The domain of rhetoric has changed dramatically since its inception as the art of persuasion. It has adapted to encompass many forms of digital media, including, for example, data visualization and coding as a form of literature, but the approach has frequently been that of an outsider looking in. The use of comprehensive computational tools as a part of rhetorical analysis has largely been lacking. In this report, we attempt to address this lack by means of three case studies in natural language processing tasks, all of which can be used as part of a computational approach to rhetoric. At this same moment in time, it is becoming all the more important to transition to renewable energy in order to keep global warming under 1.5 degrees Celsius and ensure that countries meet the conditions of the Paris Agreement. Thus, we make use of speech data on climate policy from the Swedish parliament to ground these three analyses in semantic textual similarity, topic modeling, and political party attribution. We find that speeches are, to a certain extent, consistent within parties, given that a slight majority of most semantically similar speeches come from the same party. We also find that some of the most common topics discussed in these speeches are nuclear energy and the Swedish Green party, purported environmental risks due to renewable energy sources, and the job market. Finally, we find that though pairs of speeches are semantically similar, party rhetoric on the whole is generally not unique enough for speeches to be distinguishable by party. These results then open the door for a broader exploration of computational rhetoric for Swedish political science in the future.

Sedan början av 2000-talet har Försvarsmakten genomfört en mängd förändringar med sin organisation och personal. Nedskärningarna av förband tillsammans med ett ökat fokus på internationella insatser i början av 00-talet har under slutet av 10-talet förändrats till en tillväxt av förband samt en återgång till ett nationellt fokus. Yrkesofficerarnas profession har förändrats genom införandet av ett tvåbefälssystem under slutet av 00-talet från ett enbefälssystem, likväl som att värnplikten lades vilande 2010 och anställda soldater introducerades i organisationen. Sedan 2018 är värnplikten återinförd med utbildning av soldater och gruppbefäl, 9 respektive 11 månader. Från och med sommaren 2021 återinförs 15 månaders värnpliktiga, som övertar befattningar i krigsorganisationen som tidigare bemannats av yrkesofficerare. Studien utforskar vad detta införande innebär för yrkesofficersprofessionen, vilka konsekvenser som uppstår likväl som hur införandet bör implementeras. Genom intervjuer av åtta respondenter från Trängregementet i Skövde utforskas införandet ur yrkesofficerarnas perspektiv. Studien tar hjälp av Bolman och Deal’s fyra perspektiv på organisation och ledarskap samt Runsten och Werr’s modell för kunskapsintegration för att förstå och tolka organisationsförändringen. Införandet av 15 månaders värnpliktiga omfattar yrkesofficerarnas förväntningar på förändringen, som baseras från tidigare erfarenheter men innehåller även farhågor och positiva förväntningar. Erfarenheten att utbilda dessa värnpliktiga saknas idag på grundutbildningskompanierna, denna kunskap återfinns hos äldre yrkesofficerare i högre nivåer - varvid ett utbyte av gamla erfarenheter i dagens kontext är en framgångsfaktor. Införandet påverkar även utbildning och övningar för förbanden, då värnplikten sker under en längre tid samt att de värnpliktiga ställer ett ökat ansvarstagande hos plutonchefer. Professionen för främst specialistofficerare förändras, genom att värnpliktiga innehar samma grad som specialistofficerare likväl som att expertisen för specialistofficerare kommer ta längre tid att uppnå. En maktförskjutning sker från yrkesofficerare till värnpliktiga, vilket främst påverkar unga specialistofficerare. Studien rekommenderar Försvarsmakten att överse nuvarande styrdokument och uppdatera dessa, samt vidare utveckla yrkesofficersprofessionen. Studien rekommenderar Försvarsmakten att med nuvarande indelning i trebefälssystemet att sergeantsgraden är lämplig för de värnpliktiga och att en förstegsutbildning är bäst lämpad för att utbilda och öva sig till ställföreträdande plutonchef. Om Försvarsmakten inför värnpliktiga plutonchefer i framtiden bör detta föregås av en analys utifrån den militära professionen och trebefälssystemet. / Since the early 2000’s the Swedish Armed Forces has conducted several changes concerning its organization and personnel. The downsizings of units as well as an increased focus on international operations in the early 2000’s