A leishmaniose é considerada uma doença tropical extremamente negligenciada, que afeta regiões de extrema pobreza. Trata-se de doença emergente com alta morbidade e mortalidade. Aproximadamente 20 diferentes espécies de parasitas intracelulares obrigatórios do gênero Leishmania têm sido identificadas como patogênicas. Estes protozoários são transmitidos ao hospedeiro humano por meio da picada de insetos hematófagos, conhecidos como flebotomíneos. A quimioterapia é escassa, limitada e muito cara. Os fármacos disponíveis apresentam elevada toxicidade, bem como suscitam número elevado de casos de resistência. Considerando-se que a descoberta e o desenvolvimento de novos agentes leishmanicidas são extremamente necessários o objetivo deste trabalho foi contribuir com essa busca utilizando-se o método de modificação molecular, a latenciação. Tendo em vista que os dendrímeros têm sido muito utilizados como transportadores de fármacos, propôs-se a síntese de fármacos dirigidos dendriméricos de primeira geração de hidroximetilnitrofural (NFOH), composto que apresenta potencial atividade leishmanicida. Assim, pretendeu-se desenvolver fármacos dirigidos de NFOH, que apresentem ação seletiva nos macrófagos, devido à presença de manose na estrutura, além de fármacos dirigidos, que apresentem ação seletiva no parasita, devido à presença de inositol em sua estrutura. Variedade de métodos sintéticos foi utilizada com o intuito de obter os dendrímeros dirigidos propostos. Os estudos sintéticos desenvolvidos indicam a provável obtenção dos dendrímeros dirigidos de manose e de inositol, embora os produtos estejam impuros e sem o agente bioativo incorporado. A maior dificuldade encontrada foi a purificação dos compostos obtidos. Em adição, métodos de modelagem molecular foram empregados para compreender os mecanismos de liberação dos fármacos dendriméricos dirigidos de primeira geração. Características moleculares, tais como disponibilidade espacial e potencial eletrostático, foram avaliadas para predizer a região mais suscetível à ação enzimática com vistas à liberação do composto ativo. / Leishmaniasis is considered a superneglected tropical disease and affects primarily areas of extreme poverty. It represents an emergent illness with high morbidity and mortality. About 20 different species of intracellular parasites of Leishmania spp. have been identified as pathogenic. Those protozoans are transmitted to human hosts by means of the female phlebotomine sandflies bite. The chemotherapy is scarce, limited, and expensive. The drugs available can cause undesirable side effects and there are already reports of the increased number of drug resistance. Considering that, the discovery and the development of new leishmanicide agents are urgently needed, the objective of this work as to contribute to this search using the method of molecular modification, prodrug design. Taken into account that dendrimers can be used as drug carriers, the purpose of this work was the synthesis of first generation dendrimer targeted drugs of hydroxymethylnitrofurazone (NFOH), that shows potential leishmanicide activity. Thus, we designed NFOH targeted drugs with selective action in macrophages, due to the presence of mannose in the structure, and also NFOH targeted drugs that present selectivity for the parasite, due to the presence of inositol in the structure. Several synthetic methods have been used with the aim of synthesizing the targeted dendrimers with NFOH. Synthetic studies suggested the targeted dendrimers of D-mannose and of myo-inositol have been obtained, although impure and without the bioactive agent. The major difficulty was the purification of the compounds. In addition, molecular modeling methods were applied to understand the bioactive compound releasing from the first generation leishmanicide targeted dendrimers. Molecular features as spatial hindrance and electrostatic potential were evaluated to predict which region would be the most available to an enzymatic action regarding the bioactive compound release.

