<p>ABSTRACT</p><p>The Swedish Human Proteome Resource (HPR) is a project about mapping of human genes and proteins. It aims to describe the function and distribution of all human genes and corresponding proteins, using in-house produced antibodies and tissue microarrays (TMA) for enzyme based immunohistochemistry. The mono-specific antibodies are used for immunostaining of human tissue. Specific predicted antigens named Protein Epitope Signature Tag (PrEST) are needed to produce mono-specific antibodies. PrEST are produced using recombinant technology from predicted genes and used as immunogens to produce (mono-specific) antibodies in rabbits. In this study, 84 mono-specific antibodies with known specificity to human proteins were used for immunohistochemical analysis of mouse tissues to determine the cross reactivity between mouse and human.</p><p>For 6 of the 84 antibodies the results differed between mouse and human tissue. A comparison between the PrEST used with the mouse proteome using database search programs showed homologies that were less than 100% in these six cases.</p><p>Thus, future studies on cross reactivity will focus on how to increase the accuracy in its prediction at the in silico stage of the experiment.</p>

ABSTRACT The Swedish Human Proteome Resource (HPR) is a project about mapping of human genes and proteins. It aims to describe the function and distribution of all human genes and corresponding proteins, using in-house produced antibodies and tissue microarrays (TMA) for enzyme based immunohistochemistry. The mono-specific antibodies are used for immunostaining of human tissue. Specific predicted antigens named Protein Epitope Signature Tag (PrEST) are needed to produce mono-specific antibodies. PrEST are produced using recombinant technology from predicted genes and used as immunogens to produce (mono-specific) antibodies in rabbits. In this study, 84 mono-specific antibodies with known specificity to human proteins were used for immunohistochemical analysis of mouse tissues to determine the cross reactivity between mouse and human. For 6 of the 84 antibodies the results differed between mouse and human tissue. A comparison between the PrEST used with the mouse proteome using database search programs showed homologies that were less than 100% in these six cases. Thus, future studies on cross reactivity will focus on how to increase the accuracy in its prediction at the in silico stage of the experiment.

The goal of this Master's thesis is to perform valuation of the company PREST PŘEROV a.s. and determine its market value on the date of valuation 20th of December 2011. The theoretical part describes basic terms, general description of the valuation process, strategic and financial analysis methodology and the most commonly used valuation methods. In the practical part of this work, the theoretical foundations are applied on the specific conditions of the valuated company PREST PŘEROV a.s. Strategic and financial analysis of the company is performed. After the formulation of its financial plan the company is valuated using the discounted cash flow method in the entity version, because pre-conditions of the going concern principle are met.