have changed to a growth of units and a return to a national focus. The professions of professional officers have changed with the transformation from a single-command system to a two-officer system in the late 00s, as well as the pause of national conscript service in 2010 and introduction of professional soldiers within the organization. The national conscript service was reintroduced in 2018, for soldiers and squad leaders between 9 and 11 months. During the summer of 2021, 15 months conscript service will be reintroduced. These new conscripts will take over the positions previously manned by professional officers in the wartime organization. The study explores the reintroduction’s affects for professional officers, what consequences that occur as well as how the reintroduction should be implemented. Through interviews with eight respondents from the Logistics regiment in Skövde the reintroduction is explored through the professional officers’ view. The study seeks guidance from Bolman and Deal’s four perspectives on organization and leadership and also Runsten and Werr’s model for integration of knowledge, to be able to understand and interpret the organizational change. The reintroduction of 15 months conscripts includes professional officers’ expectations of the change, which is based from earlier experiences but also includes concerns and hopes. There is a lack of experience to train this new category of conscripts today at company level, the ability to do so is found in older officers in higher echelons – therefore an exchange of old experiences in the context of today is a criterion for success. The reintroduction also affects the training and the field exercises, because the conscript service is prolonged and the conscripts require an increased responsibility from the platoon leader. The profession for non-commissioned officers (NCO) changes - through the use of the same rank by conscripts and NCO’s. It will also take longer time for NCO’s to achieve expertise within their profession. A shift of power occurs from professional officers to conscripts, which mostly affects junior NCO’s. The study recommends the Swedish Armed Forces to overlook its current steering documents and to update these, and also to further develop its military profession for all officers. The study recommends the Armed Forces that with the current officer system, the rank of sergeant is well suited for these conscripts and the conscripts service should be based upon a pre-training concept to be able to succeed as deputy platoon leader. If the Armed Forces introduce conscripts as platoon leaders in the future, an analysis should be conducted prior to the change, concerning the military profession and the three-officer system.

Colorectal cancer (CRC) is the third leading cause of cancer related deaths in the United States. The nuclear factor κB (NF-κB) is an important family of transcription factors whose aberrant activation has been found in many types of cancer, including CRC. Therefore, understanding the regulation of NF-κB is of ultimate importance for cancer therapy. Using a novel validation-based insertional mutagenesis (VBIM) strategy, our lab has identified the novel adenomatous polyposis coli (APC) like protein (ALP) gene as a negative regulator of NF-κB. Preliminary studies from our lab demonstrated that overexpression of ALP led to decreased NF-κB activity by κB reporter assay and electrophoresis mobility gel shift assay (EMSA). The current project aims to further evaluate the role of ALP in the regulation of NF-κB signaling in CRC cells. We found that overexpression of ALP in human CRC HT29 cells greatly reduced both the number and the size of colonies that were formed in a soft agar assay. ALP overexpression also decreased the cell growth rate and cell migration ability, while shRNA mediated knockdown of ALP showed opposite effects, confirming that ALP is a tumor suppressor in CRC HT29 cells. Overexpression of ALP led to decreased NF-κB activity by κB reporter assay and condition media assay in CRC HT29 cells. Furthermore, immunohistochemical analysis with human colon vii tissues revealed that there is a gradual loss of ALP protein with tumor progression. We also found that ALP predominantly localizes in the cytoplasm, and binds to the p65 subunit of NF-κB, and might be functioning downstream of IκB kinase (IKK). In summary, in this study, we provide evidence regarding the tumor suppressor role of ALP in CRC by functioning as novel negative regulator of NF-κB. This discovery could lead to the establishment of ALP as a potential biomarker and therapeutic target in CRC.

No description available.

