This master´s thesis describes the concept of Bin-picking technology, image recognition and the posibility of grasping parts of various shapes. It deals with the interaction between industrial robot, end-effector and recognition cell system in the industrial cell in process of random collection of parts from the box. The practical part is concerned with construction of end effector for a given part of a specific shape in a random selection/collection of parts from industrial pallets. For this operation is used the KUKA KR16 industrial robot and for safety reasons senzor FTC/collision OPS. Further work is designed working cell which deals with the design and layout of the working elements including security.

The goal of this diploma thesis on the topic of „Automation of workplaces for laser welding“ is the design of a robotic manipulation device for a four position laser welder which has been designed for a real project for automotive parts company. The introduction part of this work analyses the assignment and describes the current state of the workplace, describes each work cycle operation and customer requirements at the workplace. The work then deals with solving individual key nodes at the workplace, including introducing this robot and end effector design. The conclusion of this work explains safety solutions at the workplace, and financial and technical evaluation.

This master’s thesis deals with a description of Laser Welded Common Rail, which is produced by the company Bosch Diesel, s. r. o. in Jihlava. The theoretical part is focused primarily on a proposal for an industrial robot which would be manipulated with Common Rail during the laser welding process. The practical section is concerned with changes in the current paths of the robot to reduce production times, and to avoid using the same parts of the robot for the whole duration it is being manipulated with the Common Rail. At the end of this master's thesis is a techno-economic evaluation of previous alternatives, and it determines the appropriate option for robotic manipulation with the Common Rail during the laser welding process.

This diploma thesis deals with the problem of designing an automatic robotic workstation for automatic tightening screws connections seatback – seat and belt buckle. The research part of this thesis deals with current trends in the automotive industry, especially with the problem of assembling seats part, with focus on screwing. The design part of this thesis maps the process of designing two variants of an automatic robotic workstation. Special attention was given to the process of choosing the right industrial robot for the task, as well as to the construction of the end-effector and of the resulting 3D data. The right version of the workstation for the consecutive implementation was then chosen based on multi-criteria evaluation. The finishing touch of this thesis is a risk analysis for the implemented workstation.

Reaching a target quickly and accurately with a robotic arm containing multiple joints while avoiding moving and fixed obstacles can be a daunting (and sometimes impossible) task for any user behind the remote control. Current existing solutions are often hard to use and to scale for all user body types and robotic arm configurations. In this work, we propose a vision-based gesture recognition approach to naturally control the overall posture of a robotic arm using human hand gestures and an inverse kinematic exploration approach using the FABRIK algorithm. Three different methods are investigated to intuitively control a robotic arm's posture in real-time using depth data collected by a Kinect sensor. Each of the posture control methods are users scalable and compatible with most existing robotic arm configurations. In the first method, the user's right index fingertip position is mapped to compute the inverse kinematics on the robot. The inverse kinematics solutions are displayed in a graphical interface. Using this interface and the left hand, the user can intuitively browse and select a desired robotic arm posture. In the second method, the user's right index fingertip position and finger direction are respectively used to determine the end-effector position and an attraction point position. The latter enables the control of the robotic arm posture. In the third method, the user's right index finger is mapped to compute the inverse kinematics on the robot. Using static gesture with the same hand, the user's right index finger can be transformed into a virtual pen that can trace the form of the desired robotic arm posture. The trace can be visualized in real-time on a graphical interface. A search is then performed using an inverse kinematic exploration and the Dynamic Time Warping algorithm to select the closest matching possible posture. In the last two proposed methods, different search strategies to optimize the speed and the inverse kinematic exploration coverage are proposed. Using a combination of Greedy Best First search and an efficient selection of input postures based on the FABRIK's algorithm characteristics, these optimizations allow for smoother and more accurate posture control of the robotic arm. The performance of these real-time natural human control approaches is evaluated for precision and speed against static (i.e. fixed) and dynamic (i.e. moving) obstacles in a simulated experiment. An adaptation of the vision-based gesture recognition system to operate the AL5D robotic arm was also implemented to conduct further evaluation in a real-world environment. The results showed that the first and third methods were better suited for obstacle avoidance in static environments not requiring continuous posture changes. The second method gave excellent results in the dynamic environment experience and was able to complete a challenging pick and place task in a difficult real-world environment with static constraints.

Transcription Activator-Like Effectors (TALEs) can be used as adaptable DNAbinding modules to create site-specific chimeric nucleases or synthetic transcriptional regulators. The central repeat domain mediates specific DNA binding via hypervariable repeat di-residues (RVDs). This DNA-Binding Domain can be engineered to bind preferentially to any user-selected DNA sequence if engineered appropriately. Therefore, TALEs and their derivatives have become indispensable molecular tools in site-specific manipulation of genes and genomes. This thesis revolves around two problems: in silico design and improved binding site prediction of TALEs. In the first part, a study is shown where TALEs are successfully designed in silico and validated in laboratory to yield the anticipated effects on selected genes. Software is developed to accompany the process of designing and prediction of binding sites. I expanded the functionality of the software to be used as a more generic set of tools for the design, target and offtarget searching. Part two contributes a method and associated toolkit developed to allow users to design in silico optimized synthetic TALEs with user-defined specificities for various experimental purposes. This method is based on a mutual relationship of three consecutive tandem repeats in the DNA-binding domain. This approach revealed positional and compositional bias behind the binding of TALEs to DNA. In conclusion, I developed methods, approaches, and software to enhance the functionality of synthetic TALEs, which should improve understanding of TALEs biology and will further advance genome-engineering applications in various organisms and cell types.

