Development and characterization of a scaffold-free 3D spheroid model of iPSC-derived human cardiomyocytes

Purpose: Cardiomyocytes are terminally differentiated cells in the adult heart and ischemia and cardiotoxic compounds can lead to cell death and irreversible decline of cardiac function. As testing platforms, isolated organs and primary cells from rodents have been the standard in research and toxicology, but there is a need for better models that more faithfully recapitulate native human biology. Hence, a new in vitro model comprising the advantages of 3D cell culture and the availability of induced pluripotent stem cells (iPSC) from human origin was developed and characterized. Methods: Human cardiomyocytes (CMs) derived from induced pluripotent stem cells (iPSCs) were studied in standard 2D culture and as cardiac microtissues (MTs) formed in hanging drops. 2D cultures were examined using immunofluorescence microscopy and Western blotting while the cardiac MTs were subjected to immunofluorescence, contractility, and pharmacological investigations. Results: iPSC-derived CMs in 2D culture showed well-formed myofibrils, cell-cell contacts positive for connexin-43, and other typical cardiac proteins. The cells reacted to pro-hypertrophic growth factors with a substantial increase in myofibrils and sarcomeric proteins. In hanging drop cultures, iPSC-derived cardiomyocytes formed spheroidal MTs within 4 days showing a homogeneous tissue structure with well-developed myofibrils extending throughout the whole spheroid without a necrotic core. MTs showed spontaneous contractions for more than 4 weeks that were recorded by optical motion tracking, sensitive to temperature, and responsive to electrical pacing. Contractile pharmacology was tested with several agents known to modulate cardiac rate and viability. Calcium-transients underlay the contractile activity and were also responsive to electrical stimulation, caffeine-induced Ca2+-release, extracellular calcium levels. Conclusions: 3D culture using iPSC-derived human cardiomyocytes provides an organoid human-based cellular platform that is free of necrosis and recapitulates vital cardiac functionality, thereby providing new and promising relevant model for the evaluation and development of new therapies and detection of cardiotoxicity.

Comparison of 3D kinetic and hydrodynamic models to ROSINA-COPS measurements of the neutral coma of 67P/Churyumov-Gerasimenko

67P/Churyumov-Gerasimenko (67P) is a Jupiter-family comet and the object of investigation of the European Space Agency mission Rosetta. This report presents the first full 3D simulation results of 67P’s neutral gas coma. In this study we include results from a direct simulation Monte Carlo method, a hydrodynamic code, and a purely geometric calculation which computes the total illuminated surface area on the nucleus. All models include the triangulated 3D shape model of 67P as well as realistic illumination and shadowing conditions. The basic concept is the assumption that these illumination conditions on the nucleus are the main driver for the gas activity of the comet. As a consequence, the total production rate of 67P varies as a function of solar insolation. The best agreement between the model and the data is achieved when gas fluxes on the night side are in the range of 7% to 10% of the maximum flux, accounting for contributions from the most volatile components. To validate the output of our numerical simulations we compare the results of all three models to in situ gas number density measurements from the ROSINA COPS instrument. We are able to reproduce the overall features of these local neutral number density measurements of ROSINA COPS for the time period between early August 2014 and January 1 2015 with all three models. Some details in the measurements are not reproduced and warrant further investigation and refinement of the models. However, the overall assumption that illumination conditions on the nucleus are at least an important driver of the gas activity is validated by the models. According to our simulation results we find the total production rate of 67P to be constant between August and November 2014 with a value of about 1 × 10²⁶ molecules s⁻¹.

High depth-resolution laser ablation chemical analysis of additive-assisted Cu electroplating for microchip architectures

Laser ablation/ionisation mass spectrometry with a vertical resolution at a nanometre scale was applied for the quantitative characterisation of the chemical composition of additive-assisted Cu electroplated deposits used in the microchip industry. The detailed chemical analysis complements information gathered by optical techniques and allows new insights into the metal deposition process.

