A probabilistic approach for the optimisation of ultrasonic array inspection techniques

AbstractUltrasonic arrays are now used routinely for the inspection of engineering structures in order to maintain their integrity and assess their performance. Such inspections are usually optimised manually using empirical measurements and parametric studies which are laborious, time-consuming, and may not result in an optimal approach. In this paper, a general framework for the optimisation of ultrasonic array inspection techniques in NDE is presented. Defect detection rate is set as the main inspection objective and used to assess the performance of the optimisation framework. Statistical modelling of the inspection is used to form the optimisation problem and incorporate inspection uncertainty such as crack type and location, material properties and geometry, etc. A genetic algorithm is used to solve the global optimisation problem. As a demonstration, the optimisation framework is used with two objective functions based on array signal amplitude and signal-to-noise ratio (SNR). The optimal use of plane B-scan and total focusing method imaging algorithms is also investigated. The performance of the optimisation scheme is explored in simulation and then validated experimentally. It has been found that, for the inspection scenarios considered, TFM provides better detectability in a statistical sense than plane B-scan imaging in scenarios where uncertainty in the inspection is expected

Developmental delays and subcellular stress as downstream effects of sonoporation

Posters: no. 2Control ID: 1672434OBJECTIVES: The biological impact of sonoporation has often been overlooked. Here we seek to obtain insight into the cytotoxic impact of sonoporation by gaining new perspectives on anti-proliferative characteristics that may emerge within sonoporated cells. We particularly focused on investigating the cell-cycle progression kinetics of sonoporated cells and identifying organelles that may be stressed in the recovery process. METHODS: In line with recommendations on exposure hardware design, an immersion-based ultrasound platform has been developed. It delivers 1 MHz ultrasound pulses (100 cycles; 1 kHz PRF; 60 s total duration) with 0.45 MPa peak negative pressure to a cell chamber that housed HL-60 leukemia cells and lipid-shelled microbubbles at a 10:1 cell-tobubble ratio (for 1e6/ml cell density). Calcein was used to facilitate tracking of sonoporated cells with enhanced uptake of exogenous molecules. The developmental trend of sonoporated cells was quantitatively analyzed using BrdU/DNA flow cytometry that monitors the cell population’s DNA synthesis kinetics. This allowed us to measure the temporal progression of DNA synthesis of sonoporated cells. To investigate whether sonoporation would upset subcellular homeostasis, post-exposure cell samples were also assayed for various proteins using Western blot analysis. Analysis focus was placed on the endoplasmic reticulum (ER): an important organelle with multi-faceted role in cellular functioning. The post-exposure observation time spanned between 0-24 h. RESULTS: Despite maintaining viability, sonoporated cells were found to exhibit delays in cell-cycle progression. Specifically, their DNA synthesis time was lengthened substantially (for HL-60 cells: 8.7 h for control vs 13.4 h for the sonoporated group). This indicates that sonoporated cells were under stress: a phenomenon that is supported by our Western blot assays showing upregulation of ER-resident enzymes (PDI, Ero1), ER stress sensors (PERK, IRE1), and ER-triggered pro-apoptotic signals (CHOP, JNK). CONCLUSIONS: Sonoporation, whilst being able to facilitate internalization of exogenous molecules, may inadvertently elicit a cellular stress response. These findings seem to echo recent calls for reconsideration of efficiency issues in sonoporation-mediated drug delivery. Further efforts would be necessary to improve the efficiency of sonoporation-based biomedical applications where cell death is not desirable.postprin