Supply chain operations of construction industry including road projects in disaster regions results in exceeding project budget and timelines. In road construction projects, supply chain with poor performance can affect efficiency and completion time of the project. This is also the case of the road projects in disaster areas. Disaster areas consider both natural and man-made disasters. Few examples of disaster zones are; Pakistan, Afghanistan, Iraq, Sri Lanka, India, Japan, Haiti and many other countries with similar environments. The key factors affecting project performance and execution are insecurity, uncertainties in demand and supply, poor communication and technology, poor infrastructure, lack of political and government will, unmotivated organizational staff, restricted accessibility to construction materials, legal hitches, multiple challenges of hiring labour force and exponential construction rates due to high risk environment along with multiple other factors. The managers at all tiers are facing challenges of overrunning time and budget of supply chain operations during planning as well as execution phase of development projects. The aim of research is to develop a Multiple Linear Regression Model (MLRM) and a Rule Based Decision Support System by incorporating various factors affecting supply chain management of road projects in disaster areas in the order of importance. This knowledge base (KB) (importance / coefficient of each factor) will assist infrastructure managers (road projects) and practitioners in disaster regions in decision making to minimize the effect of each factor which will further help them in project improvement. Conduct of Literature Review in the fields of disaster areas, supply chain operational environments of road project, statistical techniques, Artificial Intelligence (AI) and types of research approaches has provided deep insights to the researchers. An initial questionnaire was developed and distributed amongst participants as pilot project and consequently results were analysed. The results’ analysis enabled the researcher to extract key variables impacting supply chain performance of road project. The results of questionnaire analysis will facilitate development of Multiple Linear Regression Model, which will eventually be verified and validated with real data from actual environments. The development of Multiple Linear Regression Model and Rule Based Decision Support System incorporating all factors which affect supply chain performance of road projects in disastrous regions is the most vital contribution to the research. The significance and novelty of this research is the methodology developed that is the integration of those different methods which will be employed to measure the SCM performance of road projects in disaster areas.

Cette thèse a pour objectif d'évaluer un programme de formation visant à doter les professionnels de la santé d'outils de courtage de connaissances afin d'assurer l'intégration des meilleures pratiques dans les établissements de santé et de services sociaux. Elle vise à évaluer la fidélité de sa mise en œuvre et les effets du programme de formation sur les participants et leur cadre professionnel. Le programme est composé de huit ateliers de 2 heures, dispensés sur une période de trois semaines. Il a été construit et offert par le Consortium InterS4, dont l'objectif est de développer et d'intégrer les meilleures connaissances et pratiques dans les établissements de santé et de services sociaux de cinq régions administratives du Québec, par le biais d'activités de courtage de connaissances. Le programme de formation a été évalué auprès de quatre de ces régions en utilisant le modèle de Kirkpatrick. La collecte de données de ce projet d'évaluation comprend deux parties menées en parallèle, employant des méthodes mixtes pour recueillir des informations complètes. Les données ont été collectées au moyen d'une combinaison de questionnaires quantitatifs et d'entretiens qualitatifs, en tirant parti du modèle d'efficacité de la formation de Kirkpatrick, du Theoretical Domains Framework (TDF) et du Behavior Change Wheel (BCW) pour évaluer à la fois l'impact du programme de formation et les facteurs plus larges influençant le transfert de connaissances et le changement de comportement. Tout au long du processus d'évaluation de 24 mois, trois animateurs et 46 participants répartis en 7 cohortes ont pris part au projet de recherche. La mise en œuvre du programme a été évaluée à travers le contenu utilisé pour chaque cohorte ainsi qu'à travers les discussions avec les animateurs suite au dernier atelier de formation de chaque cohorte. Concernant les effets du programme, les réactions des participants, les apprentissages, les comportements et les résultats ont été mesurés à quatre moments différents ; avant le