Thesis (M.Sc.(Chemistry)) -- University of Limpopo, 2020 / Breast cancer is a malignant tumour that starts in the cells of the breast. Many studies revealed aromatase (CYP19A1) and cyclin-dependent kinase 2 (CDK2) as possible therapeutic targets regarding breast cancer treatment, because they play crucial roles in anti-apoptotic processes during cell proliferation. Quinoxaline derivatives have attracted a great deal of attention due to their biological activities against fungi, virus, bacteria and cancer. Computer modelling was employed in order to reduce time and cost by searching the library of molecules and identifying those which are likely to bind to the drug target. A library of new one hundred (100) nitro and amino quinoxaline alkyne derivatives were successfully designed and screened against target proteins (CYP19A1 and CDK2) using virtual screening technique and thirteen (13) molecules were identified to be hit compounds against both targets with the docking score ranging from -6.143 to -8.372 kcal/mol as a measure of binding affinity. The hit compounds were subjected to IFD in order to identify tight binding through intermolecular interactions with active site residues of the binding pocket of the target proteins. All identified nitro and amino quinoxaline alkyne derivatives were successfully synthesised in a multi-step reaction sequence and their spectroscopic analysis (NMR, FTIR and MS) were in good agreement with the proposed structures in a good to moderate yield. The newly synthesised novel amino and nitro-quinoxaline derivatives were evaluated for anti-proliferative activity against breast cancer (MCF-7). Compound 59 showed to possess good inhibition against MCF-7 with an IC50 of 9.102 μM, whereas compounds 34, 54, 56 and 61 showed promising activity against MCF-7 with an IC50 value of < 50 μM. However, the MTT assay results showed that 59 was found to be toxic with an IC50 value of 0.205 μM against Raw 264.7 cell line. The dose response investigations showed that 31 and 34 have the promising anti-cancer activity against CYP19A and the correlation between molecular modelling (in-silico) and CYP19A inhibition activities (in- vitro), was established as compounds 31 and 34 were identified to bind to the drug target (CYP19A) with the docking score of -8.372 and 7.630 kcal/mol respectively. All the synthesized compounds were evaluated for the antitubercular activity against Mtb H37Rv strain as a secondary study. Compounds 57-62 with nitro-quinoxaline derivatives exhibited stronger inhibitory effects on Mtb H37Rv strain. In addition, compounds 60 and 62 were found to be most active against Mtb H37Rv with the high activity at MIC90 of <0.65 and <0.64 μM respectively. All active compounds are currently investigated for their cytotoxicity which have not been investigated before / National Research Foundation (NRF) and SASOL Inzalo Foundation

Diploma theses represents project of marketing a communication strategy of manufacturing company. This work includes situation analysis of present situation including evaluation of weaknesses and streghts of the company, opportunities and threaths and on the basis of this analysis particular marketing and communication strategy, budget and timing has been designed.

Le développement de thérapies ciblées est un enjeu majeur en santé et l’essor des nanovecteurs permet de répondre à ces besoins cliniques. Le premier axe de cette thèse est consacré à l’étude du potentiel thérapeutique de nanoparticules multifonctionnelles pour l’imagerie médicale, la thérapie photothermique et la délivrance de drogue pour le traitement du cancer. Le deuxième axe de recherche s’oriente vers le ciblage thérapeutique actif. L’entreprise NanoMedSyn a pour objectif de développer un ciblage actif du récepteur du mannose 6-phosphate, permettant un meilleur adressage des médicaments et des traitements plus efficaces. Ce type de ciblage peut avoir des retombées multiples pour la thérapie anticancéreuse mais également pour la thérapie des maladies lysosomales qui sont des maladies génétiques rares. NanoMedSyn développe des dérivés synthétiques de glycovecteurs innovants, appelés AMFA, qu’elle exploite en exclusivité. Les AMFA ont une bonne affinité pour le récepteur du mannose 6-phosphate et ont été greffés sur des nanoparticules multifonctionnelles dans le but d’améliorer l’adressage et la thérapie photodynamique biphotonique d’un cancer pédiatrique : le rhabdomyosarcome ; et sur des enzymes lysosomales pour le traitement de maladies lysosomales telle que la maladie de Pompe. / The development of targeted therapies is a major health issue and the rise of nanovectors makes it possible to meet these clinical needs. The first approach of this thesis is dedicated to the study of the therapeutic potential of multifunctional nanoparticles for medical imaging, photothermal therapy and drug delivery in cancer treatment. The second line of research focuses on active therapeutic targeting. The NanoMedSyn company aims to develop an active targeting of the mannose 6-phosphate receptor, allowing a better addressing of drugs and more effective treatments. This type of targeting may have multiple benefits for cancer therapy but also for the treatment of the lysosomal diseases which are rare genetic diseases. NanoMedSyn develops innovative synthetic derivatives of glycovectors, called AMFA, which it exploits exclusively. AMFA have a good affinity for the mannose 6-phosphate receptor and have been grafted on multifunctional nanoparticles in order to improve addressing and two-photon photodynamic therapy of a pediatric cancer: the rhabdomyosarcoma; and on lysosomal enzymes for the lysosomal diseases treatment such as for Pompe disease.

