Pp2a: At The Crossroads Between Growth and Defence In Plants

Living organisms manage their resources in well evolutionary-preserved manner to grow and reproduce. Plants are no exceptions, beginning from their seed stage they have to perceive environmental conditions to avoid germination at wrong time or rough soil. Under favourable conditions, plants invest photosynthetic end products in cell and organ growth to provide best possible conditions for generation of offspring. Under natural conditions, however, plants are exposed to a multitude of environmental stress factors, including high light and insufficient light, drought and flooding, various bacteria and viruses, herbivores, and other plants that compete for nutrients and light. To survive under environmental challenges, plants have evolved signaling mechanisms that recognise environmental changes and perform fine-tuned actions that maintain cellular homeostasis. Controlled phosphorylation and dephosphorylation of proteins plays an important role in maintaining balanced flow of information within cells. In this study, I examined the role of protein phosphatase 2A (PP2A) on plant growth and acclimation under optimal and stressful conditions. To this aim, I studied gene expression profiles, proteomes and protein interactions, and their impacts on plant health and survival, taking advantage of the model plant Arabidopsis thaliana and the mutant approach. Special emphasis was made on two highly similar PP2A-B regulatory subunits, B’γ and B’ζ. Promoters of B’γ and B’ζ were found to be similarly active in the developing tissues of the plant. In mature leaves, however, the promoter of B’γ was active in patches in leaf periphery, while the activity of B’ζ promoter was evident in leaf edges. The partially overlapping expression patterns, together with computational models of B’γ and B’ζ within trimeric PP2A holoenzymes suggested that B’γ and B’ζ may competitively bind into similar PP2A trimmers and thus influence each other’s actions. Arabidopsis thaliana pp2a-b’γ and pp2a-b’γζ double mutants showed dwarfish phenotypes, indicating that B’γ and B’ζ are needed for appropriate growth regulation under favorable conditions. However, while pp2a-b’γ displayed constitutive immune responses and appearance of premature yellowings on leaves, the pp2a-b’γζ double mutant supressed these yellowings. More detailed analysis of defense responses revealed that B’γ and B’ζ mediate counteracting effects on salicylic acid dependent defense signalling. Associated with this, B’γ and B’ζ were both found to interact in vivo with CALCIUM DEPENDENT PROTEIN KINASE 1 (CPK1), a crucial element of salicylic acid signalling pathway against pathogens in plants. In addition, B’γ was shown to modulate cellular reactive oxygen species (ROS) metabolism by controlling the abundance of ALTERNATIVE OXIDASE 1A and 1D in mitochondria. PP2A B’γ and B’ζ subunits turned out to play crucial roles in the optimization of plant choices during their development. Taken together, PP2A allows fluent responses to environmental changes, maintenance of plant homeostasis, and grant survivability with minimised cost of redirection of resources from growth to defence.

A conjugate of an anti-midkine single-chain variable fragment to doxorubicin inhibits tumor growth

Doxorubicin (DOX) was conjugated to a single-chain variable fragment (scFv) against human midkine (MK), and the conjugate (scFv-DOX) was used to target the chemotherapeutic agent to a mouse solid tumor model in which the tumor cells expressed high levels of human MK. The His-tagged recombinant scFv was expressed in bacteria, purified by metal affinity chromatography, and then conjugated to DOX using oxidative dextran (Dex) as a linker. The molecular formula of this immunoconjugate was scFv(Dex)1.3(DOX)20. In vitro apoptosis assays showed that the scFv-DOX conjugate was more cytotoxic against MK-transfected human adenocarcinoma cells (BGC823-MK) than untransfected cells (55.3 ± 2.4 vs 22.4 ± 3.8%) for three independent experiments. Nude mice bearing BGC823-MK solid tumors received scFv-DOX or equivalent doses of scFv + DOX for 2 weeks and tumor growth was more effectively inhibited by the scFv-DOX conjugate than by scFv + DOX (51.83% inhibition vs 40.81%). Histological analysis of the tumor tissues revealed that the highest levels of DOX accumulated in tumors from mice treated with scFv-DOX and this resulted in more extensive tumor cell death than in animals treated with the equivalent dose of scFv + DOX. These results show that the scFv-DOX conjugate effectively inhibited tumor growth in vivo and suggest that antigen-specific scFv may be competent drug-carriers.

