As Neoplasias mieloproliferativas (NMP) representam um vasto grupo de doenças clonais hematológicas malignas com três elementos principais: Policitemia vera (PV), Trombocitemia essencial (TE), e Mielofibrose Primária (MFP). JAK2 é uma proteína citoplasmática com atividade de tirosina quinase com função na transdução de várias vias na hematopoiese. A identificação da mutação do gene JAK2 (JAK2V617F) nas PV, TE e MFP representa um importante avanço para a compreensão da biologia destas NMPs. Variações marcantes na frequência desta mutação são observadas entre os diferentes estudos e acredita-se que um dos fatores responsáveis por estas diferenças seja a sensibilidade do método utilizado. Atualmente, diversas técnicas para detecção de JAK2V617F têm sido utilizadas, testadas e validadas quanto à sua sensibilidade e especificidade, entre elas: PCR RFLP (Restriction Fragment Lenght Polymorphysm), ARMS PCR (Amplification-Refractory Mutation System), HRM (High-Resolution Melt Analysis) e Sequenciamento pela técnica de Sanger. Neste estudo foram realizadas todas as metodologias citadas anteriormente para a detecção da mutação de JAK2V617F em amostras de sangue de 136 pacientes (PV=20; MFP=20; TE=28; suspeita de NMP=68). Os resultados obtidos foram concordantes para as quatro técnicas empregadas nos pacientes com PV e MFP, já nos pacientes com TE as metodologias PCR-ARMS e PCR-HRM detectaram a mutação JAK2V617F em 67,8% enquanto o PCR-RFLP e o Sequenciamento pela técnica de Sanger foi 71,4% e 64,2% respectivamente. Nos casos onde houve suspeita diagnóstica de NMP também foram encontradas discordâncias entre as metodologias PCR-RFLP (4,4%) e PCR-HRM (1,5%) quando comparadas ao PCR-ARMS (3%) e o Sequenciamento (3%). O PCR-ARMS foi considerado nesse estudo como a melhor técnica para a detecção da mutação JAK2V617F, devido o menor risco de contaminação cruzada durante a reação, baixo tempo de execução, além da sua capacidade de determinação da carga alélica de JAK2, importante para o acompanhamento do paciente / Myeloproliferative neoplasms (MPN) represent a large group of clonal hematologic malignant diseases with three main members: Polycythemia Vera (PV), Essential Thrombocythemia (ET), and Primary Mielofibroses (PMF). JAK2 is a cytoplasmic tyrosine kinase protein and is important in different signal transduction pathways. Identification of JAK2V617F mutation in PV, ET and PMF is an important advance for understanding the biology of MPN. Differences in the frequency of this mutation are reported among different studies and it is believed that technical sensitivity could be the major reason for this variability. Currently, several techniques for detection of JAK2V617F have been developed, tested and validated for their sensitivity and specificity, including: PCR-RFLP (Restriction Fragment Lenght Polymorphysm), PCR-ARMS (Amplification Refractory Mutation System), PCR-HRM (High-Resolution Melt analysis) and Sanger Direct Sequencing. The present study, evaluated all four molecular diagnostic methods mentioned above blood samples from 136 patients (PV=20; MFP=20; ET=28 and other MPN=68). Comparable results were observed for PV and PMF when all technics were applied. Patients with diagnosis of ET JAK2V617F mutations were detected in 67.8% when PCR-ARMS and PCR-HRM were used whilst PCR-RFLP and direct sequencing detected 71.4% and 64.2% respectively. In 68 patients with suspicion of MPN discordant results were seen between PCR-RFLP (4.4%) and PCR-HRM (1.5%) when compared to PCR-ARMS (3%) and direct sequencing (3%) related to JAK2V617F frequency. In conclusion PCR-ARMS was considered the most reliable methodology for JAK2V617F detection by presenting the lowest risk for cross contamination, less laborious, and the ability in determining allele burden that is becoming an important tool for risk stratification

