Identification and characterisation of a novel adhesin of Streptococcus suis and its use as a target of adhesion inhibition and bacterial detection

Streptococcus suis is an important pig pathogen but it is also zoonotic, i.e. capable of causing diseases in humans. Human S. suis infections are quite uncommon but potentially life-threatening and the pathogen is an emerging public health concern. This Gram-positive bacterium possesses a galabiose-specific (Galalpha1−4Gal) adhesion activity, which has been studied for over 20 years. P-fimbriated Escherichia coli−bacteria also possess a similar adhesin activity targeting the same disaccharide. The galabiose-specific adhesin of S. suis was identified by an affinity proteomics method. No function of the protein identified was formerly known and it was designated streptococcal adhesin P (SadP). The peptide sequence of SadP contains an LPXTG-motif and the protein was proven to be cell wall−anchored. SadP may be multimeric since in SDS-PAGE gel it formed a protein ladder starting from about 200 kDa. The identification was confirmed by producing knockout strains lacking functional adhesin, which had lost their ability to bind to galabiose. The adhesin gene was cloned in a bacterial expression host and properties of the recombinant adhesin were studied. The galabiose-binding properties of the recombinant protein were found to be consistent with previous results obtained studying whole bacterial cells. A live-bacteria application of surface plasmon resonance was set up, and various carbohydrate inhibitors of the galabiose-specific adhesins were studied with this assay. The potencies of the inhibitors were highly dependent on multivalency. Compared with P-fimbriated E. coli, lower concentrations of galabiose derivatives were needed to inhibit the adhesion of S. suis. Multivalent inhibitors of S. suis adhesion were found to be effective at low nanomolar concentrations. To specifically detect galabiose adhesin−expressing S. suis bacteria, a technique utilising magnetic glycoparticles and an ATP bioluminescence bacterial detection system was also developed. The identification and characterisation of the SadP adhesin give valuable information on the adhesion mechanisms of S. suis, and the results of this study may be helpful for the development of novel inhibitors and specific detection methods of this pathogen.

Essays on auditor independence

Auditor independence is a cornerstone of the auditing profession and the basic principle that underpins the reputation of the auditing profession in the public eye. Indeed, it is the attribute most demanded from auditors by the public. Therefore, the sustainability of the auditing profession depends on how auditors can protect this principle. This dissertation consists of four interrelated essays concerned with auditor independence. Specifically, it examines situations that can threaten and impair auditor independence. In addition, this dissertation also examines several variables that may enhance and protect auditor independence. The first essay aims to examine the impact of social pressures occurring within audit firms on auditors’ judgment in the setting of a society with “high power distance” and “low individualism” cultural dimensions. The social pressures consisted of obedience pressure exerted by an auditor’s superior and conformity pressure exerted by an auditor’s colleague. Moreover, two moderating variables—a multi-dimensional professional commitment and locus of control—were included as moderator variables in the relationship between the social pressures faced by auditors and their judgment. The findings show that obedience and conformity pressures influence auditor judgment. Auditors who face the social pressures will make a judgment that may be even diametrically opposite to the independence principle. The findings also indicate that a multi-dimensional professional commitment and locus of control may potentially influence auditor judgment in a situation with social pressures. The second essay aims to investigate the association of advocacy and familiarity threats caused by auditor fee dependence and auditor tenure on auditor independence based on Finnish data, law, and auditing environment. This essay was motivated by the Green Paper on Audit Policy, published by the European Commission in 2010 that questions whether the maximum fee collected from a client should be regulated and whether consecutive assignments should be limited, among others. Contrary to popular belief, this essay does not find evidence that audit fees and long auditor tenure will jeopardise auditor independence. Therefore, the findings do not support policies to regulate auditor fees or limit auditor tenure in Finland. The third essay aims to examine the effect of client intimidation on auditor independence in an audit-client conflict situation. Intimidation threat is one of five independence threats that are explicitly referenced in the IFAC’s independence framework. Client intimidation was manifested in the client threatening to replace the auditor if the auditor did not adopt the client’s position. In addition, this essay examines the role of auditor’s perceived pressure and multi-dimensions of professional commitment as moderator variables. The findings suggest that auditors who experience client intimidation in an audit conflict situation are more likely to have their independence impaired than those who are in a similar situation but without client intimidation. Moreover, auditors who experience client intimidation perceive higher pressure than those who do not experience intimidation. Finally, auditors’ affective and continuance professional commitment dimensions moderate the relationship between auditors’ perceived pressures and auditor independence. The aim of the fourth essay is twofold. First, it aims to develop a scale for measuring auditors’ reputation awareness. Second, it aims to examine the correlation between the levels of auditor reputation awareness and auditor independence. A seven-item scale was developed as the reputation awareness scale. The findings indicate that the scale consists of one dimension. It also has a level of satisfactory reliability and a high level of validity. The findings show that there is a positive correlation between the level of auditors’ reputation awareness and auditor independence.