CIAPIN1 gene silencing enhances chemosensitivity in a drug-resistant animal model in vivo

Overexpression of cytokine-induced apoptosis inhibitor 1 (CIAPIN1) contributes to multidrug resistance (MDR) in breast cancer. This study aimed to evaluate the potential of CIAPIN1 gene silencing by RNA interference (RNAi) as a treatment for drug-resistant breast cancer and to investigate the effect of CIAPIN1 on the drug resistance of breast cancer in vivo. We used lentivirus-vector-based RNAi to knock down CIAPIN1 in nude mice bearing MDR breast cancer tumors and found that lentivirus-vector-mediated silencing of CIAPIN1 could efficiently and significantly inhibit tumor growth when combined with chemotherapy in vivo. Furthermore, Western blot analysis showed that both CIAPIN1 and P-glycoprotein expression were efficiently downregulated, and P53 was upregulated, after RNAi. Therefore, we concluded that lentivirus-vector-mediated RNAi targeting of CIAPIN1 is a potential approach to reverse MDR of breast cancer. In addition, CIAPIN1 may participate in MDR of breast cancer by regulating P-glycoprotein and P53 expression.

Putative role of ischemic postconditioning in a rat model of limb ischemia and reperfusion: involvement of hypoxia-inducible factor-1α expression

Hypoxia-inducible factor-1α (HIF-1α) is one of the most potent angiogenic growth factors. It improves angiogenesis and tissue perfusion in ischemic skeletal muscle. In the present study, we tested the hypothesis that ischemic postconditioning is effective for salvaging ischemic skeletal muscle resulting from limb ischemia-reperfusion injury, and that the mechanism involves expression of HIF-1α. Wistar rats were randomly divided into three groups (n=36 each): sham-operated (group S), hindlimb ischemia-reperfusion (group IR), and ischemic postconditioning (group IPO). Each group was divided into subgroups (n=6) according to reperfusion time: immediate (0 h, T0), 1 h (T1), 3 h (T3), 6 h (T6), 12 h (T12), and 24 h (T24). In the IPO group, three cycles of 30-s reperfusion and 30-s femoral aortic reocclusion were carried out before reperfusion. At all reperfusion times (T0-T24), serum creatine kinase (CK) and lactate dehydrogenase (LDH) activities, as well as interleukin (IL)-6, IL-10, and tumor necrosis factor-α (TNF-α) concentrations, were measured in rats after they were killed. Histological and immunohistochemical methods were used to assess the skeletal muscle damage and HIF-1α expression in skeletal muscle ischemia. In groups IR and IPO, serum LDH and CK activities and TNF-α, IL-6, and IL-10 concentrations were all significantly increased compared to group S, and HIF-1α expression was up-regulated (P<0.05 or P<0.01). In group IPO, serum LDH and CK activities and TNF-α and IL-6 concentrations were significantly decreased, IL-10 concentration was increased, HlF-1α expression was down-regulated (P<0.05 or P<0.01), and the pathological changes were reduced compared to group IR. The present study suggests that ischemic postconditioning can reduce skeletal muscle damage caused by limb ischemia-reperfusion and that its mechanisms may be related to the involvement of HlF-1α in the limb ischemia-reperfusion injury-triggered inflammatory response.

Effect of physical training on liver expression of activin A and follistatin in a nonalcoholic fatty liver disease model in rats

Nonalcoholic fatty liver disease (NAFLD) is characterized by fat accumulation in the liver and is associated with obesity and insulin resistance. Activin A is a member of the transforming growth factor beta (TGF)-β superfamily and inhibits hepatocyte growth. Follistatin antagonizes the biological actions of activin. Exercise is an important therapeutic strategy to reduce the metabolic effects of obesity. We evaluated the pattern of activin A and follistatin liver expression in obese rats subjected to swimming exercise. Control rats (C) and high-fat (HF) diet-fed rats were randomly assigned to a swimming training group (C-Swim and HF-Swim) or a sedentary group (C-Sed and HF-Sed). Activin βA subunit mRNA expression was significantly higher in HF-Swim than in HF-Sed rats. Follistatin mRNA expression was significantly lower in C-Swim and HF-Swim than in either C-Sed or HF-Sed animals. There was no evidence of steatosis or inflammation in C rats. In contrast, in HF animals the severity of steatosis ranged from grade 1 to grade 3. The extent of liver parenchyma damage was less in HF-Swim animals, with the severity of steatosis ranging from grade 0 to grade 1. These data showed that exercise may reduce the deleterious effects of a high-fat diet on the liver, suggesting that the local expression of activin-follistatin may be involved.