Denna rapport behandlar det arbete som under våren 2020 utfördes av en grupp på sju civilingenjörsstudenter i data- och mjukvaruteknik vid Linköpings universitet som en del av kursen TDDD96 – Kandidatprojekt i programvaruutveckling. Projektet utfärdades på efterfrågan av företaget Airpelago, och gick ut på att utveckla en stabil grund för ett molnbaserat videoströmningssystem. Denna rapport beskriver den färdiga produkten, förklarar de beslut som togs, beskriver vilka problem som uppstått och hur dessa lösts, samt diskuterar det slutgiltiga arbetet. Rapporten innehåller även sju individuella bidragskrivna av gruppens enskilda medlemmar / <p>Presentationen genomfördes på distans</p>

Les institutions de la République romaine (509-27 av. J. C.) prévoyaient théoriquement que les magistratures électives et annuelles ne devaient jamais être vacantes. Ce principe juridique fondamental avait pour conséquence une continuité dans la détention du pouvoir exécutif qui reposait sur le ius, les auspicia et l'imperium. Cependant, il arriva à maintes reprises que les magistratures supérieures - consulat, tribunat militaire à pouvoir consulaire - fussent suspendues en raison soit d'entrave à la tenue des comices électoraux soit de scrupules religieux entraînant l'expiation rituelle et la renouatio auspiciorum. La légitimité et la légalité de la solution à la vacance du pouvoir exécutif reposait alors sur les patres auctores, détenteurs de l'auspicia patrum. Le rite de l'interregnum, procédure juridique archaïque, était l'émanation la plus claire de cette auctoritas patrum, apanage exclusif des sénateurs patriciens. Les vénérables pères, descendants des plus illustres familles de Rome, étaient les seuls à même de remédier à la vacance des magistratures par ce rite qui remontait, d'après la tradition, à l'époque royale latino-sabine et renvoyait au mythe bien connu du démembrement et de l'apothéose de Romulus. Le partage du corps souverain constitue à ce titre un symbole fondamental pour la représentation de l'auctoritas patrum et de la magistrature républicaine dont il serait peut-être vain de rechercher l'historicité. L'objet de cette étude est donc d'analyser la fort ancienne institution de l'interregnum qui, comme bien d'autres, était caractérisée par un passage progressif du sacral au juridique. Le droit public prévoit dans tout système institutionnel des recours d'exception révélant la représentation psychologique du pouvoir souverain. Rome n'échappe pas à la règle voire même elle put, d'une certaine façon, l'inventer.