Les mastocytoses systémiques (SM) constituent un groupe hétérogène de maladies rares, caractérisées par l’accumulation anormale de mastocytes malins dans la moelle osseuse et dans d’autres organes extra-cutanés. La majorité des patients avec SM ont une mutation activatrice du gène KIT, le plus souvent la mutation KIT D816V, retrouvée chez plus de 90% de tous les patients. Cette mutation induit l’activation constitutive du récepteur KIT en déclenchant de façon aberrante une cascade de voies de signalisation, dont la voie PI3K/AKT et JAK/STAT5, aboutissant à l’inhibition de l’apoptose et à l’augmentation de la prolifération et de la survie des mastocytes malins. Cependant, l’efficacité des inhibiteurs de tyrosines kinases (ITKs) sur cette mutation est limitée à cause de la résistance et/ou de toxicité liée à un manque de spécificité. Il est donc nécessaire de trouver de nouvelles approches thérapeutiques afin de contourner cette résistance au cours des SM KIT D816V+ avancées. Nous avons utilisé une approche consistant à cibler de façon combinée des molécules activées en aval de KIT D816V, comme AKT et STAT5, par des inhibiteurs pharmacologiques. Ceci nous a permis d’identifier une combinaison synergique entre un inhibiteur d’AKT (GSK690693) et un inhibiteur de STAT5 (BP-1-102). Ces composés sont capables d’inhiber la prolifération des cellules KIT D816V+ seuls ou en combinaison, mais à de très fortes concentrations, malheureusement non utilisables en thérapeutique. Néanmoins, ces premiers résultats ont permis de valider AKT et STAT5 comme cibles potentielles dans le traitement des SM avancées. La seconde approche employée a été de cibler directement le récepteur KIT D816V par des inhibiteurs pharmacologiques. A l’issu d’un criblage, nous avons identifié trois composés - BLU2317, BLU2718 et DCC-2618 - capables d’inhiber sélectivement la phosphorylation de KIT D816V. Ces composés inhibent la prolifération des cellules ROSAKIT D816V et HMC-1.2, et induisent l’apoptose des cellules de façon dose-dépendante. Bien que les effets de ces trois composés soient similaires, DCC-2618 agit à des concentrations plus faibles par rapport aux composés BLU2317 et BLU2718. Afin d’apprécier l’efficacité in vivo de DCC-2618, nous avons d’abord établi un nouveau modèle de SM basé sur l’injection intraveineuse des cellules ROSAKIT D816V-Gluc exprimant la Gaussia luciferase (Gluc) dans des souris NSG. La présence de la Gluc sécrétée par les cellules ROSAKIT D816V-Gluc facilite la mise en évidence de prise de greffe et permet un contrôle précis de la progression de la maladie. Ce modèle reproduit, au bout de 4 semaines, chez toutes les souris greffées, une SM avancée similaire à celle retrouvée chez l’homme, avec atteinte de la moelle osseuse, du sang, de la rate et du foie, tandis que la dégradation de l’état général des souris n’est observée qu’à partir de 12 semaines. Ce nouveau modèle offre suffisamment de temps pour explorer la cinétique de la progression de la maladie et surtout pour effectuer des études pharmacologiques précliniques. L’évaluation de l’effet de DCC-2618 in vivo a été réalisée sur ce modèle. Etonnamment, DCC-2618 n’a pas été capable d’inhiber la progression de la maladie chez les souris traitées, bien qu’atteignant des concentrations élevées dans la moelle osseuse et le plasma des souris traitées. Néanmoins, DCC-2618 s’est montré capable d’inhiber la phosphorylation de KIT dans les cellules issues de la moelle osseuse des souris traitées. En revanche, contrairement aux effets observés in vitro, DCC-2618 a induit une surexpression de phospho-ERK1/2 dans les cellules malignes des souris greffées. Ceci suggère qu’ERK1/2 joue un rôle important dans la résistance au composé DCC-2618 et éventuellement à d’autres ITKs, indépendamment du récepteur KIT. ERK1/2 pourrait donc être une nouvelle cible thérapeutique d’intérêt dans le traitement des SM résistantes aux ITKs / Systemic mastocytosis (SM) is a heterogeneous group of rare diseases characterized by abnormal accumulation of malignant mast cells (MCs) in the bone marrow (BM) and other extra-cutaneous organs. The majority of SM patients have an activating mutation in the KIT gene, usually the D816V point mutation, which is found in more than 90% of all patients. This mutation induces constitutive activation of the KIT receptor by triggering a cascade of signaling pathways, including the PI3K/AKT and the JAK/STAT5 pathways, resulting in the inhibition of apoptosis and increased survival and proliferation of malignant mast cells. However, the efficacy of the tyrosine kinase inhibitors (TKIs) on this mutation is limited due to resistance