Production Streamlining through Design, Implementation, and Analysis of Lean Manufacturing Methods

Through the correct implementation of lean manufacturing methods, a company can greatly improve their business. Over a period of three months at TTM Technologies, I utilized my knowledge to fix existing problems ans streamline production. In addition, other trouble areas in their production process were discovered and proper lean methods were used to address them. TTM Technologies saw many changed in the right direction over this time period.

Development of a 3D Game Engine

This paper presents the development history and specification of a 3D game engine titled "Spark Engine". The term "engine" is used to describe a complex graphics software suite that streamlines application development and provides efficient rendering functionality. A game engine specifically provides tools to simplify game development. Spark Engine is fully shader driven and is built on top of Microsoft's XNA Framework. It is a reusable and flexible platform that can be used to build any type of graphics application ranging from gaming to simulation. The engine was released as open source software under the New BSD License with an interest in furthering its development.

Calculation Methodology and Fabrication Procedures for Particle Accelerator Strip-Line Kickers: Application to the CTF3 Combiner Ring Extraction Kicker and TL2 Tail Clippers

A particle accelerator is any device that, using electromagnetic fields, is able to communicate energy to charged particles (typically electrons or ionized atoms), accelerating and/or energizing them up to the required level for its purpose. The applications of particle accelerators are countless, beginning in a common TV CRT, passing through medical X-ray devices, and ending in large ion colliders utilized to find the smallest details of the matter. Among the other engineering applications, the ion implantation devices to obtain better semiconductors and materials of amazing properties are included. Materials supporting irradiation for future nuclear fusion plants are also benefited from particle accelerators. There are many devices in a particle accelerator required for its correct operation. The most important are the particle sources, the guiding, focalizing and correcting magnets, the radiofrequency accelerating cavities, the fast deflection devices, the beam diagnostic mechanisms and the particle detectors. Most of the fast particle deflection devices have been built historically by using copper coils and ferrite cores which could effectuate a relatively fast magnetic deflection, but needed large voltages and currents to counteract the high coil inductance in a response in the microseconds range. Various beam stability considerations and the new range of energies and sizes of present time accelerators and their rings require new devices featuring an improved wakefield behaviour and faster response (in the nanoseconds range). This can only be achieved by an electromagnetic deflection device based on a transmission line. The electromagnetic deflection device (strip-line kicker) produces a transverse displacement on the particle beam travelling close to the speed of light, in order to extract the particles to another experiment or to inject them into a different accelerator. The deflection is carried out by the means of two short, opposite phase pulses. The diversion of the particles is exerted by the integrated Lorentz force of the electromagnetic field travelling along the kicker. This Thesis deals with a detailed calculation, manufacturing and test methodology for strip-line kicker devices. The methodology is then applied to two real cases which are fully designed, built, tested and finally installed in the CTF3 accelerator facility at CERN (Geneva). Analytical and numerical calculations, both in 2D and 3D, are detailed starting from the basic specifications in order to obtain a conceptual design. Time domain and frequency domain calculations are developed in the process using different FDM and FEM codes. The following concepts among others are analyzed: scattering parameters, resonating high order modes, the wakefields, etc. Several contributions are presented in the calculation process dealing specifically with strip-line kicker devices fed by electromagnetic pulses. Materials and components typically used for the fabrication of these devices are analyzed in the manufacturing section. Mechanical supports and connexions of electrodes are also detailed, presenting some interesting contributions on these concepts. The electromagnetic and vacuum tests are then analyzed. These tests are required to ensure that the manufactured devices fulfil the specifications. Finally, and only from the analytical point of view, the strip-line kickers are studied together with a pulsed power supply based on solid state power switches (MOSFETs). The solid state technology applied to pulsed power supplies is introduced and several circuit topologies are modelled and simulated to obtain fast and good flat-top pulses.