Bone marrow mesenchymal stem cells overexpressing human basic fibroblast growth factor increase vasculogenesis in ischemic rats

Administration or expression of growth factors, as well as implantation of autologous bone marrow cells, promote in vivo angiogenesis. This study investigated the angiogenic potential of combining both approaches through the allogenic transplantation of bone marrow-derived mesenchymal stem cells (MSCs) expressing human basic fibroblast growth factor (hbFGF). After establishing a hind limb ischemia model in Sprague Dawley rats, the animals were randomly divided into four treatment groups: MSCs expressing green fluorescent protein (GFP-MSC), MSCs expressing hbFGF (hbFGF-MSC), MSC controls, and phosphate-buffered saline (PBS) controls. After 2 weeks, MSC survival and differentiation, hbFGF and vascular endothelial growth factor (VEGF) expression, and microvessel density of ischemic muscles were determined. Stable hbFGF expression was observed in the hbFGF-MSC group after 2 weeks. More hbFGF-MSCs than GFP-MSCs survived and differentiated into vascular endothelial cells (P<0.001); however, their differentiation rates were similar. Moreover, allogenic transplantation of hbFGF-MSCs increased VEGF expression (P=0.008) and microvessel density (P<0.001). Transplantation of hbFGF-expressing MSCs promoted angiogenesis in an in vivo hind limb ischemia model by increasing the survival of transplanted cells that subsequently differentiated into vascular endothelial cells. This study showed the therapeutic potential of combining cell-based therapy with gene therapy to treat ischemic disease.

Promoting inflammatory lymphangiogenesis by vascular endothelial growth factor-C (VEGF-C) aggravated intestinal inflammation in mice with experimental acute colitis

Angiogenesis and lymphangiogenesis are thought to play a role in the pathogenesis of inflammatory bowel diseases (IBD). However, it is not understood if inflammatory lymphangiogenesis is a pathological consequence or a productive attempt to resolve the inflammation. This study investigated the effect of lymphangiogenesis on intestinal inflammation by overexpressing a lymphangiogenesis factor, vascular endothelial growth factor-C (VEGF-C), in a mouse model of acute colitis. Forty eight-week-old female C57BL/6 mice were treated with recombinant adenovirus overexpressing VEGF-C or with recombinant VEGF-C156S protein. Acute colitis was then established by exposing the mice to 5% dextran sodium sulfate (DSS) for 7 days. Mice were evaluated for disease activity index (DAI), colonic inflammatory changes, colon edema, microvessel density, lymphatic vessel density (LVD), and VEGFR-3mRNA expression in colon tissue. When acute colitis was induced in mice overexpressing VEGF-C, there was a significant increase in colonic epithelial damage, inflammatory edema, microvessel density, and neutrophil infiltration compared to control mice. These mice also exhibited increased lymphatic vessel density (73.0±3.9 vs 38.2±1.9, P<0.001) and lymphatic vessel size (1974.6±104.3 vs 1639.0±91.5, P<0.001) compared to control mice. Additionally, the expression of VEGFR-3 mRNA was significantly upregulated in VEGF-C156S mice compared to DSS-treated mice after induction of colitis (42.0±1.4 vs 3.5±0.4, P<0.001). Stimulation of lymphangiogenesis by VEGF-C during acute colitis promoted inflammatory lymphangiogenesis in the colon and aggravated intestinal inflammation. Inflammatory lymphangiogenesis may have pleiotropic effects at different stages of IBD.