and/or toxicity associated with a lack of specificity. It is therefore critical to find new therapeutic approaches to overcome this resistance to TKIs, particularly for advanced KIT D816V+ SM. In the present thesis, we have used an approach consisting in targeting molecules activated downstream of KIT D816V, such as AKT and STAT5, using pharmacological inhibitors in combination. This allowed us to identify a synergistic combination of an AKT inhibitor (GSK690693) and an inhibitor of STAT5 (BP-1-102). These compounds are able to inhibit proliferation of KIT D816V+ cells, alone or in combination, but at very high concentrations, unfortunately not useful therapeutically. Nevertheless, these initial results have validated STAT5 and AKT as potential targets for the treatment of advanced SM. The second approach used was to target directly the KIT D816V receptor by pharmacological inhibitors. After a large screening, we identified three compounds - BLU2317, BLU2718 and DCC-2618 - which selectively inhibit the phosphorylation of KIT D816V. These compounds inhibit the proliferation of ROSAKIT D816V and HMC-1.2 cells, and induce apoptosis of these cells in a dose-dependent manner. Although the effects of these three compounds are similar, the DCC-2618 compound acts at lower concentrations relative to BLU2317 and BLU2718 compounds. In order to assess the in vivo efficacy of DCC-2618, we first established a new model of SM based on intravenous injection of cells expressing Gaussia luciferase (Gluc), ROSAKIT D816V-Gluc cells, in NSG mice. The presence of the secreted Gluc in ROSAKIT D816V-Gluc cells facilitates the detection of engraftment and allows precise monitoring of disease progression. This model reproduced within four weeks, in all grafted mice, an advanced SM similar to the one found in humans, with neoplastic MCs infiltration in BM, blood, spleen and liver, while the terminal deterioration of the clinical condition of the mice was observed after 12 weeks. Thus, this new in vivo model allows modulating the aggressiveness of the disease by varying the number of injected cells. It provides sufficient time to explore the kinetics of disease progression and especially to conduct preclinical pharmacological studies. We then evaluated the effect of DCC-2618 compound in vivo on this model. Surprisingly, DCC-2618 was not able to inhibit disease progression in treated mice, although it reached high concentrations in the BM and the plasma of treated mice. Nevertheless, we showed that the compound was able to inhibit the phosphorylation of the KIT receptor in cells derived from the BM of treated mice. In addition, contrasting to the effects observed in vitro, DCC-2618 induced an over-expression of phospho-ERK1/2 in the malignant cells of transplanted mice. This suggests that ERK1/2 may play a critical role in the resistance to DCC-2618, and possibly to other TKIs, independently of the KIT receptor. ERK1/2 could thus be a new interesting therapeutic target in the treatment of advanced SM resistant to TKIs

Cette thèse porte sur la problématique de l’application de la politique monétaire en République Slovaque (RS) depuis 1990. Malgré sa courte histoire la Banque Nationale de la Slovaquie (BNS), possédait et possède toujours une position importante dans le processus de transformation, dans la stabilisation de la situation macroéconomique et dans l’accomplissement des ambitions d’intégration de la RS. Par conséquent, la BNS doit réagir aux tendances actuelles dans le domaine des cibles, instruments et stratégies monétaires. La BNS a significativement avancé dans ce domaine dans une direction positive. La mise en oeuvre d‘une stratégie monétaire optimale, la combinaison appropriée de la règle monétaire et de la politique monétaire discretionnaire peuvent influencer favorablement l’efficacite de la BNS. L’application de la règle monétaire en tant que fonction de réaction de la banque centrale augmente la transparence et la clarté des mesures monétaires de la cette dernière. Une règle monétaire permet aux investisseurs, banquiers, analystes financiers et aux autres agents économiques de prévoir plus précisément la direction et le volume des mesures de la BNS. En conséquence de quoi la transmission et l’impact des démarches est plus efficace. Etant donné que la version officielle de la règle monétaire appliquée par la BNS n’est pas encore connue, il est dans l’intérêt de nombreux analystes de calculer cette formule. Cette thèse offre des processus éventuels pour élaboration d’une règle monétaire dans le contexte de la RS et en propose une forme possible. / The objective of this thesis is to characterise