Dissertations / Theses on the topic 'MAB PURIFICATION'

Counter current chromatography (CCC) is a form of liquid liquid chromatography, which the Brunel Institute for Bioengineering (BIB) team have developed to process scale. In this thesis, its application has been successfully extended to the rapid, scalable purification of monoclonal antibodies (mAb) from mammalian cell culture, using aqueous two-phase systems (ATPS) of inorganic salts and polymer. A polyethylene glycol (PEG) and sodium citrate system was found to be the most appropriate by robotic phase system selection. The search for an economical alternative to protein A HPLC is a substantial bioprocessing concern; in this work CCC has been investigated. Initial studies showed that unpredictably, despite separation from impurities being achieved, some loss in the IgG‘s ability to bind to Protein A was seen, as confirmed by Protein A BiaCore analysis. CCC machines were seen to adversely affect IgG functionality. This led to a systematic investigation of the effect of CCC phase mixing on IgG functionality in a number of different CCC instruments, allowing direct comparisons of modes of CCC (hydrodynamic and hydrostatic CCC) and their associated mixing (wave-like and cascade, respectively). The varying g forces produced within the CCC column were determined using a recently developed model to calculate g force range. The effect of interfacial tension was also studied using a custom built 'g' shaker. The optimum CCC mode was identified to be the non synchronous CCC, operated in a hydrodynamic mode but allowing bobbin to rotor speed (Pr ratio) to be controlled independently. In a normal synchronous J type centrifuge a Pr of 1 is fixed, this is where the bobbin and rotor speed are identical I.e. one bobbin rotation (where mixing occurs) to one rotor revolution (where settling occurs). Constraints were seen with this 1:1 ratio and the separation of mAb using ATPS. This work has shown with the use of the non synchronous CCC at a Pr of 0.33, mixing is reduced and rotor rotations increased. Consequently the associated g force range is decreased. Furthermore, by the extension of settling time, the clear separation of the mAb from impurities has been achieved with retention of biological activity. This thesis demonstrates the importance of settling time for ATPS in phase separation and documents the fundamental requirements for the successful separation of biologics. Purified non synchronous CCC samples have additionally undergone rigorous quality control testing at Lonza Biologics by their purification scientists. This work has ultimately showed that with optimisation, the non synchronous CCC can be used to produce biological samples that are of industry standard.

Le cytosquelette des microtubules (MTs) est constitué de filaments dynamiques impliqués dans une multitude de fonctions telles que la division cellulaire, le maintien de forme des cellules, les battements ciliaires ou encore la différenciation neuronale. Une régulation stricte des fonctions des MTs est donc d'une grande importance pour l'homéostasie cellulaire, et toute perturbation pourrait potentiellement conduire à des maladies comme le cancer, les ciliopathies ou la neurodégénérescence. Dans un contexte cellulaire, les propriétés des MTs peuvent être contrôlées par leurs interactions avec une grande variété de protéines associées (MT-associated proteins ; MAPs). Notre connaissance de ces interacteurs s'est continuellement enrichie au cours des dernières décennies, mais il n'existe à ce jour aucune étude systématique visant à décrire et à classer ces protéines en fonction de leurs mécanismes de liaison et de leurs effets structuraux sur les MTs. Dans mon projet de thèse, j’ai mis au point un essai permettant une analyse rapide et systématique à la base des lysats clarifiés de cellules humaines surexprimant une multitude des différents MAPs. Le comportement dynamique des MT en présence d'environ 50 MAPs différentes a été imagé à l'aide de la microscopie TIRF. Cela nous permet d'étudier le comportement des MAP dans une situation proche de leur environnement naturel, mais en éliminant la complexité de l'espace intracellulaire, telle que l'encombrement par des organelles et des filaments du cytosquelette à l'intérieur de l'espace intracellulaire confiné. En effet, la plupart des MAPs étaient bien solubles dans notre approche d'extraction, tandis que les approches de purification pour plusieurs d'entre elles ont conduit à leur précipitation, rendent les expériences de reconstitution in vitro classique impossible. Ma nouvelle approche m’a permis de définir plusieurs nouvelles protéines comme de véritables MAP. J’ai montré que des MAPs non-caractérisées auparavant ont des effets étonnamment différents sur la polymérisation et la structure des MTs, créant ainsi une variété de réseaux de MT distincts. J’ai également démontré que mon approche permet d'étudier les structures des MAPs associées aux MTs par cryo-microscopie électronique, ou d'étudier le dynamique des MTs porteuses de mutations trouvées dans les pathologies humaines. J’ai également démontré que mon approche permet à tester la sensibilité des MAPs aux modifications post-traductionnelles de la tubuline, ou d'étudier le rôle des MAPs dans les interactions entre l'actine et les MTs. Mon approche expérimentale permet donc de mieux comprendre comment les MAP et les MT contrôlent ensemble le fonctionnement du cytosquelette