Acquisition target selection of a serial acquirer : case KONE Oyj

Acquisitions are a way for a company to grow, enter new geographical areas, buy out competition or diversify. Acquisitions have recently grown in both size and value. Despite of this, only approximately 25 percent of acquisitions reach their targets and goals. Companies making serial acquisitions seem to be exceptionally successful and succeed in the majority of their acquisitions. The main research question this study aims to answer is: “What issues impact the selection of acquired companies from the point of view of a serial acquirer? The main research question is answered through three sub questions: “What is a buying process for a serial acquirer like?”, “What are the motives for a serial acquirer to buy companies?” and “What is the connection between company strategy and serial acquisitions?”. The case company KONE is a globally operating company which mainly produces and maintains elevators and escalators. Its headquarter is located in Helsinki, Finland. The company has a long history of making acquisitions and does 20- 30 acquisitions a year. By a key person interview, the acquisition process of the case company is compared with the literature about successful serial acquirers. The acquisition motives in this case are reflected upon three of the acquisition motive theories by Trautwein: efficiency theory, monopoly theory and valuation theory. The linkage between serial acquisitions and company strategy is studied through the key person interview. The main research findings are that the acquisition process of KONE is compatible with a successful acquisition process recognized in literature (RAID). This study confirms the efficiency theory as an acquisition motive and more closely the operational synergies. The monopoly theory can only vaguely be supported by this study, but cannot be totally rejected because of the structure of the industry. The valuation theory does not get any support in this study and can therefore be rejected. The linkage between company strategy and serial acquisitions is obvious and making acquisitions can be seen as growth strategy and a part of other company strategies.

A Modified phosphate-carrier protein theory is proposed as a non-target site mechanism For glyphosate resistance in weeds

Glyphosate is an herbicide that inhibits the enzyme 5-enolpyruvyl-shikimate-3-phosphate synthase (EPSPs) (EC 2.5.1.19). EPSPs is the sixth enzyme of the shikimate pathway, by which plants synthesize the aromatic amino acids phenylalanine, tyrosine, and tryptophan and many compounds used in secondary metabolism pathways. About fifteen years ago it was hypothesized that it was unlikely weeds would evolve resistance to this herbicide because of the limited degree of glyphosate metabolism observed in plants, the low resistance level attained to EPSPs gene overexpression, and because of the lower fitness in plants with an altered EPSPs enzyme. However, today 20 weed species have been described with glyphosate resistant biotypes that are found in all five continents of the world and exploit several different resistant mechanisms. The survival and adaptation of these glyphosate resistant weeds are related toresistance mechanisms that occur in plants selected through the intense selection pressure from repeated and exclusive use of glyphosate as the only control measure. In this paper the physiological, biochemical, and genetic basis of glyphosate resistance mechanisms in weed species are reviewed and a novel and innovative theory that integrates all the mechanisms of non-target site glyphosate resistance in plants is presented.

Resistance to ACCase inhibitors in Eleusine indica from Brazil involves a target site mutation

Eleusine indica (goosegrass) is a diploid grass weed which has developed resistance to ACCase inhibitors during the last ten years due to the intensive and frequent use of sethoxydim to control grass weeds in soybean crops in Brazil. Plant dose-response assays confirmed the resistant behaviour of one biotype obtaining high resistance factor values: 143 (fenoxaprop), 126 (haloxyfop), 84 (sethoxydim) to 58 (fluazifop). ACCase in vitro assays indicated a target site resistance as the main cause of reduced susceptibility to ACCase inhibitors. PCR-generated fragments of the ACCase CT domain of the resistant and sensitive reference biotype were sequenced and compared. A point mutation was detected within the triplet of aspartate at the amino acid position 2078 (referred to EMBL accession no. AJ310767) and resulted in the triplet of glycine. These results constitute the first report on a target site mutation for a Brazilian herbicide resistant grass weed.

