Anchor or Accelerate – A Study on Cancer Cell Adhesion and Motility

Cell migration and adhesion to the extracellular matrix (ECM) are crucial in many biological and pathological processes such as morphogenesis, tissue repair, inflammatory responses, survival, and cancer. Cell-matrix adhesion is mediated by the integrin family of transmembrane receptors, which not only anchor cells to their surroundings, but also transmit bidirectional signalling at the cell surface and couple the ECM to the cytoskeleton. Another group of adhesion receptors are the syndecan proteoglycans, which engage the ECM and possess signalling activity in response to a variety of ligands. Cell migration is a complex process that requires spatial and temporal coordination of adhesion, cell contractility, intracellular traffic of integrins, and matrix turnover by matrix metalloproteinases (MMPs). Thus, integrins and syndecans, as well as MMPs, play essential roles in cancer cell migration and invasion. The understanding of the cooperation of syndecans and integrins was broadened in this thesis study. The results reveal that syndecan-1 functions in concert with 21 integrin in cell adhesion to collagen, whereas syndecan-4 is essential in 21 integrin-mediated matrix contraction. Finally, oncogenic K-Ras was shown to regulate 21 integrin, membrane-type 1 MMP, and syndecan-1 and -4 expression and their cooperation in cell invasion. Epithelial-mesenchymal transition (EMT) is fundamental during embryogenesis and organ development. Activation of EMT processes, including the upregulation of mesenchymal intermediate filament protein vimentin, has also been implicated in the acquisition of a malignant phenotype by epithelial cancer cells. Members of the protein kinase C (PKC) superfamily are involved in cell migration and various integrindependent cellular functions. One aim of this work was to shed light on the role of vimentin in the regulation of integrin traffic and cell motility. In addition, the mechanism by which vimentin participates in EMT was investigated. The results show that integrin recycling and motility are dependent on the PKC–mediated phosphorylation of vimentin. In addition, vimentin was found to be a positive regulator of EMT and regulate the expression of several migratory genes. Specifically, vimentin governs the expression of receptor tyrosine kinase Axl, which is implicated in tumour growth and metastasis. Taken together, the findings described in this thesis reveal novel aspects of the complex interplay between distinct cellular components: integrins, syndecans, and the vimentin cytoskeleton, which all contribute to the regulation of human cancer cell adhesion, migration, and invasion.

Matrix- based logic for avoiding paradoxes and its paraconsistent alternative

The present article shows that there are consistent and decidable many- valued systems of propositional logic which satisfy two or all the three criteria for non- trivial inconsistent theories by da Costa (1974). The weaker one of these paraconsistent system is also able to avoid a series of paradoxes which come up when classical logic is applied to empirical sciences. These paraconsistent systems are based on a 6- valued system of propositional logic for avoiding difficulties in several domains of empirical science (Weingartner (2009)).

Nuclear Matrix in Apoptotic Cell Death and Cell Proliferation. The role of Nuclear Mitotic Apparatus (NuMA) Protein

The nucleus is a membrane enclosed organelle containing most of the genetic information of the cell in the form of chromatin. The nucleus, which can be divided into many sub-organelles such as the nucleoli, the Cajal bodies and the nuclear lamina, is the site for several essential cellular functions such as the DNA replication and its regulation and most of the RNA synthesis and processing. The nucleus is often affected in disease: the size and the shape of the nucleus, the chromatin distribution and the size of the nucleoli have remained the basis for the grading of several cancers. The maintenance of the vertebrate body shape depends on the skeleton. Similarly, in a smaller context, the shape of the cell and the nucleus are mainly regulated by the cytoskeletal and nucleoskeletal elements. The nuclear matrix, which by definition is a detergent, DNase and salt resistant proteinaceous nuclear structure, has been suggested to form the nucleoskeleton responsible for the nuclear integrity. Nuclear mitotic apparatus protein, NuMA, a component of the nuclear matrix, is better known for its mitotic spindle organizing function. NuMA is one of the nuclear matrix proteins suggested to participate in the maintenance of the nuclear integrity during interphase but its interphase function has not been solved to date. This thesis study concentrated on the role of NuMA and the nuclear matrix as structural and functional components of the interphase nucleus. The first two studies clarified the essential role of caspase-3 in the disintegration of the nuclear structures during apoptosis. The second study also showed NuMA and chromatin to co-elute from cells in significant amounts and the apoptotic cleavage of NuMA was clarified to have an important role in the dissociation of NuMA from the chromatin. The third study concentrated on the interphase function of NuMA showing NuMA depletion to result in cell cycle arrest and the cytoplasmic relocalization of NuMA interaction partner GAS41. We suggest that the relocalization of the transcription factor GAS41 may mediate the cell cycle arrest. Thus, this study has given new aspects in the interactions of NuMA, chromatin and the nuclear matrix.

