Objetivo: Evaluar los factores asociados a NASH severo según algortimos FLIP/Clasificación SAF en pacientes obesos intervenidos en un centro bariátrico de Lima, Perú. Diseño: El diseño del estudio es de tipo transversal analítico, en el cual se analizará una base de datos secundaria.

Additive manufacturing is revolutionizing the aerospace, transportation, energy, healthcare and various consumer product industries, replacing centralized manufacturing plants with more localized fabrication. 3D printing has become ubiquitous within these industries for prototyping and production. Currently, the smallest 3D printed features are on the order of a micron. While sufficient for some academic and industry applications, nanoscale features are required for the electronics industry and research endeavors. Optical lithography is still the workhorse for industrial nanofabrication utilizing large expensive commercial foundries. Here, an atomic scale 3D printer is presented with many of the features found in a complex semiconductor fabrication plant. This process is reproduced using three separate die with microelectromechanical systems (MEMS), which are bonded together to create an integrated 3D printer with the capability to print at the atomic scale. Due to the microscale size and surface areas of MEMS devices, they are extremely sensitive with rapid response times. These onboard MEMS devices replicate the functions of a thermal evaporator, patterning mask, mass sensor, heaters, temperature sensors and Van de Pauw setups. The assembled 3D printer dimensions are 3.8 mm x 2.5 mm x 1.8 mm (LxWxH) and it is therefore ideal for cryogenic environments. Quenched condensed thin film metals can be deposited using the atomic scale thermal evaporators in varying thicknesses up to approximately 50 nm. Replacing the atomic scale evaporators with microscale evaporators, the deposited film thickness can reach 3.5 microns. Evaporated films are monitored during and after the deposition with the embedded MEMS devices. While this particular 3D printing assembly is designed for research-scale investigations, the same technology could be extended to wafer-scale 3D printing with high resolution, rapid throughput, and reduced cost. / 2023-06-01T00:00:00Z

Intermediate lipid structures such as inverted micelles and interlamellar attachment are thought to play a crucial role in different biological processes like viral infection. Lysophosphatidylcholine has been shown to inhibit membrane fusion at stabilizing concentrations (between 1 and 10% with respect to membrane lipids). Studies in this thesis looked at the effects of Lysophosphatidylcholine (LPC) properties on the inhibition of Sendai viral fusion. The effects of bilayer sidedness preference as well as flip-flop of Lysophosphatidylcholine (lyso PC) were examined. Octadecylrhodamine (R₁₈) lipid mixing assays were used to measure the fusion of Sendai virus with different biological, erythrocyte ghosts, and artificial systems consisting of different lipids and different viral receptor compositions. The data showed that external addition of LPC exhibits a dependency between the incubation time of the lysolipids and the inhibition of viral fusion. The results also demonstrate a relationship between the location of LPC in the bilayer and its ability to inhibit lipid mixing. LPC present only on the outer monolayer plays a role in the inhibition of viral fusion. Reorientation of LPC was also measured for the same incubation periods. A method using Bovine Serum Albumin (BSA) and radioactivity labelled LPC, was applied to measure the flip-flop. Significant transbilayer reorientation of Lyso PC in the bilayer was shown to take place. The rate of flip-flop was measured at 0.32 ± 0.08% LPC /min. Such reorientation can explain the time dependency observed earlier. The conclusions of this thesis lend support to stalk intermediate mechanism of viral membrane fusion. The ability of LPC to inhibit only when present on one side of the bilayer supports the idea of a negatively curved stalk intermediate. Moreover, it showed that the shape and curvature tendencies of the bilayer stabilizer determine its effects on viral fusion. / Thesis / Master of Science (MS)