Imazapyr herbicide efficacy on floating macrophyte control and ecotoxicology for non-target organisms

the aims of this study were to determine imazapyr efficacy for floating macrophyte control and ecotoxicology for non-target organisms. For the floating macrophyte control efficacy tests were used the doses of 0,5; 1,0; 2,0; 2,5; 3,0; 3,5 and 4,0 L ha-1 and a control with 10 replicates. The acute toxicology for non-target organisms was estimated by lethal concentration 50% (LC50 and EC50). The floating macrophyte control efficacy was over 90%. Imazapyr was classified as moderately toxic for the following biomarkers: L. minor, H. eques, B. rerio, P. caudimaculatus, P. canaliculata, and P. mesopotamicus and lightly toxic for A. caroliniana. Thus, imazapyr herbicide is a tool with great potential to be used on floating macrophyte control (E. crassipes, P. stratiotes e S. molesta) in Brazil and this practice can be evaluated by the use of application biomarkers.

Target audience analysis in cyber domain

This thesis aims to find an effective way of conducting a target audience analysis (TAA) in cyber domain. There are two main focal points that are addressed; the nature of the cyber domain and the method of the TAA. Of the cyber domain the object is to find the opportunities, restrictions and caveats that result from its digital and temporal nature. This is the environment in which the TAA method is examined in this study. As the TAA is an important step of any psychological operation and critical to its success, the method used must cover all the main aspects affecting the choice of a proper target audience. The first part of the research was done by sending an open-ended questionnaire to operators in the field of information warfare both in Finland and abroad. As the results were inconclusive, the research was completed by assessing the applicability of United States Army Joint Publication FM 3-05.301 in the cyber domain via a theory-based content analysis. FM 3- 05.301 was chosen because it presents a complete method of the TAA process. The findings were tested against the results of the questionnaire and new scientific research in the field of psychology. The cyber domain was found to be “fast and vast”, volatile and uncontrollable. Although governed by laws to some extent, the cyber domain is unpredictable by nature and not controllable to reasonable amount. The anonymity and lack of verification often present in the digital channels mean that anyone can have an opinion, and any message sent may change or even be counterproductive to the original purpose. The TAA method of the FM 3-05.301 is applicable in the cyber domain, although some parts of the method are outdated and thus suggested to be updated if used in that environment. The target audience categories of step two of the process were replaced by new groups that exist in the digital environment. The accessibility assessment (step eight) was also redefined, as in the digital media the mere existence of a written text is typically not enough to convey the intended message to the target audience. The scientific studies made in computer sciences and both in psychology and sociology about the behavior of people in social media (and overall in cyber domain) call for a more extensive remake of the TAA process. This falls, however, out of the scope of this work. It is thus suggested that further research should be carried out in search of computer-assisted methods and a more thorough TAA process, utilizing the latest discoveries of human behavior. ---------------------------------------------------------------------------------------------------------------------------------- Tämän opinnäytetyön tavoitteena on löytää tehokas tapa kohdeyleisöanalyysin tekemiseksi kybertoimintaympäristössä. Työssä keskitytään kahteen ilmiöön: kybertoimintaympäristön luonteeseen ja kohdeyleisöanalyysin metodiin. Kybertoimintaympäristön osalta tavoitteena on löytää sen digitaalisesta ja ajallisesta luonteesta juontuvat mahdollisuudet, rajoitteet ja sudenkuopat. Tämä on se ympäristö jossa kohdeyleisöanalyysiä tarkastellaan tässä työssä. Koska kohdeyleisöanalyysi kuuluu olennaisena osana jokaiseen psykologiseen operaatioon ja on onnistumisen kannalta kriittinen tekijä, käytettävän metodin tulee pitää sisällään kaikki oikean kohdeyleisön valinnan kannalta merkittävät osa-alueet. Tutkimuksen ensimmäisessä vaiheessa lähetettiin avoin kysely informaatiosodankäynnin ammattilaisille Suomessa ja ulkomailla. Koska kyselyn tulokset eivät olleet riittäviä johtopäätösten tekemiseksi, tutkimusta jatkettiin tarkastelemalla Yhdysvaltojen armeijan kenttäohjesäännön FM 3-05.301 soveltuvuutta kybertoimintaympäristössä käytettäväksi teorialähtöisen sisällönanalyysin avulla. FM 3-05.301 valittiin koska se sisältää kokonaisvaltaisen kohdeyleisöanalyysiprosessin. Havaintoja verrattiin kyselytutkimuksen tuloksiin ja psykologian uusiin tutkimuksiin. Kybertoimintaympäristö on tulosten perusteella nopea ja valtava, jatkuvasti muuttuva ja kontrolloimaton. Vaikkakin lait hallitsevat kybertoimintaympäristöä jossakin määrin, on se silti luonteeltaan ennakoimaton eikä sitä voida luotettavasti hallita. Digitaalisilla kanavilla usein läsnäoleva nimettömyys ja tiedon tarkastamisen mahdottomuus tarkoittavat että kenellä tahansa voi olla mielipide asioista, ja mikä tahansa viesti voi muuttua, jopa alkuperäiseen tarkoitukseen nähden vastakkaiseksi. FM 3-05.301:n metodi toimii kybertoimintaympäristössä, vaikkakin jotkin osa-alueet ovat vanhentuneita ja siksi ne esitetään päivitettäväksi mikäli metodia käytetään kyseisessä ympäristössä. Kohdan kaksi kohdeyleisökategoriat korvattiin uusilla, digitaalisessa ympäristössä esiintyvillä ryhmillä. Lähestyttävyyden arviointi (kohta 8) muotoiltiin myös uudestaan, koska digitaalisessa mediassa pelkkä tekstin läsnäolo ei sellaisenaan tyypillisesti vielä riitä halutun viestin välittämiseen kohdeyleisölle. Tietotekniikan edistyminen ja psykologian sekä sosiologian aloilla tehty tieteellinen tutkimus ihmisten käyttäytymisestä sosiaalisessa mediassa (ja yleensä kybertoimintaympäristössä) mahdollistavat koko kohdeyleisöanalyysiprosessin uudelleenrakentamisen. Tässä työssä sitä kuitenkaan ei voida tehdä. Siksi esitetäänkin että lisätutkimusta tulisi tehdä sekä tietokoneavusteisten prosessien että vielä syvällisempien kohdeyleisöanalyysien osalta, käyttäen hyväksi viimeisimpiä ihmisen käyttäytymiseen liittyviä tutkimustuloksia.