A study on the change in plasma membrane potential during sonoporation

Posters: no. 4Control ID: 1680329OBJECTIVES: There has been validated that the correlation of sonoporation with calcium transients is generated by ultrasound-mediated microbubbles activity. Besides calcium, other ionic flows are likely involved in sonoporation. Our hypothesis is the cell electrophysiological properties are related to the intracellular delivery by ultrasound and microbubbles. In this study, a real-time live cell imaging platform is used to determine whether plasma membrane potential change is related to the sonoporation process at the cellular level. METHODS: Hela cells were cultured in DMEM supplemented with 10% FBS in Opticell Chamber at 37 °C and 5% CO2, and reached 80% confluency before experiments. The Calcein Blue-AM, DiBAC4(3) loaded cells in the Opticell chamber filled with PI solution and Sonovue microbubbles were immerged in a water tank on a inverted fluorescence microscope. Pulsed ultrasound (1MHz freq., 20 cycles, 20Hz PRF, 0.2-0.5MPa PNP) was irradiated at the angle of 45° to the region of interest for 1s.The real-time fluorescence imaging for different probes was acquired by a cooled CCD camera every 20s for 10min. The time-lapse fluorescence images were quantitatively analyzed to evaluate the correlation of cell viability, intracellular delivery with plasma membrane potential change. RESULTS: Our preliminary data showed that the PI fluorescence, which indicated intracellular delivery, was immediately accumulated in cells adjacent to microbubbles after exposure, suggesting that their membranes were damaged by ultrasound-activated microbubbles. However, the fluorescence reached its highest level within 4 to 6 minutes and was unchanged thereafter, indicating the membrane was gradually repaired within this period. Furthermore, using DIBAC4(3), which detected the change in the cell membrane potential, we found that the loss of membrane potential might be associated with intracellular delivery, because the PI fluorescence accumulation was usually accompanied with the change in DIBAC4 (3) fluorescence. CONCLUSIONS: Our study suggests that there may be a linkage between the cell membrane potential change and intracellular delivery mediated by ultrasound and microbubbles. We also suggest that other ionic flows or ion channels may be involved in the cell membrane potential change in sonoporation. Further efforts to explore the cellular mechanism of this phenomenon will improve our understanding of sonoporation.postprin

How sonoporation disrupts cellular structural integrity: morphological and cytoskeletal observations

Posters: no. 1Control ID: 1672429OBJECTIVES: In considering sonoporation for drug delivery applications, it is essential to understand how living cells respond to this puncturing force. Here we seek to investigate the effects of sonoporation on cellular structural integrity. We hypothesize that the membrane morphology and cytoskeletal behavior of sonoporated cells under recovery would inherently differ from that of normal viable cells. METHODS: A customized and calibrated exposure platform was developed for this work, and the ZR-75-30 breast carcinoma cells were used as the cell model. The cells were exposed to either single or multiple pulses of 1 MHz ultrasound (pulse length: 30 or 100 cycles; PRF: 1kHz; duration: up to 60s) with 0.45 MPa spatial-averaged peak negative pressure and in the presence of lipid-shelled microbubbles. Confocal microscopy was used to examine insitu the structural integrity of sonoporated cells (identified as ones with exogenous fluorescent marker internalization). For investigations on membrane morphology, FM 4-64 was used as the membrane dye (red), and calcein was used as the sonoporation marker (green); for studies on cytoskeletal behavior, CellLight (green) and propidium iodide (red) were used to respectively label actin filaments and sonoporated cells. Observation started from before exposure to up to 2 h after exposure, and confocal images were acquired at real-time frame rates. Cellular structural features and their temporal kinetics were quantitatively analyzed to assess the consistency of trends amongst a group of cells. RESULTS: Sonoporated cells exhibited membrane shrinkage (decreased by 61% in a cell’s cross-sectional area) and intracellular lipid accumulation (381% increase compared to control) over a 2 h period. The morphological repression of sonoporated cells was also found to correspond with post-sonoporation cytoskeletal processes: actin depolymerization was observed as soon as pores were induced on the membrane. These results show that cellular structural integrity is indeed disrupted over the course of sonoporation. CONCLUSIONS: Our investigation shows that the biophysical impact of sonoporation is by no means limited to the induction of membrane pores: e.g. structural integrity is concomitantly affected in the process. This prompts the need for further fundamental studies to unravel the complex sequence of biological events involved in sonoporation.postprin

Real-time imaging of cellular dynamics during low-intensity pulsed ultrasound exposure