and to evaluate evolution of monetary policy in Slovakia as well as to suggest an optimal monetary strategy for the future. Theoretical part treats beginning and evolution of the monetary policy in the world and main approaches in its application within central banking. In the conclusion of theoretical part is review of literature dealing with similar problematic. The thesis analyses evolution of National Bank of Slovakia (NBS) independence, instruments and objectives. Evaluation of NBS monetary policy application, respectively State Bank of Czechoslovakia monetary policy application, is divided into five basic periods since 1990 to 2005. It is crucial to choose an optimal monetary strategy in order to increase central bank effectiveness and at the same time in respect to the Slovak Republic integration procedures. The thesis compares and analyses advantages and disadvantages of several alternative monetary strategies as for intermediate objectives and as for decisional process. According to our conclusions a Taylor-type rule should be the integral part of the optimal NBS monetary strategy. The rule originates from basic Taylor rule that enables to determinate direction and volume of central bank measurements. Our Taylor rule is applied and adopted to Slovak macroeconomic conditions. Time ranks of chosen macroeconomic indicators since 1993 to 2005 were applied in determination and calculation of the rule. Achieved results are significantly consistent with theory, initial pre-assumptions and current NBS activity.

Philosophiae Doctor - PhD / Introduction: Tuberculosis (TB) is a serious bacterial infections caused by the Mycobacterium Tuberculosis (MTB) organism affecting mainly the lungs. Occasionally, MTB bacilli may be transported out of the pulmonary region and infect peripheral organs causing extra-pulmonary tuberculosis. Many therapeutic agents were developed over the years to combat TB, however the rapid emergence of resistant strains hampered their use. Furthermore, most of the current anti-TB drugs experience many challenges, which can be summarized in treatment regimen factors, drug-drug interactions, and physicochemical characteristics factors (such as hydrophobicity and low permeability into alveolar macrophages). These challenges have a significant role in treatment failure and the emergence of resistant TB. Due to the lack of newly discovered anti-TB drugs, and the absence of effective vaccines, many scientists have suggested the use of novel modalities for the current anti-TB drugs to enhance their efficacy and overcome some of the drawbacks. One of these modalities is nanotechnology-based drug delivery systems. Most of the anti-TB drugs experience low drug distribution to the lung and particularly alveolar macrophages within which the MTB resides, leading to treatment failure. Employing nanoparticles as drug delivery systems can have a significant impact on improving the pharmacokinetic profile of anti-TB drugs, the feasibility of different routes of administration, enhancing drug permeability, controlled/sustained drug release, and targeting specific disease sites. Collectively, these impacts will aid in enhancing drug concentration at the site of infection and reduce dosing and regimen duration. Dendrimers, such as polyamidoamine (PAMAM) dendrimers, are synthetic polymeric nanoparticles that have unique features that afford a dendrimer-conjugate complex the possibility to overcome the most common hurdles associated with drug delivery and treatment of diseases. Obstacles associated with solubility, permeability, inadequate biodistribution associated side effects may be enhanced. Manipulating the outermost surface functional groups with various ligands and polymers, will enhance the dendrimer properties and targeting potential. Aim: This study aims to develop a novel pulmonary delivery system for the anti-TB drug rifampicin using surface-modified G4 PAMAM dendrimer nanoparticles (polyethylene glycol (PEG) or mannose moieties), to improve drug solubility, prolong-release, enhance permeability into the macrophages, and decrease the toxicity of the drug-dendrimer conjugates. Methods: PAMAM dendrimers having increasing concentrations of poly(ethylene glycol) (PEG) 2 kDa or mannose residues were synthesized. The 4-nitrophenyl chloroformate was used as an activator in the case of PEG functionalization, while for the mannose conjugation the 4-isothiocyanatophenyl alpha-D-mannopyranoside (4-ICPMP) directly interacted with the primary amines of the dendrimer. The conjugated PEG polymers and mannose moieties on the dendrimer periphery were confirmed using FTIR and 