Choosing the target loci : heat shock factors HSF1 and HSF2 as regulators of cell stress and development

Heat shock factors (HSFs) are an evolutionarily well conserved family of transcription factors that coordinate stress-induced gene expression and direct versatile physiological processes in eukaryote organisms. The essentiality of HSFs for cellular homeostasis has been well demonstrated, mainly through HSF1-induced transcription of heat shock protein (HSP) genes. HSFs are important regulators of many fundamental processes such as gametogenesis, metabolic control and aging, and are involved in pathological conditions including cancer progression and neurodegenerative diseases. In each of the HSF-mediated processes, however, the detailed mechanisms of HSF family members and their complete set of target genes have remained unknown. Recently, rapid advances in chromatin studies have enabled genome-wide characterization of protein binding sites in a high resolution and in an unbiased manner. In this PhD thesis, these novel methods that base on chromatin immunoprecipitation (ChIP) are utilized and the genome-wide target loci for HSF1 and HSF2 are identified in cellular stress responses and in developmental processes. The thesis and its original publications characterize the individual and shared target genes of HSF1 and HSF2, describe HSF1 as a potent transactivator, and discover HSF2 as an epigenetic regulator that coordinates gene expression throughout the cell cycle progression. In male gametogenesis, novel physiological functions for HSF1 and HSF2 are revealed and HSFs are demonstrated to control the expression of X- and Y-chromosomal multicopy genes in a silenced chromatin environment. In stressed human cells, HSF1 and HSF2 are shown to coordinate the expression of a wide variety of genes including genes for chaperone machinery, ubiquitin, regulators of cell cycle progression and signaling. These results highlight the importance of cell type and cell cycle phase in transcriptional responses, reveal the myriad of processes that are adjusted in a stressed cell and describe novel mechanisms that maintain transcriptional memory in mitotic cell division.