programme,3 après le dernier atelier, trois mois et six mois après le dernier atelier. Les facteurs contribuant au changement de comportement, à savoir la motivation, l’opportunité et la capacité, ont été évalués à ces multiples moments afin de déterminer leur influence sur la mise en œuvre des activités de courtage de connaissances. Les résultats mettent en évidence la satisfaction des participants concernant l’utilisation du courtage de connaissances dans leur travail. Tous les participants recommandent le programme à leurs pairs et parfois à d'autres professionnels de leur domaine. L'analyse thématique des verbatims suggère l’importance de conditions spécifiques pour la mise en place efficace de pratiques de courtage de connaissances au sein de ces établissements. Concernant les facteurs liés au changement de comportement, les données recueillies lors de l'évaluation font état d'une perception de gains de capacité par les participants à l'issue de la formation. Les participants avaient des convictions positives concernant le courtage de connaissances et ses effets. Cependant, beaucoup d’entre eux n’ont pas eu l’occasion de mettre en pratique les apprentissages en raison de priorités différentes dans leur rôle professionnel et de leur environnement de travail. Les recommandations consistent notamment à garantir que la direction connaît et exploite les capacités en courtage de connaissances, que les objectifs sont bien définis et que les parties prenantes sont alignées dans l'exécution des mandats. / This thesis has for objective to evaluate a training program aimed at equipping health professionals with KB tools to ensure that best practices can be integrated into health and social services institutions. It aims to assess the fidelity of its implementation and the effects of the training program on the participants and their professional setting. The program is made up of eight 2-hour workshops, administered over a period of three weeks. The training program was built and offered by the InterS4 Consortium, whose objective is to develop and integrate best knowledge and practices in the health and social services institutions of five Québec administrative regions, of which four were evaluated in this study, through KB activities. The research design of this evaluation project includes a two-part investigation conducted in parallel, employing mixed methods to gather comprehensive insights. Data was collected through a combination of quantitative questionnaires and qualitative interviews, leveraging the Kirkpatrick Training Effectiveness Model, the Theoretical Domains Framework (TDF), and the Behavior Change Wheel (BCW) to evaluate both the training program's impact and the broader factors influencing knowledge transfer and behavior change. Throughout the 24-month evaluation process, three animators and 46 participants spread into 7 cohorts took part in the research project. The implementation of the program was evaluated through the content used for each cohort as well as through discussions with the animators following the last workshop for each cohort training. Regarding the effects of the program, the reaction, learnings, behaviors and results were measured at four different times; before the program, following the last workshop, three months and six months after the last workshop. Factors contributing to behavior change—namely motivation, opportunity, and capability—were assessed at these multiple time points to capture their influence on the implementation of KB activities. The results highlight the satisfaction of the participants concerning the use of KB in their work. All participants recommend the program to their peers and sometimes to other professionals in their field. The thematic analysis of the verbatims suggests the importance of specific conditions for the effective implementation of knowledge brokering practices within these establishments. Concerning the factors related to the behavior change, the data collected during the evaluation reports a perception of gains in capability by the participants at the end of the training. Participants held positive beliefs regarding KB and its effects. However, many of them did not have the opportunity to put the learnings into application due to differing priorities in their professional role and due to their work environment. Recommendations include ensuring the management is aware of and leveraging the KB capabilities, goals are well-defined and stakeholders are aligned in the execution of the mandates.

