JNK, a versatile regulator of kinesin-1 transport and cytoskeleton dynamics

C-Jun N-terminal kinase (JNK) is traditionally recognized as a crucial factor in stress response and inducer of apoptosis upon various stimulations. Three isoforms build the JNK subfamily of MAPK; generally expressed JNK1 and JNK2 and brain specific JNK3. Degenerative potency placed JNK in the spotlight as potential pharmacological option for intervention. Unfortunately, adverse effects of potential drugs and observation that expression of only JNK2 and JNK3 are induced upon stress, restrained initial enthusiasm. Notably, JNK1 demonstrated atypical high constitutive activity in neurons that is not responsive to cellular stresses and indicated existence of physiological activity. This thesis aimed at revealing the physiological functions of JNK1 in actin homeostasis through novel effector MARCKS-Like 1 (MARCKSL1) protein, neuronal trafficking mediated by major kinesin-1 motor protein and microtubule (MT) dynamics via STMN2/SCG10. The screen for novel physiological JNK substrates revealed specific phosphorylation of C-terminal end of MARCKSL1 at S120, T148 and T183 both ex vivo and in vitro. By utilizing site-specific mutagenesis, various actin dynamics and migrations assays we were able to demonstrate that JNK1 phosphorylation specifically facilitates F-actin bundling and thus filament stabilisation. Consecutively, this molecular mechanism was proved to enhance formation of filopodia; cell surface projections that allow cell sensing surrounding environment and migrate efficiently. Our results visualize JNK dependent and MARCKSL1 executed induction of filopodia in neurons and fibroblast indicating general mechanism. Subsequently, inactivation of JNK action on MARCKSL1 shifts cellular actin machinery into lamellipodial dynamic arrangement. Tuning of actin cytoskeleton inevitably melds with cell migration. We observed that both active JNK and JNK pseudo-phosphorylated form of MARCKSL1 reduce actin turnover in intact cells leading to overall diminished cell motility. We demonstrate that tumour transformed cells from breast, prostate, lung and muscle-derived cancers upregulate MARCKSL1. We showed on the example of prostate cancer PC-3 cell line that JNK phosphorylation negatively controls MARCKSL1 ability to induce migration, which precedes cancer cell metastasis. The second round of identification of JNK physiological substrates resulted in detection of predominant motor protein kinesin-1 (Kif5). Mass spectrometry detailed analysis showed evident endogenous phosphorylation of kinesin-1 on S176 within motor domain that interacts with MT. In vitro phosphorylation of bacterially expressed kinesin heavy chain by JNK isoforms displayed higher specificity of JNK1 when compared to JNK3. Since, JNK1 is constitutively active in neurons it signified physiological aspect of kinesin-1 regulation. Subsequent biochemical examination revealed that kinesin-1, when not phosphorylated on JNK site, exhibits much higher affinity toward MTs. Expression of the JNK non-phosphorable kinesin-1 mutant in intact cells as well as in vitro single molecule imaging using total internal reflection fluorescence microscopy indicated that the mutant loses normal speed and is not able to move processively into proper cellular compartments. We identify novel kinesin-1 cargo protein STMN2/SCG10, which along with known kinesin-1 cargo BDNF is showing impaired trafficking when JNK activity is inhibited. Our data postulates that constitutive JNK activity in neurons is crucial for unperturbed physiologically relevant transport of kinesin-1 dependant cargo. Additionally, my work helps to validate another novel physiological JNK1 effector STMN2/SCG10 as determinant of axodendritic neurites dynamics in the developing brain through regulation of MT turnover. We show successively that this increased MT dynamics is crucial during developmental radial migration when brain layering occurs. Successively, we are able to show that introduction of JNK phosphorylation mimicking STMN2/SCG10 S62/73D mutant rescues completely JNK1 genetic deletion migration phenotype. We prove that STMN2/SCG10 is predominant JNK effector responsible for MT depolymerising activity and neurite length during brain development. Summarizing, this work describes identification of three novel JNK substrates MARCKSL1, kinesin-1 and STMN2/SCG10 and investigation of their roles in cytoskeleton dynamics and cargo transport. This data is of high importance to understand physiological meaning of JNK activity, which might have an adverse effect during pharmaceutical intervention aiming at blocking pathological JNK action.

Trust in inter-organisational dyadic relationships

Understanding the trust in organisations and the business relationships have been an emerging trend in past years. Researchers and executives and subject matter experts in companies have recognised the importance of managing the trust in inter-organisational relationships. Further, the interest both in academia and in business has raised an interest to under-stand dyadic relationships. The objective of this thesis was to examine trust in inter-organisational and inter-personal dyadic relationships. Trust and the dyadic relationships were analysed in information technolo-gy services and outsourcing industry. Dyads were established between the supplier company operating in the industry and its customers. The empiric part of the study was carried out by an email survey that was sent to 331 respondents resulting with effective response rate of 48.7 per cent. Statistical analyses were applied to analyse the data sample. In general, results indicated reputation, trustworthiness, capability and collaboration in dyadic trust relationships. Experience grown in duration of the relationship affected the trust positively. No drastic differences in trust were recognised when comparing supplier and customer responses.