Apoptose, der programmierte Zelltod, spielt eine wichtige Rolle für das Gleichgewicht zwischen Proliferation und Sterben von Zellen und ist außerdem an der Beseitigung von infizierten und geschädigten Zellen beteiligt. Apoptose kann durch Stimulation von Rezeptoren aus der Familie der TNF-Rezeptoren wie CD95, ausgelöst werden. Nach Liganden-induzierter Trimerisierung der Rezeptoren bindet FADD an den zytoplasmatischen Teil des Rezeptors und rekrutiert Caspase-8 und/oder -10. Die räumliche Nähe der Caspasen in diesem als DISC bezeichneten Komplex führt zu ihrer auto- und transkatalytischen Spaltung und damit Aktivierung. Dadurch wird das apoptotische Programm gestartet, welches zum Tod der Zelle führt. Kontrolliert wird dieser Vorgang von einer Vielzahl anti-apoptotischer Proteine. Störungen in diesem System sind an der Entstehung einer Reihe von Krankheiten beteiligt. Die Blockade der Apoptoseinduktion kann zur Entstehung von Tumoren beitragen. Das klassische Hodgkin Lymphom ist eine maligne Erkrankung des lymphatischen Systems. Die Tumorzellen sind große, einkernige Hodgkin- oder mehrkernige Reed/Sternbergzellen (HRS-Zellen). Sie leiten sich von Keimzentrum-B-Zellen ab. In HRS-Zellen fehlt die Expression einer Vielzahl von typischen B-Zellmarkern, darunter die des B-Zellrezeptors. Solche B-Zellen sterben normalerweise während der Keimzentrumsreaktion durch Apoptose. An diesem Prozess ist CD95 beteiligt. In einer Reihe von malignen Erkrankungen wurden eine Herunterregulation der CD95-Expression oder Mutationen im CD95-Gen beobachtet. Es wird daher vermutet, dass CD95-induzierte Apoptose zur Entfernung von Tumorzellen beiträgt. Im Gegensatz dazu exprimieren sowohl primäre HRS-Zellen als auch etablierte HRS-Zelllinien in der Regel Wildtyp-CD95, sind aber trotzdem CD95-resistent. In dieser Arbeit konnte gezeigt werden, dass Komponenten des CD95-Systems, im Gegensatz zu anderen malignen Erkrankungen, in den HRS-Zellen hochreguliert sind, darunter CD95 selbst. In immunpräzipitierten DISCs von CD95-stimulierten HRS-Zellen wurde neben FADD und Caspase-8/-10 auch c-FLIP nachgewiesen. c-FLIP ist ein Caspase-8/-10-Homolog, das ebenfalls an FADD bindet, aber aufgrund fehlender katalytischer Aktivität die Aktivierung der Caspasen im DISC und damit die Apoptoseinduktion verhindert. Eine starke c-FLIP-Expression konnte in allen HRS-Zelllinien und in den HRS-Zellen nahezu aller untersuchter primärer Hodgkinfälle (55/59) gezeigt werden. Durch siRNA-vermittelte Herunterregulation von c-FLIP war es möglich, HRS-Zelllinien gegenüber CD95-induzierter Apoptose zu sensitivieren. Dies zeigt, dass die CD95-Rezeptor-induzierte Apoptose in den HRS-Zellen nicht strukturell, sondern funktionell inhibiert ist und c-FLIP stark zu dieser Inhibition beiträgt. Darüber hinaus konnte gezeigt werden, dass die c-FLIP-Expression in den HRS-Zellen von der konstitutiven Aktivität des Transkriptionsfaktors NF-κB abhängt, die charakteristisch für diese Zellen ist. Normalerweise wird NF-κB von Inhibitorproteinen, den IκBs, im Zytoplasma zurückgehalten. Diverse Stimuli können den IKK-Komplex aktivieren, der die IκBs an bestimmten Serinresten phosphoryliert. Dies hat die Ubiquitinylierung und den Abbau der IκBs zur Folge, wodurch NF-κB frei wird, in den Kern wandert und dort seine Zielgene aktiviert. Es wird angenommen, dass in HRS-Zellen ein konstitutiv aktiver IKK-Komplex und teilweise Mutationen der IκB-Proteine zur konstitutiven NF-κB-Aktivität beitragen. Zu den NF-κB-abhängigen Genen in den HRS-Zellen gehören solche mit anti-apoptotischer und Zellzyklus-treibender Wirkung. Die Inhibition der NF-κB-Aktivität in den HRS-Zellen führt zu Apoptose und eingeschränkter Proliferation. Von dreiwertigem Arsen ist bekannt, dass es die Induzierbarkeit des IKK-Komplexes inhibieren kann und damit letztendlich die Aktivierung von NF-κB. In dieser Arbeit konnte gezeigt werden, dass Arsen in HRS-Zellen den konstitutiv aktiven IKK-Komplex inhibiert. In Zelllinien mit intakten IκB-Proteinen führte dies zur NF-κB-Inhibition und Apoptoseinduktion. Die Reduktion der NF-κB-Aktivität ging mit der Herunterregulation von anti-apoptotischen und Proliferations-fördernden Zielgenen einher. Die ektope Überexpression von NF-κB hob die Apoptose-induzierende Wirkung von Arsen teilweise auf. Durch Arsen-Behandlung von Mäusen konnte das Tumorwachstum xenotransplantierter HRS-Zellen stark verlangsamt werden. In explantierten Tumorzellen konnte ebenfalls eine NF-κB-Inhibition nachgewiesen werden. Die NF-κB-Inhibition durch Arsen trägt also stark zur Apoptoseinduktion in den HRS-Zellen bei. Zusammengefasst zeigen die Ergebnisse dieser Arbeit, dass die Modulation der Apoptoseresistenz neue therapeutische Ansätze für die Behandlung des Hodgkin Lymphoms bieten könnte. Der Einsatz von Arsen ist dabei besonders interessant, da Arsen schon für