Ischaemia-reperfusion (IR) injury makes a major contribution to graft damage during kidney transplantation and increases the risks of primary non-function, delayed graft function and rejection. Oxidative damage to mitochondria is a key early event in IR injury. The aim of this project was to examine the safety and efficacy of the mitochondria-targeted antioxidant MitoQ in reducing pig and human kidney IR injury using an ex vivo normothermic perfusion (EVNP) system. Over a range of 500 nM to 250 µM using a 150 pig kidneys and 80 declined deceased human kidneys, MitoQ was successfully taken up by pig and human kidneys in a concentration-dependent manner, resulting in stable tissue concentrations over 24 hours of cold storage followed by 6 hours of EVNP. The uptake of MitoQ was increased approximately 2-fold when MitoQ was administered to warm (rather than cold) kidneys and when kidneys were preserved using hypothermic machine perfusion (rather than cold static storage). 50 µM MitoQ, administered to pig kidneys at the end of warm ischaemia, significantly increased renal blood and urine output flow at the end of 6 h EVNP compared to the control group. Creatinine clearance was numerically higher in the 50 µM MitoQ group compared to the control group but the difference did not reach statistical significance. To test the safety and efficacy of MitoQ in human kidney IRI, pairs of declined deceased human kidneys were used, with one kidney in each pair used as control. The total urine output, creatinine clearance and percentage fall of serum creatinine were numerically higher in the 50 µM MitoQ group compared to the control group, although the differences did not reach statistical significance during 3 h of EVNP. There was a significant difference in the renal blood flow between the 50 µM MitoQ group and the control group at the end of the first hour of EVNP. The renal blood flow remained relatively stable during the first hour of EVNP in the 50 µM MitoQ group compared to a significant decrease in renal blood flow in the control group. There was no effect on fractional excretion of sodium or oxidative injury markers (protein carbonyl formation, lipid peroxidation) in pig or human kidneys, which is consistent with previous studies that demonstrated the requirement of >24 hour after reperfusion for manifestation of changes in these parameters. In this thesis, I was able to successfully demonstrate the safety and potential efficacy of MitoQ in ameliorating renal IRI using pig kidneys. While more declined deceased human kidneys need to be analysed to fully explore the potential efficacy of MitoQ in ameliorating renal IRI, this study provides important data that will help inform future studies and ultimately a clinical trial for assessing the efficacy of the mitochondria-targeted antioxidant MitoQ in human kidney transplantation. My findings suggest that MitoQ has the potential to increase the use of marginal kidneys and to improve graft and patient outcomes.

The demand for labour force in the European building industry has increased in recent years due to the constant migrations and the need to provide housing, schools and hospitals to accommodate this growing population. Due to this scenario and to governmental policies that were created to supply this need, Swedish building companies want to recruit more female employees in particular. The purpose of this case study is to investigate whether, and if so how, the building company NCC uses its external communication to strategically attract more women to the industry. Our research was of a qualitative nature. We analysed, with the help of a Multimodal Critical Discourse Analysis approach, recruitment materials and job advertisements published on the company’s homepage, in order to demonstrate how men and women are represented in NCC’s recruitment communication and in which way the company communicates specifically to women. We have also analysed which aspects of the company’s recruitment communication were appealing to women, through focus group interviews made with female engineering students. As theoretical ground for this case study we have chosen previous research and literature in the fields of organizational communication, employer branding, decoding in communication processes, gender studies, stereotyping and gender aware recruitment. The findings in this study show that both men and women are represented in NCC’s recruitment communication, but that a bigger focus is placed on the visual representation of women.  NCC strategically aims at attracting women both in the way female workers are visually portrayed on the homepage but even in the wording found in the company’s recruitment texts and job advertisements. This targeted communication does not reach its full potential in terms of attractiveness to women though, perhaps due to the rather cosmetic visual representation of the company’s work environment, as well as the lack of personal connection that is made between the portrayed employees and the potential job candidates.