Towards Inter-organizational Working Capital Management

Working capital is an investment which is tied up into the inventories and accounts receivable and which is released with accounts payable. Due to the current business landscape with tightened financial conditions and finance markets, organizations emphasize efficient working capital management. With efficient working capital management, a company can reduce the need of finance, free up cash, increase profitability, improve liquidity, increase the efficiency of operations, and decrease (financing) costs. From the perspective of an individual company, efficient working capital management means decreasing inventory levels by shortening the cycle time of inventories, decreasing accounts receivable by shortening the trade credit terms and effective collection procedures, and increasing the level of accounts payable by paying the suppliers later. From an inter-organizational perspective, however, working capital should not be sub-optimized by a single company but holistic view to working capital management through the supply chain should be adopted to create value and improve performance together. The purpose of this research is to take academic research as well as practical management towards inter-organizational working capital management. The thesis discusses the benefits as well as mechanisms of working capital management in the inter-organizational context and has two main objectives: (1) to examine the effect of inter-organizational working capital management on performance in the value chain context and (2) to develop models of working capital management for internal as well as inter-organizational value chains. The results of the archival research conducted in the value chain of the pulp and paper industry and the value chain of the automotive industry indicate that companies can increase relative profitability by managing working capital comprehensively by taking into account all three components, and holistically though the value chain. Companies in the value chain benefit from different strategies in working capital management depending on the position of the company in the value chain. This can be taken into account in inter-organizational working capital management. The effects of inter-organizational working capital management actions on the financing costs of working capital were studied via simulations. Simulations also show that the value chain and individual companies benefit from an inter-organizational view to working capital management. Inter-organizational working capital management actions include for example: shortening the cycle time of inventories, reducing product costs, shifting inventories, shortening payment terms, and considering the cost of capital. The thesis also provides solutions for the practical requirements for tools to control working capital. The design science part of the research introduces the adjusted cash conversion cycle (ACCC) model for internal value chains, as well as models for working capital management in the inter-organizational value chain context: the working capital management model (WCMM) and the financial cycle time model (FCTM) designed in corporation and product levels respectively. This research contributes to literature on working capital management and interorganizational accounting. The research gives a holistic, inter-organizational view to the management of working capital. It advances the knowledge in working capital management on operational level, increases knowledge in the recently risen theme of supply chainoriented, collaborative working capital management, combines management accounting research with supply chain management research, and contributes to the demand of practical inter-organizational accounting methods. In addition, the research has strong practical focus as new managerial methods are introduced.

A global transport packaging guideline for selected business areas

In this Master’s Thesis a global transport packaging guideline for selected business areas was compiled for the Fiskars the company, which provides branded consumer goods for home, garden and outdoor use. The business areas included were Home and Garden business areas. The aim of the guideline was to be a comprehensive guide for the suppliers, product development, operations and external vendors of the company. The guideline consists of written instructions, tables and illustrations that provide useful information for players working with transport packages from sourcing through to shipments. As the role of corporate responsibility and sustainability has grown, a part of responsible manufacturing strategy includes using materials that are re-usable, recyclable or recoverable as energy or through composting. Hence packaging waste management implementations of different regions were also inspected. The resulting guide covers a range of topics concerning packaging and its transport. The topics include legal requirements, restricted materials and substances, preferred materials, markings, labeling of boxes, logistics and distribution center requirements, physical testing and an inspection checklist.

Deregulation’s Impact on the Railway Freight Transport Sector’s Future in the Baltic Sea Region

