Mantle cell lymphoma (MCL) is a non-Hodgkin B-cell lymphoma with very bad prognosis. The genetic hallmark of MCL, is the translocation t(11;14)(q13;q32) which leads to overexpression of cyclin D1, a D-type cyclin that is not usually expressed at high levels in normal B lymphocytes.   Previous studies indicate that cannabinoid receptors are expressed in lymphoma and have shown that lymphoma cell death is induced as a result of exposure to cannabinoids (ligands).   The aim of this diploma work was to combined cytostatics with the cannabinoid receptor ligand R (+)-Methanandmide (R-MA). Our data suggest that combination treatment with cytostatics and R-MA induces synergistic effects in most cases.

The in vitro cell culture models of human pluripotent stem cells (hPSC)-derived cardiomyocytes (CMs) have gained a predominant value in the field of drug discovery and is considered an attractive tool for cardiovascular disease modellings. However, despite several reports of different protocols for the hPSC-differentiation into CMs, the development of an efficient, controlled and reproducible 3D differentiation remains challenging. The main aim of this research study was to understand the changes in the gene expression as an impact of spatial orientation ofhPSC-derived CMs in 2D(two-dimensional) and 3D(three-dimensional) culture conditions and to identify the topologically important Hub and Hub-Bottleneck proteins using centrality measures to gain new knowledge for standardizing the pre-clinical models for the regeneration of CMs. The above-mentioned aim was achieved through an extensive bioinformatic analysis on the list of differentially expressed genes (DEGs) identified from RNA-sequencing (RNA-Seq). Functional annotation analysis of the DEGs from both 2D and 3D was performed using Cytoscape plug-in ClueGO. Followed by the topological analysis of the protein-protein interaction network (PPIN) using two centrality parameters; Degree and Betweeness in Cytoscape plug-in CenTiScaPe. The results obtained revealed that compared to 2D, DEGs in 3D are primarily associated with cell signalling suggesting the interaction between cells as an impact of the 3D microenvironment and topological analysis revealed 32 and 39 proteins as Hub and Hub-Bottleneck proteins, respectively in 3D indicating the possibility of utilizing those identified genes and their corresponding proteins as cardiac disease biomarkers in future by further research.

A constant flow of various pathogens enters the respiratory system on daily basis through the involuntary mechanism of breathing. Respiratory viral infections are common yet can be fatal in vulnerable populations. Respiratory syncytial virus (RSV) is one of the first and most common viruses that the human population acquire in the first two years of life. Despite the ability of most infants to recover from a RSV infection, many require hospitalization and, in few cases, die from such an infection. The pattern of seasonality of respiratory viruses also applies to RSV. In this work the temperature dependence of infectivity was studied in Hep-2 cells infected with RSV that had been incubated with bronchoalveolar lavage (BAL) fluid. The results indicate a temperature dependence of infectivity. Inhibition of the viral infectivity was observed at three different temperatures 37 ̊C, 40 ̊C and 42 ̊C. The inhibition appears to be linked to the appearance of large agglutinates that appear to reduce the infectivity of RSV. Such a study found that viral neutralization is dependent on a temperature-dependent agglutination reaction. The causality of agglutination formation requires further investigation in order to conclusively confirm the immunological component(s) of this reaction, and how temperature is contributing to this reaction.

Inflammation is a biological defence mechanism applied by living organisms against foreign invaders. In the response to DAMPs and PAMPs, organisms use inflammatory multi-protein complexes to fight the attackers. The most studied inflammasome proteins are NLRP3, ASC and Caspase-1. This study is aimed at understanding the role of NEK7 protein in the NLRP3 inflammasome’s activation, using CRISPR/Cas9 system. To determine the effect of CRISPR/Cas9 and transfection, mRNA expression was analyzed. The results obtained suggest that neither the transfection nor the NEK7 protein knockout have sufficiently worked. This study could not experimentally establish that NEK7 triggers NLRP3 inflammasome activation because ELISA was not conducted to verify the levels of cytokines emitted, due to there being no statistical differences between the samples. Above all, the research question in this thesis project was not answered because the instability of the ACTB reference gene negatively influenced the results. However, previous related studies conclude that NEK7 plays a crucial role in the activation of the NLRP3 inflammasome.