Pinta-aktiiviaineiden vaikutus kuitulinjan toimintaan

Työssä tutkittiin kahden eri pinta-aktiiviaineen A ja B käytön vaikutusta kahdella eri kuitulinjalla. Molempien pinta-aktiiviaineiden pääfunktiona oli vaikuttaa nes-teen pintajännitykseen, jolloin nesteen tunkeutumisnopeus kuitumatriisiin ja kui-tujen huokosiin paranee. Lisäksi pinta-aktiiviaineilla A ja B oli kyky stabiloida nesteeseen liuenneita uuteainekomponentteja, jolloin uudelleensaostumisen riski pienenee. Koeajoilla pyrittiin vaikuttamaan happidelignifiointivaiheen uuteainetasoihin, alkali- ja happiannoksiin sekä kappareduktioon. Lisäksi tarkasteltiin annostelun vaikutusta kuitulinjan pesurien toimintaan, uuteainetasojen kehittymiseen koko kuitulinjalla sekä valkaisun kemikaalikulutukseen. Työssä kokeiltiin myös pinta-aktiiviaineen B vaikutusta EOP-vaiheen uuteainetasoon. Työn kirjallisuusosassa perehdyttiin puun uuteaineisiin ja niiden käyttäytymiseen kuitulinjalla, pinta-aktiiviaineisiin, pesun teoriaan sekä kuitulinjan kemikaaliannosteluihin vaikuttaviin tekijöihin. Molemmissa koeajoissa pinta-aktiiviaineet A ja B annosteltiin massan joukkoon juuri ennen happivaihetta. EOP-vaiheeseen pinta-aktiiviaine B annostelu tehtiin D0-vaiheen pesurin jälkeiseen massaan. Asetoniuutemääritykset tehtiin massasta ennen happivaihetta, happivaiheen jälkeen ja valkaistulle massalle sekä pinta-aktiiviaine B koeajon yhteydessä EOP-vaiheen jälkeen. Pinta-aktiiviaineiden käytöllä havaittiin olevan positiivinen vaikutus kuitulinjan asetoniuutetasojen kehittymisessä. Verrattuna referenssijaksoon, oli happivaiheen alkali- ja happiannokset pienemmät pinta-aktiiviaineiden käytön aikana ilman, että saavutettu kappareduktio heikkeni. Pesurien toiminnan kannalta pinta-aktiiviaineiden käytöllä oli myönteisiä vaikutuksia pesutehokkuuden ja pesun ta-saisuuden kannalta. Lisäksi havaittiin valkaisussa merkittävä muutos D0- ja D1-vaiheiden klooridioksiveden käytössä ja EOP-vaiheen alkaliannoksessa.

Determining the concrete stiffness matrix through ultrasonic testing

The determination of the modulus tangent (Eci ) and of the modulus secant (Ecs) of the concrete can be done using compression test but, to be simpler, it is used relations with characteristic strength (f ck). Relations are also used to determine the transversal modulus (Gc) and, in the case of the Poisson's ratio (ν), a fixed value 0.20 is established. The objective of this research was to evaluate the use of the ultrasonic propagation waves to determine these properties. For the tests were used specimens with f ck varying from 10 to 35 MPa. For the ultrasonic tests were used cylindrical and cubic specimens. The modulus of deformation obtained by ultrasound was statistically equivalent to the obtained by compression tests. The results of modules obtained using the relations with f ck was far away from those obtained by ultrasound or by compression tests. The Poisson's ratio obtained by ultrasound was superior to the fixed value. We can conclude that the concrete characterization by ultrasound is consistent and, to this characterization the cylindrical specimen, normally used to determine f ck, can be used.