Juvenil idiopatisk artrit (JIA) är en autoimmun reumatisk sjukdom som drabbar barn före 16 års ålder. Den vanligaste varianten är oligoartikulär JIA, vilket innebär att den påverkar två till fyra leder med symptom så som svullnad, ömhet, stelhet, rodnad eller nedsatt motorik. Oligoartikulär JIA är även associerad med uveit, vilket är en autoimmun ögonsjukdom som förstör synen. Sjukdomen är i nuläget dåligt förstådd och forskning behövs för att kunna identifiera markörer som kan användas för diagnostik. Enzymkopplad immunadsorberande analys (ELISA) är inom vårdens laborativa arbete en mycket vanlig metod för att undersöka patienters serum och identifiera specifika markörer som tyder på sjukdom. En ELISA kan utföras på olika sätt beroende på om den letar efter antigen eller antikroppar. Syftet med denna studie var att optimera en indirekt ELISA för detektion av autoantikroppar och genomföra en metodjämförelse med en redan väl undersökt kulbaserad autoantikroppsmetod. Studien omfattar 14 serumprov från olika patienter och använder sig av rekombinant framställda protein epitope signature tag (PrEST)-antigener för att identifiera autoantikroppar. PrEST-antigener består av ett humant variabelt proteinsegment och en protein-tag som består av ett albuminbindande protein (ABP) med en N-terminal HIS-tag (His6ABP). För att förhindra ospecifik inbindning till His6ABP-delen av PrEST-antigenerna så framställdes His6ABP, med vilket serum inkuberades innan de undersöktes med ELISA. Då resultatet tolkades binärt kunde stor likhet ses mellan de två olika metoderna för samtliga PrEST-antigener som undersöktes, vilket innebär att metoden kan fortsätta utforskas. Resultatet visade också att His6ABP-delen skulle kunna orsaka korsreaktivitet, vilket dock inte är något negativt för metodens validitet. / Juvenile idiopathic arthritis (JIA) is an autoimmune rheumatic disease that affects children before the age of 16. The most common variant is oligoarticular JIA, which means it affects two to four joints with symptoms such as swelling, tenderness, stiffness, redness, or reduced mobility. Oligoarticular JIA is also associated with uveitis, an autoimmune eye disease which damage eyesight. The disease is currently poorly understood, and research is needed to identify markers that can be used for diagnostics. Enzyme-linked immunosorbent assay (ELISA) is a very common method in healthcare laboratory work to examine patients' serum and identify specific markers that indicate disease. An ELISA method can be performed in different ways depending on whether it is looking for antigen or antibodies. The aim of this study was to optimize an indirect ELISA for detection of autoantibodies and conduct a method comparison with an already well established bead-based autoantibody array method. The study included 14 sera samples from different patients and used recombinantly produced protein epitope signature tag (PrEST) antigens to identify autoantibodies. PrEST antigens consist of a human variable protein segment and a protein tag consisting of albumin binding protein (ABP) with an N-terminal HIS-tag (His6ABP). To prevent non-specific binding to the His6ABP portion of the PrEST antigens, His6ABP were recombinantly produced and used for pre-absorption of serum before being examined by ELISA. When the result was interpreted binary, great similarity could be seen between the two different methods for all PrEST antigens examined, which calls for further exploring of the method. The result also showed that the His6ABP part could possibly cause cross-reactivity, which is interesting but doesn’t affect the validity of the method.

With the full sequence of the human genome now available, anexciting era in biomedical research has started. The sequenceprovides information about all our genes and greatly increasesthe scope to compare genetic activities in different cells, toanalyze genetic variation between individuals and betweendifferent species and, most importantly, to investigatesystematically the whole genome in a gene-by-gene manner, andthus increase our understanding of gene function. This thesis describes studies in which developments weremade in several areas of functional genomics. Messenger RNAlevels were analyzed by the use of an amplification procedure,in which the 3´-ends of the transcripts were selected inorder to amplify the mRNA population in an unbiased fashion. Bysonicating cDNA originating from expressed mRNA, uniformlysized representatives of the transcripts,“signaturetags”, were obtained. The mRNA levels in the original mRNApopulation correlated well with the levels in the amplifiedmaterial, as verified by microarray analysis and realtimequantitative PCR. The expressed transcripts can be identifiedusing pyrosequencing, by comparing the obtained sequenceinformation from the signature tags to information contained invarious sequence databases. In one of the articles, the use ofpyrosequencing is illustrated by efforts to find genes involvedin the disease progression of atherosclerosis. More challenging than the study of mRNA levels is to analyzewhen, where and how proteins fulfill their wide-ranging rolesin all the various cellular processes. Proteins are morecomplex biomolecules than mRNA, each having unique properties.Current techniques for studying proteins need much improvement,and are often limited to investigations of a specific portionof the proteome. One approach for studying the whole proteomeis to systematically generate reagents with specific affinityfor the proteins encoded by the genome, one by one. Theaffinity reagents can be used as flags for their targets,providing a flag-specific detection system, so that the targetproteins can be sub-cellularly localized in the majority ofhuman tissues in an array format. One of the articles includedin the thesis presents a pilot project for large-scale affinityreagent production. The aim was to provide a sound basis forwhole proteome studies, but as a pilot study this investigationwas limited to the proteins encoded by human chromosome 21. Allputative genes on the chromosome were subjected to antibodygeneration in a systematic manner. Small, uniform, and easilyproduced representative portions of the full-length proteinswere expressed. These were denoted“Protein EpitopeSignature Tags”and were designed to be unique for theirfull-length counterparts. The antibodies were produced inrabbits and two of the articles in the thesis discuss differentapproaches for affinity purification of the antibodies toachieve the highest possible specificity towards the targets.The resulting“mono-specific”, but still“multi-epitope”, antibodies can be used for a widerange of additional biochemical studies, such as protein arrayand protein pull-out analyses. <b>Keywords:</b>functional genomics, 3´-end signaturetags, pyrosequencing, amplification, PrEST, chromosome 21,polyclonal antibodies, dual expression, affinitypurification.