En France, le cancer colorectal (CCR) est la 2ème cause de décès par cancer. Chez les patients, lorsque la tumeur est localisée, elle est réséquée par chirurgie. Toutefois, 50% des patients sont diagnostiqués à un stade métastatique, ces patients sont alors traités par chimiothérapie (FOLFOX/FOLFIRI), souvent en combinaison avec des thérapies ciblées incluant des anticorps tel que le Cetuximab (anti-EGFR) ou le Bevacizumab (anti-VEGF). Cependant, il reste encore environ 40% des patients qui ne répondent pas au traitement et l’une des causes les mieux établies est l'influence des mutations de la voie RAS en aval du récepteur à l'EGF sur la réponse au Cetuximab. C’est pourquoi l’identification de nouvelles cibles moléculaires accessibles aux anticorps permettrait le développement de nouveaux modèles de thérapie ciblée. Depuis peu, les Claudines ont suscité un intérêt pour le ciblage tumoral comme la claudin-4 dans les cancers de l’endomètre ou de la prostate. Très récemment, les résultats d'une étude clinique ont démontré que la combinaison d'un anticorps anti-Claudin-18.2, l'Acm IMAB362, avec la chimiothérapie prolongeait nettement la survie chez des patients atteints de cancer gastrique avancé.C’est dans cette optique que nous nous sommes focalisés sur les claudines dans le CCR. Tout d’abord en étudiant leur expression dans des échantillons de patients atteints de CCR métastatique. En analysant l’expression des claudines dans les nouveaux sous-types moléculaires, nous avons montré que certaines pouvaient avoir une valeur pronostique. Nous avons également identifié des claudines comme cibles thérapeutiques potentielles dans le CCR, parmi elles la CLDN1. En effet, nous avons montré que la forme membranaire de la CLDN1 était surexprimée dans les tumeurs primaires et métastases du CCR. Nous avons développé un anticorps monoclonal (Acm) ciblant les parties extracellulaires de la CLDN1. Nous avons montré que le ciblage thérapeutique de la CLDN1 par Acm provoquait un ralentissement significatif de la croissance, la migration et l'invasion des cellules tumorales aussi bien in vitro que in vivo. Afin d’améliorer l’efficacité du ciblage de la CLDN1 par Acm, nous avons couplé ce dernier avec une toxine, générant ainsi un Antibody Drug conjugated (ADC). Nous avons montré que le ciblage de la CLDN1 par un ADC diminuait la survie cellulaire in vitro en culture cellulaire 2D, mais également celle des sphéroïdes via un effet cytotoxique. Ce travail a permis d'établir la preuve de concept du ciblage de la CLDN1 aussi bien par Acm que par ADC. Afin de finaliser ce travail, ces résultats doivent être confirmés in vivo. A terme, nous espérons que ce ciblage puisse trouver sa place au sein de l’arsenal thérapeutique du CCR métastatique, en particulier chez les patients résistants aux traitements. / Colorectal cancer (CRC) is one of the major causes of cancer-related deaths in the Western world. When localized, CRC is often curable by surgery. However, 50% of patients are diagnosed at a metastatic stage, these patients are then treated with chemotherapy (FOLFOX / FOLFIRI), often in combination with targeted therapies including antibodies such as Cetuximab (anti-EGFR) or Bevacizumab (anti -VEGF). Despite these treatment, almost 40% of patients develop resistance. One of the most known resistance mechanisms of resistance is due to the RAS pathway downstream of the EGF receptor in response to Cetuximab. Therefore, more therapeutic options are required particularly by identifying new molecular targets that can be reached by antibodies. Recently, Claudines have generated interest as targets in cancer, such as claudin-4 in endometrial or prostate cancer. More recently, the results of a clinical study demonstrated that the combination of an anti-Claudin-18.2 antibody, IMAB362 mAb, with chemotherapy significantly prolonged survival in patients with advanced gastric cancer.Therefore, one of our aims was to focus on claudins in CCR. First, by studying their expression in metastatic CRC patient samples. We demonstrated the prognostic value of some claudins, after analyzing their gene expression in the new molecular subtype of CRC. Beside, we identified some claudins as potential therapeutic targets in CCR, among them claudin-1 (CLDN1). Indeed, we showed that the membrane form of CLDN1 is overexpressed in primary tumors and metastases of CRC. Therefore, we developed a monoclonal antibody (mAb) targeting the extracellular parts of CLDN1. We showed that therapeutic targeting of CLDN1 by mAb significantly decreased tumor cell growth, migration and invasion both in vitro and in vivo. In order to improve the efficiency of targeting CLDN1 by mAb, we conjugated it with a toxin, thus generating an Antibody Drug Conjugate (ADC). We showed that CLDN1 targeting by an ADC decreased cell survival in vitro in 2D cell culture, but also spheroids growth via a cytotoxic effect.This work demonstrated the proof of concept of CLDN1 targeting by both mAb and ADC. In order to achieve this work, the next steps remains on testing ADC affect in vivo models. CLDN1 is a good therapeutic target in the CCR. In the long term, we hope that its targeting can find its place within the therapeutic arsenal of metastatic CRC, particularly in treatment of resistant patients.