Fungos do complexo de espécies do gênero Paracoccidioides são agentes causadores da Paracoccidioidomicose (PCM), uma micose sistêmica e endêmica na América Latina. Lesões orais ocorrem em aproximadamente 80% dos pacientes com a forma crônica e são caracterizadas pela presença de granulomas que podem indicar a extensão da doença, de acordo com sua organização. A evolução da doença depende não somente da virulênica do fungo, mas também da integridade da resposta imune e de fatores genéticos do hospedeiro. Dentre este último, destacam-se os polimorfismos de nucleotídeo único (SNPs) em vários componentes da resposta imunológica. Dentre os SNPs, cujas associações já foram descritas para diversas doenças infecciosas, destacam-se os presentes em genes como TNF-α, JAK1, VDR, DC-SIGN e FcγRIIa. Dessa forma, este trabalho teve como objetivo avaliar a frequência dos SNPs DC-SIGN (rs4804803), FcγRIIa (rs1801274), JAK1 (rs11208534), TNF-α (rs1800629) e VDR (rs7975232) em amostras de DNA de pacientes com PCM oral e verificar se há associações com o padrão de organização dos granulomas nas lesões. Para isso, foram utilizadas 66 amostras de DNA obtidas a partir de biópsias de lesões orais e suas respectivas lâminas para a classificação do granuloma e 106 amostras de DNA de indivíduos saudáveis. A genotipagem foi realizada pelo método de discriminação alélica por PCR em tempo real e os resultados demonstraram que no SNP VDR (rs7975232), o genótipo CC (p-valor<0,001, OR = 5,94, IC 95% 2,07 – 17,05) ou indivíduos que carregam o alelo C (p-valor = 0,027, OR = 2,71 IC 95% 1,07 – 6,86 para CC+AC) e o genótipo GG no DC-SIGN (rs4804803) (p-valor = 0,032, OR = 3,76, IC 95% 1,06 – 13,38) estão associados ao aumento do risco de desenvolvimento da PCM oral. Porém, não foram observadas correlações com o tipo de granuloma. Esses dados indicam que as variações derivadas dos SNPs VDR (rs7975232) e DC-SIGN (rs4804803) estão relacionadas à susceptibilidade da PCM oral nas amostras analisadas. / Fungi of the complex of species of the genus Paracoccidioides are agents that cause Paracoccidioidomycosis (PCM), a systemic mycosis and endemic in Latin America. Oral lesions occur in approximately 80% of patients with the chronic form and are characterized by the presence of granulomas that can indicate the level of the inflammatory reaction and the extension of the PCM. The disease evolution depend not only of the fungi strain, but also on the integrity and type of the immune response and on host genetic factors. Among the latter, single nucleotide polymorphisms (SNPs) are highlighted in several components of the immune response. Among the SNPs whose associations have already been described for various infectious diseases, those present in genes such as TNF-α, JAK1, VDR, DC-SIGN and FcγRIIa are highlighted. In this context, this work aimed to evaluate the presence of the SNPs CD209 (rs4804803), FcγRIIa (rs1801274), JAK1 (rs11208534), TNF-α (rs1800629) and VDR (rs7975232) in the DNA samples from patients with oral PCM and its associations with the organization patterns of the granulomas in the lesions. For this, 66 samples of DNA obtained from oral lesions biopsies and respective slides of the biopsies for granuloma type classification and a control group of 106 DNA samples of healthy individuals were used. Genotyping was performed by real-time PCR and allele discrimination method. The results demonstrate that in the VDR (rs7975232) SNP, CC genotype(p-value <0.001, OR = 5.94, 95% CI 2.07 - 17.05), or individuals carrying the C allele (p-value = 0.027, OR = 2.71 CI 95% 1.07 - 6.86) and the GG genotype on DC-SIGN (rs4804803) (p-value = 0.032, OR = 3.76, 95% CI, 1,06 - 13,38) are associated with an increased risk of developing oral PCM, but not to the type of granuloma. Thus, the data presented in this work indicate that the variations derived from VDR (rs7975232) and DC-SIGN (rs4804803) SNPs are related to oral PCM susceptibility in the south/southwest of the state of Minas Gerais.

Des colloïdes à base de silice et de polystyrène ont été synthétisés. Les particules d'oxyde ont d'abord été élaborées et modifiées en surface, puis ont servi de germes au cours d'une étape de polymérisation du styrène. Deux procédés de polymérisation en phase hétérogène ont été utilisés (émulsion ou dispersion) menant à des colloïdes aux morphologies originales et contrôlées. Une étude morphologique par tomographie électronique a permis de mieux comprendre les mécanismes de croissance et d'organisation des particules de latex autour des germes de silice. <br />La synthèse de particules Janus pour l'imagerie biomédicale est aussi décrite. Ces particules de silice ont été modifiées en surface par un chromophore biphotonique et un agent de reconnaissance de certaines cellules tumorales. Des études spectroscopiques et des tests de cytotoxicité ont été entrepris.