Cylindrospermopsin (CYN) is a toxic alkaloid produced by several freshwater cyanobacterial species, the most prevalent in Australian waters being Cylindrospermopsis raciborskii. The occurrence of CYN-producing cyanobacteria in drinking water sources worldwide poses a potential human health risk, with one well-documented case of human poisoning attributed to the toxin. While extensive characterisation of CYN-induced toxicity has been conducted in rodents both in vivo and in primary cell cultures, little is known about mechanisms of toxicity in human cell types. This thesis describes studies undertaken to further define the molecular mechanisms of CYN toxicity in human cells. Concentration-response relationships were determined in various cultured human cell types using standard toxicity assays. As expected, CYN caused dose-dependent decreases in the growth of three cell lines, HepG2, Caco-2 and HeLa, and one primary cell type, human dermal fibroblasts, according to tetrazolium reduction assays. CYN treatment did not disrupt cellular membranes according to the lactate dehydrogenase release assay in HepG2 or Caco-2 cells after 24, 48 or 72 h exposure, but did cause membrane disruption in fibroblasts after 72 h exposure to relatively high concentrations of the toxin. Apoptosis occurred more readily in HeLa cells than HepG2 cells or fibroblasts, with 72 h exposure to 1 &mug/mL required before statistically significant rates of apoptosis occurred in the latter cell types. CYN did not appear to directly affect the structure of actin filaments or microtubules under the conditions used in the present study. The major portion of the work presented in this thesis comprises a large-scale interrogation of changes in gene expression induced by the toxin in cultured cells. To assess the effects of CYN on global gene expression, relative messenger RNA (mRNA) levels in human dermal fibroblasts and HepG2 cells after 6 h and 24 h exposure to 1 &mug/mL CYN were determined using oligonucleotide microarrays representing approximately 19 000 genes. Overall, the number of transcripts significantly altered in abundance was greater in fibroblasts than in HepG2 cells. In both cell types, mRNA levels for genes related to amino acid biosynthesis, carbohydrate metabolism, and protein folding and transport were reduced after CYN treatment, while transcripts representing genes for apoptosis, RNA biosynthesis and RNA processing increased in abundance. More detailed data analyses revealed the modulation of a number of stress response pathways—genes regulated by NF-&kappaB were induced, DNA damage response pathways were up-regulated, and a large number of genes involved in endoplasmic reticulum stress were strongly down-regulated. Genes for the synthesis and processing of mRNA, tRNA and rRNA were strongly up-regulated, indicating that CYN treatment may increase the turnover of all forms of cellular RNA. A small group of genes were differentially expressed in HepG2 cells and fibroblasts, revealing cell-specific responses to the toxin. Selected changes in transcript level were validated using real-time quantitative reverse transcriptase PCR (qRT-PCR). The modulation of stress response pathways by CYN, indicated by microarray analysis, was further investigated using other methods. The role of tumour suppressor protein p53 in CYN-mediated gene expression was confirmed by measuring the expression of known p53-regulated genes following CYN treatment of HepG2 cells and human dermal fibroblasts using qRT-PCR. Western blotting of protein extracts from CYNtreated cells showed that p53 protein accumulation occurred in HepG2 cells, providing additional evidence of the activation of the p53 pathway by CYN in this cell line. The immediate-early genes JUN and FOS were found to be induced by CYN in a concentration-dependent manner, and MYC was induced to a lesser extent. The mitogen-activated protein kinase c-Jun NH2-terminal kinase, implicated in the ribotoxic stress response initiated by damage to ribosomal RNA, appeared to become phosphorylated in HeLa cells after CYN exposure, suggesting that ribotoxic stress may occur in response to CYN in at least some cell types. The expression of a reporter gene under the control of a response element specific for NF-&kappaB was induced at the mRNA level but inhibited at the protein level. This shows that while transcription factors such as p53 and NF-&kappaB are apparently activated in response to the toxin, transactivation of target genes may not necessarily manifest a corresponding increase at the protein level. The current work contributes significantly to the current understanding of cylindrospermopsin toxicity in human-derived cell types, and provides further insight into putative modes of action.