die Behandlung anderer maligner Erkrankungen eingesetzt wird. / Apoptosis, the programmed cell death, is important for the balance between proliferation and dying of cells. It is also involved in the removal of infected and damaged cells. Apoptosis can be induced by stimulation of receptors of the TNF-receptor family like CD95. After ligand-induced trimerisation of these receptors, FADD binds to the cytoplasmic part of the receptor and recruits Caspase-8 and/or -10. The induced proximity of the caspases in this complex, called DISC, leads to their auto- and transcatalytic cleavage and subsequently to their activation. This starts the apoptotic program which leads to the death of the cell. A number of anti-apoptotic proteins control this process. The deregulation of this system is involved in a variety of diseases. The disruption of the apoptotic program can contribute to the development of tumors. Classical Hodgkin Lymphoma is a malignant disease of the lymphatic system, characterized by mononucleated Hodgkin or multinucleated Reed/Sternberg (HRS) cells. These tumor cells are derived from germinal-center B-cells. However, HRS cells lack the expression of typical B cell markers, such as the B-cell receptor. Usually, B-cells without B-cell receptor expression die by apoptosis during the germinal-center reaction. CD95 is involved in this process. It has been shown previously that in many malignant diseases CD95 is down-regulated or mutated, indicating that CD95 is involved in the removal of tumor cells. In opposite to these findings, primary HRS cells and Hodgkin-derived cell lines usually express wild type CD95, but are resistant to CD95 induced apoptosis. In this work, it could be demonstrated that in contrast to other malignant diseases components of the CD95 system are up-regulated in HRS cells, including CD95 itself. By immunoprecipitation it was shown that, in addition to FADD and Caspase-8/-10, c-FLIP is a component of the DISC in CD95-stimulated cells. c-FLIP is a caspase homolog which, like caspases, binds to FADD, but lacks proteolytic activity. It inhibits the activation of caspases in the DISC and thus prevents apoptosis induction. A strong c-FLIP expression was shown in all HRS cell lines and in HRS cells of nearly all investigated primary cases of Hodgkin Lymphoma (55/59). siRNA-mediated (small interfering RNA) down-regulation of c-FLIP sensitized HRS cell lines to CD95-induced apoptosis. This shows that the CD95 receptor-induced apoptosis in HRS cells is not structurally but functionally inhibited and that c-FLIP strongly contributes to this inhibition. In addition, it was shown that c-FLIP expression depends on the constitutive activity of the transcription factor NF-κB which is characteristic for HRS cells. Usually, NF-κB is sequestered in the cytoplasm by inhibitor proteins, the IκBs. A variety of stimuli can activate the IKK-complex which subsequently phosphorylates the IκBs, leading to their ubiquitinylation and degradation. The released NF-κB translocates to the nucleus where it activates the transcription of target genes. It is supposed that a constitutively activated IKK complex and, in some cases, mutated IκB proteins contribute to the constitutive NF-κB activity in HRS cells. To the NF-κB dependent genes in HRS cells belong those with anti-apoptotic and cell cycle promoting activities. Inhibition of the NF-κB activity in HRS cells leads to apoptosis and decreased proliferation. Trivalent arsenic is known to inhibit the induction of the IKK complex and thus the activation of NF-κB. In this work, it was shown that arsenic inhibits the constitutively active IKK complex in HRS cells. This led to an inhibition of NF-κB and induction of apoptosis in HRS cell lines with non-mutated IκB proteins. The NF-κB inhibition was accompanied by the down-regulation of anti-apoptotic and cell cycle promoting genes. Ectopic overexpression of NF-κB partially reverted the apoptotic effect of arsenic. Treatment of mice with arsenic reduced the growth of subcutaneously xenotransplanted HRS cells. In explanted tumor cells, a reduced NF-κB activity could be demonstrated following treatment with arsenic. Thus, the inhibition of NF-κB by arsenic contributes to the induction of apoptosis in HRS cells. Taken together, the results indicate that modulation of the apoptosis resistance may offer new strategies for the treatment of Hodgkin Lymphoma. Of particular interest is the application of arsenic because it is already used in the treatment of other malignant disorders.