No description available.

Este trabalho analisa a retórica presente nas capas da revista Mente e Cérebro, buscando repre-sentações sociais do cérebro recorrentes em revistas de divulgação científica. É um estudo de caso que descreve como o cérebro é apresentado na articula-ção simbólica que denominamos simbiose ima-gem/palavra no sentido de identificar as estratégias de convencimento a partir dos elementos simbólicos presentes nesta articulação, e não em função do va-lor de verdade discursiva. Procurou-se avaliar como o cérebro é representado para a sociedade e se ele se configura, no periódico, não apenas como um órgão do corpo, mas também como núcleo central do sujei-to humano, comandando suas emoções e controlan-do-lhe a saúde e o viver, podendo ser medicalizado e condicionado para maximizar a vida biológica. Pre-tendeu-se com esta análise, contribuir para o debate sobre o papel relevante das Biociências no imaginário social como dispositivo de produção de verdades so-bre a saúde, o adoecimento e na normatização da vida e do viver na sociedade contemporânea. / This paper analyzes the rhetoric on the covers of Mente e Cérebro Magazine (Brazilian version of Scientific American Mind magazine), to seek social representa-tions of brain on scientific publications. This study describes how the brain is present-ed in a symbolic articulation that we called symbiosis picture/word, to identify the persuasion strategies from the symbolic elements present in this joint, not according to value of truth in the discursive. Sought to evaluate how the brain is represented to society and if it is configured in this magazine chosen, not only as an organ of the body, but also as the core of the human subject, commanding emotions and con-trolling health and living, may be medicalized and conditioned to maximize the bio-logical life. It was intended contribute to the debate on the role of Biosciences in the social imaginary as truths producing device on health, illness and standardization of life and living in contemporary society. / Este trabajo analiza la retórica en portadas de ediciones brasileñas de la re-vista Mente y Cerebro, buscando las representaciones sociales del cerebro recurren-tes en las revistas de divulgación científica. Es un estudio de caso que describe có-mo se presenta el cerebro en la articulación simbólica que llamamos simbiosis ima-gen/palabra para identificar las estrategias de persuasión de los elementos simbóli-cos presentes en esta junta, y no según el valor de verdad discursiva. Buscamos eva-luar cómo se representa el cerebro para la sociedad y si este está configurado, en la revista, no sólo como un órgano del cuerpo, sino también como el núcleo central del sujeto humano, al mando de sus emociones y controlándole su salud y la vida, como puede ser medicalizado y acondicionado para maximizar la vida biológica. Se pretende con este análisis, contribuir al debate sobre el papel de Biociencias en el imaginario social como dispositivo de producción de verdades a respecto de la salud, de la enfermedad y de la normalización de la vida y del viver en la sociedad contemporánea.

Identification, monitoring and quantification of biomolecules in the CNS is a field of growing interest for identifying biomarkers of neurological diseases. In this thesis, silicon needle-shaped multi-molecules sensing microprobes were developed. Our microelectrode array design comprises a needle length of 6mm with 100x50 µm2 cross-section bearing three platinum electrodes with a size of 40x200 µm and 200µm spacing between them. We have used these microprobes for simultaneous glucose and lactate monitoring, using the third electrode for control of non-specific current variations. Local microdroplet protein deposition on the electrode surface was achieved using a pneumatic picopump injection system. Enzyme immobilization on the electrode surface is a key step in microelectrode biosensor fabrication. We have developed a simple, low cost, non-toxic enzyme immobilization method employing poly(ethyleneglycol) diglycidyl ether (PEGDE). Successful biosensor fabrication was demonstrated with glucose oxidase, D-amino acid oxidase, and glutamate oxidase. We found that these biosensors exhibited high sensitivity and short response time sufficient for observing biological events in vivo on a second-by-second timescale. PEGDE-based biosensors demonstrated an excellent long-term stability and reliably monitored changes in brain glucose levels induced by sequential administration of insulin and glucose solution. We then carried out a comparative study of five enzyme immobilization procedures commonly used in Neuroscience: covalent immobilization by cross-linking using glutaraldehyde, PEGDE, or a hydrogel matrix and enzyme entrapment in a sol-gel or polypyrrole-derived matrices. Enzymatic microelectrodes prepared using these different procedures were compared in terms of sensitivity, response time, linear range, apparent Michaelis-Menten constant, stability and selectivity. We conclude that PEGDE and sol-gel techniques are potentially promising procedures for in vivo laboratory studies. The comparative study also revealed that glutaraldehyde significantly decreased enzyme selectivity while PEGDE preserved it. The effects that immobilization can have on enzyme substrate specificity, produce dramatic consequences on glutamate detection in complex biological samples and in the CNS. Our biosensor's results were systematically controlled by HPLC or capillary electrophoresis. The highly selective PEGDE-based biosensors allowed accurate measurements glutamate concentrations in the anesthetized and awaked rats at physiological conditions and under pharmacological and electrical stimulations. The microfabricated multielectrodes based on silicon needles coupled to the simple, non-toxic and mild immobilization method based on PEGDE, open new possibilities for specific neurotransmitter detection in the central nervous system and the study of cell-cell communication in vivo.

