The spindle assembly checkpoint as a drug target - Novel small-molecule inhibitors of Aurora kinases

Cell division (mitosis) is a fundamental process in the life cycle of a cell. Equal distribution of chromosomes between the daughter cells is essential for the viability and well-being of an organism: loss of fidelity of cell division is a contributing factor in human cancer and also gives rise to miscarriages and genetic birth defects. For maintaining the proper chromosome number, a cell must carefully monitor cell division in order to detect and correct mistakes before they are translated into chromosomal imbalance. For this purpose an evolutionarily conserved mechanism termed the spindle assembly checkpoint (SAC) has evolved. The SAC comprises a complex network of proteins that relay and amplify mitosis-regulating signals created by assemblages called kinetochores (KTs). Importantly, minor defects in SAC signaling can cause loss or gain of individual chromosomes (aneuploidy) which promotes tumorigenesis while complete failure of SAC results in cell death. The latter event has raised interest in discovery of low molecular weight (LMW) compounds targeting the SAC that could be developed into new anti-cancer therapeutics. In this study, we performed a cell-based, phenotypic high-throughput screen (HTS) to identify novel LMW compounds that inhibit SAC function and result in loss of cancer cell viability. Altogether, we screened 65 000 compounds and identified eight that forced the cells prematurely out of mitosis. The flavonoids fisetin and eupatorin, as well as the synthetic compounds termed SACi2 and SACi4, were characterized in more detail utilizing versatile cell-based and biochemical assays. To identify the molecular targets of these SAC-suppressing compounds, we investigated the conditions in which SAC activity became abrogated. Eupatorin, SACi2 and SACi4 preferentially abolished the tensionsensitive arm of the SAC, whereas fisetin lowered also the SAC activity evoked by lack of attachments between microtubules (MTs) and KTs. Consistent with the abrogation of SAC in response to low tension, our data indicate that all four compounds inhibited the activity of Aurora B kinase. This essential mitotic protein is required for correction of erratic MT-KT attachments, normal SAC signaling and execution of cytokinesis. Furthermore, eupatorin, SACi2 and SACi4 also inhibited Aurora A kinase that controls the centrosome maturation and separation and formation of the mitotic spindle apparatus. In line with the established profound mitotic roles of Aurora kinases, these small compounds perturbed SAC function, caused spindle abnormalities, such as multi- and monopolarity and fragmentation of centrosomes, and resulted in polyploidy due to defects in cytokinesis. Moreover, the compounds dramatically reduced viability of cancer cells. Taken together, using a cell-based HTS we were able to identify new LMW compounds targeting the SAC. We demonstrated for the first time a novel function for flavonoids as cellular inhibitors of Aurora kinases. Collectively, our data support the concept that loss of mitotic fidelity due to a non-functional SAC can reduce the viability of cancer cells, a phenomenon that may possess therapeutic value and fuel development of new anti-cancer drugs.

Application of social media and ICT-tools in new and small business ventures.

The current research emphasizes on various questions raised and deliberated upon by different entrepreneurs. It provides a valuable contribution to comprehend the importance of social media and ICT-applications. Furthermore, it demonstrates how to support and implement the management consulting and business coaching start-ups with the help of social media and ICT-tools. The thesis presents a literary review from different information systems science, SME and e-business journals, web articles, as well as, survey analysis reports on social media applications. The methodology incorporated into a qualitative research method in which social anthropological approaches were used to oversee the case study activities in order to collect data. The collaborative social research approach was used to shelter the action research method. The research discovered that new business start-ups, as well as small businesses do not use social media and ICT-tools, unlike most of the large corporations use. At present, the current open-source ICT-technologies and social media applications are equally available for new and small businesses as they are available for larger companies. Successful implementation of social media and ICT-applications can easily enhance start-up performance and overcome business hassles. The thesis sheds some light on effective and innovative implementation of social media and ICT-applications for new business risk takers and small business birds. Key words

Working capital management in the small and medium-sized enterprises Case: Project-based business environment in Flowrox Oy