Microtubules (MTs) are dynamic filaments involved in a plethora of functions such as cell division, cell shape, ciliary beating, neuronal differentiation. Strict regulation of MT functions is therefore of high importance for the cellular homeostasis, and any perturbations could potentially lead to diseases like cancer, ciliopathies and neurodegeneration. At the protein level, there are accumulating studies showing that MT properties can be controlled via interaction with a large variety of MT-associated proteins (MAPs). Our knowledge of MAPs has been enriched over time, but up to this date no systematic studies exist that aim to describe and categorize these proteins according to their binding mechanisms and structural effects on MTs. In my PhD project, I have developed an assay for rapid and systematic analysis of MAPs using cleared lysates of cultured human cells in which I overexpress a variety of different MAPs. The dynamic behaviour of growing MTs in the presence of those MAPs were imaged using TIRF microscopy. This allows me to study the behaviour of around 50 MAP candidates in a situation close to their natural environment, but eliminating complexity coming from different organelles and crammed cytoskeleton filaments inside the confined intracellular space. Indeed, most MAPs were nicely soluble in the extract approach, while purification attempts of several of them led to protein precipitation, thus making classical invitro reconstitution approaches impossible. This novel approach allowed me to compare many MAPs under similar experimental conditions, and helped to define several novel proteins as bona-fide MAPs. I demonstrate that previously uncharacterized MAPs have strikingly different effects on MT polymerization and MT structure, thus creating a variety of distinct MT arrays. I further extended this cell-free pipeline to study structures of MAPs bound to MTs by cryo-electron microscopy, or to study the MT interactions of MAPs carrying patient mutations. Finally, I demonstrated that my approach can be used to test the sensitivity of MAPs to tubulin PTMs, as well as to study the role of MAPs in actin-MT crosstalk. In the future, this novel approach will allow for a better mechanistic understanding of how MAPs and MTs together control cytoskeleton functions

La vía de señalización celular MEK5-ERK5 juega un papel importante en sistema nervioso, dado que se activa frente a neurotrofinas, frente al estrés oxidativo producido por reactive oxygen species (ROS), tras la isquemia cerebral y tiene un papel en la especificación de fenotipo neuronal. Sin embargo, poco se conoce sobre los sustratos y proteínas que interaccionan con ERK5. En este trabajo se han identificado proteínas que interaccionan con ERK5 en células de neuroblastoma SH-SY5Y mediante el método TAP (Tandem-Affinity Purification). Se han obtenido diversas proteínas que incluyen proteínas de citoesqueleto, chaperonas y proteínas de metabolismo celular. Entre las proteínas identificadas destacan la proteína chaperona Hsp90β y la piruvato kinasa PKM2.
Por otro lado, se ha determinado el papel que juega ERK5 en la isquemia cerebral, utilizando el modelo de privación de oxígeno y glucosa (OGD) en cultivos mixtos de neuronas corticales. ERK5 se degrada rápidamente en respuesta a la OGD en cultivos mixtos de neuronas corticales. Esta degradación es mediada por calpaína. ERK5 también se degrada rápidamente in vivo (4 horas) en córtex de cerebros de ratas Sprague-Dawley sometidas a isquemia por oclusión de la arteria cerebral media. En este trabajo se establece que tras la isquemia, la entrada masiva de calcio a través de los receptores de NMDA y la consiguiente activación de calpaína conlleva la degradación de ERK5, produciéndose así una disminución de los niveles totales de esta kinasa.
MEK5-ERK5 pathway plays an important role in nervous system. This pathway is activated in response to neutrophins, oxidative stress induced by reactive oxygen species (ROS), after cerebral ischemia and plays an important role in neuronal fate determination. However, little is known about substrates and proteins that interact with ERK5. This work describes the search for proteins which interact with ERK5 in the cell line SH-SY5Y using Tandem-Affinity Purification (TAP). Using this technique, we obtained different proteins that include citosqueleton components, chaperons and cell metabolism proteins, among which it is worth highlighting Hsp90β and piruvate kinase PKM2.