Transcriptional regulation of CD8+ T cell differentiation during acute and chronic viral infections is an intricate web made up of many of transcription factors. While several transcription factors have been elucidated in this process, there are still many more that remain elusive. In this work, we look into the role of two transcription factors, IRF4 and Runx2, and their role in CD8+ T cell terminal effector cells and memory precursor cells during acute LCMV-Armstrong infection. We found that IRF4 expression was regulated by TCR signal strength during infection, and that IRF4 expression levels directly correlated with the magnitude of the effector cell response. IRF4 was also shown to regulate T-bet and Eomes, two transcription factors critical for CD8+ T cell differentiation into effector and memory cells. From these results, we were interested in the potential role of IRF4 during chronic LCMV-clone 13 infection, where ratios of T-bet and Eomes are critical for viral clearance. We found that haplodeficiency of IRF4 in the T cell compartment lead to an increase in the ratio of Eomes to T-bet in T cells, which in turn affected the proportion of Eomeshi versus T-bethi cells and resulted in a loss in ability to clear viral infection. Irf4+/-Eomes+/- compound heterozygous mice were generated to test if decreasing Eomes expression would rescue the Irf4+/- phenotype. Irf4+/-Eomes+/- mice were phenotypically similar to WT mice in terms of Eomes to T-bet ratios, and were able to clear viral infection, demonstrating a critical role of IRF4 in regulating T-bet and Eomes during chronic viral infection. Next we looked into the role of Runx2 during acute LCMV-Armstrong infection and found that Runx2-deficient pathogen-specific CD8+ T cells had a defect in the total number of memory precursor cells compared to WT controls. We further showed that Runx2 was inversely correlated with TCR signal strength, and that Runx2 expression was repressed by IRF4. From these work, we have introduced two more transcription factors that are critical for CD8+ T cells differentiation during acute and chronic viral infection. Given the sheer number of transcription factors known to regulate these processes, having a full understanding of the transcriptional network will allow us to find the best targets for therapeutic intervention for treatments ranging from vaccine development and autoimmunity to cancer immunotherapy and treatment of chronic viral infections.

Salmonella enterica serovar Typhimurium (S. Typhimurium) is a Gram-negative facultative anaerobe that induces severe inflammation resulting in gastroenteritis. In the case of S. Typhimurium infection, induction of an inflammatory response has been linked to its primary virulence mechanism, the type III secretion system (T3SS). The T3SS secretes protein effectors that exploit the host’s cell biology to facilitate bacterial entry and intracellular survival, and to modulate the host immune response. One such effector, SifA, is a bi-functional T3SS effector protein that plays an important role in Salmonella virulence. The N-terminal domain of SifA binds SifA-Kinesin-Interacting-Protein (SKIP), and via an interaction with kinesin, forms tubular membrane extensions called Sif filaments (Sifs) that emanate from the Salmonella Containing Vacuole (SCV). The C-terminal domain of SifA harbors a WxxxE motif that functions to mimic active host cell GTPases. Taken together, SifA functions in inducing endosomal tubulation in order to maintain the integrity of the SCV and promote bacterial dissemination. Since SifA performs multiple, unrelated functions, the objective of this study was to determine how each functional domain of SifA becomes processed. In the present study, we demonstrate that a linker region containing a caspase-3 cleavage motif separates the two functional domains of SifA. To test the hypothesis that processing of SifA by caspase-3 at this particular site is required for function and proper localization of the effector protein domains, we developed two tracking methods to analyze the intracellular localization of SifA. We first adapted a fluorescent tag called phiLOV that allowed for T3SS mediated delivery of SifA and observation of its intracellular colocalization with caspase-3. Additionally, we created a dual-tagging strategy that permitted tracking of each of the SifA functional domains following caspase-3 cleavage to different subcellular locations. The results of this study reveal that caspase-3 cleavage of SifA is required for the proper localization of functional domains and bacterial dissemination. Considering the importance of these events in Salmonella pathogenesis, we conclude that caspase-3 cleavage of effector proteins is a more broadly applicable effector processing mechanism utilized by Salmonella to invade and persist during infection.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第20798号 / 医博第4298号 / 新制||医||1025(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 辻川 明孝, 教授 清水 章, 教授 戸口田 淳也 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

<p>Minimally Invasive Robotic Surgery (MIRS) has revolutionized the way modern surgery is conducted by allowing for smaller incisions, finer control, reduced pain, and faster recovery. The state-of-the-art end-effector technology used for MIRS are tools based off of the rigid-body instruments used in traditional ‘open’ surgery. The rigid nature of the end-effectors, specifically the grasping jaws, leads to a lack of force feedback when implemented in a robotic system. </p> <p>Without additional feedback from active sensing, the blanching that occurs from restricted blood flow around a grasping site is the only indication a surgeon can use to assess the force applied to a tissue. Ongoing efforts to develop active force sensing solutions are currently faced with two major obstacles: miniaturization and sterilization. The lack of force feedback causes a gap between intention and result during robotic surgery. </p> <p>This work proposes the introduction of Visual Force Feedback (VFF) through the integration of a compliant end-effector design. Visual Force Feedback is an intuition, developed through practice, that allows a surgeon to estimate the reaction force of a compliant mechanism by the deflection of the outer flexures. An understanding of the relationship between opening size, flexure deformation, and pinch force allows for rapid estimation of the force applied to a manipulated object. </p> <p>Force and dimensional data were gathered through finite element simulation and the finite element model was validated with physical experimentation on a custom test bench. Multiple functions relating the flexure deformation to the reactionary force, referred to as pinch force, for specific opening sizes were resolved. Notable observations made through the analysis of these results were: (1) a closely linear relationship between outer flexure deformation and pinch force in both experimental and computational results and (2) a higher rate of pinch force increase due to draw displacement as an effect of wider jaw opening. These findings are intended to help shrink the gap between intention and result in the field of MIRS.</p>