1H NMR analytical techniques. Thereafter, rifampicin was loaded into the native and surface-modified dendrimers via a simple dissolution solvent evaporation method. Rifampicin-loaded dendrimers were then characterized using several analytical techniques namely; FTIR, DSC, NMR, SEM, and DLS. The polymer encapsulation efficiency (EE%) and percentage of drug loading (DL%) were determined directly using a validated HPLC method. In vitro drug release was studied at pH 7.4 and pH 4.5. The MTT technique was used to assess the cytotoxicity of the dendrimer formulations against raw 264.7 cell lines. Finally, the uptake of dendrimer nanoparticles by raw macrophages was studied using a flow cytometer and fluorescence microscopy techniques. Results: The percentage coverage of 4.0 G PAMAM dendrimer peripheral with PEG was achieved in a range of 38% - 100%, while for mannose moieties was from 44% - 100%. The EE% of unmodified dendrimer was 7.5% (w/w). The EE% of PEGylated dendrimers ranged from 65.0% - 78.75% (w/w), whereas for mannosylated dendrimers was from 43.43% - 57.91% (w/w). The size of the unloaded dendrimer nanoparticles was less than 25 nm, a gradual increase in the size after drug conjugation followed. The zeta potential of dendrimers was positive with values greater than 12 mV, the nanoparticle's zeta potential decreased upon increasing the density of PEG/mannose and after drug loading. FTIR and NMR data showed that rifampicin molecules were conjugated to the dendrimer at three sites; at the surface amines via electrostatic linkages, within the PEG/mannose, and into the dendrimer interior. SEM images of dendrimer nanoparticles confirmed the spherical shape of particles, and DSC data verified drug entrapment. Drug release was found to be affected by the pH of the medium and the extent of dendrimer functionalization. At the physiologic pH, surface-modified dendrimers showed a slower release rate compared to the unmodified dendrimer and free drug. Among surface-modified dendrimers, the release rate was inversely associated with the density of PEG/mannose molecules. At pH 4.5, a relatively higher drug release from all formulations was observed which suggests a burst release inside the alveolar macrophages. Toxicity studies showed that the unmodified dendrimer experienced time-dependent and concentration-dependent cytotoxicity against raw 264.7 cells. The toxicity gradually decreased upon increasing the density of PEG/mannose, and negligible toxicity was detected for formulations with 100% functionalization. Dendrimer nanoparticles were successfully internalized into raw cells after 24 hrs of incubation. The order of nanoparticles permeability was PEG 100% < PEG 85% < PEG 70% < PEG 49% < PEG 38% < unmodified dendrimer < mannose 44% < mannose 69% < mannose 93% < mannose 100%. The significant increase in the uptake of mannosylated dendrimers was due to the interaction with lectin receptors at the surface of raw macrophages, whereas the lower internalization of PEGylated dendrimers was due to the shielding of the surface positive charges. Conclusion: The in-vitro and ex-vivo data studies suggested that the developed novel surface-modified G4 PAMAM dendrimers are suitable drug carriers in terms of biocompatibility, release behaviour, and site-specific delivery of the anti-TB drug rifampicin.

Karel Husník: Economic and Legal Position of the Czech National Bank in the Environment of Inflation and Deflation The aim of this thesis is to analyse the position of the Czech National Bank (CNB) from the legal and economic point of view and to analyse the monetary policy instruments which the central bank can use efficiently in inflation targeting. The primary criterium is macroeconomic environment of inflation, respectively deflation. The thesis newly divides the monetary policy instruments on conventional and non-conventional. The thesis uses multidisciplinary approach, it combines economic and legal point of view. The methodology of this thesis is analysis, description in historical projection and comparation. The laws of the Czech Republic and the documents of CNB, which are available online, are used. Author also worked with data from the Czech Statistical Office and with various economic studies and papers. The first chapter of the thesis analyses position of independent Czech National Bank in its constitutional and lawful position. The Law of the Czech National Bank and the Constitution of the Czech Republic defines the purpose of the CNB and her relationship with other components of state power. The CNB's monetary policy objective is set forth in Article 98 of the Constitution of the...