Efficient Model-based 3D Tracking by Using Direct Image Registration

This thesis deals with the problem of efficiently tracking 3D objects in sequences of images. We tackle the efficient 3D tracking problem by using direct image registration. This problem is posed as an iterative optimization procedure that minimizes a brightness error norm. We review the most popular iterative methods for image registration in the literature, turning our attention to those algorithms that use efficient optimization techniques. Two forms of efficient registration algorithms are investigated. The first type comprises the additive registration algorithms: these algorithms incrementally compute the motion parameters by linearly approximating the brightness error function. We centre our attention on Hager and Belhumeur’s factorization-based algorithm for image registration. We propose a fundamental requirement that factorization-based algorithms must satisfy to guarantee good convergence, and introduce a systematic procedure that automatically computes the factorization. Finally, we also bring out two warp functions to register rigid and nonrigid 3D targets that satisfy the requirement. The second type comprises the compositional registration algorithms, where the brightness function error is written by using function composition. We study the current approaches to compositional image alignment, and we emphasize the importance of the Inverse Compositional method, which is known to be the most efficient image registration algorithm. We introduce a new algorithm, the Efficient Forward Compositional image registration: this algorithm avoids the necessity of inverting the warping function, and provides a new interpretation of the working mechanisms of the inverse compositional alignment. By using this information, we propose two fundamental requirements that guarantee the convergence of compositional image registration methods. Finally, we support our claims by using extensive experimental testing with synthetic and real-world data. We propose a distinction between image registration and tracking when using efficient algorithms. We show that, depending whether the fundamental requirements are hold, some efficient algorithms are eligible for image registration but not for tracking.

Optimal Multiresolution 3D Level-Set Method for Liver Segmentation incorporating Local Curvature Constraints

Advanced liver surgery requires a precise pre-operative planning, where liver segmentation and remnant liver volume are key elements to avoid post-operative liver failure. In that context, level-set algorithms have achieved better results than others, especially with altered liver parenchyma or in cases with previous surgery. In order to improve functional liver parenchyma volume measurements, in this work we propose two strategies to enhance previous level-set algorithms: an optimal multi-resolution strategy with fine details correction and adaptive curvature, as well as an additional semiautomatic step imposing local curvature constraints. Results show more accurate segmentations, especially in elongated structures, detecting internal lesions and avoiding leakages to close structures

Virtual 3D Support Contents Oriented to Interactive Self-Learning

In the educational project described in this paper, new virtual 3D didactical contents have been developed to achieve specific outcomes, within the frame of a new methodology oriented to objectives of the European Higher Education Area directives. The motivation of the project was to serve as a new assessment method, to create a link between new programs of study with the older ones. In this project, new rubrics have been developed to be employed as an objective method of evaluation of specific and transversal outcomes, to accomplish the certification criteria of institutions like ABET (Accreditation Board for Engineering and Technology).

Dynamic Response of Wind Turbines to Theoretical 3D Seismic Motions Taking into Account the Rotational Component

We study the dynamic response of a wind turbine structure subjected to theoretical seismic motions, taking into account the rotational component of ground shaking. Models are generated for a shallow moderate crustal earthquake in the Madrid Region (Spain). Synthetic translational and rotational time histories are computed using the Discrete Wavenumber Method, assuming a point source and a horizontal layered earth structure. These are used to analyze the dynamic response of a wind turbine, represented by a simple finite element model. Von Mises stress values at different heights of the tower are used to study the dynamical structural response to a set of synthetic ground motion time histories

Integrated learning of computer applications for production engineering

A high productivity rate in Engineering is related to an efficient management of the flow of the large quantities of information and associated decision making activities that are consubstantial to the Engineering processes both in design and production contexts. Dealing with such problems from an integrated point of view and mimicking real scenarios is not given much attention in Engineering degrees. In the context of Engineering Education, there are a number of courses designed for developing specific competencies, as required by the academic curricula, but not that many in which integration competencies are the main target. In this paper, a course devoted to that aim is discussed. The course is taught in a Marine Engineering degree but the philosophy could be used in any Engineering field. All the lessons are given in a computer room in which every student can use each all the treated software applications. The first part of the course is dedicated to Project Management: the students acquire skills in defining, using Ms-PROJECT, the work breakdown structure (WBS), and the organization breakdown structure (OBS) in Engineering projects, through a series of examples of increasing complexity, ending up with the case of vessel construction. The second part of the course is dedicated to the use of a database manager, Ms-ACCESS, for managing production related information. A series of increasing complexity examples is treated ending up with the management of the pipe database of a real vessel. This database consists of a few thousand of pipes, for which a production timing frame is defined, which connects this part of the course with the first one. Finally, the third part of the course is devoted to the work with FORAN, an Engineering Production package of widespread use in the shipbuilding industry. With this package, the frames and plates where all the outfitting will be carried out are defined through cooperative work by the studens, working simultaneously in the same 3D model. In the paper, specific details about the learning process are given. Surveys have been posed to the students in order to get feed-back from their experience as well as to assess their satisfaction with the learning process. Results from these surveys are discussed in the paper