Este trabalho analisa a retórica presente nas capas da revista Mente e Cérebro, buscando repre-sentações sociais do cérebro recorrentes em revistas de divulgação científica. É um estudo de caso que descreve como o cérebro é apresentado na articula-ção simbólica que denominamos simbiose ima-gem/palavra no sentido de identificar as estratégias de convencimento a partir dos elementos simbólicos presentes nesta articulação, e não em função do va-lor de verdade discursiva. Procurou-se avaliar como o cérebro é representado para a sociedade e se ele se configura, no periódico, não apenas como um órgão do corpo, mas também como núcleo central do sujei-to humano, comandando suas emoções e controlan-do-lhe a saúde e o viver, podendo ser medicalizado e condicionado para maximizar a vida biológica. Pre-tendeu-se com esta análise, contribuir para o debate sobre o papel relevante das Biociências no imaginário social como dispositivo de produção de verdades so-bre a saúde, o adoecimento e na normatização da vida e do viver na sociedade contemporânea. / This paper analyzes the rhetoric on the covers of Mente e Cérebro Magazine (Brazilian version of Scientific American Mind magazine), to seek social representa-tions of brain on scientific publications. This study describes how the brain is present-ed in a symbolic articulation that we called symbiosis picture/word, to identify the persuasion strategies from the symbolic elements present in this joint, not according to value of truth in the discursive. Sought to evaluate how the brain is represented to society and if it is configured in this magazine chosen, not only as an organ of the body, but also as the core of the human subject, commanding emotions and con-trolling health and living, may be medicalized and conditioned to maximize the bio-logical life. It was intended contribute to the debate on the role of Biosciences in the social imaginary as truths producing device on health, illness and standardization of life and living in contemporary society. / Este trabajo analiza la retórica en portadas de ediciones brasileñas de la re-vista Mente y Cerebro, buscando las representaciones sociales del cerebro recurren-tes en las revistas de divulgación científica. Es un estudio de caso que describe có-mo se presenta el cerebro en la articulación simbólica que llamamos simbiosis ima-gen/palabra para identificar las estrategias de persuasión de los elementos simbóli-cos presentes en esta junta, y no según el valor de verdad discursiva. Buscamos eva-luar cómo se representa el cerebro para la sociedad y si este está configurado, en la revista, no sólo como un órgano del cuerpo, sino también como el núcleo central del sujeto humano, al mando de sus emociones y controlándole su salud y la vida, como puede ser medicalizado y acondicionado para maximizar la vida biológica. Se pretende con este análisis, contribuir al debate sobre el papel de Biociencias en el imaginario social como dispositivo de producción de verdades a respecto de la salud, de la enfermedad y de la normalización de la vida y del viver en la sociedad contemporánea.