<p>With the full sequence of the human genome now available, anexciting era in biomedical research has started. The sequenceprovides information about all our genes and greatly increasesthe scope to compare genetic activities in different cells, toanalyze genetic variation between individuals and betweendifferent species and, most importantly, to investigatesystematically the whole genome in a gene-by-gene manner, andthus increase our understanding of gene function.</p><p>This thesis describes studies in which developments weremade in several areas of functional genomics. Messenger RNAlevels were analyzed by the use of an amplification procedure,in which the 3´-ends of the transcripts were selected inorder to amplify the mRNA population in an unbiased fashion. Bysonicating cDNA originating from expressed mRNA, uniformlysized representatives of the transcripts,“signaturetags”, were obtained. The mRNA levels in the original mRNApopulation correlated well with the levels in the amplifiedmaterial, as verified by microarray analysis and realtimequantitative PCR. The expressed transcripts can be identifiedusing pyrosequencing, by comparing the obtained sequenceinformation from the signature tags to information contained invarious sequence databases. In one of the articles, the use ofpyrosequencing is illustrated by efforts to find genes involvedin the disease progression of atherosclerosis.</p><p>More challenging than the study of mRNA levels is to analyzewhen, where and how proteins fulfill their wide-ranging rolesin all the various cellular processes. Proteins are morecomplex biomolecules than mRNA, each having unique properties.Current techniques for studying proteins need much improvement,and are often limited to investigations of a specific portionof the proteome. One approach for studying the whole proteomeis to systematically generate reagents with specific affinityfor the proteins encoded by the genome, one by one. Theaffinity reagents can be used as flags for their targets,providing a flag-specific detection system, so that the targetproteins can be sub-cellularly localized in the majority ofhuman tissues in an array format. One of the articles includedin the thesis presents a pilot project for large-scale affinityreagent production. The aim was to provide a sound basis forwhole proteome studies, but as a pilot study this investigationwas limited to the proteins encoded by human chromosome 21. Allputative genes on the chromosome were subjected to antibodygeneration in a systematic manner. Small, uniform, and easilyproduced representative portions of the full-length proteinswere expressed. These were denoted“Protein EpitopeSignature Tags”and were designed to be unique for theirfull-length counterparts. The antibodies were produced inrabbits and two of the articles in the thesis discuss differentapproaches for affinity purification of the antibodies toachieve the highest possible specificity towards the targets.The resulting“mono-specific”, but still“multi-epitope”, antibodies can be used for a widerange of additional biochemical studies, such as protein arrayand protein pull-out analyses.</p><p><b>Keywords:</b>functional genomics, 3´-end signaturetags, pyrosequencing, amplification, PrEST, chromosome 21,polyclonal antibodies, dual expression, affinitypurification.</p>

This thesis argues that the twin concept of separate personality and limited liability from its historical beginnings, has entrenched corporate irresponsibility. It assesses the role that these concepts have played in tackling corporate irresponsibility from their historical origins to the present day, commenting on the lessons learnt. Whilst the institution of the company as a legal person is unquestionably the bedrock of modern company law,1 this thesis examines these concepts not necessarily from the position of disputing the philosophical, economic, or political imperatives, all of which are incredibly important – but from the viewpoint that historically, the principle of separate personality and limited liability entrenches corporate irresponsibility. As such, this thesis suggests a partial abandonment of the separate personality principle because it provides a mechanism for dishonest directors to escape liability for their fraudulent conduct. It also argues that the existing judicial evasion and concealment2 principles and the statutory fraudulent and wrongful trading provisions under the Insolvency Act 19863 are too restrictive, and ambiguous in combating corporate abuse. It is concluded that the existing common law and statutory rules geared towards combating abuse of limited liability provides no coherent format upon which the courts and legislature may effectively curb abuse of the corporate form. As such, these laws in light of their inability to make dishonest directors personally liable for their fraudulent conducts ought to be challenged. There is a need to challenge the existing rules in order to show the effect abuse of limited liability has on creditors, the public and the economy. This research indicates that there ought to be an adequate and effective alternative law which provides balance and support for genuine enterprise whilst providing a robust system whereby those who abuse the corporate form can be easily made liable for corporate debts.

