Characterization of adiposity and inflammation genetic pleiotropy underlying cardiovascular risk factors in Hispanics.

The observed overlap between genetic variants associated with both adiposity and inflammatory markers suggests that changes in both adiposity and inflammation could be partially mediated by common pathways. The pervasive but sparsely characterized “pleiotropic” genetic variants associated with both adiposity and inflammation have been hypothesized to provide insight into the shared biology. This study explored and characterized the genetic pleiotropy underpinning adiposity and inflammation using genetic and phenotypic observations from the Cameron County Hispanic Cohort (CCHC). A total of 3,313 samples and \u3e9 million single nucleotide polymorphisms (SNPs) were examined in this study. Mixed model genome-wide association studies (GWAS) were performed for 9 phenotypes including C-reactive protein (CRP), Interleukin (IL)-6, IL-8, fibrinogen, body mass index (BMI), waist circumference (WC) in males and females, and waist to hip ratio (WHR) in males and females (separately). GWAS for WHR and WC were meta-analyzed to obtain sex-combined results. Pleiotropy assessment was completed using adaptive Sum of Powered Score (aSPU) test. Three genetic loci with evidence of pleiotropy on chromosome 3, 12 and 18 were fine-mapped to distinguish the set of likely vi causal variants. Causal mediation analysis was used to assess whether likely causal variants were independently associated with both inflammation and adiposity. At least 3 signals, on chromosomes 3, 12, and 12, were identified that suggested the presence of SNPs with strong pleiotropic p-values (\u3c 5 × 10−6 ). The fine-mapping of these three suspected pleiotropic regions distinguished 22 variants with posterior causality probabilities greater than 50%. The mediation analysis indicated that rs60505812, on chromosome 3, was independently associated with both an inflammatory marker (IL-6) and an adiposity measure (BMI). For the variant rs73093474, on chromosome 12, results indicated both a direct association with CRP and an indirect association (via WHR). The identification of likely pleiotropic variants indicated that 1) a considerable degree of overlapping genetic pleiotropy exists between adiposity and inflammation, and 2) evidence exists to support both the direct and indirect pleiotropy. The results showed the potential of these genetic variants to provide biological insight, intended to improve the cardiovascular health of the Hispanics, and by extension all populations

Establishment of a fully automatized microfluidic platform for the screening and characterization of novel Hepatitis B virus capsid assembly modulators

El procés de descobriment de fàrmacs s'enfronta a importants desafiaments a causa de la constant disminució dels guanys per medicament atesa la disminució en les noves aprovacions de la FDA combinada amb el constant augment dels costos i el temps de desenvolupament. Les plataformes integrades de detecció usant microfluídica van sorgir com a possibles solucions per accelerar el desenvolupament de molècules actives i reduir els requisits de temps i costos. El projecte VIRO-FLOW té com a objectiu identificar nous agents curatius per al virus de l'hepatitis B (VHB), integrant els avantatges de la química de flux continu amb tecnologies de bioassaigs in vitro en microfluídica. Durant aquesta tesi es va construir un sistema microfluídic aplicant dispositius modulars automatitzats. Es van redactar protocols d'avaluació per a les dades de fluorescència i reflexió, permetent el càlcul del factor Z, les desviacions estàndard, les corbes de dilució i els valors de concentracions efectives mitjanes màximes (EC50). La proteïna central del VHB (HBc) es va seleccionar com a objectiu principalEl proceso de descubrimiento de fármacos se enfrenta a importantes desafíos debido a la constante disminución de las ganancias por medicamento dada la disminución en las nuevas aprobaciones de la FDA combinada con el constante aumento de los costes y el tiempo de desarrollo. Las plataformas integradas de detección usando microfluídica surgieron como posibles soluciones para acelerar el desarrollo de moléculas activas y reducir los requisitos de tiempo y costes. El proyecto VIRO-FLOW tiene como objetivo la identificación de nuevos agentes curativos para el virus de la hepatitis B (VHB), integrando las ventajas de la química de flujo continuo con tecnologías de bioensayos in vitro en microfluídica. Durante la presente tesis se construyó un sistema microfluídico aplicando dispositivos modulares automatizados. Se redactaron protocolos de evaluación para los datos de fluorescencia y reflexión, permitiendo el cálculo del factor Z, desviaciones estándar, curvas de dilución y valores de concentraciones efectivas medias máximas (EC50). La proteína central del VHB (HBc) se seleccionó como objetivo principal.Drug Discovery as known today faces major challenges due to the constant decrease of earnings per drug given the decrease in new FDA approvements combined with the steadily rising development costs and time. Integrated microfluidic screening platforms emerged as possible solutions by accelerating the hit-to-lead development cycle and reducing time and cost requirements. The VIRO-FLOW project aims at the fast and efficient identification of novel curative agents for the Hepatitis B Virus (HBV), integrating the advantages of continuous flow chemistry with in vitro microfluidic bioassay technologies. During the present thesis a microfluidic system was built, applying automatized modular devices. Evaluation protocols were written for the fluorescence and reflection data, allowing the Z´-factor calculation, standard deviations, dilution curves, and half‐maximal effective concentrations (EC50) values. HBV core protein (HBc) was selected as primary target due to the ongoing demand for a functional cure to reduce the economic and social challenges imposed by the chronic diseas