This Master´s thesis illustrates how growing a business ties up the company´s working capital and what the cost of committed capital. In order to manage a company´s working capital in rapid business growth phase, the thesis suggests that by monitoring and managing the operating and cash conversion cycles of customers´ projects, a company can find ways to secure the required amount of capital. The research method of this thesis was based on literature reviews and case study research. The theoretical review presents the concepts of working capital and provides the background for understanding how to improve working capital management. The company in subject is a global small and medium-sized enterprise that manufactures pumps and valves for demanding process conditions. The company is expanding, which creates lots of challenges. This thesis concentrates to the company´s working capital management and its efficiency through the supply chain and value chain perspective. The main elements of working capital management are inventory management, accounts receivable management and accounts payable management. Prepayments also play a significant role, particularly in project-based businesses. Developing companies´ working capital management requires knowledge from different kind of key operations´ in the company, like purchasing, production, sales, logistics and financing. The perspective to develop and describe working capital management is an operational. After literature reviews the thesis present pilot projects that formed the basis of a model to monitor working capital in the case company. Based on analysis and pilot projects, the thesis introduces a rough model for monitoring capital commitments in short time period. With the model the company can more efficiently monitor and manage their customer projects.

Puuelementtien teollisen tuotannon kehittäminen

Tavoitteena tässä diplomityössä oli selvittää puuelementtien teollisen tuotannon mahdollisuuksia ja kehityksen tilaa. Millaiset valmiudet puuelementtien valmistajilla on valmistaa rakennuttajien ja rakentajien edellyttämiä suur- ja tilaelementtejä puukerrostaloja varten, millaista tuotekehitystä ja tuotantoinvestointeja tarvitaan kaiken tämän mahdollistamiseksi, mikä on yritysten halukkuus ja resurssit investoida uuteen teknologiaan sekä kuinka voidaan parantaa tuotannon tehokkuutta. Ensin diplomityössä luotiin katsaus puukerrostalorakentamisen historiaan, nykytilaan, kerrostalojärjestelmiin, palomääräyksiin ja ruotsalaiseen puuelementtien valmistamisen kehityksen tilaan. Puuelementtien valmistukseen tutustumisen jälkeen laadittiin kyselylomake tutkimusta varten, joka lähettiin 64:lle puuelementtien valmistajalle. Tutkimukseen vastasi kaikkiaan 20 puuelementtien valmistajaa. Vastausprosentiksi muodostui 31 %. Tutkimuksen perusteella voidaan todeta puuelementtien valmistajien olevan pieniä ja keskisuuria pk-yrityksiä jotka valmistavat puuelementtejä pääasiassa pientaloteollisuudelle. Suomalaisten valmistajien automaation taso ja tekninen osaaminen ei juuri eroa ruotsalaisesta puuelementtien valmistuksesta. Suomessa puuelementtien valmistus puukerrostaloja varten ei ole vielä vakiintunutta samalla tavalla kuten Ruotsissa. Suomessa Puukerrostalokohteita lähtee liikkeelle tällä hetkellä vielä aivan liian vähän. Kilpailukyky ja tehokkuus tulevat kehittymään nopeasti kun teollinen valmistus saadaan riittävän korkealle tasolle.

Peritoneal fluid changes in horses subjected to small colon distension

Intestinal devitalization in cases of small colon obstruction may be difficult to detect based only in clinical signs. The purpose was to serially evaluate blood and peritoneal fluid of horses subjected to small colon distension. Seventeen adult horses were allotted in three groups. In the small colon-distended group (DG, n=7) a surgically-implanted latex balloon was inflated to promote intraluminal small colon distension. In the shamoperated group (SG, n=5), the balloon was implanted but not inflated, and no surgery was done in the control group (CG, n=5). Blood and peritoneal fluid were sampled before and after (6 samples with a 30-minute interval) intestinal obstruction for cytological and biochemical analyses. No significant changes in clinical signs occurred within groups or across time during the experimental period. There were no statistical differences among SG and SG groups in hematologic and blood chemistry variables. Although total protein concentration and lactate dehydrogenase (LDH) activity in peritoneal fluid remained most of the time within reference values during the experimental period in all groups, increases from baseline values were detected in SG and DG groups. Such increases occurred earlier, progressively and with greater magnitude in the DG when compared with the SG (P<0.05). Increases from baselines values were also observed in total nucleated cells and neutrophils counts in the DG (P<0.05). In conclusion, distension of the equine small colon induced progressive subtle increases in total protein and LDH concentrations in the peritoneal fluid during the first hours. Serial evaluation of these variables in peritoneal fluid may be useful for early detection of intestinal devitalization in clinical cases of equine small colon obstruction.