On the other hand, we have established the role of ERK5 in cerebral ischemia, using oxygen and glucose deprivation model (OGD) in rat mixed neuronal cortical cultures. Using this model we observed that ERK5 is quickly degraded in response to OGD, and that calpain is responsible for ERK5 degradation. ERK5 is also quickly (4 hours) degraded in vivo in cerebral cortex from Sprague-Dawley rats subjected to middle cerebral artery occlusion. On this work we establish that after cerebral ischemia, massive income of calcium mediated by NMDA receptors and resulting activation of calpain entails ERK5 degradation, producing a decrease in total levels of this kinase.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
O trabalho investiga como se dá a (re)configuração corporal pela cirurgia plástica estética, a partir da perspectiva de um grupo específico de mulheres, que realizaram plásticas de lipoaspiração e mamoplastia de aumento. Trata-se de um grupo relativamente homogêneo, no que diz respeito à faixa etária e classe social, e que compartilha um mesmo ideal de mulher bela, bem sucedida e independente. A partir da afirmação da identidade de poderosa, uma concepção específica de feminilidade é valorizada pelo grupo, diretamente vinculada a um suposto poder de sedução feminino. Dentro do grupo, compartilham-se também valores advindos da moral da saúde, que a partir da designação de “bons” e “maus” hábitos, fornece critérios de avaliação individual à sociedade moderna contemporânea. Assim, a hexis corporal da poderosa é portadora de marcas distintivas que a remetem diretamente ao estilo de vida saudável, tido como moralmente superior. Nesse contexto, onde a gordura corporal localizada é vista como impura, a plástica de lipoaspiração é (re)significada como parte de um rito de purificação. Ao se retirar a gordura acumulada no corpo, busca-se apagar a memória de um passado de transgressões à moral da saúde e, mais especificamente, às normas instituídas sobre a alimentação e o trabalho. Por sua vez, a mamoplastia de aumento também não se mostra como um fato isolado nas trajetórias das pesquisadas, mas como uma prática vinculada a uma ascensão na estrutura social, vivida a partir da conquista de independência financeira, e mudança para territórios urbanos mais desenvolvidos. O poder aquisitivo, somado ao cumprimento de requisitos implícitos nas normas de purificação corporal do grupo, direciona essas mulheres à prática de “colocar silicone”, que pode ser entendida como parte de um rito de instituição, na medida em que vem para consagrar a inclusão no grupo valorizado das poderosas, marcando as diferenças entre essa categoria de mulher, tipicamente moderna, e as que ficam de fora: a “gorda”, a “feia” e a “pobre”.
The research investigates how the body (re)configuration by cosmetic plastic surgeries happens, according to the perspective of a specific group of women, who have undergone through plastic surgeries as liposuction and breast augmentation. It is a relatively homogeneous group, regarding to age and social class, and that shares the same ideal of beautiful, successful and independent woman. By the identity affirmation of poderosa (powerful woman), a specific conception of femininity is prized by the group, which is directly linked to the presumed female seductiveness. Within the group, are also shared values related to the health’s moral, or healthism, which defines "good" and "bad" habits, providing standards for individual evaluation in the contemporary modern society. Thus, the poderosa’s body hexis carries distinctive marks that relate it directly to a healthy lifestyle, seen as morally superior. In this context, where the concentrated body fat is seen as impure, the liposuction is (re)interpreted as part of a purification rite. By eliminating the accumulated fat in the body, it’s intended to erase the memory of a past of transgressions against the health’s moral, or healthism, and more specifically, against the rules that control food and work. At its side, the breast augmentation shows not to be an isolated incident in the trajectories studied, but a practice linked to a rise in the social structure, described like financial independence achievement, and moving to more developed urban areas. The purchasing power, combined with the fulfillment of requirements implied in the group standards of body purification, leads these women to the practice of "putting silicone" which can be understood as part of a rite of institution, once it comes to consecrate the inclusion into the valued group of poderosas, marking the differences between this category of woman, typically modern, and those left out: the "fat", the "ugly" and the "poor".