I takt med den digitala utvecklingen har förutsättningarna för politisk kommunikation förändrats. Vidare möjliggör den kraftigt ökade internetanvändningen för nya digitala verktyg. Genom ett experiment ämnar denna uppsats undersöka vilken roll verktyget microtargeting kan fylla för att stärka svenska förstagångsväljares engagemang och delaktighet i den samhälleiga- och politiska diskussionen. Studiens formulerade forskningsfrågor ämnar belysa uppsatsen huvudsyfte. (1) Vilket samband kan urskiljas mellan effekten på upplevt engagemang och inramning av budskap med hjälp av verktyget microtargeting, då svenska förstagångsväljare exponeras för politiska annonser? (2) Vilken roll kan microtargeting fylla som ett verktyg för politisk kommunikation i den svenska demokratiska offentligheten? Med mål att replikera processen av microtargeting har information om studiens testpersoner, vilka samtliga är svenska förstagångsväljare, samlats in med hjälp av ett socialpsykologiskt personlighetstest. Därefter har manipulerade annonser skapats för att bekräfta deras individuella värderingar och motiv. Testpersonerna har vidare vid en exponering fått värdera till vilken grad de manipulerade respektive icke manipulerade annonserna anses väcka deras engagemang.  Uppsatsen ingår i forskningsfältet politisk kommunikation och utgår från ett teoretiskt ramverk innefattande Entmans (1993) teori om framing samt Habermas (1998) teori om offentligheten. Som analysmetod används en kvalitativ innehållsanalys med deduktiv ansats, för att undersöka hur resultatet kan relateras till det teoretiska ramverket. Det sammanställda resultatet visar att de manipulerade annonserna genererade ett högre upplevt engagemang hos testpersonerna i jämförelse med de originalannonser de exponerats för i studien. Således kunde ett samband mellan inramningen och det väckta politiska engagemanget i studiens experimentgrupp urskiljas. Analysen bidrar till argumentet att användningen av microtargeting kan tänkas minska digitala kunskapsklyftor och öka unga medborgares engagemang och därmed deras deltagande i den offentliga diskussionen. På så vis kan verktyget microtargeting anses vara en anpassning till den digitala offentligheten och vidare potentiellt gynna framtidens politiska kommunikation i en digital, demokratisk kontext i Sverige.

Nanoparticles are currently finding increasing use as drug delivery systems in the treatment of cancer and other disorders. When nanoparticles are introduced into body fluids, they adsorb proteins forming a coating called protein corona. The protein corona is vital since it controls biological responses of nanoparticles through interactions with cells and biological barriers. Due to the dynamic behaviour of protein-protein and protein-nanoparticle interactions, the protein corona evolves during circulation in the body. This results in difficulties to predict the biological behaviour and outcome of nanoparticles. In this work, it is hypothesised that a nanoparticle-protein corona (NP-PC) enriched in specific proteins could serve as a model to determine if the design and formation of a patient-specific nanodrug-protein corona could offer a novel approach to control nanodrug-protein corona evolution. Through usage of a model nanoparticle and model plasmas and by applying shotgun proteomics and SUrface proteomics, Safety, Targeting, and Uptake (SUSTU), NP-PC proteins were identified and quantified. The results indicate that desirable proteins are maintained in the protein corona surface when nanoparticles with a pre-made corona are introduced into model plasma. This implies that a designed NP-PC would be a strategy to control nanodrug-protein corona evolution, offering a route to improve nanodrug targeting and uptake by cells.

Mitochondria carry out several important functions in eukaryotic cells such as energy metabolism, iron-sulfur cluster assembly, apoptosis, signaling pathways, protein quality control etc. Most mitochondrial proteins are synthesized on the cytosolic ribosomes and transported to the organelles by the cytosolic chaperones and mitochondrial protein import machinery based on specific targeting signals. Although, the basic principles of protein import have been explained, many questions remain unanswered, particularly for highly modified mitochondria such as hydrogenosomes. The aim of the study was to investigate protein translocation into hydrogenosomes of a human parasite, Trichomonas vaginalis (Tv) with a focus on the composition, function and structure of protein translocases and the role of targeting signals. The translocase of the outer membrane (TOM) is responsible for the import of most proteins into the organelle. Even though, the presence of a TOM complex in trichomonad hydrogenosomes was predicted, its components were not known. Moreover, the generic structure of the mitochondrial TOM complex was not resolved. This study showed that the TvTOM complex is highly divergent consisting of two modified core subunits - channel- forming TvTom40 isoforms and a Tom22-like protein, and two...