I dagens informationssamhälle pågår en växande trend av molnbaserade tjänster. Vid implementering av molnbaserade tjänster används metoden Virtualisering. Denna metod minskar behovet av antal fysiska datorsystem i ett datacenter. Vilket har en positiv miljöpåverkan eftersom energikonsumtionen minskar när hårdvaruresurser kan utnyttjas till sin fulla kapacitet. Molnbaserade tjänster skapar samhällsnytta då nya aktörer utan teknisk bakgrundskunskap snabbt kan komma igång med verksamhetsberoende tjänster. För tillämpning av Virtualisering används en så kallad Hypervisor vars uppgift är att distribuera molnbaserade tjänster. Efter utvärdering av vetenskapliga studier har vi funnit att det finns skillnader i prestanda och funktionalitet mellan olika Hypervisors. Därför väljer vi att göra en prestanda- samt funktionsanalys av Hypervisors som kommer från de största aktörerna på marknaden. Dessa är Microsoft Hyper-V Core Server 2012, Vmware ESXi 5.1.0 och Citrix XenServer 6.1.0 Free edition. Vår uppdragsgivare är försvarsmakten som bekräftade en stor efterfrågan av vår undersökning. Rapporten innefattar en teoretisk grund som beskriver tekniker bakom virtualisering och applicerbara användningsområden. Genomförandet består av två huvudsakliga metoder, en kvalitativ- respektive kvantitativ del. Grunden till den kvantitativa delen utgörs av ett standardsystem som fastställdes utifrån varje Hypervisors begränsningar. På detta standardsystem utfördes prestandatester i form av dataöverföringar med en serie automatiserade testverktyg. Syftet med testverktygen var att simulera datalaster som avsiktligt påverkade CPU och I/O för att avgöra vilka prestandaskillnader som förekommer mellan Hypervisors. Den kvalitativa undersökningen omfattade en utredning av funktionaliteter och begränsningar som varje Hypervisor tillämpar. Med tillämpning av empirisk analys av de kvantitativa mätresultaten kunde vi fastställa orsaken bakom varje Hypervisors prestanda. Resultaten visade att det fanns en korrelation mellan hur väl en Hypervisor presterat och vilken typ av dataöverföring som den utsätts för. Den Hypervisor som uppvisade goda prestandaresultat i samtliga dataöverföringar är ESXi. Resultaten av den kvalitativa undersökningen visade att den Hypervisor som offererade mest funktionalitet och minst begränsningar är Hyper-V. Slutsatsen blev att ett mindre datacenter som inte planerar en expansion bör lämpligtvis välja ESXi. Ett större datacenter som både har behov av funktioner som gynnar molnbaserade tjänster och mer hårdvaruresurser bör välja Hyper-V vid implementation av molntjänster. / A growing trend of cloud-based services can be witnessed in todays information society. To implement cloud-based services a method called virtualization is used. This method reduces the need of physical computer systems in a datacenter and facilitates a sustainable environmental and economical development. Cloud-based services create societal benefits by allowing new operators to quickly launch business-dependent services. Virtualization is applied by a so-called Hypervisor whose task is to distribute cloud-based services. After evaluation of existing scientific studies, we have found that there exists a discernible difference in performance and functionality between different varieties of Hypervisors. We have chosen to perform a functional and performance analysis of Hypervisors from the manufacturers with the largest market share. These are Microsoft Hyper-V Core Server 2012, Vmware ESXi 5.1.0 and Citrix XenServer 6.1.0 Free edition. Our client, the Swedish armed forces, have expressed a great need of the research which we have conducted. The thesis consists of a theoretical base which describes techniques behind virtualization and its applicable fields. Implementation comprises of two main methods, a qualitative and a quantitative research. The basis of the quantitative investigation consists of a standard test system which has been defined by the limitations of each Hypervisor. The system was used for a series of performance tests, where data transfers were initiated and sampled by automated testing tools. The purpose of the testing tools was to simulate workloads which deliberately affected CPU and I/O to determine the performance differences between Hypervisors. The qualitative method comprised of an assessment of functionalities and limitations for each Hypervisor. By using empirical analysis of the quantitative measurements we were able to determine the cause of each Hypervisors performance. The results revealed that there was a correlation between Hypervisor performance and the specific data transfer it was exposed to. The Hypervisor which exhibited good performance results in all data transfers was ESXi. The findings in the qualitative research revealed that the Hypervisor which offered the most functionality and least amount of constraints was Hyper-V. The conclusion of the overall results uncovered that ESXi is most suitable for smaller datacenters which do not intend to expand their operations. However a larger datacenter which is in need of cloud service oriented functionalities and requires greater hardware resources should choose Hyper-V at implementation of cloud-based services.

No description available.