Target audience and message alignment for integrated marketing communication of luxury products: entering the Chinese market

The purpose of this study is to identify opportunities to match marketing communication message strategies with the target audience characteristics in the Chinese luxury market entry context. Therefore, consumer behaviour and psychographic marketing segmentation fields are being reviewed in a holistic view in order to identify the similarities and connection points. Through the analysis of the messages in advertisements placed in a certain luxury and fine living magazine, message creation strategies are being anticipated.

Na,K-ATPase: a molecular target for Leptospira interrogans endotoxin

On the basis of our report that a glycolipoprotein fraction (GLP) extracted from Leptospira interrogans contains a potent inhibitor of renal Na,K-ATPase, we proposed that GLP-induced inhibition of Na,K-ATPase might be the primary cellular defect in the physiopathology of leptospirosis. The present study was designed to test this hypothesis by determining whether or not 1) GLP inhibits all the isoforms of Na,K-ATPase which are expressed in the tissues affected by leptospirosis, 2) Na,K-ATPase from leptospirosis-resistant species, such as the rat, is sensitive to GLP, 3) GLP inhibits Na,K-ATPase from intact cells, and 4) GLP inhibits ouabain-sensitive H,K-ATPase. The results indicate that in the rabbit, a leptospirosis-sensitive species, GLP inhibits with similar efficiency (apparent IC50: 120-220 µg protein GLP/ml) all isoforms of Na,K-ATPase known to be expressed in target tissues for the disease. Na,K-ATPase from rat kidney displays a sensitivity to GLP similar to that of the rabbit kidney enzyme (apparent IC50: 25-80 and 50-150 µg protein GLP/ml for rat and rabbit, respectively), indicating that resistance to the disease does not result from the resistance of Na,K-ATPase to GLP. GLP also reduces ouabain-sensitive rubidium uptake in rat thick ascending limbs (pmol mm-1 min-1 ± SEM; control: 23.8 ± 1.8; GLP, 88 µg protein/ml: 8.2 ± 0.9), demonstrating that it is active in intact cells. Finally, GLP had no demonstrable effect on renal H,K-ATPase activity, even on the ouabain-sensitive form, indicating that the active principle of GLP is more specific for Na,K-ATPase than ouabain itself. Although the hypothesis remains to be demonstrated in vivo, the present findings are compatible with the putative role of GLP-induced inhibition of Na,K-ATPase as an initial mechanism in the physiopathology of leptospirosis

Inhibitory effects in the detection of 1 cpd jo targets superimposed to angular frequency stimuli or sinewave gratings

Independence among channels processing different aspects of spatial information, including orthogonal stimuli, has been generally assumed in the literature. We tested independence between the processing of jo targets and the processing of either vertical sinusoidal gratings or angular frequency stimuli with suprathreshold summation. We found the detection of a jo target at 1 cpd to be affected in an inhibitory fashion by either background angular frequencies in the range of 3-96 cycles or sinewave gratings in the range of 0.8-3.0 cpd. These results demonstrate interactions both among orthogonal stimuli and among channels processing vertical sinewave gratings and jo target stimuli. Our discussion focuses on the hypothesis of frequency decomposition in polar coordinates

Context-specific independence in graphical models

The theme of this thesis is context-speci c independence in graphical models. Considering a system of stochastic variables it is often the case that the variables are dependent of each other. This can, for instance, be seen by measuring the covariance between a pair of variables. Using graphical models, it is possible to visualize the dependence structure found in a set of stochastic variables. Using ordinary graphical models, such as Markov networks, Bayesian networks, and Gaussian graphical models, the type of dependencies that can be modeled is limited to marginal and conditional (in)dependencies. The models introduced in this thesis enable the graphical representation of context-speci c independencies, i.e. conditional independencies that hold only in a subset of the outcome space of the conditioning variables. In the articles included in this thesis, we introduce several types of graphical models that can represent context-speci c independencies. Models for both discrete variables and continuous variables are considered. A wide range of properties are examined for the introduced models, including identi ability, robustness, scoring, and optimization. In one article, a predictive classi er which utilizes context-speci c independence models is introduced. This classi er clearly demonstrates the potential bene ts of the introduced models. The purpose of the material included in the thesis prior to the articles is to provide the basic theory needed to understand the articles.