Concepts of GPCR-controlled navigation in the immune system

G-protein-coupled receptor (GPCR) signaling is essential for the spatiotemporal control of leukocyte dynamics during immune responses. For efficient navigation through mammalian tissues, most leukocyte types express more than one GPCR on their surface and sense a wide range of chemokines and chemoattractants, leading to basic forms of leukocyte movement (chemokinesis, haptokinesis, chemotaxis, haptotaxis, and chemorepulsion). How leukocytes integrate multiple GPCR signals and make directional decisions in lymphoid and inflamed tissues is still subject of intense research. Many of our concepts on GPCR-controlled leukocyte navigation in the presence of multiple GPCR signals derive from in vitro chemotaxis studies and lower vertebrates. In this review, we refer to these concepts and critically contemplate their relevance for the directional movement of several leukocyte subsets (neutrophils, T cells, and dendritic cells) in the complexity of mouse tissues. We discuss how leukocyte navigation can be regulated at the level of only a single GPCR (surface expression, competitive antagonism, oligomerization, homologous desensitization, and receptor internalization) or multiple GPCRs (synergy, hierarchical and non-hierarchical competition, sequential signaling, heterologous desensitization, and agonist scavenging). In particular, we will highlight recent advances in understanding GPCR-controlled leukocyte navigation by intravital microscopy of immune cells in mice

Circuit Techniques for Low-Power and Secure Internet-of-Things Systems

The coming of Internet of Things (IoT) is expected to connect the physical world to the cyber world through ubiquitous sensors, actuators and computers. The nature of these applications demand long battery life and strong data security. To connect billions of things in the world, the hardware platform for IoT systems must be optimized towards low power consumption, high energy efficiency and low cost. With these constraints, the security of IoT systems become a even more difficult problem compared to that of computer systems. A new holistic system design considering both hardware and software implementations is demanded to face these new challenges. In this work, highly robust and low-cost true random number generators (TRNGs) and physically unclonable functions (PUFs) are designed and implemented as security primitives for secret key management in IoT systems. They provide three critical functions for crypto systems including runtime secret key generation, secure key storage and lightweight device authentication. To achieve robustness and simplicity, the concept of frequency collapse in multi-mode oscillator is proposed, which can effectively amplify the desired random variable in CMOS devices (i.e. process variation or noise) and provide a runtime monitor of the output quality. A TRNG with self-tuning loop to achieve robust operation across -40 to 120 degree Celsius and 0.6 to 1V variations, a TRNG that can be fully synthesized with only standard cells and commercial placement and routing tools, and a PUF with runtime filtering to achieve robust authentication, are designed based upon this concept and verified in several CMOS technology nodes. In addition, a 2-transistor sub-threshold amplifier based "weak" PUF is also presented for chip identification and key storage. This PUF achieves state-of-the-art 1.65% native unstable bit, 1.5fJ per bit energy efficiency, and 3.16% flipping bits across -40 to 120 degree Celsius range at the same time, while occupying only 553 feature size square area in 180nm CMOS. Secondly, the potential security threats of hardware Trojan is investigated and a new Trojan attack using analog behavior of digital processors is proposed as the first stealthy and controllable fabrication-time hardware attack. Hardware Trojan is an emerging concern about globalization of semiconductor supply chain, which can result in catastrophic attacks that are extremely difficult to find and protect against. Hardware Trojans proposed in previous works are based on either design-time code injection to hardware description language or fabrication-time modification of processing steps. There have been defenses developed for both types of attacks. A third type of attack that combines the benefits of logical stealthy and controllability in design-time attacks and physical "invisibility" is proposed in this work that crosses the analog and digital domains. The attack eludes activation by a diverse set of benchmarks and evades known defenses. Lastly, in addition to security-related circuits, physical sensors are also studied as fundamental building blocks of IoT systems in this work. Temperature sensing is one of the most desired functions for a wide range of IoT applications. A sub-threshold oscillator based digital temperature sensor utilizing the exponential temperature dependence of sub-threshold current is proposed and implemented. In 180nm CMOS, it achieves 0.22/0.19K inaccuracy and 73mK noise-limited resolution with only 8865 square micrometer additional area and 75nW extra power consumption to an existing IoT system.PHDElectrical EngineeringUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttps://deepblue.lib.umich.edu/bitstream/2027.42/138779/1/kaiyuan_1.pd