L’attractivité des territoires vis-à-vis des investissements directs étrangers constitue, avec l’ouverture accrue des espaces économiques et politiques, un élément clé de l’intensification des flux d’échanges et d’investissements.Pour tous les pays et, plus particulièrement, pour ceux ne disposant pas d’un marché national important, l’établissement d’un diagnostic d’attractivité et la détermination d’axes prioritaires de développement constituent une démarche fondamentale applicable particulièrement à l’exemple du Luxembourg, une économie considérée comme une petite économie mature ouverte (PEMO).A travers cette recherche, on s’attachera à appliquer et à adapter un modèle d’analyse des fondements de l’attractivité, puis un modèle de détermination des orientations économiques et institutionnelles à privilégier.Cette recherche conduit à définir une démarche prospective structurée, appliquée à la PEMO considérée :- en partant de l’établissement d’une liste de déterminants de compétitivité et d’attractivité ayant potentiellement trait à une PEMO en général, validée ensuite pour le cas concret du Luxembourg par un panel de décideurs ;- en permettant l’établissement d’un diagnostic d’attractivité portant sur la situation actuelle de la PEMO considérée ;- pour déboucher sur la détermination d’activités prioritaires et de profilsd’investisseurs directs étrangers à privilégier, dans un « espace de référence » géo - sectoriel reconsidéré.Cette recherche devrait permettre d’identifier les secteurs d’investissement les plus attractifs pour les entreprises étrangères dans une PEMO comme le Luxembourg. / Together with increased economic and political openness, a location’s or territory’s attractiveness and competitiveness for foreign direct investment (FDI) are key elements for intensifying investment and trade flows.For countries, especially those without a national market of significant scale, establishing a way to identify and analyse priorities for economic development is fundamental.This research defines a general analytical framework applicable to the concepts of attractiveness and competitiveness, and, building on this framework, develops a specific model that could assist key institutions as to identify their preferred orientations in the specific context of an economy considered as a small open mature economy (SOME), in particular, Luxembourg.This research will lead to the definition of a prospective structured approach by:- providing a list of factors which potentially influence the competitiveness and attractiveness of SOMEs; this list having been validated with respect to Luxembourgby national decision makers;- establishing a method for analysing the current situation regarding attractiveness and competitiveness in Luxembourg; and- identifying priority sectors for FDI that could be targeted in a specific location/territory of reference such as Luxembourg.

L'attractivité des territoires vis-à-vis des investissements directs étrangers constitue, avec l'ouverture accrue des espaces économiques et politiques, un élément clé de l'intensification des flux d'échanges et d'investissements.Pour tous les pays et, plus particulièrement, pour ceux ne disposant pas d'un marché national important, l'établissement d'un diagnostic d'attractivité et la détermination d'axes prioritaires de développement constituent une démarche fondamentale applicable particulièrement à l'exemple du Luxembourg, une économie considérée comme une petite économie mature ouverte (PEMO).A travers cette recherche, on s'attachera à appliquer et à adapter un modèle d'analyse des fondements de l'attractivité, puis un modèle de détermination des orientations économiques et institutionnelles à privilégier.Cette recherche conduit à définir une démarche prospective structurée, appliquée à la PEMO considérée :- en partant de l'établissement d'une liste de déterminants de compétitivité et d'attractivité ayant potentiellement trait à une PEMO en général, validée ensuite pour le cas concret du Luxembourg par un panel de décideurs ;- en permettant l'établissement d'un diagnostic d'attractivité portant sur la situation actuelle de la PEMO considérée ;- pour déboucher sur la détermination d'activités prioritaires et de profilsd'investisseurs directs étrangers à privilégier, dans un " espace de référence " géo - sectoriel reconsidéré.Cette recherche devrait permettre d'identifier les secteurs d'investissement les plus attractifs pour les entreprises étrangères dans une PEMO comme le Luxembourg.