Environmental stress tests, or accelerated life tests, apply stresses to electronic packages that exceed the stress levels experienced in the field. In theory, these elevated stress levels are used to generate the same failure mechanisms that are seen in the field, only at an accelerated rate. The methods of assessing reliability of electronic packages can be classified into two categories: a statistical failure based approach and a physics of failure based approach. This research uses a statistical based methodology to identify the critical factors in reliability performance of a flip chip on board component level assembly and a physics of failure based approach to develop a low cycle strain based fatigue equation for flip chip component level assemblies. The critical factors in determining reliability performance were established via experimental investigation and their influence quantified via regression analysis. This methodology differs from other strain based fatigue approaches because it is not an empirical fit to experimental data; it utilizes regression analysis and least squares to obtain correction factors, or correction functions, and constants for a strain based fatigue equation, where the total inelastic strain is determined analytically. The end product is a general flip chip on board equation rather than one that is specific to a certain test vehicle or material set.

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Avatrel 8000P's excellent photo-definition properties and mechanical strength make it an ideal polymer collar material. Avatrel 8000P is a high contrast, I-line sensitive mixture that can be developed in traditional aqueous-base developers. The great photolithographical performance of this photopolymer can be partly contributed to the minimal amount of light absorbed by the base norbornene polymer. The processing conditions noted in this work are an optimized version, which have been shown to give superior photolithographical performance. The simple baking procedures make Avatrel 8000P easier to process than SU-8. The ability to develop Avatrel 8000P in aqueous base can reduce chemical waste. As shown by SEM images, high fidelity structures with aspect ratios of 7:1 can be fabricated in thick films with vertical sidewalls. Bonding between two copper surfaces over various gap sizes was achieved by electroless deposition without the addition of surfactants or inhibitors in the bath. The effect of anneal temperature on the electroless bond formed was analyzed. The electroless bond strength increased with anneal temperature. However, the bond strength estimation for samples annealed at 80°C to 120°C is a minimum value due to the failure location of most of the pillars and the resulting area used in the calculation of bond strength. Grain growth from copper recrystallization and removal of small defects improve the bond strength. Large voids at the interface of the two pillars were related to rough starting surfaces for the electroplated pillars.

Flip Chip in Package (FCIP) has been developed to achieve the assembly process with area array interconnects. Particularly, a high I/O count coupled with finer pitch area array interconnects structured FCIP can be achieved using no-flow underfill assembly process. Using the assembly process, a high, stable yield assembly process recently reported with eutectic lead-tin solder interconnections, 150 µm pitch, and I/O counts in excess of 3000. The assembly process reported created a large number of voids among solder interconnects in FCIP. The voids formed among solder interconnections can propagate, grow, and produce defects such as solder joint cracking and solder bridging. Moreover, these voids can severely reduce reliability performance. Indeed, many studies were conducted to examine the void formation in FCIP. Based on the studies, flip chip geometric design, process conditions, and material formulation have been considered as the potential causes of void formation. However, the present research won't be able to identify the mechanism of void formation, causing a lot of voids in assembly process without consideration of chemical reaction in the assembly process with a fine-pitch, high I/O density FCIP. Therefore, this research will present process technology necessary to achieve high yields in FCIP assemblies using no-flow underfills and investigate the underlying problem of underfill void formation in these assemblies. The plausible causes of void formation will be investigated using experimental techniques. The techniques will identify the primary source of the void formation. Besides, theoretical models will be established to predict the number of voids and to explain the growth behavior of voids in the FCIP. The established theoretical models will be verified by experiments. These models will validate with respect to the relationship between process parameters to achieve a high yield and to minimize voids in FCIP assemblies using no-flow underfill materials regarding process as well as material stand points. Eventually, this research provides design guideline achieving a high, stable yield and void-free assembly process.