A platform for anti-biofilm assays combining viability, biomass and matrix quantifications in susceptibility assessments of antimicrobials against Staphylococcus aureus biofilms

Bacteria can exist as planktonic, the lifestyle in which single cells exist in suspension, and as biofilms, which are surface-attached bacterial communities embedded in a selfproduced matrix. Most of the antibiotics and the methods for antimicrobial work have been developed for planktonic bacteria. However, the majority of the bacteria in natural habitats live as biofilms. Biofilms develop dauntingly fast high resistance towards conventional antibacterial treatments and thus, there is a great need to meet the demands of effective anti-biofilm therapy. In this thesis project it was attempted to fill the void of anti-biofilm screening methods by developing a platform of assays that evaluate the effect that screened compounds have on the total biomass, viability and the extracellular polysaccharide (EPS) layer of the biofilms. Additionally, a new method for studying biofilms and their interactions with compounds in a continuous flow system was developed using capillary electrochromatography (CEC). The screening platform was utilized with a screening campaign using a small library of cinchona alkaloids. The assays were optimized to be statistically robust enough for screening. The first assay, based on crystal violet staining, measures total biofilm biomass, and it was automated using a liquid handling workstation to decrease the manual workload and signal variation. The second assay, based on resazurin staining, measures viability of the biofilm, and it was thoroughly optimized for the strain used, but was then a very simple and fast method to be used for primary screening. The fluorescent resazurin probe is not toxic to the biofilms. In fact, it was also shown in this project that staining the biofilms with resazurin prior to staining with crystal violet had no effect on the latter and they can be used in sequence on the same screening plate. This sequential addition step was indeed a major improvement on the use of reagents and consumables and also shortened the work time. As a third assay in the platform a wheat germ agglutinin based assay was added to evaluate the effect a compound has on the EPS layer. Using this assay it was found that even if compounds might have clear effect on both biomass and viability, the EPS layer can be left untouched or even be increased. This is a clear implication of the importance of using several assays to be able to find “true hits” in a screening setting. In the pilot study of screening for antimicrobial and anti-biofilm effects using a cinchona alkaloid library, one compound was found to have antimicrobial effect against planktonic bacteria and prevent biofilm formation at low micromolar concentration. To eradicate biofilms, a higher concentration was needed. It was also shown that the chemical space occupied by the active compound was slightly different than the rest of the cinchona alkaloids as well as the rest of the compounds used for validatory screening during the optimization processes of the separate assays.

Immunohistochemical evaluation of fibrillar components of the extracellular matrix of transversalis fascia and anterior abdominal rectus sheath in men with inguinal hernia

OBJECTIVE: to evaluate the role of fibrillar extracellular matrix components in the pathogenesis of inguinal hernias. METHODS: samples of the transverse fascia and of the anterior sheath of the rectus abdominis muscle were collected from 40 men aged between 20 and 60 years with type II and IIIA Nyhus inguinal hernia and from 10 fresh male cadavers (controls) without hernia in the same age range. The staining technique was immunohistochemistry for collagen I, collagen III and elastic fibers; quantification of fibrillar components was performed with an image analysis processing software. RESULTS: no statistically significant differences were found in the amount of elastic fibers, collagen I and collagen III, and the ratio of collagen I / III among patients with inguinal hernia when compared with subjects without hernia. CONCLUSION: the amount of fibrillar extracellular matrix components did not change in patients with and without inguinal hernia.

Loss of PTEN expression and AKT activation in HER2-positive breast carcinomas

PURPOSE: To examine the expression of AKT and PTEN in a series of HER2-positive primary invasive breast tumors using immunohistochemistry, and to associate these expression profiles with classic pathologic features such as tumor grade, hormone receptor expression, lymphatic vascular invasion, and proliferation.METHODS: A total of 104 HER2-positive breast carcinoma specimens were prepared in tissue microarrays blocks for immunohistochemical detection of PTEN and phosphorylated AKT (pAKT). Original histologic sections were reviewed to assess pathological features, including HER2 status and Ki-67 index values. The associations between categorical and numeric variables were identified using Pearson's chi-square test and the Mann-Whitney, respectively.RESULTS: Co-expression of pAKT and PTEN was presented in 59 (56.7%) cases. Reduced levels of PTEN expression were detected in 20 (19.2%) cases, and these 20 tumors had a lower Ki-67 index value. In contrast, tumors positive for pAKT expression [71 (68.3%)] were associated with a higher Ki-67 index value.CONCLUSION: A role for AKT in the proliferation of HER2-positive breast cancers was confirmed. However, immunohistochemical detection of PTEN expression did not correlate with an inhibition of cellular proliferation or control of AKT phosphorylation, suggesting other pathways in these mechanisms of control.