Pre- and postnatal nutrition – target for allergy risk reduction

A rapid increase in allergic diseases in Western societies has led to the conclusion that our modern lifestyle is a risk factor for immune dysregulation. Potential culprits and benefactors are searched among early dietary and microbial exposures, which may act to program later allergic disease. The aim of this thesis was to investigate the role of early maternal and child nutrition in reducing the risk of child allergy. The study population comprised of 256 mother – child pairs from families with a history of allergy participating in a randomized controlled dietary counseling and probiotic intervention (Lactobacillus rhamnosus GG and Bifidobacterium lactis Bb12) study from early pregnancy onwards. The dietary counseling aimed for a diet complying with dietary recommendations for pregnant and lactating women, with special attention to fat quality. Maternal dietary counseling was reflected in cord blood fatty acids suggesting better essential fatty acid status in infants in the counseling group. Dietary counseling with probiotics or placebo had no effect on child allergy risk, but associations between maternal diet during pregnancy and breastfeeding and child allergic outcomes were found in secondary analyses. During pregnancy, milk intake was related to decreased and cheese intake to increased risk of child atopic eczema. During breastfeeding, intake of vitamin C was related to increased risk of asthma and intake of egg was related to decreased risk of atopic eczema. The timing of introduction of complementary foods to infant’s diet was not associated with risk of atopic eczema, when adjusted with parental opinion of child allergic symptoms (i.e., potential reverse causality). In conclusion, the results demonstrate that infant fatty acid supply can be modified via maternal dietary changes. In addition, interesting associations of maternal diet with child allergy risk were discovered. However, no difference in the incidence of allergic diseases with dietary counseling was observed. This suggests that more potent dietary interventions might be necessitated to induce clinical risk reduction of allergy. Highrisk families can safely adhere to dietary recommendations for pregnant and lactating women, and the results support the current conception that no additional benefit is gained with delaying introduction of complementary feeding.

Coronary endothelial dysfunction after ischemia and reperfusion: a new therapeutic target?

Although cardiac ischemia is usually characterized as a disease of the myocyte, it is clear that the vasculature, and especially endothelial cells, is also a major target of this pathology. Indeed, using a rat model of ischemia/reperfusion, we were able to detect severe endothelial dysfunction (assessed as a decreased response to acetylcholine) after acute or chronic reperfusion. Given the essential role of the endothelium in the regulation of vascular tone, as well as platelet and leukocyte function, such a severe dysfunction could lead to an increased risk of vasospasm, thrombosis and accelerated atherosclerosis. This dysfunction can be prevented by free radical scavengers and by exogenous nitric oxide. Endothelial dysfunction can also be prevented by preconditioning with brief periods of intermittent ischemia, thus extending to coronary endothelial cells the concept of endogenous protection previously described at the myocyte level. Experiments performed on cultured cells showed that the endothelial protection induced by free radical scavengers or by preconditioning was due to a lesser expression of endothelial adhesion molecules such as intercellular adhesion molecule-1, leading to a lesser adhesion of neutrophils to endothelial cells. Identification of the mechanisms of this protection may lead to the development of new strategies aimed at protecting the vasculature in ischemic heart diseases.

The early facilitatory effect of a peripheral spatially noninformative prime stimulus depends on target stimulus features

We investigated the dependency of the early facilitatory effect of a prime stimulus (S1) on the physical characteristics of the target stimulus (S2). A go-no go reaction time paradigm was used. The S1 was a gray ring and the S2s were a white vertical line, a white horizontal line, a white cross and a white small ring, all inside a white ring with the same dimensions as the S1. S1 onset-S2 onset asynchrony was 100 ms. The stimuli appeared randomly in any one of the quadrants of a monitor screen. The S2 could occur at the same position as the S1 or at a different one. We observed a strong facilitatory effect when the vertical line or the horizontal line was the go stimulus and no effect when the cross was the go stimulus. These results show that the features of the target stimulus can be decisive for the appearance of the facilitatory effect of a peripheral spatially noninformative prime stimulus.