Embora o melanoma represente apenas 4% das neoplasias malignas da pele, é considerado a mais grave por ser altamente etal. Em virtude da via MAPK (Mitogen activated protein kinase) estar intimamente ligada ao descontrole da proliferação celular, especialmente em melanoma, esta via se tornou um alvo para o desenvolvimento de terapias direcionadas a oncogenes, como os potentes quimioterápicos Vemurafenibe (inibidor da mutação V600E em BRAF - BRAFV600E) e Trametinibe (inibidor de MEK). Cada vez mais, melhores taxas de respostas vêm sendo alcançadas com os novos medicamentos, porém a maioria dos pacientes está sujeita a recidivas após 7 meses de tratamento devido ao desenvolvimento de quimioresistência, justificando a constante busca por novos compostos terapêuticos. Dados de nosso laboratório indicam que 4-nerolidilcatecol (4-NC) induz aumento na expressão de p53, produção de ROS e dano ao DNA, culminando em apoptose dependente de caspase-3 em células de melanoma por ser um inibidor proteassomal. Além disto, o 4-nerolidilcatecol (4- NC) demonstrou efeito inibitório na proliferação de células de melanoma em modelo de cultura organotípica de pele. Desta forma, este projeto visa avaliar a possibilidade de superação da quimioresistência aos inibidores de BRAF e de MEK, utilizando terapias combinatórias com 4-NC em células de melanoma humano resistentes a estes inibidores. Primeiramente, linhagens celulares de melanoma resistentes aos inibidores de BRAF (R) e BRAF/MEK (DR) foram geradas a partir de células parentais BRAF mutadas (P) e caracterizadas por MTT, microscopia de fluorescência e western blotting. Estas células foram submetidas ao tratamento com 4-NC que apresentou citotoxicidade na concentração de 30µM, inibição de formação de colônias e diminuição na invasão em modelos in vitro de culturas 2D e 3D em todas as linhagens estudadas (P, R e DR). O 4-NC foi ainda capaz de induzir estresse de retículo endoplasmático com indução de apoptose. Visando a explorar o efeito terapêutico in vivo do 4-NC, outro estudo foi conduzido em animais submetidos a enxerto xenográfico com células parentais de melanoma NRAS mutadas. Após desenvolvimento do tumor, os animais foram tratados 3 vezes por semana durante 3 semanas com 4-NC (10 mg/kg) por via i.p. O 4-NC foi capaz de inibir em até 4 vezes o crescimento dos tumores xenográficos (4/10) quando comparado com os controles, com remissão completa do tumor em um animal. A expressão de p53 e PARP clivada foi aumentada nos tumores dos animais tratados, sugerindo apoptose. A expressão gênica de MMP2 e MMP14 estava diminuída nas mesmas amostras, demonstrando o papel do 4-NC na inibição da invasão do melanoma in vivo. Finalmente, a toxicidade sistêmica do 4-NC foi avaliada nas mesmas doses empregadas no ensaio in vivo de tumorigênese. A baixa toxicidade observada nos ensaios toxicológicos com tratamentos sub-crônicos com 4-NC e a citotoxicidade demonstrada em modelos xenográficos nos leva a considerar este composto como promissor para estudos futuros e sua aplicação no tratamento do melanoma cutâneo humano, incluindo pacientes resistentes aos inibidores de BRAF e MEK. / Melanoma accounts for only 4% of malignant neoplasms of the skin, but is considered the most serious because it is highly deadly. Because the MAPK (Mitogen activated protein kinase) pathway is closely linked to the lack of control of cell proliferation, especially in melanoma, this pathway has become a target for the development of oncogene-targeted therapies, such as the potent chemotherapeutic agents Vemurafenib (V600E mutation inhibitor in BRAF - BRAFV600E) and Trametinib (MEK inhibitor). Increasingly, better response rates have been achieved with the new drugs, but most patients are subject to relapses after 7 months of treatment due to several mechanisms, which justify the constant search for new therapeutic compounds. Data from our laboratory indicate that 4-nerolidylcatechol (4-NC) induces increased p53 expression, ROS production and DNA damage, culminating in caspase-3 dependent apoptosis in melanoma cells. The 4-NC compound demonstrated an inhibitory effect on melanoma cell proliferation in an organotypic skin culture model. Thus, this project aims to evaluate the possibility of overcoming the existing chemoresistance to BRAF and MEK inhibitors, using 4-NC combinatory therapies in human melanoma cells resistant to these inhibitors. Firstly, melanoma cell lines resistant to BRAF (R) and BRAF / MEK (DR) inhibitors were generated from naive cells mutated BRAF (N) and characterized by MTT, fluorescence microscopy and western blotting. These cells were submitted to 4-NC treatment that showed cytotoxicity with 30 µM, inhibition of colony formation and decrease in invasion in 2D and 3D in vitro models in all cell line studied (N, R and DR). Furthermore, 4-NC was able to induce endoplasmic reticulum stress with apoptosis induction. In order to explore the in vivo therapeutic effect of 4-NC, an additional study was conducted using xenograft model with NRAS-mutated melanoma cell line. After tumor development, the animals were treated 3 times per week for 3 weeks with 4-NC (10 mg / kg) by i.p. injection. 4-NC was able to inhibit up to 4- fold the growth of xenograft tumors (4/10) when compared to controls, with complete tumor remission in one animal. Cleaved PARP and p53 expression were increased in the tumors of treated animals, suggesting apoptosis. MMP2 and MMP14 gene expression were decreased in the same samples, demonstrating the role of 4-NC in inhibiting melanoma invasion in vivo. Finally, the systemic toxicity of 4-NC was evaluated at the same doses employed in the in vivo tumorigenesis assay. The low toxicity observed in the toxicological assays with sub-chronic 4- NC treatments and the demonstrated cytotoxicity in xenograft models leads us to consider this compound as promising for future studies and its application in the treatment of cutaneous human melanoma, including patients resistants to BRAF and MEK inhibitors.