This thesis conducts numerical and experimental studies of the nonlinear electrokinetic motion of heterogeneous particles in microfluidic systems and their corresponding applications in laboratory-on-a-chip (LOC) systems. Induced-charge electrokinetic (ICEK) phenomena flow is generated by applying an external electric field to a conducting particle immersed in an aqueous solution. As a result of this field, micro-vortices form around the conducting particle. Using this phenomenon, many shortcomings of classical electrokinetics (e.g. poor mixing, leakage, back flow problem) can be improved. This thesis proposes and investigates a complete 3-D numerical multi-physics method to calculate the induced zeta potential on the conducting surface of a heterogeneous object. To model the ICEK motion of a heterogeneous particle in a DC electric field, the moving grid technique is used to conduct the particle-fluid simulation. It was numerically shown that the vortices form near the conducting surface of a particle. Both transitional and rotational motions of heterogeneous particles are investigated. A set of novel experiments are designed and conducted to investigate several aspecs of ICEK. It is demonstrated for the first time that four vortices form around a conducting sphere in contact with an aqueous solution while the DC electric field is applied. The motions of heterogeneous particles are experimentally studied. The speed of a heterogeneous particle is compared with the same size non-conducting particle under the same experimental conditions and it is shown that the heterogeneous particle moves significantly faster than the non-conducting particle. It is also shown that the micro-vortices on the conducting section of the heterogeneous particle act like an engine and push the particle to move faster. These experiments verify the results of our simulation studies. We introduce three applications for induced-charge electrokinetic phenomena in ths thesis: ICEK micro-valve, ICEK micro-mixer, and ICEK micro-motor, which can be used in microfluidics and lab-on-a-chip devises. This ICEK micro-valve significantly improves many shortcomings of other micro-valves reported in the literature (such as leakage, considerable dead volume and complicated fabrication processes). Our ICEK micro-mixers take the advantages of induced micro-vortices and boost the mixing process in a micro-channel. As a result well mixed homogeneous (100%) mixture could be obtained at the downstream of the mixer. Our proposed no-contact ICEK micro-motor rotates as long as the DC electric field is being applied. This thesis develops a new understanding of several ICEK phenomena and applications related to heterogeneous particles. The 3D numerical model developed in this thesis along with the experimental studies are capable of describing the ICEK motion of a heterogeneous particle and is a considerable step to calculate the ICEK phenomena for real-world applications. This thesis, for the first time, experimentally visualized and verified the induced micro-vortices around conducting particles under applied DC electric field. The proposed ICEK micro-mixers, valve and motor can be used in various LOC devices and applications.

<p>This research presents the development of a phase-based lithographic system for three-dimensional micropatterning and manipulation. The system uses a spatial light modulator (SLM) to display specially designed phase holograms. The use of holograms with the SLM provides a novel approach to photolithography that offers the unique ability to operate in both serial and single-shot modes. In addition to the lithographic applications, the optical system also possesses the capability to optically trap microparticles. New advances include the ability to rapidly modify pattern templates for both serial and single-shot lithography, individually control three-dimensional structural properties, and manipulate Janus particles with five degrees of freedom.</p><p>A number of separate research investigations were required to develop the optical system and patterning method. The processes for designing a custom optical system, integrating a computer aided design/computer aided manufacturing (CAD/CAM) platform, and constructing series of phase holograms are presented. The resulting instrument was used primarily for the photonic excitation of both photopolymers and proteins and, in addition, for the manipulation of Janus particles. Defining research focused on the automated fabrication of complex three-dimensional microscale structures based on the virtual designs provided by a custom CAD/CAM interface. Parametric studies were conducted to access the patterning transfer rate and resolution of the system.</p><p>The research presented here documents the creation of an optical system that is capable of the accurate reproduction of pre-designed microstructures and optical paths, applicable to many current and future research applications, and useable by anyone interested in researching on the microscale.</p> / Dissertation