Gearing Towards Omnichannel Grocery Retail Through Business Model Innovation

Technological innovations and the advent of digitalization have led retail business into one of its biggest transformations of all time. Consumer behaviour has changed rapidly and the customers are ever more powerful, demanding, tech-savvy and moving on various plat-forms. These attributes will continue to drive the development and robustly restructure the architecture of value creation in the retail business. The largest retail category, grocery yet awaits for a real disruption, but the signals for major change are already on the horizon. The first wave of online grocery retail was introduced in the mid 1990’s and it throve until millennium. Many overreactions, heavy investments and the burst IT-bubble almost stag-nated the whole industry for a long period of time. The second wave started with a venge-ance around 2010. Some research was carried out during the first wave from a single-viewpoint of online grocery retail, but without a comprehensive approach to online-offline business model integration. Now the accelerating growth of e-business has initiated an increased interest to examine the transformation from traditional business models towards e-business models and their integration on the companies’ traditional business models. This research strove to examine how can we recognize and analyze how digitalization and online channels are affecting the business models of grocery retail, by using business mod-el canvas as an analysis tool. Furthermore business model innovation and omnichannel retail were presented and suggested as potential solutions for these changes. 21 experts in online grocery industry were being interviewed. The thoughts of the informants were being qualitatively analysed by using an analysis tool called the business model canvas. The aim of this research was to portray a holistic view on the Omnichannel grocery retail business model, and the value chain, in which the case company Arina along with its partners are operating. The key conclusions exhibited that online grocery retail business model is not an alterna-tive model nor a substitute for the traditional grocery retail business model, though all of the business model elements are to some extent affected by it, but rather a complementary business model that should be integrated into the prevailing, conventional grocery retail business model. A set of business model elements, such as value proposition and distribu-tion channels were recognized as the most important ones and sources of innovation within these components were being illustrated. Segments for online grocery retail were empiri-cally established as polarized niche markets in contrast of the segmented mass-market of the conventional grocery retail. Business model innovation was proven to be a considera-ble method and a conceptual framework, by which to come across with new value proposi-tions that create competitive advantage for the company in the contemporary, changing business environment. Arina as a retailer can be considered as a industry model innovator, since it has initiated an entire industry in its market area, where other players have later on embarked on, and in which the contributors of the value chain, such as Posti depend on it to a great extent. Consumer behaviour clearly affects and appears everywhere in the digi-talized grocery trade and it drives customers to multiple platforms where retailers need to be present. Omnichannel retail business model was suggested to be the solution, in which the new technologies are being utilized, contemporary consumer behaviour is embedded in decision-making and all of the segments and their value propositions are being served seamlessly across the channels.

Theoretical modelling of Staphylococcus aureus growth in a cooked meat product kept at ambient temperature using temperature profiles of selected Mexican cities

A theoretical model is used to predict the growth of Staphylococcus aureus in a pasteurized meat product kept at ambient temperatures for several hours. For this purpose, the temperature profiles of some cities of Mexico were combined with literature data on the kinetics of S. aureus growth. As shown by theoretical predictions, if the food is kept at ambient temperature, the average daily temperature may not give accurate predictions.

Mathematical modeling of microbial growth in milk

A mathematical model to predict microbial growth in milk was developed and analyzed. The model consists of a system of two differential equations of first order. The equations are based on physical hypotheses of population growth. The model was applied to five different sets of data of microbial growth in dairy products selected from Combase, which is the most important database in the area with thousands of datasets from around the world, and the results showed a good fit. In addition, the model provides equations for the evaluation of the maximum specific growth rate and the duration of the lag phase which may provide useful information about microbial growth.

The role of business model in computer game development organizations

The computer game industry has grown steadily for years, and in revenues it can be compared to the music and film industries. The game industry has been moving to digital distribution. Computer gaming and the concept of business model are discussed among industrial practitioners and the scientific community. The significance of the business model concept has increased in the scientific literature recently, although there is still a lot of discussion going on on the concept. In the thesis, the role of the business model in the computer game industry is studied. Computer game developers, designers, project managers and organization leaders in 11 computer game companies were interviewed. The data was analyzed to identify the important elements of computer game business model, how the business model concept is perceived and how the growth of the organization affects the business model. It was identified that the importance of human capital is crucial to the business. As games are partly a product of creative thinking also innovation and the creative process are highly valued. The same applies to technical skills when performing various activities. Marketing and customer relationships are also considered as key elements in the computer game business model. Financing and partners are important especially for startups, when the organization is dependent on external funding and third party assets. The results of this study provide organizations with improved understanding on how the organization is built and what business model elements are weighted.

Germany's socio-economic model and the Euro crisis

Germany's socio-economic model, the "social market economy", was established in West Germany after World War II and extended to the unified Germany in 1990. During a prolonged recession after the adoption of the Euro in 1998, major reforms (Agenda 2010) were introduced which many consider as the key of Germany's recent success. The reforms had mixed results: employment increased but has consisted to a large extent of precarious low-wage jobs. Growth depended on export surpluses based on an internal real devaluation (low unit labour costs) which make Germany vulnerable to global recessions as in 2009. Overall inequality increased substantially.