Effects of starter diet supplementation with arginine on broiler production performance and on small intestine morphometry

The effects of starter diet (days 1 to 21) supplemented with arginine (Arg) on the production performance and duodenum and jejunum mucosa morphometry of broilers were studied. Male Cobb broiler chickens (990) were randomly assigned to one of five treatments in a complete random design. Measurements of 33 chicks per treatment were made in six repetitions. The treatments consisted of a basal diet with 1.390% digestible Arg (no supplementation) and four dietary levels (1.490%, 1.590%, 1.690%, and 1.790%), providing a relationship with lysine of 1.103; 1.183; 1.262; 1.341 and 1.421%, respectively. From the age of 22 days on, all birds received conventional grower diet. The data were submitted to regression analysis by polynomial decomposition of the degrees of freedom in relation to the levels of Arg. The Arg supplementation increased (P<0.05) the live weight and the feed conversion ratio without increasing the feed intake of the birds. However, no effect was observed (P>0.05) in the growth phase (days 22 to 42) in the absence of the Arg supplementation. The supplementation of Arg over of NRC recommendation during the starter phase may be necessary for the expression of the maximal weight gain potential in birds. No effect (P<0.05) of Arg dietary supplementation was observed either on small intestine weight and length at any age. However, the duodenum villus:crypt ratio increased and the crypt depth decreased in the first week in response to increasing dietary Arg. It is concluded that broiler Arg dietary supplementation in the starter diet improved production performance and small intestine morphometry, especially in the first week.

Antimicrobial resistance of Staphylococcus spp. from small ruminant mastitis in Brazil

The study aimed to determine the antimicrobial resistance patterns and to identify molecular resistance markers in Staphylococcus spp. (n=210) isolated from small ruminant mastitis in Brazil. The antimicrobial resistance patterns were evaluated by the disk diffusion test and by detection of the presence of mecA, blaZ, ermA, ermB, ermC and msrA genes by PCR. The efflux pump test was performed using ethidium bromide and biofilm production was determined by Congo red agar test along with PCR for detection of the icaD gene. The isolates were most resistant to amoxicillin (50.0%), streptomycin (42.8%), tetracycline (40.4%), lincomycin (39.0%) and erythromycin (33.8%). Pan-susceptibility to all tested drugs was observed in 71 (33.8%) isolates and 41 Staphylococcus isolates were positive for the efflux pump. Although phenotypic resistance to oxacillin was observed in 12.8% of the isolates, none harbored the mecA gene. However, 45.7% of the isolates harbored blaZ indicating that beta-lactamase production was the main mechanism associated with staphylococci resistance to beta-lactams in the present study. The other determinants of resistance to antimicrobial agents ermA, ermB, ermC, and msrA were observed in 1.4%, 10.4%, 16.2%, and 0.9% of the isolates, respectively. In addition, the icaD gen was detected in 32.9% of the isolates. Seventy three isolates (54 from goats and 19 from sheep) were negative for all resistance genes tested and 69 isolates presented two or more resistance genes. Association among blaZ, ermA, ermB, ermC and efflux pump were observed in 17 isolates, 14 of which originated from goats and three from sheep. The data obtained in this study show the resistance of the isolates to beta-lactamics, which may be associated with the use of antimicrobial drugs without veterinary control.

QT interval in healthy dogs: which method of correcting the QT interval in dogs is appropriate for use in small animal clinics?