Understanding the dynamic regulation of SOCS3

In the past two decades, it has become evident that signal transduction pathways are more than two dimensional pathways consisted of proteins that are just activated or supressed in response to distinct cues. Instead, the dynamic nature of key proteins regulates the strength and quality of the signal. Several key signal transduction pathways are controlled by negative feedback loops that are highly dynamic and demonstrate oscillatory behaviours. Negative feedback regulation of the JAK/STAT pathway by Suppressors of Cytokine Signalling (SOCS) is an example of oscillatory signalling. We sought to investigate the oscillatory capacity of the tumour suppressor protein SOCS3 and its role in important cellular functions using whole-cell population and single-cell analysis. An important aspect of cell biology using experimental cell-population techniques is to produce a synchronized cell culture. Serum starvation and subsequent shock is able to capture the oscillatory behaviour of SOCS3 protein to some extent. However, the average response in whole-cell population systems demonstrated to be ‘noisy’ leading to establishment of a single-cell analysis system. To investigate SOCS3 oscillation at the single cell level, we first attempted to generate cell clones stably expressing SOCS3 C-terminal GFPSpark fusion protein from its respective endogenous promoter to monitor its expression in real time with confocal microscopy. Despite careful optimization of each step of CRISPR/Cas9 strategy, the generation of GFPSpark knockin cell line was not successful. Finally, we utilised the tandem fluorescent protein timer (tFT) strategy to investigate localisation and trafficking of SOCS3 protein and monitor its promoter activity in response to different stimuli. The use of tFT provided us the ability to analyse SOCS3 dynamics across spatial and temporal dimensions under either normal culture conditions or different treatments that are known to influence on SOCS3 half-life and degradation rates

The 1992 4th NASA SERC Symposium on VLSI Design

Papers from the fourth annual NASA Symposium on VLSI Design, co-sponsored by the IEEE, are presented. Each year this symposium is organized by the NASA Space Engineering Research Center (SERC) at the University of Idaho and is held in conjunction with a quarterly meeting of the NASA Data System Technology Working Group (DSTWG). One task of the DSTWG is to develop new electronic technologies that will meet next generation electronic data system needs. The symposium provides insights into developments in VLSI and digital systems which can be used to increase data systems performance. The NASA SERC is proud to offer, at its fourth symposium on VLSI design, presentations by an outstanding set of individuals from national laboratories, the electronics industry, and universities. These speakers share insights into next generation advances that will serve as a basis for future VLSI design

Deriving a mathematical framework for data-driven analyses of immune cell dynamics

