inhibit or promote caspase-8 activation, which is required to induce apoptosis. Not only has c-FLIP opposing effects on initiating apoptosis, but it also regulates various pro-survival signaling pathways in the cell. It is well known that protein expression level is a determinant of how c-FLIP can regulate different signaling pathways, but other regulatory mechanisms potentially affecting the role of c-FLIP are less well understood. This work addresses novel insights into the mechanisms of c-FLIP post-translational modifications and their functional consequences. We have identified that phosphorylation is an important inception for subcellular localization of c-FLIP, thereby dictating which apoptotic and non-apoptotic signaling pathways c-FLIP could regulate to promote cell survival. Furthermore, we have constructed mathematical models to unite independent studies to establish more systematic c-FLIP signaling pathways to understand the dynamics of extrinsically-induced apoptosis.

Quality of radiotherapy care by development of e-feedback knowledge

The aim of this three phase study was to develop quality of radiotherapy care by the e-Feedback knowledge of radiotherapy -intervention (e-Re-Know). In Phase I, the purpose was to describe the quality of radiotherapy care and its deficits experienced by cancer patients. Based on the deficits in patient education in Phase II, the purpose was to describe cancer patients’ e-knowledge expectations in radiotherapy. In Phase III, the purpose was to develop and evaluate the outcomes of the e-Re-Know among breast cancer patients. The ultimate aim was to develop radiotherapy care to support patients’ empowerment with patient e-education. In Phase I (2004-2005), the descriptive design was used, and 134 radiotherapy patients evaluated their experiences by Good Nursing Care Scale for Patients (GNCS-P) in the middle of RT period. In Phase II (2006-2008), the descriptive longitudinal design was used and 100 radiotherapy patients’ e-knowledge expectations of RT were evaluated using open-ended questionnaire developed for this study before commencing first RT, in the middle of the treatment, and concluding RT period. In Phase III, firstly (2009-2010), the e-Re-Know intervention, i.e. knowledge test and feedback, was developed in terms of empowering knowledge and implemented with e-feedback approach based on literature and expert reviews. Secondly (2011-2014), the randomized controlled study was used to evaluate the e-Re-Know. Breast cancer patients randomized to either the intervention group (n=65) receiving the e-Re-Know by e-mail before commencing first RT and standard education or the control group (n=63) receiving standard education. The data were collected before commencing first RT, concluding last RT and 3 months after last RT using RT Knowledge Test, Spielberger’s State Trait Inventory (STAI) and Functional Assessment of Cancer Therapy - Breast (FACT-B) –instruments. Data were analyzed using statistical methods and content analysis. The study showed radiotherapy patients experienced quality of care high. However, there were deficits in patient education. Furthermore, radiotherapy patients’ multidimensional e-knowledge expectations through Internet covered mainly bio-physiological and functional knowledge. Thus, the e-Re-Know was developed and evaluated. The study showed when breast cancer patients’ carried out the e-Re-Know their knowledge of side effects self-care was significantly increased and quality of life (QOL) significantly improved in line with decrease in anxiety from time before radiotherapy period to three months after. In addition, the e-Re-Know has potential to have positive effects on anxiety and QOL, regardless of patient characteristics or knowledge level. The results support the theory of empowering patient education suggesting that empowerment can be supported by confirming patients’ understanding of own knowledge level. In summary, the e-Feedback knowledge of radiotherapy (e-Re-Know) intervention can be recommended in development of quality of radiotherapy care experienced by breast cancer patients. Further research is needed to assess and develop patient-centred quality of care by patient education among cancer patients.

How to increase brand quality perception with audiovisual content? : Exploring the role of emotions in B2C and B2B contexts

This study discusses how audiovisual content can influence brand quality perceptions. The purpose of this study is to explore how audiovisual content creation can increase brand quality perceptions. This research problem is addressed with three sub questions, which aim at clarifying the role of emotions between content marketing and brand quality perception, explaining how different functions of audiovisual content can increase brand quality perception, and by identifying and comparing the key differences in content creation in business-to-consumer and business-to-businesscontexts. The theoretical background of the study is in brand personality, consumer emotions, consumerbrand relationships, content marketing and B2B branding literature. The empirical research part includes a single-case study. The case company was a Swiss startup that wished to build a highquality brand for both B2C and B2B segments. The empirical data was collected in September 2014. Eight interviews were conducted; seven with target segment representatives and one with an existing customer of the case company. The empirical findings were analyzed with thematic analysis and finally a 5-stage framework was created based on the findings of the research, offering a guideline for high-quality content creation. This study finds that emotions play an important role in brand quality perceptions. Psychological processes, emotion, cognition and conation, influence the engagement process of the target segment which ultimately can lead to activation and electronic word-of-mouth. Brand quality perception is the result of the overall emotion of the brand. The overall emotion derives from brand personality, brand concept, product attributes and utilitarian benefits of the brand. The entertaining and educational functions of the audiovisual content can target and evoke these emotional processes, and result in increased quality perceptions. In the B2B context, emotions are found to play a relatively smaller role in the quality perception processes. However, the significance of emotions cannot be ignored, since they can emphasize the value for the buying organization, and build on the trust and loyalty among the potential customers. The final framework presents five stages of content creation that ultimately improve brand quality perceptions. These stages help marketers to design and implement their content and evoke positive emotions in their target segment as part of a quality-based marketing strategy. Further research is warranted to quantitatively test the generalizability of the framework. Further research is also suggested to make the framework adaptable to different stages of the brand life cycle.