The European transport market has confronted several changes during the last decade. Due to European Union legislative mandates, the railway freight market was deregulated in 2007. The market followed the trend started by other transport modes as well as other previously regulated industries such as banking, telecommunications and energy. Globally, the first country to deregulate the railway freight market was the United States, with the introduction of the Staggers Rail Act in 1980. Some European countries decided to follow suit already before regulation was mandated; among the forerunners were the United Kingdom, Sweden and Germany. The previous research has concentrated only on these countries, which has provided an interesting research gap for this thesis. The Baltic Sea Region consists of countries with different kinds of liberalization paths, including Sweden and Germany, which have been on the frontline, whereas Lithuania and Finland have only one active railway undertaking, the incumbent. The transport market of the European Union is facing further challenges in the near future, due to the Sulphur Directive, oil dependency and the changing structure of European rail networks. In order to improve the accessibility of this peripheral area, further action is required. This research focuses on topics such as the progression of deregulation, barriers to entry, country-specific features, cooperation and internationalization. Based on the research results, it can be stated that the Baltic Sea Region’s railway freight market is expected to change in the future. Further private railway undertakings are anticipated, and these would change the market structure. The realization of European Union’s plans to increase the improved rail network to cover the Baltic States is strongly hoped for, and railway freight market counterparts inside and among countries are starting to enhance their level of cooperation. The Baltic Sea Region countries have several special national characteristics which influence the market and should be taken into account when companies evaluate possible market entry actions. According to thesis interviews, the Swedish market has a strong level of cooperation in the form of an old-boy network, and is supported by a positive attitude of the incumbent towards the private railway undertakings. This has facilitated the entry process of newcomers, and currently the market has numerous operating railway undertakings. A contrary example was found from Poland, where the incumbent sent old rolling stock to the scrap yard rather than sell it to private railway undertakings. The importance of personal relations is highlighted in Russia, followed by the railway market’s strong political bond with politics. Nonetheless, some barriers to entry are shared by the Baltic Sea Region, the main ones being acquisition of rolling stock, bureaucracy and needed investments. The railway freight market is internationalizing, which is perceived via several alliances as well as the increased number of mergers and acquisitions. After deregulation, markets seem to increase the number of railway undertakings at a rather fast pace, but with the passage of time, the larger operators tend to acquire smaller ones. Therefore, it is expected that in a decade’s time, the number of railway undertakings will start to decrease in the deregulation pioneer countries, while the ones coming from behind might still experience an increase. The Russian market is expected to be totally liberalized, and further alliances between the Russian Railways and European railway undertakings are expected to occur. The Baltic Sea Region’s railway freight market is anticipated to improve, and, based on the interviewees’ comments, attract more cargoes from road to rail.

Estudo de competição inter e intraespecífica envolvendo Glycine max (L. ) Merril e Cyperus rotundus (L.), em condições de casa de vegetação

O presente experimento teve por objetivo estudar os efeitos da competição inter e intraespecífica envolvendo Glycine max (L.) Merril e Cyperus rotundus L. sobre as características das plantas e acúmulos de N, P, K, Ca e Mg pelas espécies envolvidas. Para tanto, estipulou-se um tratamento em que se desenvolveram três plantas de Glycine max e outro em que se desenvolveram 3 tubérculos de Cyperus rotundus por vaso. Para estudar os efeitos da competição intraespecífica, em dois outros tratamentos dobraram-se as populações por vaso. No estudo da competição interespecífica permitiu-se o desenvolvimento de três plantas de Glycine max e de três tubérculos de Cyperus rotundus num mesmo vaso. De uma maneira geral, pôde-se observar que principalmente devido às diferenças no hábito de crescimento das duas espécies, a expressão da competição inter e intraespecífica, em cada uma delas, assume aspectos distintos. No caso de Cyperus rotundus, o efeito do dobro da densidade pôde ser compensado, em parte, pela maior produção de manifestações epígeas e de tubérculos, aliados ao maior desenvolvimento dos tubérculos na menor densidade de plantio. No caso da competição interespecífica, os resultados sugerem um efeito decisivo do sombreamento de Leguminosae sobre o comportamento da Cyperaceae. Os padrões de efeitos da competição pelos nutrientes foi determinado também pelas diferenças no recrutamento dos elementos do solo pelas espécies envolvidas.

State of logistics and transport in the Baltic Sea region in 2025

The world’s pace of change is accelerating and new innovations, inventions and technologies come about every day. Change is unavoidable. It is difficult to keep up and even more difficult to prepare for the future. Even though it is not possible to know exactly what will happen in the future, by studying futures people can better anticipate what might lie ahead. By making decisions and realizing the consequences of their choices today, people and governments are able to actively decide how they will act in the future. Both opportunities and pitfalls lie ahead, which encourages actors to make more farsighted decisions. The Baltic Sea region is an interesting area for futures studies. It comprises 11 nations and more than 100 million inhabitants and entails countries with advanced, high-income economies, like Finland, Germany and Denmark, and developing economies, like Russia, Latvia and Lithuania. The western, eastern, northern and southern parts of the region are separated by the Baltic Sea, which at the same time represents a barrier and a facility for trade and travel between the countries belonging to the region The purpose of this study was to uncover the most probable future of transport and logistics in the Baltic Sea region in 2025 by using the Delphi method. Altogether 109 responses were collected in two separate instances from experts in all the Baltic Sea region countries, 56 of whom were defined as academic respondents and 53 of whom business respondents. Only minor differences in the opinions of academic and business experts were discovered, and the larger differences lie between eastern and western response groups. The Baltic Sea region is a very heterogeneous region and the division is clearest between East and West, which differ in political, economic, social, technological and environmental aspects. The probable future of the Baltic Sea region presented in this study is coherent with previous studies on the same subject. The future of the Baltic Sea region in terms of logistics and transport looks quite bright according to the experts who participated in the study. Trade volumes will grow and the importance of logistics and transport to the competitiveness of the region will increase. Respondents from eastern countries seemed to be more optimistic about the future in general. Most differences between opinions could be explained by the gap in technological and infrastructural development between the East and West. As eastern countries are less-developed in some parts of their economies, it is easier for them to improve the technical condition of infrastructure by merely catching up with the western countries.