Cancer, which is characterised by aggressiveness and increased capacity for metastatic spread still requires basic researchers and clinicians to direct enormous efforts toward the development of novel therapeutic targets. Potential novel targets can be identified and exploited in combination with currently existing therapeutic approaches to improve their efficacy and overcome treatment resistance of tumour cells, protecting the patient from recurrence. To achieve this, different strategies and techniques can be proposed to identify the most promising candidate molecules for further exploitation as therapeutic targets. Human epidermal growth factor receptors (HERs) and NF-E2-related factor 2 (NRF2) are regulators of cellular proliferation and determinants of cancer initiation and progression. NRF2 and HERs confer cancers with resistance to several therapeutic agents. Nevertheless, there is limited understanding of the regulation of HER expression and activation, and the link between NRF2 and HER signalling pathways. This research has demonstrated that pharmacological activation of NRF2 by tert-butyl hydroquinone (tBHQ) upregulates the expression of HER family receptors, HER1 and HER4, elevates phospho protein kinase B (pAKT) levels, and enhances the proliferation of ovarian cancer cells. Pharmacological inhibition using retinoic acid (RA) and bexarotene and genetic inhibition using small interfering RNA (siRNA), did the opposite. Further, tBHQ caused transcriptional induction of HER1 and HER4 with different levels of expression, while siRNA-mediated knockdown of NRF2 prevented this and further caused transcriptional repression. A panel of potent NRF2 inhibitors were screened with the hope of finding the most potent for further investigation. Bexarotene was found to be the most potent and was used either alone, or in combination with lapatinib or erlotinib. The use of these drugs in combination with bexarotene resulted in the repression of HER1, HER2, HER3 and HER4 expression, inhibition of NRF2, elevation of ROS, depletion of glutathione and enhanced cytotoxicity in PEO1, OVCAR3, SKOV3 and MCF7-AREc32 cell lines. This explained the crosstalk mechanism between HER receptor family and NRF2 and the role of NRF2 in drug resistance and as a relevant anti-cancer target which opens up novel avenues of targeting HER receptor kinase family and NRF2 pathways for improving cancer therapy.