The electrocardiography (ECG) QT interval is influenced by fluctuations in heart rate (HR) what may lead to misinterpretation of its length. Considering that alterations in QT interval length reflect abnormalities of the ventricular repolarisation which predispose to occurrence of arrhythmias, this variable must be properly evaluated. The aim of this work is to determine which method of correcting the QT interval is the most appropriate for dogs regarding different ranges of normal HR (different breeds). Healthy adult dogs (n=130; German Shepherd, Boxer, Pit Bull Terrier, and Poodle) were submitted to ECG examination and QT intervals were determined in triplicates from the bipolar limb II lead and corrected for the effects of HR through the application of three published formulae involving quadratic, cubic or linear regression. The mean corrected QT values (QTc) obtained using the diverse formulae were significantly different (ρ<0.05), while those derived according to the equation QTcV = QT + 0.087(1- RR) were the most consistent (linear regression). QTcV values were strongly correlated (r=0.83) with the QT interval and showed a coefficient of variation of 8.37% and a 95% confidence interval of 0.22-0.23 s. Owing to its simplicity and reliability, the QTcV was considered the most appropriate to be used for the correction of QT interval in dogs.

Entrepreneurial orientation in small and medium-sized enterprises during economic crisis

The economic importance of small and medium-sized enterprises (SMEs) and entrepreneurship has increased significantly in recent decades and entrepreneurial activity and SMEs are deemed vital to economic progress. Therefore, it is justifiable to study how small firms and entrepreneurs can enhance their performance and emergence in the turbulent economic environment. The concept of entrepreneurial orientation (EO) has recently attracted considerable attention in the field of entrepreneurship research. EO generally refers to a firm’s propensity to be innovative, to be proactive and to take risks. A majority of EO studies so far have found that adopting EO associated entrepreneurial behaviors will help firms to create or sustain a high level of performance. This dissertation explores the main drivers and performance implications of EO for SMEs in time of economic crisis. Hence the first objective of this dissertation is to examine the performance implications of EO and to test the role of EO on how firms are treated by the crisis at operative level. The second objective is to expand the prevailing understanding of determinants of EO by exploring the relationship between owner's work related values, attitudes, demographic characteristics, firm’s financial resources and EO. EO was found to be a significant and positive factor behind a firm’s long run growth. Hence it can be said that EO has positive implications for firm performance. But on the other hand, during a time of economic crisis the different dimensions of EO had both positive and negative effects on performance of SMEs. The performance implications varied across different stages of the crisis and were also dependent on what measure was used for measuring the performance. The main drivers of EO in SMEs were the personal work related values of the entrepreneur and his/her prior experience as an entrepreneur. The intrinsic work values related to interest, responsibility, challenge, self-development or intellectual stimulation and values related to status, power, achievement and recognition had a positive effect on the level of EO. On the other hand, extrinsic values related to high income, material possessions, benefits such as generous holidays, job security, and comfort through good working conditions decreased the level of EO

Licensing Model Development for Small Modular Reactors (SMRs) - Focusing on the Finnish Regulatory Framework

Recently, Small Modular Reactors (SMRs) have attracted increased public discussion. While large nuclear power plant new build projects are facing challenges, the focus of attention is turning to small modular reactors. One particular project challenge arises in the area of nuclear licensing, which plays a significant role in new build projects affecting their quality as well as costs and schedules. This dissertation - positioned in the field of nuclear engineering but also with a significant section in the field of systems engineering - examines the nuclear licensing processes and their suitability for the characteristics of SMRs. The study investigates the licensing processes in selected countries, as well as other safety critical industry fields. Viewing the licensing processes and their separate licensing steps in terms of SMRs, the study adopts two different analysis theories for review and comparison. The primary data consists of a literature review, semi-structured interviews, and questionnaire responses concerning licensing processes and practices. The result of the study is a recommendation for a new, optimized licensing process for SMRs. The most important SMR-specific feature, in terms of licensing, is the modularity of the design. Here the modularity indicates multi-module SMR designs, which creates new challenges in the licensing process. As this study focuses on Finland, the main features of the new licensing process are adapted to the current Finnish licensing process, aiming to achieve the main benefits with minimal modifications to the current process. The application of the new licensing process is developed using Systems Engineering, Requirements Management, and Project Management practices and tools. Nuclear licensing includes a large amount of data and documentation which needs to be managed in a suitable manner throughout the new build project and then during the whole life cycle of the nuclear power plant. To enable a smooth licensing process and therefore ensure the success of the new build nuclear power plant project, management processes and practices play a significant role. This study contributes to the theoretical understanding of how licensing processes are structured and how they are put into action in practice. The findings clarify the suitability of different licensing processes and their selected licensing steps for SMR licensing. The results combine the most suitable licensing steps into a new licensing process for SMRs. The results are also extended to the concept of licensing management practices and tools.