Three dimensional (3D) microstructure-based modeling of interfacial decohesion in particle reinforced metal matrix composites

Modeling and prediction of the overall elastic–plastic response and local damage mechanisms in heterogeneous materials, in particular particle reinforced composites, is a very complex problem. Microstructural complexities such as the inhomogeneous spatial distribution of particles, irregular morphology of the particles, and anisotropy in particle orientation after secondary processing, such as extrusion, significantly affect deformation behavior. We have studied the effect of particle/matrix interface debonding in SiC particle reinforced Al alloy matrix composites with (a) actual microstructure consisting of angular SiC particles and (b) idealized ellipsoidal SiC particles. Tensile deformation in SiC particle reinforced Al matrix composites was modeled using actual microstructures reconstructed from serial sectioning approach. Interfacial debonding was modeled using user-defined cohesive zone elements. Modeling with the actual microstructure (versus idealized ellipsoids) has a significant influence on: (a) localized stresses and strains in particle and matrix, and (b) far-field strain at which localized debonding takes place. The angular particles exhibited higher degree of load transfer and are more sensitive to interfacial debonding. Larger decreases in stress are observed in the angular particles, because of the flat surfaces, normal to the loading axis, which bear load. Furthermore, simplification of particle morphology may lead to erroneous results.

Detection and Isolation of Simultaneous Additive and Parametric Faults in Nonlinear Stochastic Dynamical Systems

This paper presents a new fault detection and isolation scheme for dealing with simultaneous additive and parametric faults. The new design integrates a system for additive fault detection based on Castillo and Zufiria, 2009 and a new parametric fault detection and isolation scheme inspired in Munz and Zufiria, 2008 . It is shown that the so far existing schemes do not behave correctly when both additive and parametric faults occur simultaneously; to solve the problem a new integrated scheme is proposed. Computer simulation results are presented to confirm the theoretical studies.

Rapid prototyping for biomedical engineering: current capabilities and Challenges

A new set of manufacturing technologies has emerged in the past decades to address market requirements in a customized way and to provide support for research tasks that require prototypes. These new techniques and technologies are usually referred to as rapid prototyping and manufacturing technologies, and they allow prototypes to be produced in a wide range of materials with remarkable precision in a couple of hours. Although they have been rapidly incorporated into product development methodologies, they are still under development, and their applications in bioengineering are continuously evolving. Rapid prototyping and manufacturing technologies can be of assistance in every stage of the development process of novel biodevices, to address various problems that can arise in the devices' interactions with biological systems and the fact that the design decisions must be tested carefully. This review focuses on the main fields of application for rapid prototyping in biomedical engineering and health sciences, as well as on the most remarkable challenges and research trends.

Activity based costing, time-driven activity based costing and lean accounting: differences among three accounting systems' approach to manufacturing

Choosing an appropriate accounting system for manufacturing has always been a challenge for managers. In this article we try to compare three accounting systems designed since 1980 to address problems of traditional accounting system. In the first place we are going to present a short overview on background and definition of three accounting systems: Activity Based costing, Time-Driven Activity Based Costing and Lean Accounting. Comparisons are made based on the three basic roles of information generated by accounting systems: financial reporting, decision making, and operational control and improvement. The analysis in this paper reveals how decisions are made over the value stream in the companies using Lean Accounting while decisions under the ABC Accounting system are taken at individual product level, and finally we will show how TD-ABC covers both product and process levels for decision making. In addition, this paper shows the importance of nonfinancial measures for operational control and improvement under the Lean Accounting and TD-ABC methods whereas ABC relies mostly on financial measures in this context.