Les cellules phagocytaires constituent la première ligne de défense contre les pathogènes. Leur migration dirigée vers le site infectieux et leurs fonctions microbicides sont l'aboutissement de voies de signalisation intracellulaires sollicitées par la stimulation de récepteurs couplés aux protéines G, les récepteurs de chimioattractants. Après fixation du ligand et transmission du signal par la protéine G, les récepteurs sont phosphorylés et interagissent avec les b-arrestines, protéines d'échafaudage concourrant à l'internalisation des récepteurs. Plusieurs exemples récents suggèrent que les b-arrestines pourraient également participer à la signalisation. Le travail présenté dans ce mémoire concerne les récepteurs de la famille FPR (Formyl Peptide Receptor) et plus spécialement le récepteur FPRL1 (FPR-like 1), pour lesquels de nouveaux agonistes dérivant de protéines bactériennes ou mitochondriales humaines ont été identifiés. La phosphorylation du récepteur FPRL1 a été caractérisée. Il a été montré que les -arrestines interagissent avec celui-ci et qu'elles sont indispensables à son internalisation. Diverses approches ont conclu que l'activation rapide des MAP kinases ERK1/2, enclenchée par la stimulation du récepteur FPRL1, est majoritairement dépendante de la protéine G héterotrimérique et qu'il n'y pas de signalisation transmise par les b-arrestines. Enfin, une analyse protéomique des complexes multi-protéiques bâtis autour du couple FPRL1/b-arrestine a été menée par la méthode TAP (Tandem Affinity Purification). Le complexe adaptateur AP3, homologue d'AP2 a été identifié comme partenaire des b-arrestines après stimulation du récepteur FPRL1
Phagocytes are the first host defense line against pathogens. Their directed migration to infection sites and their microbicidal functions result from intracellular signaling cascades elicited by specific G protein-coupled receptors, the chemoattractant receptors. After ligand binding and G protein-mediated signaling events, receptors are phosphorylated and interact with the b-arrestins, which are scaffolding proteins participating to receptor internalization. Recent examples suggest that b-arrestins could also participate to signaling events. The work presented here is focused on the FPR family (Formyl Peptide Receptor) and specially the FPRL1 receptor (FPR-like 1), for which new agonists from bacterial origin or derived from human mitochondrial proteins have been identified. The phosphorylation of the FPRL1 receptor has been characterised and it has been shown that b-arrestins interact with this receptor and are essential to its internalization. Various approaches have concluded that the FPRL1-elicited rapid and transient activation of the MAP kinases ERK1/2 is mainly dependant on G protein and that there is no signaling events mediated by the b-arrestins. Finally, a proteomic study of the multiprotein complexes surrounded the couple FPRL1/b-arrestin has been conducted by the TAP method (Tandem Affinity Purification). The adaptor complex AP3, an homolog of AP2, has been identified as partner of b-arrestins after FPRL1 stimulation

La recherche et les questions abordées dans cette thèse sont de type inverse : la reconstitution d'une source ponctuelle ou la complétion d'une donnée à la limite inconnue à l'extrémité du domaine dans les modèles paraboliques de transport de contaminants. La modélisation mathématique des problèmes de pollution des eaux fait intervenir deux traceurs, l'oxygène dissous (OD) et la demande biochimique en oxygène (DBO) qui est la quantité d'oxygène nécessaire à la biodégradation de la matière organique. En effet, au cours des procédés d'autoépuration, certaines bactéries aérobies jouent un rôle principal. Ces micro-organismes décomposent les matières organiques polluantes en utilisant l'oxygène dissous dans le milieu. Afin de compenser ces données manquantes, les champs, solutions du problème, sont observés directement ou indirectement. Les problèmes inverses qui en résultent sont quasi certainement mal-posés voire même sévèrement mal-posés pour la plupart. Dans cette thèse, nous proposons justement une analyse aussi poussée que possible sur la question de l'identifiabilité pour les deux problèmes inverses décrits ci-dessus. Nous avons démontré un résultat d'unicité pour des sources fixes dans le cas d'observations décalées. La réalité pour l'observation est nuancée et l'idéal n'est pas acquis ; des mesures directes sur la DBO sont difficiles à obtenir. En revanche collecter des données sur l'OD est possible en temps réel et avec un faible coût. La DBO est donc observée de façon indirecte, grâce au couplage dans le système de Streeter et Phelps, l'information passe de l'OD à la DBO. Pour ce problème aussi, nous avons produit un résultat d'unicité pour la reconstruction de la source ou puits ponctuel qui serait présent dans l'équation de transport sur l'OD. Nous avons ensuite examiné des questions annexes à l'identifiabilité telles que le degré d'instabilité des équations à résoudre. De ce type d'informations dépendent le comportement des méthodes numériques et des algorithmes de calcul à utiliser