How Cloud Services may Enable SMEs to Overcome International Growth Challenges: Finnish SMEs in the IT Industry

Digitalization and technology megatrends such as Cloud services have provided SMEs with a suitable atmosphere and conditions to internationalize and seek for further business growth. There is a limited amount of research on Cloud services from the business perspective and the limitations and challenges SMEs encounter when pursuing international business growth. Thus, the main research question of this study was how Cloud services may enable Finnish SMEs to overcome international growth challenges. The research question was further divided into three sub-questions dealing with matters related to features and characteristics of Cloud services, limitations and challenges Finnish SMEs experience when pursuing international growth of business, and benefits and advantages of utilizing Cloud services to mitigate and suppress international growth challenges. First, the theoretical framework of this study was constructed based on the existing literature on Cloud services, SMEs, and international growth challenges. After this, qualitative research approach and methodology were applied for this study. The data was collected through six semi-structured expert interviews in person with representatives of IBM, Exidio, Big Data Solutions, and Comptel. After analyzing the collected data by applying thematic analysis method, the results were compared with the existing theory and the original framework was modified and complemented accordingly. Resource scarcity, customer base expansion and retention, and lack of courage to try new things and take risks turned out to be major international growth challenges of Finnish SMEs. Due to a number of benefits and advantages of utilizing Cloud services including service automation, consumption-based pricing model, lack of capital expenditures (capex) and huge upfront investments, lightened organization structure, cost savings, speed, accessibility, scalability, agility, geographical expansion potential, global reaching and covering, credibility, partners, enhanced CRM, freedom, and flexibility, it can be concluded that Cloud services can help directly and indirectly Finnish SMEs to mitigate and overcome international growth challenges and enable further business growth.

SME Growth Strategy - Exploring New Market Segments

Tämän tutkimuksen tarkoituksena oli tunnistaa uusia markkinoita ja markkinasegmenttejä yrityksen kasvustrategiaa varten. Yritys on kasvanut erittäin voimakkaasti viime vuosina nykyisillä markkinoillaan ja haluaa nyt jatkaa kasvuaan uusilla markkinoilla. Työ koostuu kirjallisuuskatsauksesta ja empiirisestä osuudesta. Kirjallisuuskatsauksessa on tunnistettu useita pk-yrityksen kasvuun vaikuttavia tekijöitä, uusien markkinoiden kartoittamiseen ja arvioimiseen vaikuttavia tekijöitä, ja lisäksi liiketoimintastrategiaan ja markkinasegmentin valintaan vaikuttavia tekijöitä. Empiirisen osuuden data kerättiin 60 asiantuntijahaastattelusta, joilla kartoitettiin kokonaismarkkinoita ja saatiin tarkkaa markkinainformaatiota kohdemarkkinoista. Tutkimuksessa tunnistettiin kymmenen potentiaalista toimialaa ja 54 potentiaalista markkinasegmenttiä. Kirjallisuudesta tunnistetuilla kriteereillä, teknologisin perustein ja yrityksen omien näkökulmien perusteella näistä valittiin kohdemarkkinoiksi kahdeksan. Tutkimuksessa kehitettiin markkinasegmenttien arviointiin malli. Mallin avulla valitut kahdeksan markkinasegmenttiä listattiin järjestykseen niiden houkuttelevuuden perusteella. Jatkotoimenpiteenä yrityksessä tullaan kehittämään kasvustrategia perustuen tutkimuksen tuloksille.

Teacher professional growth in an authentic learning environment

The last several decades have been marked by tremendous changes in education - technological, pedagogical, administrative, and social. These changes have led to considerable increments in the budgets devoted to professional development for teachers ~ with the express purpose of helping them accommodate their practices to the new realities oftheir classrooms. However, research has suggested that, in spite of the emphasis placed on encouraging sustained change in teaching practices, little has been accomplished. This begs the question of what ought to be done to not only reverse this outcome, but contribute to transformational change. The literature suggests some possibilities including: a) considering teachers as learners and applying what, is known about cognition and learning; b) modifying the location and nature ofprofessional development so that it is authentic, based in the classroom and focusing on tasks meaningful to the teacher; c) attending to the infrastructure underlying professional development; and d) ensuring opportunities for reflective practice. This dissertation looks at the impact of each ofthese variables through an analysis ofthe learning journeys of a group ofteachers engaged in a program called GrassRoots in one midsized school board in Ontario. Action research was conducted by the researcher in his role as consultant facilitating teacher professional growth around the use of Web sites as culminating performance tasks by students. Research focused on the pedagogical approach to the learning of the teachers involved and the infrastructure underlying their learning. Using grounded theory, a model for professional development was developed that can be used in the future to inform practices and, hopefully, lead to sustained transformational school change.