TRAIL (TNF-related apoptosis inducing ligand) est une protéine du système immunitaire appartenant à la famille du TNF (Tumor necrosis factor). L'intérêt de TRAIL en thérapie anti-cancéreuse réside dans sa capacité à induire la mort par apoptose des cellules tumorales, sans exercer de toxicité envers les cellules saines. Le principal frein à l’utilisation de TRAIL est la survenue courante de résistances dans les tumeurs, limitant ainsi son efficacité. Mon travail de thèse a consisté à étudier l’intérêt de l’hyperthermie (ou choc thermique) en tant qu’adjuvant à TRAIL et a décrire ses capacités à contourner les mécanismes de résistance.Dans un premier temps, l’activité et la portée de cette combinaison a été évaluée dans de nombreuses et diverses lignées cellulaires cancéreuses sensibles ou résistantes à TRAIL. Un choc thermique (1h 42°C) permet de sensibiliser efficacement les lignées devenues résistantes à TRAIL et cette association s’est avérée efficace dans toutes les lignées testées.Dans un deuxième temps, mon travail s’est focalisé sur les mécanismes induits par l’hyperthermie, responsables de la sensibilisation et de l’apoptose des cellules. Les analyses des complexes initiateurs de la mort (DISC - Death-inducing signaling complex) ont révélé de nombreuses disparités suivant les conditions thermiques. Les différences majeures impliquent avant tout l’absence de FLIP dans le DISC, celui-ci étant l’inhibiteur principal de la voie, ainsi qu’un retard de la formation du complexe en condition d’hyperthermie. Ceci est associé à l’activation des caspases initiatrices d’une meilleure qualité, une fois la température revenue à la normale.L’absence de FLIP est expliquée par son inactivation due à l’agrégation de cette protéine suite à l’augmentation de la température. Ce phénomène est indépendant d’une quelconque modification post-traductionnelle connue, mais peut être inhibé par la présence de glycérol qui stabilise les protéines dénaturées.L’ensemble de ce travail met en lumière l’intérêt de la combinaison de TRAIL avec une hyperthermie et présente un point de vue nouveau sur les mécanismes expliquant son efficacité. / The TNF-family member TRAIL (TNF-related apoptosis inducing ligand) is a cytokine involved in the immune anti-tumour surveillance. TRAIL is a promising agent currently under investigation for its anti-cancer properties with limited side effects on healthy cells. However, the use of TRAIL in oncology has been limited due to its lack of efficiency, mainly associated with cell resistance to apoptosis. The aim of this project was to study the interest of hyperthermia (or heat shock - HS) as an adjuvant for TRAIL therapy and the mechanisms involved in this sensitization.We have first evaluated the significance of this combination in a large variety of cancer cell lines known to be sensitive or resistant to TRAIL. We could demonstrate that hyperthermia was able to efficiently sensitize resistant cancer cells to TRAIl-induced apoptosis in almost every cell lines tested.We next, focused our work on the molecular mechanisms responsible for the sensitization, during hyperthermia. Analyses of the DISC (Death-Inducing Signaling Complex) revealed a lack of recruitment of FLIP in the DISC, the main inhibitor of the extrinsic pathway, and a delay in the formation of the complex under hyperthermic conditions. Inhibition of FLIP recruitment was associated with enhanced initiator caspases activation when cells were reincubated at 37°C after the HS.The absence of FLIP within the TRAIL DISC was due to its aggregation during HS and was independent of post-translational modifications. Inhibition of FLIP aggregation by glycerol, which stabilizes denaturerated proteins, restored FLIP recruitment within the TRAIL DISC and consequently inhibited TRAIL-induced cell death. Taken together, these results highlight the interest of combining TRAIL with hyperthermia and highlight new mechanisms explaining its efficiency.