Este trabalho analisa a retórica presente nas capas da revista Mente e Cérebro, buscando repre-sentações sociais do cérebro recorrentes em revistas de divulgação científica. É um estudo de caso que descreve como o cérebro é apresentado na articula-ção simbólica que denominamos simbiose ima-gem/palavra no sentido de identificar as estratégias de convencimento a partir dos elementos simbólicos presentes nesta articulação, e não em função do va-lor de verdade discursiva. Procurou-se avaliar como o cérebro é representado para a sociedade e se ele se configura, no periódico, não apenas como um órgão do corpo, mas também como núcleo central do sujei-to humano, comandando suas emoções e controlan-do-lhe a saúde e o viver, podendo ser medicalizado e condicionado para maximizar a vida biológica. Pre-tendeu-se com esta análise, contribuir para o debate sobre o papel relevante das Biociências no imaginário social como dispositivo de produção de verdades so-bre a saúde, o adoecimento e na normatização da vida e do viver na sociedade contemporânea. / This paper analyzes the rhetoric on the covers of Mente e Cérebro Magazine (Brazilian version of Scientific American Mind magazine), to seek social representa-tions of brain on scientific publications. This study describes how the brain is present-ed in a symbolic articulation that we called symbiosis picture/word, to identify the persuasion strategies from the symbolic elements present in this joint, not according to value of truth in the discursive. Sought to evaluate how the brain is represented to society and if it is configured in this magazine chosen, not only as an organ of the body, but also as the core of the human subject, commanding emotions and con-trolling health and living, may be medicalized and conditioned to maximize the bio-logical life. It was intended contribute to the debate on the role of Biosciences in the social imaginary as truths producing device on health, illness and standardization of life and living in contemporary society. / Este trabajo analiza la retórica en portadas de ediciones brasileñas de la re-vista Mente y Cerebro, buscando las representaciones sociales del cerebro recurren-tes en las revistas de divulgación científica. Es un estudio de caso que describe có-mo se presenta el cerebro en la articulación simbólica que llamamos simbiosis ima-gen/palabra para identificar las estrategias de persuasión de los elementos simbóli-cos presentes en esta junta, y no según el valor de verdad discursiva. Buscamos eva-luar cómo se representa el cerebro para la sociedad y si este está configurado, en la revista, no sólo como un órgano del cuerpo, sino también como el núcleo central del sujeto humano, al mando de sus emociones y controlándole su salud y la vida, como puede ser medicalizado y acondicionado para maximizar la vida biológica. Se pretende con este análisis, contribuir al debate sobre el papel de Biociencias en el imaginario social como dispositivo de producción de verdades a respecto de la salud, de la enfermedad y de la normalización de la vida y del viver en la sociedad contemporánea.

Fragment-screening by X-ray crystallography (XFS) is an expensive and low throughput fragment drug discovery screening method, and it requires a lot of optimization for each protein target. The advantages with this screening method are that it is very sensitive, it directly gives the three-dimensional structure of the protein-fragment complexes, and false positives are rarely obtained. The aim of this project was to help Sprint Bioscience assess if the advantages with XFS outweigh the disadvantages, and if this method should be used as a complement to their differential scanning fluorimetry (DSF) screening method. An XFS campaign was run using the oncoprotein vaccinia related kinase 1 (VRK1) as a target protein to evaluate this screening method. During the development of the XFS campaign, a diverse fragment library was created which consisted of 298 fragments that were all soluble in DMSO at 1 M concentration. The crystallization of the protein VRK1 was also optimized in this project to get a robust, high throughput crystallization set up which generated crystals that diffracted at higher resolution than 2.0 Å when they were not soaked with fragments. The soaking protocol was also optimized in order to reduce both the steps during the screening procedure and mechanical stress caused to the crystals during handling. Lastly, the created fragment library was used in screening VRK1 at 87.5 mM concentration with XFS. 23 fragment hits could be obtained from the X-ray crystallography screening campaign, and the mean resolution of the crystal structures of the protein-fragment complexes was 1.87Å. 11 of the 23 fragment hits were not identified as hits when they were screened against VRK1 using DSF. XFS was deemed as a suitable and efficient screening method to complement DSF since the hit rate was high and fragments hits could be obtained with this method that could not be obtained with DSF. However, in order to use this screening method a lot of time needs to be spent in optimizing the crystal system so it becomes suitable for fragment screening. Sprint Bioscience would therefore need to evaluate the cost/benefit ratio of using this screening method for each new project.