GROWTH AND YIELD RESPONSE OF MAIZE (Zea mays) TO INTER AND INTRA-ROW WEED COMPETITION UNDER DIFFERENT FERTILIZER APPLICATION METHODS

A field experiment was conducted for two consecutive years to study the effect of fertilizer application methods and inter and intra-row weed-crop competition durations on density and biomass of different weeds and growth, grain yield and yield components of maize. The experimental treatments comprised of two fertilizer application methods (side placement and below seed placement) and inter and intra-row weed-crop competition durations each for 15, 30, 45, and 60 days after emergence, as well as through the crop growing period. Fertilizer application method didn't affect weed density, biomass, and grain yield of maize. Below seed fertilizer placement generally resulted in less mean weed dry weight and more crop leaf area index, growth rate, grain weight per cob and 1000 grain weight. Minimum number of weeds and dry weight were recorded in inter-row or intra-row weed-crop competition for 15 DAE. Number of cobs per plant, grain weight per cob, 1000 grain weight and grain yield decreased with an increase in both inter-row and intra-row weed-crop competition durations. Maximum mean grain yield of 6.35 and 6.33 tha-1 were recorded in inter-row and intra-row weed competition for 15 DAE, respectively.

Inter-annual variations in the floristic and population structure of an herbaceous community of "caatinga" vegetation in Pernambuco, Brazil

The influence of climatic variations on the herbaceous component of the "caatinga" vegetation was examined in the state of Pernambuco, Brazil. A total of 105 1 × 1 m-plots were established, of which 35 were in a level micro-habitat, 35 in a riparian micro-habitat, and 35 in a stony microhabitat. During two consecutive years all herbaceous plants in these plots were counted, measured (height and diameter), and collected for identification. The Shannon-Wiener diversity index and the equitability were calculated for each year, as well as the density, frequency, dominance, total basal area and importance index for each species. The total annual pluviometric was 819.5 and 448.8 mm in 2002 and 2003, respectively. The herbaceous flora in the study area was composed of 71 species, of which 58 were sampled in the plots. The families with the greatest species richness were Malvaceae (8 species), Euphorbiaceae (7), Poaceae (6), Convolvulaceae (4), Fabaceae (4), and Portulacaceae (4). The diversity indices were 2.66 and 3.01 nats ind-1 in 2002 and 2003, respectively. The density, frequency, dominance and importance value of herbaceous populations, as well as, the height and diameter of plants were low in the dryer year. The riparian group was the most isolated of the microhabitats examined, both in terms of its floristic and its population structure. Annual seasonal climatic variations greatly modified these populations structure during the course of this study, emphasizing the fact that long-term studies are needed in order to better understand the dynamics of the herbaceous component of the "caatinga" vegetation.

Modal testing and numerical modeling of the dynamic properties of layered sheet-steel structure

Recently, due to the increasing total construction and transportation cost and difficulties associated with handling massive structural components or assemblies, there has been increasing financial pressure to reduce structural weight. Furthermore, advances in material technology coupled with continuing advances in design tools and techniques have encouraged engineers to vary and combine materials, offering new opportunities to reduce the weight of mechanical structures. These new lower mass systems, however, are more susceptible to inherent imbalances, a weakness that can result in higher shock and harmonic resonances which leads to poor structural dynamic performances. The objective of this thesis is the modeling of layered sheet steel elements, to accurately predict dynamic performance. During the development of the layered sheet steel model, the numerical modeling approach, the Finite Element Analysis and the Experimental Modal Analysis are applied in building a modal model of the layered sheet steel elements. Furthermore, in view of getting a better understanding of the dynamic behavior of layered sheet steel, several binding methods have been studied to understand and demonstrate how a binding method affects the dynamic behavior of layered sheet steel elements when compared to single homogeneous steel plate. Based on the developed layered sheet steel model, the dynamic behavior of a lightweight wheel structure to be used as the structure for the stator of an outer rotor Direct-Drive Permanent Magnet Synchronous Generator designed for high-power wind turbines is studied.