The myc family of oncogenic transcription factors, which includes c-myc, N-myc and L-myc, control major cellular processes such as proliferation and differentiation by integrating upstream signals and orchestrating global gene transcription. They do this largely through dimerising with Max, which together bind to enhancer (E)-box elements in DNA. Myc proteins function similarly but differ in potency and tissue distribution. For instance, N-myc is expressed predominantly during development in undifferentiated cells of the nervous system, whereas c-myc is ubiquitously expressed in all proliferating cells. Myc proteins, when deregulated, are major drivers of tumourigenesis. Myc deregulation occurs in up to 70% of all human cancers and is often associated with the most aggressive forms. For example, MYCN, the gene encoding N-myc, is amplified in 20-30% of neuroblastomas, and amplification strongly correlates with advanced stage and poor prognosis. Myc proteins are therefore considered “most wanted” targets for cancer therapy, but have long been considered undruggable mainly due to challenges in nuclear drug delivery and physically targeting myc directly given that it is a largely disordered protein that lacks discernible clefts and pockets for small molecules to inhabit. Furthermore, c-myc is important in normal tissue maintenance so the effect of its inhibition in humans is difficult to predict. However, recent in vivo studies showed that systemic myc inhibition (using the peptide pan myc inhibitor Omomyc) has mild and reversible side effects and induces tumour regression. This has alleviated concerns about the side effects that myc inhibition might have, and reinforced the promise of myc as a powerful drug target. However, the translation of Omomyc into the clinic has been hindered by poor cellular delivery. In fact, no direct myc inhibitor has yet been approved, indicating that novel approaches are needed. Moreover, inhibitors in development tend to inhibit all myc family proteins. An inhibitor that could specifically target N-myc might improve safety through bypassing c-myc inhibition. This could be used for the treatment of N-myc-driven cancers such as MYCN-amplified neuroblastoma. Nanobodies, camelid-derived single-domain antibodies, are a relatively new drug class. Whilst some are already in clinical trials for a wide range of diseases, these are specific for cell-surface or extracellular targets. However, their properties also make them ideal for use as intracellular antibodies or ‘intrabodies’. For example, they are small (just 12-15 kDa) and highly soluble due to naturally occurring hydrophobic to hydrophilic amino acid substitutions. Their small size and convex shape makes them advantageous in capturing structures in intrinsically disordered proteins and allows them to reach hidden epitopes not accessible to conventional antibodies, which could improve biological activity. Importantly, nanobodies retain the high specificities and affinities of conventional antibodies. Their small, single-domain nature also means they can be engineered with ease to modify aspects of their localisation and/or function. For example, they can be coupled to carrier molecules to facilitate cellular entry, and a nuclear localisation signal (NLS) can be added to drive them into the nucleus. Also, it was recently shown that an F-box domain could also be incorporated into nanobodies to recruit degradation machinery to its antigen, which depletes the antigen from cells via the proteasomal degradation pathway. Due to their highly advantageous properties, nanobodies raised against N-myc might overcome the barriers to targeting N-myc, providing potent and specific means of directly inhibiting N-myc therapeutically, which has not yet been achieved. In this thesis, nine unique nanobodies were raised against N-myc. These included three against the basic helix-loop-helix leucine zipper (bHLH-LZ) domain where Max dimerises, and six against the transactivation domain where numerous regulatory and cofactor proteins bind, such as the E3 ubiquitin ligase Skp2. Nanobodies against the transactivation domain were more specific for N-myc and were shown to inhibit its Skp-2-mediated ubiquitylation. This could provide novel means of eradicating tumours based on a study showing that inhibition of ubiquitylation at this domain triggers a transcriptional ‘switch’ that induces a non-canonical target gene Egr1, leading to p53-independent apoptosis. A nanobody against the bHLH-LZ (Nb C2) was shown to bind both N- and c-myc to similar magnitudes. Its affinity for N-myc bHLH-LZ was superior to that of the small molecule myc inhibitor 10058-F4, which prolongs survival in a MYCN-dependent mouse model of high-risk neuroblastoma. Nb C2 spontaneously transduced cell membranes and its coupling to a novel small molecule carrier (SMoC) enhanced its cellular uptake. Furthermore, the addition of a NLS increased its nuclear localisation. Preliminary experiments showed that Nb C2 might slow proliferation and induce apoptosis in cancer cell lines expressing c-myc, suggesting that Nb C2 might also be effective against cancers characterised by deregulated c-myc. Taken together, data generated in this thesis have revealed intriguing findings that provide a basis for the development of these nanobodies for the treatment of N-myc- and c-myc-driven cancers.

The goal of the diploma thesis is to analyze the situation of behavioral targeted advertising in the Czech Internet market, assess its potential in the future and evaluate the effectiveness of behavioral targeting on the Czech market. This thesis is focused first on the Internet advertising market, media planning, explanation of selected targeting approaches and on detailed description of the behaviorally targeted ads including the online privacy issue. In the practical part analysis of the behavioral targeted advertising on the Czech market through expert interviews and evaluation of the effectiveness of behavioral targeting on an example of an advertising campaign of selected company is to be found.