La synthèse de colloïdes dont les propriétés peuvent être ajustées par modification de l'organisation spatiale de leurs constituants permet d'envisager la formation de matériaux uniques et révolutionnaires. Au cours de ce travail, nous nous sommes intéressés à la fabrication de particules hybrides constituées d'un coeur de silice décoré en surface par des latex de polystyrène élaborés in situ par un procédé original de polymérisation en émulsion.<br />Le contrôle du nombre et de la taille des nodules organiques en croissance peut être réalisé via l'ajustement de certains paramètres expérimentaux tels que les caractéristiques physicochimiques de la silice et/ou de la réaction de polymérisation en émulsion. Des colloïdes remarquables par leurs formes (polyèdres réguliers) et par leurs tailles (quelques centaines de nanomètres) ont été obtenus.<br />Nous avons également synthétisé des nanoparticules dissymétriques ou « Janus », en référence au dieu romain à deux visages. Nous avons pour cela développé deux voies de synthèse, basées sur le masquage partiel d'un des hémisphères de particules minérales suivi d'une étape de fonctionnalisation de surface régiosélective. Ainsi, des dimères hybrides silice/polystyrène, où le nodule polymère sert de masque temporaire du coeur minéral, se sont avérés de bons candidats dans la réalisation de particules « Janus ». Nous avons également exploité une interface liquide/liquide (émulsions de Pickering) pour générer ce type d'objets.<br />L'étude des propriétés de ces colloïdes en vue de diverses applications telles que la réalisation de cristaux colloïdaux ou de nanostructures multifonctionnelles complexes a été abordée.

This study is focused on the fabrication of plasma polymerized ultrathin films and the elucidation of their unique properties with an emphasis on the solvent-less, dry polymerization process to introduce post-deposition functionality, robustness, and shape preservation. Two major classes of materials are the subject of this study: biological monomers, specifically the amino acids tyrosine and histidine and synthetic organic and inorganic monomers including acrylonitrile, 2-hydroxyethyl methacrylate, N-isopropylacrylamide, titanium isopropoxide and ferrocene. The unique chemical and physical properties of highly cross-linked ultrathin plasma polymerized amino acid and synthetic polymer films are demonstrated along with their functional response and robustness on both planar and complex surface structures. The work emphasizes the facile ability of plasma polymerization to create unique, tailored ultrathin coatings. Chemical functionality retention (OH, NH₂) of the tyrosine and histidine amino acids is demonstrated by the subsequent mineralization of gold or titania nanoparticles on the plasma polymerized ultrathin films using a wet chemical approach. Inorganic nanoparticle mineralization is further investigated as a method to modify the optical properties of composite nanocoatings. Plasma co-polymerization of tyrosine and synthetic monomers is used to create nanocomposite coatings with unique surface functionalities, responsive behavior, optical characteristics and a high level of integration between monomers. The fabrication of novel plasma polymerized Janus microspheres, micropatterned substrates and free-standing films also demonstrate numerous plasma polymerized materials which exhibit unique structural properties. Overall, facile plasma polymerization of novel, functional ultrathin films and complex topological coatings having potential biocompatible and optical applications is established.

Bipolar electrochemistry deals with the exposure of an isolated conducting substrate that has no direct connection with a power supply except via an electric field. Therefore it can be considered as a "wireless technique". The polarization of the substrate with respect to the surrounding medium generates a potential difference between its opposite ends which can support localized electrochemical oxidation reduction reactions and break the surface symmetry of the substrate. The method was applied in the present thesis to materials science and biological systems. In the frame of designing asymmetric particles, also called "Janus" particles, bipolar electrochemistry was adapted for the bulk preparation of these objects. Conductive substrates with different nature, sizes and shapes have been modified with various materials such as metals, ionic and inorganic compounds using this approach. Moreover, a control over the deposit topology could be achieved for substrates at different length scales. Bipolar electrodeposition is also a good tool for investigating the generation of different metal morphologies. Further developments in the bipolar setup allowed us to use the technology for microstructuration of conductive objects. Furthermore the concept has shown to be very useful in the field of the induced motion of particles. The asymmetric objects that have been prepared by bipolar electrodeposition were employed as microswimmers which could show both translational and rotational motion. The application of electric fields in the bipolar setup can be used for the direct generation of motion of isotropic objects through bubble generation. A levitation motion of objects combined with light emission was possible using this concept. Finally, bipolar electrochemistry was also used for studying the intrinsic conductivity of biological molecules (DNA), which is of great importance in the nanotechnology.