The research and the questions approached on this thesis are inverse type : the reconstruction of point-wise source or the data completion problem in parabolic models of transport of contaminants. The mathematical modelling of the problems of water pollution includes two tracers, the dissolved oxygen (DO) and the biochemical demand in oxygen (BDO) which is the quantity of oxygen necessary for the biodegradation of organic matter. Indeed, during the biodegradation process, aerobic bacteria play a leading part. These micro-organisms decompose polluting organic matters by using the dissolved oxygen in the middle. To compensate these missing data, fields, solutions of the problem, are observed directly or indirectly. The resulting inverse problems are ill-posed. Their mathematical study rises big complications and their numerical treatment isn't easy. We demonstrated a uniqueness result for fixed sources in the case of moved observations. The reality for the observation is qualified and the ideal is not acquired; direct measures on the BOD are difficult to obtain. On the Other hand to collect data on the DO is possible in real time With a moderate cost. The BOD is thus observed in indirect way, thanks to the coupling in the system of Streeter and Phelps, the information passes from the DO to the BOD. For this problem, we produced a uniqueness result for the reconstruction of source. Then, we examined the degree of instability of the equation to be solved. The behaviour of numerical methods depend on this type of information

Master of Science
Department of Chemical Engineering
James Edgar
The bio-pharmaceutical industry began over 30 years ago with the production of human insulin and has shown incredible growth ever since. With forecasted annual worldwide sales of over $450B in 2025 for biopharmaceuticals, they are expected to be at least 25% of the entire pharmaceutical market. Therapeutics based on monoclonal antibodies (mAbs) make up roughly a third of all biopharmaceutical sales with indications from asthma, to cancer, to Parkinson’s disease. The recent approval of the first biosimilar mAb products in the US and Europe has exposed up to 20 of the top grossing biologic products to competition for the first time, while 75% of the US market is expected to lose patent exclusivity by 2020. With the increased competition from biosimilars, the costs associated with producing mAb based therapeutics will be a constraint on maintaining market share going forward. The majority of the total manufacturing costs for mAbs resides in the downstream processing where Protein A chromatography is the predominantly employed technology for the primary capture step. With Protein A’s high unit cost of up to $15,000 per liter and susceptibility to deamidification when exposed to high pH cleaning and sanitization chemicals, it is no surprise that many mAb manufacturers are considering alternatives. The objective of this work is to review the production process of mAb therapeutics, with a specific focus on the advantages, disadvantages, and alternatives to Protein A affinity chromatography as the primary capture step in downstream processing.

Today, recombinant protein therapies represent a substantial focus of the pharmaceutical industry. Most therapeutic proteins are produced in host cells of non-human origin, including Escherichia coli, yeast, and various mammalian cell lines e.g. Chinese Hamster Ovary Cell Lines. A major focus of all therapeutic protein purification process is to reduce components of host organism, including host cell proteins (HCPs), to levels considered as adequate in the final formulated drug product. HCPs can pose potential safety risks for patients, including immune reactions, decreased product stability, adjuvant activity, and (theoretically) protein-specific biological activity.(Schenauer, Flynn, & Goetze, 2012) With rapidly growing cases and increased number of cancer, autoimmune diseases, Alzheimer’s disease has become the most common death-causing diseases worldwide. Recent studies indicate that mAb is effective in treatment of these diseases and with limited number of treatment options people need to rely on these medicines. Thus, the purity of these medicines becomes an important factor. If these medicines are not pure the HCP might induce antigenic reactions in the patient also these HCPs if proteolytic may degrade the desired amount of mAb to be effective as dose, thus making the medicine ineffective over a period. Thus here, we report results of the studies relating to the most interactive HCPs which isolated along with mAb and studied their interactions to design the wash process accordingly. Using the proteome of Chinese Hamster, the hotspot for the proteins were found using Aggrescan. Structure prediction was done using threading software’s Bhageerath, Pyre 2, Multicom-Raptor-X, Robetta and I-TASSER. The model generated was further validated by Independent servers Prochek, Verify-3d, Errat and with meta servers such as Protsav and SAVES. The models were refined using 3D refine and galaxy refine. The best models were docked with mAb using cluspro to identify the interactions between HCP and mAb then find the amino acid interaction profile using PDB-Sum and further work on developing modified wash process to break these bonds and obtain pure mAb to ensure effective treatment by functionally characterizing the proteins.