Experimental poisoning by Senecio brasiliensis in calves: quantitative and semi-quantitative study on changes in the hepatic extracellular matrix and sinusoidal cells

Extracellular matrix plays an important role in chronic hepatic lesions and has been studied in experimental intoxication models. However in cattle, studies on chronic disease have focused on the hepatocellular damage and extracellular matrix (ECM) changes are usually overlooked. There are no specific studies on the hepatic ECM in either normal or chronically damaged bovine liver. Thus an experimental model of hepatic toxicity model using Senecio brasiliensis poisoned calves was designed. Senecio brasiliensis contains pyrrolizidine alkaloids which cause either acute or chronic progressive dose dependent liver damage. Five calves were orally fed with 0.38g of dry leaves of S. brasiliensis/kg/day for 24 days. Liver needle biopsy specimens were obtained every 15 days for 60 days. Clinical signs of digestive complications appeared at 3rd week. One calf died on 45th day and four were evaluated up to 60th day. Biopsy samples were processed for routine light microscopy, immuno-histochemistry and transmission electron microscopy. From 30th day on progressive liver damage characterized by hepatocellular ballooning, necrosis, apoptosis and megalocytosis, centrilobular, pericellular and portal fibrosis were seen by light microscopy. Quantitative and semi-quantitative measurements of hepatic ECM components were performed before and after the onset of lesions. Morphometric analysis of total collagen and elastic fiber system was conducted. Total collagen and I and III collagen types progressively increased in throughout the liver of affected calves. Changes in location, amount and disposition of the elastic fiber system were also observed. Then numbers of Kupffer cells were significantly increased at 30th day and total numbers of sinusoidal cells were significantly increased at 45th and 60th days. Liver damage was progressive and irreversible even after the exposure to the plant was discontinued. Severe fibrotic lesions occurred mainly in portal tracts, followed by veno-occlusive and pericellular fibrosis. Collagen types I and III s were present in every normal and damaged liver, with predominance of type I. In affected calves the increase of total collagen and elastic fibers system paralleled the number of total sinusoidal cells.

Strategic Innovation Management Based on Three Dimensions

This thesis examines innovation development needs of firms in a remote rural region. The perspective of the study is in strategic innovation management and three dimensions of innovation development: innovation environment, value delivery and innovation capability. The framework is studied with a theoretical and methodological approach in the context of the development of a regional innovation system and the defining of innovation development needs. The thesis is based on existing innovation management literature, expanding it by examining the features of the three dimensions. The empirical data of the study comprise 50 purposefully selected firms within the region of Pielinen Karelia located in Eastern Finland. Most of the firms (70%) included in the study represent manufacturing firms, and over 90% are small and medium-sized enterprises. The research data consist of two questionnaires and an interview, which were done during 2011 in the connection of a regional development project. The point of view of the research is in regional development and harnessing the innovation capability of the firms within the region. The principal research approach applies soft systems methodology. The study explores the means to foster the innovativeness of firms from the viewpoints of innovation environment, innovation capability and value delivery. In closer detail, the study examines relations between the innovation capability factors, differences in innovation development needs within the value delivery system, between sectors and between firm size categories. The thesis offers three major contributions. First, the study extends earlier research on strategic innovation management by connecting the frameworks of innovation capability, innovation environment and value delivery process to the defining of innovation development needs at the regional level. The results deepen knowledge especially concerning practice-based innovation, peripheral regions and smaller firms. Second, the empirical work, based on a case study, confirms the existence of a structural connection integrating five factors of innovation capability. Statistical evidence is provided especially for the positive impacts of the improvement of absorption capability, marketing capability and networking capability, which are the main weaknesses of firms according to the study. Third, the research provides a methodological contribution by applying the innovation matrix in the defining of the innovation development needs of firms. The study demonstrates how the matrix improves possibility to target policy instruments and innovation services more efficiently through indicating significant differences between the innovation support needs regarding various time horizons and phases of innovation process.