BIOTULI - Selvitys bioliiketoiminnan uusista liiketoimintamahdollisuuksista ja -malleista

Tämä raportti on osa BIOTULI-projektia, jossa tutkitaan biojalostamoiden uusia tuotteita ja liiketoimintamalleja. Raportin tavoitteena on selvittää, millaisilla liiketoimintamalleilla BIOTULI-projektissa löytyneitä potentiaalisia bioliiketoimintamahdollisuuksia pystytään hyödyntämään pk-yrityksen näkökulmasta. Tavoitteena on myös tutkia millaisen toimitusketjun liiketoimintamalli vaatii, ja millä edellytyksillä sen toteuttaminen on kannattavaa. Raportissa tarkastellaan torrefioinnin ja lämmöntuotannon yhdistämistä sekä BIOTULI-projektissa kehitetyn uuden erottelumenetelmän hyödyntämistä biohajoavan desinfiointiaineen valmistuksessa. Selvitystyö toteutettiin asiantuntijahaastatteluiden ja kirjallisuuskatsauksen perusteella. Molemmille casetapauksille muodostettiin liiketoimintamalli sekä arvioitiin sen toteutettavuutta ja kehitysnäkymiä. Torrefioinnin ja lämmöntuotannon yhdistäminen ei tehdyn analyysin perusteella ole tällä hetkellä kannattavaa, mutta muutokset markkinatilanteessa voivat muuttaa tilannetta tulevaisuudessa. Biohajoavan desinfiointiaineen valmistuksessa on potentiaalia kannattavaan liiketoimintaan, mutta tutkimus on vielä kesken, joten tarkkaa liiketoiminnan tai sen kannattavuuden arviointia ei vielä voi tehdä. Työn tuloksia voi käyttää pohjana tarkemmille kannattavuusarvioille.

Histochemical study of the oviducal gland and analysis of the sperm storage tubules of Mustelus schmitti Springer, 1939 (Chondrichthyes, Triakidae)

Abstract: The paired oviducal glands of immature and mature females of Mustelus schmitti were examined macro and microscopically. Findings indicate that these glands possessed the same zonation as in most chondrichthyans from anterior to posterior: club, papillary, baffle and terminal zones. The whole gland is composed by simple tubular glands that connect with transverse grooves all along the organ. The club zone presents a typical indian club shape with a simple columnar and ciliated epithelium including secretory cells PAS (+) and AB (+). The papillary zone is characterized by lamella forming small and long cones in numbers of three. The epithelium of this zone contains ciliated cells with apical nuclei and secretory cells with basal nuclei that stain AB (+)The baffle zone consists of apically flattened lamellae alternating with spinnerets which are small projections disposed by both sides of the plateau. This whole structure is present in number of 8 or 9 units. A simple columnar ciliated epithelium covers the plateau and spinnerets and no AB or PAS staining is observed. The epithelium of the terminal zone is PAS (-) and AB (+), and elongated tubules, that run adjacent to the baffle zone are the site where groups of spermatozoa are clearly observed in the lumen. The epithelium of the sperm storage tubules do not stain with any of the dyes tested. Sperm was also observed in the baffle zone, presumably in its way to the fecundation in the oviduct because it displays no aggregation pattern and was between the folds of the epithelium. By scanning electron microscopy sperm was observed in the club and baffle zones in a gland which belonged to a pregnant female.