Zelluläre Entscheidungen, wie z. B. die Differenzierung von T-Helferzellen (Th-Zellen) in spezialisierte Effektorlinien, haben großen Einfluss auf die Spezifität von Immunreaktionen. Solche Reaktionen sind das Ergebnis eines komplexen Zusammenspiels einzelner Zellen, die über kleine Signalmoleküle, so genannte Zytokine, kommunizieren. Die hohe Anzahl der Komponenten, sowie deren komplizierte und oft nichtlineare Interaktionen erschweren dabei die Vorhersage, wie bestimmte zelluläre Reaktionen erzeugt werden. Aus diesem Grund sind die globalen Auswirkungen der gezielten Beeinflussung einzelner Zellen oder spezifischer Signalwege nur unzureichend verstanden. So wirken beispielsweise etablierte Behandlungen von Autoimmunkrankheiten oft nur bei einem Teil der Patienten. Durch Einzelzellmethoden wie Live-Cell-Imaging, Massenzytometrie und Einzelzellsequenzierung, können Immunzellen heutzutage quantitativ auf mehreren Ebenen charakterisiert werden. Diese Ansammlung quantitativer Daten erlaubt die Formulierung datengetriebener Modelle zur Vorhersage von zellulären Entscheidungen, allerdings fehlen in vielen Fällen Methoden, um die verschiedenen Daten auf geeignete Weise zu integrieren und zu annotieren. Die vorliegende Arbeit befasst sich mit quantitativen Modellformulierungen für die Entscheidungsfindung von Zellen im Immunsystem mit dem Schwerpunkt auf Lymphozytenproliferation, -differenzierung und -tod.Cellular decisions, such as the differentiation of T helper (Th) cells into specialized effector lineages, largely impact the direction of immune responses. Such population-level responses are the result of a complex interplay of individual cells which communicate via small signaling molecules called cytokines. The system's complexity, stemming not only from the number of components but also from their intricate and oftentimes non-linear interactions, makes it difficult to develop intuition for how cellular responses are actually generated. Not surprisingly, the global effects of targeting individual cells or specific signaling pathways through perturbations are poorly understood. For instance, common treatments of autoimmune diseases often work for some patients, but not for others. Recently developed methods such as live-cell imaging, mass cytometry and single-cell sequencing now enable quantitative characterization of individual immune cells. This accumulating wealth of quantitative data has laid the basis to derive predictive, data-driven models of immune cell behavior, but in many cases, methods to integrate and annotate the data in a way suitable for model formulation are missing. In this thesis, quantitative workflows and methods are introduced that allow to formulate data-driven models of immune cell decision-making with a particular focus on lymphocyte proliferation, differentiation and death

in vitro Characterisation of the Complement Cascade for Predicting Patient Outcome Post-operatively

The identification of surgical patients at higher risk of infection enables targeted allocation of critical care resources to improve patient mortality. The Complement cascade of the innate immune system is known to increase risk of infection if compromised and can be tested in vitro as a potential method for stratification of high-risk patients. Existing assays of Complement function are laboratory bound and require trained personnel to operate and interpret. This thesis describes the development of novel immunoassays for C3, C5a, TCC and TNFα, based on a multiplex biosensor platform with a duty cycle of 0.05) from the serum data of 22 volunteers. The model and cohort data provide an initial estimate of effect size for future clinical studies investigating the ability of these Complement activation phenotypes to identify high-risk surgical patients or identify the onset of infection

Program Annual Technology Report: Physics of the Cosmos Program Office

From ancient times, humans have looked up at the night sky and wondered: Are we alone? How did the universe come to be? How does the universe work? PCOS focuses on that last question. Scientists investigating this broad theme use the universe as their laboratory, investigating its fundamental laws and properties. They test Einsteins General Theory of Relativity to see if our current understanding of space-time is borne out by observations. They examine the behavior of the most extreme environments supermassive black holes, active galactic nuclei, and others and the farthest reaches of the universe, to expand our understanding. With instruments sensitive across the spectrum, from radio, through infrared (IR), visible light, ultraviolet (UV), to X rays and gamma rays, as well as gravitational waves (GWs), they peer across billions of light-years, observing echoes of events that occurred instants after the Big Bang. Last year, the LISA Pathfinder (LPF) mission exceeded expectations in proving the maturity of technologies needed for the Laser Interferometer Space Antenna (LISA) mission, and the Laser Interferometer Gravitational-Wave Observatory (LIGO) recorded the first direct measurements of long-theorized GWs. Another surprising recent discovery is that the universe is expanding at an ever-accelerating rate, the first hint of so-called dark energy, estimated to account for 75% of mass-energy in the universe. Dark matter, so called because we can only observe its effects on regular matter, is thought to account for another20%, leaving only 5% for regular matter and energy. Scientists now also search for special polarization in the cosmic microwave background to support the notion that in the split-second after the Big Bang, the universe inflated faster than the speed of light! The most exciting aspect of this grand enterprise today is the extraordinary rate at which we can harness technologies to enable these key discoveries