Cardiovascular diseases are among the leading causes of death worldwide and particularly in the developed World. The search for new therapeutic approaches for improving the functions of the damaged heart is therefore a critical endeavour. Myocardial infarction, which can lead to heart failure, is associated with irreversible loss of functional cardiomyocytes. The loss of cardiomyocytes poses a major difficulty for treating the damaged heart since terminally differentiated cardiomyocytes have very limited regeneration potential. Currently, the only effective treatment for severe heart failure is heart transplantation but this option is limited by the acute shortage of donor hearts. The high incidence of heart diseases and the scarcity donor hearts underline the urgent need to find alternative therapeutic approaches for treating cardiovascular diseases. Pluripotent embryonic stem (ES) cells can differentiate into functional cardiomyocytes. Therefore the engraftment of ES cell-derived functional cardiomyocytes or cardiac progenitor cells into the damaged heart to regenerate healthy myocardial tissues may be used to treat damaged hearts. Stem cell-based therapy therefore holds a great potential as a very attractive alternative to heart transplant for treating heart failure and other cardiovascular diseases. A major obstacle to the realisation of stem cell-based therapy is the lack of donor cells and this in turn is due to the fact that, currently, the molecular mechanisms or the regulatory signal transduction mechanisms that are responsible for mediating ES cell differentiation into cardiomyocytes are not well understood. Overcoming this huge scientific challenge is absolutely necessary before the use of stem cell-derived cardiomyocytes to treat the damaged heart can become a reality. Therefore the aim of this thesis was to investigate the signal transduction pathways that are involved in the differentiation of stem cells into cardiomyocytes. The first objective was the establishment and use of cardiomyocyte differentiation models using H9c2 cells and P19 stem cells to accomplish the specific objectives of the thesis. The specific objectives of the thesis were, the investigation of the roles of (i) nitric oxide (ii) protein kinase C (PKC), (iii) p38 mitogen-activated protein kinase (p38 MAPK) (vi) phosphoinositide 3-kinase (PI3K) and (vi) nuclear factor-kappa B (NF-kB) signalling pathways in the differentiation of stem cells to cardiomyocytes and, more importantly, to identify where possible any points of convergence and potential cross-talk between pathways that may be critical for differentiation to occur. P19 cells were routinely cultured in alpha minimal essential medium (α-MEM) supplemented with 100 units/ml penicillin /100 μg/ml streptomycin and 10% foetal bovine serum (FBS). P19 cell differentiation was initiated by culturing the cells in microbiological plates in medium containing 0.8 % DMSO to form embryoid bodies (EB). This was followed by transfer of EBs to cell culture grade dishes after four days. H9c2 cells were cultured in Dulbecco’s Modified Eagle’s medium (DMEM) supplemented with 10% FBS. Differentiation was initiated by incubating the cells in medium containing 1% FBS. In both models, when drugs were employed, they were added to cells for one hour prior to initiating differentiation. Cell monolayers were monitored daily over a period of 12 or 14 days. H9c2 cells were monitored for morphological changes and P19 cells were monitored for beating cardiomyocytes. Lysates were generated in parallel for western blot analysis of changes in cardiac myosin heavy chain (MHC), ventricular myosin chain light chain 1(MLC-1v) or troponin I (cTnI) using specific monoclonal antibodies. H9c2 cells cultured in 1% serum underwent differentiation as shown by the timedependent formation of myotubes, accompanied by a parallel increase in expression of both MHC and MLC-1v. These changes were however not apparent until 4 to 6 days after growth arrest and increased with time, reaching a peak at day 12 to 14. P19 stem cells cultured in DMSO containing medium differentiated as shown by the timedependent appearance of beating cardiomyocytes and this was accompanied by the expression of cTnI. The differentiation of both P19 stem cells and H9c2 into cardiomyocytes was blocked by the PI3K inhibitor LY294002, PKC inhibitor BIM-I and the p38 MAPK inhibitor SB2035800. However when LY294002, BIM-I or SB2035800 were added after the initiation of DMSO-induced P19 stem cell differentiation, each inhibitor failed to block the cell differentiation into beating cardiomyocytes. The NF-kB activation inhibitor, CAPE, blocked H9c2 cell differentiation into cardiomyocytes. Fast nitric oxide releasing donors (SIN-1 and NOC-5) markedly delayed the onset of differentiation of H9c2 cells into cardiomyocytes while slow nitric oxide releasing donors (SNAP and NOC-18) were less effective in delaying the onset of differentiation or long term differentiation of H9c2 cells into cardiomyocytes. Akt (protein kinase B) is the key downstream target of PI3K. Our cross-talk data also showed that PKC inhibition and p38 MAPK inhibition respectively enhanced and reduced the activation of Akt, as determined by the phosphorylation of Akt at serine residue 473. In conclusion, PKC, PI3K, p38 MAPK and NF-kB are relevant for the differentiation of stem cells into cardiomyocytes. Our data also show that the PKC, PI3K and p38 MAPK signalling pathways are activated as very early events during the differentiation of stem cells into cardiomyocytes. Our data also suggest that PKC may negatively regulate Akt activation while p38 MAPK inhibition inhibits Akt activation. Our fast NO releasing donor data suggest that nitric oxide may negatively regulate H9c2 cell differentiation.