The current study evaluated the implementation of evidence-based reading interventions using a multifaceted implementation measurement approach. Multilevel modeling was used to examine how three direct measures of implementation related to each other and to student academic outcomes and to examine patterns of implementation across time. Eight instructional groups were video taped weekly for nine weeks, and pre- and post-test assessments were given to 31 at-risk kindergartners from two schools using established evidence-based practices. Each implementation measure represented a different measurement approach (i.e., discrete behavioral measurement, global ratings) and focused on different aspects of implementation (e.g., structural, process, or multicomponent elements). Overall, results of this analysis indicated that (a) the implementation tools were highly correlated with each other, (b) only the multicomponent tool independently accounted for group differences, (c) together the multicomponent and process-oriented measures appear to account for additional variance in group differences, and (d) there were no significant trends in implementation across time as measured by any of the tools, however there were significant differences in trends over time between groups when using the structural measure. Implications for research and practice are discussed, including the importance of taking a multifaceted approach to measuring implementation and aligning implementation measures with program theory.

As doenças negligenciadas infectam mais de um bilhão de pessoas no mundo, no entanto, uma fração ínfima dos fármacos registrados nos últimos anos é direcionada a essas patologias. Entre as doenças negligenciadas consideradas prioridade, encontram-se doença de Chagas e leishmaniose. A tuberculose, que recentemente não é mais classificada como negligenciada pela Organização Mundial da Saúde, também está entre as prioridades do Ministério da Saúde no Brasil. O arsenal terapêutico disponível para o tratamento destas enfermidades compreende fármacos com toxicidade elevada e resistência crescente, entre outros inconvenientes. Dessa forma, verifica-se a importância no desenvolvimento de novos fármacos para as referidas doenças. Para este fim, o presente trabalho objetivou a síntese de um fármaco dirigido de hidroximetilnitrofural (NFOH) com manana para a liberação específica em macrófagos. O processo de latenciação foi utilizado para a consecução desse objetivo por meio da estratégia de fármacos dirigidos, buscando elevar seletividade e potência do NFOH. Este composto protótipo possui ação potencial tripanomicida, leishmanicida e tuberculostática. O transportador utilizado foi a manana, polímero de manose, que direciona a liberação da molécula ativa em receptores de manose observados na superfície de macrófagos. Em razão da presença de Trypanosoma cruzi, Leishmania sp. e Mycobacterium tuberculosis no interior de macrófagos em parte de seu ciclo biológico, o referido receptor é considerado um alvo adequado à liberação específica. Na tentativa de obter o referido fármaco dirigido, diversas metodologias sintéticas foram desenvolvidas. Também, realizaram-se estudos de modelagem molecular a fim de se obter informações sobre a liberação do fármaco dirigido proposto. Face à dificuldade de obtenção do derivado de NFOH, adicionalmente, realizou-se metodologias sintéticas a fim de se sintetizar um fármaco dirigido de metronidazol, fármaco que possui atividade antichagásica conhecida. Realizaram-se, também, estudos de modelagem molecular relacionados à sua liberação. / Neglected diseases infect more than one billion people worldwide. However, a small fraction of the drugs registered in recent years is addressed to these pathologies. Leishmaniasis and Chagas\' disease are among the neglected diseases considered priority. Tuberculosis, which is no longer classified as a neglected disease by the World Health Organization, is also among the priorities of the Ministry of Health in Brazil. The therapeutic arsenal available for the treatment of those diseases comprehends drugs with high toxicity and increasing resistance, among other inconveniences. Thus, it is important the development of new drugs for those diseases. For this purpose, this study aimed at synthesizing hydroxymethylnitrofurazone (NFOH) drug targeted with mannan for the specific release in the macrophages. Prodrug design was used to achieve this goal by means of targeted drugs strategy towards increasing NFOH selectivity and potency. This lead compound has potential trypanocidal, leishmanicidal and tuberculostatic activity. The directing group used was a mannose polymer, mannan, which directs the release of the active compound to mannose receptors (MR) on macrophages surface. Due to the presence of Trypanosoma cruzi, Leishmania sp. and Mycobacterium tuberculosis inside macrophages during part of their life cycle, MR are considered a suitable target for specific release. In an attempt to obtain the proposed compound, several synthetic methods have been developed. Also, molecular modeling studies were carried out to obtain information about the targeted drug release. Due to the difficulty of obtaining NFOH derivative, the synthesis of metronidazole targeted drug was tried. Also molecular modeling studies to predict metronidazole release from mannan derivative were performed.