Ultrastructural lesions in the myocardium and kidneys of rabbits in experimental acute Amorimia exotropica poisoning

Abstract: Amorimia exotropica is an important plant associated with sudden death in cattle in Southern Brazil. In order to understand the mechanisms by which A. exotropica causes acute lesions in the heart and kidney of intoxicated animals, an experiment was conducted to determine the histopathology and ultrastructure of myocardial and renal lesions of intoxicated rabbits. After receiving 18g/kg of dried plant, six rabbits died suddenly. At necropsy, the liver was swollen and no other macroscopic lesions were observed. Histologically, centrolobular and midzonal hepatocytes were vacuolated. These vacuoles were strong PAS stained positive, suggesting that they corresponded to glycogen accumulations. In some regions of the ventricular septum and ventricles were found vacuoles of different sizes and the kidneys of two rabbits showed vacuolar degeneration on distal convoluted tubules. Ultrastructurally, the myocardium had cardiomyocytes swelling with separation of myofibrils bundles and rupture and disorganization of the sarcomeres. The mitochondria displayed swelling, disorganization, disruption of the mitochondrial cristae, and electron-dense matrix. Some mitochondria exhibited eccentric projections of their membranes with disruption of both outer and inner membranes. The sarcoplasmic reticulum had no alterations, whereas the T-tubule system was occasionally dilated and ruptured. The kidneys had mitochondrial swelling with disorganization and disruption of the mitochondrial cristae. The vacuoles result from the swelling of the endoplasmatic reticulum and usually were located between two basolateral infoldings and mitochondria, occurring preferentially around the nucleus. The myocytes and T system damages induced by A. exotropica result in acute heart failure and death. Furthermore, this mechanism of cardiotoxicity may be common to all plant containing monofluoroacetate.

Non-linear analysis of laminated metal matrix composites by an integrated micro/macro-mechanical model

The present paper describes an integrated micro/macro mechanical study of the elastic-viscoplastic behavior of unidirectional metal matrix composites (MMC). The micromechanical analysis of the elastic moduli is based on the Composites Cylinder Assemblage model (CCA) with comparisons also draw with a Representative Unit Cell (RUC) technique. These "homogenization" techniques are later incorporated into the Vanishing Fiber Diameter (VFD) model and a new formulation is proposed. The concept of a smeared element procedure is employed in conjunction with two different versions of the Bodner and Partom elastic-viscoplastic constitutive model for the associated macroscopic analysis. The formulations developed are also compared against experimental and analytical results available in the literature.

Matrix Mind : on managerial reframing in multidimensional problem solving

Det är inte ovanligt att man i organisationer ställs inför problem som inte kan hanteras inom befintlig organisationsstruktur. Skälen kan vara att frågorna har många – ibland konflikterande – dimensioner och perspektiv som samtidigt måste beaktas. I den här avhandlingen studeras vilka tankemönster och föreställningar som fanns när grupper av chefer försökte lösa komplexa problem, som inte hade en naturlig organisatorisk hemvist och hur de omsatte dessa tankemönster i handling. Vad kännetecknade det ledarskap som utövades under problemlösningsprocessen? Avhandlingens empiri hämtas från ledarutvecklingsprogram i två internationellt verksamma företag i Sverige, och omfattar 14 verkliga affärsproblem i dessa företag och den process varigenom de löstes. De 14 seminarierna utgör exempel på hur mångdimensionella frågeställningar framgångsrikt hanteras utanför den befintliga organisationsstrukturen. Studien ger, genom att adressera frågor kring tankesätt och ledningsprocesser, en djupare förståelse för förutsättningarna för detta, och lyfter särskilt fram betydelsen av ett ledarskap som inbegriper begreppen intervention, förmåga och omtolkning. Som ett samlat begrepp introduceras bilden att utöva ledarskapet utifrån ett matrix mind. Att påverka strukturer (i vid mening) och därigenom de förmågor som utvecklas, är del i detta ledarskap. Det sker genom interventioner (ingrepp som påverkar relationer i t ex en grupp) och baserades i den aktuella empirin på uppfattningar om värdet av problematisering, erfarenhetsutbyte och av ett språk, som både beskriver och anger inriktning för aktiviteter. Interventioner i strukturer (och till dem knutna processer) beskriver dock bara delvis detta ledarskap. Att leda med ett matrix mind innefattar också ett nyfiket och kreativt förhållningssätt, och att utifrån detta leda omtolkning av problem. I empirin finns flera exempel på detta. Avhandlingen avser att ge ett bidrag inom såväl organisations- som ledarskapsteori.