The increasing demand and pollution of natural water systems caused by the constant growth in population has placed maximum strain on the resources upon which people depend. The presence of water sources has ensured the establishment, development and progression of the urban environment since settlements first originated, however, these environments have grown beyond the resources capabilities to sustain them. The preservation, purification and sustainable use of water have quickly become the most important strategies ensuring the survival of the urbanized world and its inhabitants. The loss of water as a resource as well as their ecological systems and processes is not limited to larger water sources but is evident in the smaller scale river systems that are exposed to the urban environment on a daily basis. The demands by the surrounding urban environments exceed the capability of these smaller river systems which in turn has led to their neglect and steady destruction. This study investigates the potential of small scale urban river systems to be able to provide for the surrounding urban environment as a resource through the process of purification. The reintroduction of the river as a resource will instil a sense of stewardship in man over the natural environment on which they rely, focusing on the connection between man and nature. The transformation of the riverfront into an interactive public space will promote the use and interaction of people with a previously neglected environment. The strengthening of the connection between man and environment is achieved through the incorporation of the resource into the economic sector of the urban environment, using the river as a means of income and production will add considerable value to the natural environment. A broad approach was investigated that took the entire river system into consideration, while the main design intervention took place at one specific point along the river which had potential in all social, ecological and economic aspects. A decommissioned, post-industrial site was selected into which the purification of the river system could be integrated. The purification of the river would allow for various degrees of use, interaction, education and production centred around the resource. The proposal creates opportunities for the creation of public space and economic activities which benefit the surrounding communities through the rehabilitation of a polluted urban river. The incorporation of the river system into the post-industrial site is responsible for the rehabilitation and reclamation of a wasted landscape into one of provision and connectivity between various aspects of the urban environment. The purpose of the intervention is the re-introduction of an existing urban river system into the built environment through the creation of social, economic and ecological connections, re-capturing the significance, importance and potential they hold as a resource as well as the ability to inform urban spatial design.
Mini Dissertation (ML(Prof))--University of Pretoria, 2016.
tm2016
Architecture
ML(Prof)
Unrestricted

Relação, Perdão e Reconciliação. Três conceitos que envolvem uma “mixórdia de temáticas” e que implicam necessariamente um antes, um durante e um depois. Antes do perdão, vem a relação, no sentido de uma urgente necessidade do ser humano em construir laços consigo mesmo, com os outros e com Deus. Mas, porque o ser humano está longe de ser perfeito, o mal surpreende a vida, invade o seu espaço interior, deixa-o na inquietude de uma incompreensão instalada. E entre o mal que o ser humano pratica e o mal que o ser humano sofre, surge o perdão, como um abraço impensável, um sinal de esperança num futuro diferente, uma reinvenção de uma vida. É deste pressuposto que se pode falar de reconciliação, como o depois do perdão, como o resultado de uma decisão unilateral de amor, como premissa fundamental àquilo que todo o ser humano deve ser: “imagem e semelhança de Deus”.
Relationship, Forgiveness and Reconciliation. Three concepts which involve a kind of “melting pot” of ideas and which necessarily imply a before, a throughout and a after the events. Before the forgiveness comes the relationship, in the meaning of an urgent necessity of the human being in building bonds with himself, with the others and with God. However, as the human being is far from being perfect and faultless, the evil surprises life, invades its inner space and leaves the person in great concern with the feeling of misunderstood. And among the evil that is done to others and the one that is experienced comes the forgiveness, like an unthinkable hug, a sign of hope for a different future, a reinvention of life. It’s starting with this assumption that we can approach reconciliation, it’s what follows the forgiveness like the result of a unilateral decision of love, like fundamental principles of what a human being should be: “God image and likeness”.

Dunovský M. The Situation in the Political District of Strakonice in the Period between May 1946 and February 1948. Key Words: Strakonice, parliamentary elections in May 1946, renewed ONV Strakonice, renewed MNV Strakonice, the council of ONV Strakonice, the council of MNV Strakonice, plenum of MNV Strakonice, confiscations of private possessions, February 1948, people's militia, OAV-NF Strakonice, public purification. This thesis focuses primarily on the description of the political conditions in the area of the former political district of Strakonice between May 1946 and February 1948. Hence, the main focus lies on the topics such as the activity and agenda of ONV Strakonice and MNV Strakonice with regard to the personal and political structure of these authorities following the parliamentary elections in the spring of 1946 and the radical changes after the political coup on February 25th 1948. In this respect the thesis tracks the alterations in the distribution of political power in the region and offers a clear view of the transition from the unstable post- war democracy to the communistic dictatorship. Confiscations of private possessions based on the decisions of the responsible authorities are among other issues addressed in the thesis. In this respect the thesis covers the entire area of...