Control ID: 1671584Oral Session 5 - Bioeffects of therapeutic ultrasoundOBJECTIVE: Although the therapeutic potential of low-intensity pulsed ultrasound is unquestionable, the wave-matter interactions involved in the process remain to be vaguely characterized. Here we seek to undertake a series of in-situ cellular imaging studies that aim to analyze the mechanical impact of low-intensity pulsed ultrasound on attached fibroblasts from three different aspects: membrane, cytoskeleton, and nucleus. METHODS: Our experimental platform comprised an in-house ultrasound exposure hardware that was coupled to a confocal microscopy system. The waveguided ultrasound beam was geometrically aligned to the microscope’s fieldof-view that corresponds to the center of a polystyrene dish containing fibroblasts. Short ultrasound pulses (5 cycles; 2 kHz PRF) with 0.8 MPa peak acoustic pressure (0.21 W/cm2 SPTA intensity) were delivered over a 10 min period. Live imaging was performed on both membrane (CellMask) and cytoskeleton (actin-GFP, tubulin-RFP) over the entire observation period (up to 30 min after end of exposure). Also, pre- and post-exposure fixed-cell imaging was conducted on the nucleus (Hoechst 33342) and two cytoskeleton components related to stress fibers: F-actin (phalloidin-FITC) and vincullin (Alexa Fluor 647 conjugated). To study whether mechanotransduction was responsible in mediating ultrasound-cell interactions, some experiments were conducted with the addition of gadolinium that blocks stretch-sensitive ion channels. RESULTS: Cell shrinkage was evident over the course of low-intensity pulsed ultrasound exposure. This was accompanied with contraction of actin and tubulin. Also, an increase in central stress fibers was observed at the end of exposure, while the nucleus was found to have decreased in size. Interestingly, after the exposure, a significant rebound in cell volume was observed over a 30 min. period. These effects were not observed in cases with gadolinium blockage of mechanosensitive ion channels. CONCLUSIONS: Our results suggest that low-intensity pulsed ultrasound would transiently induce remodeling of a cell’s membrane and cytoskeleton, and it will lead to repression of nucleus. This indicates that ultrasound after all represents a mechanical stress on cellular membrane. The post-exposure outgrowth phenomenon is also of practical relevance as it may be linked to the stimulatory effects that have been already observed in low-intensity pulsed ultrasound treatments.postprin

Production Engineering and Management

The annual International Conference on Production Engineering and Management takes place for the sixth time his year, and can therefore be considered a well - established event that is the result of the joint effort of the OWL University of Applied Sciences and the University of Trieste. The conference has been established as an annual meeting under the Double Degree Master Program ‘Production Engineering and Management’ by the two partner universities. The main goal of the conference is to provide an opportunity for students, researchers and professionals from Germany, Italy and abroad, to meet and exchange information, discuss experiences, specific practices and technical solutions used in planning, design and management of production and service systems. In addition, the conference is a platform aimed at presenting research projects, introducing young academics to the tradition of Symposiums and promoting the exchange of ideas between the industry and the academy. Especially the contributions of successful graduates of the Double Degree Master Program ‘Production Engineering and Management’ and those of other postgraduate researchers from several European countries have been enforced. This year’s special focus is on Direct Digital Manufacturing in the context of Industry 4.0, a topic of great interest for the global industry. The concept is spreading, but the actual solutions must be presented in order to highlight the practical benefits to industry and customers. Indeed, as Henning Banthien, Secretary General of the German ‘Plattform Industrie 4.0’ project office, has recently remarked, “Industry 4.0 requires a close alliance amongst the private sector, academia, politics and trade unions” in order to be “translated into practice and be implemented now”. PEM 2016 takes place between September 29 and 30, 2016 at the OWL University of Applied Sciences in Lemgo. The program is defined by the Organizing and Scientific Committees and clustered into scientific sessions covering topics of main interest and importance to the participants of the conference. The scientific sessions deal with technical and engineering issues, as well as management topics, and include contributions by researchers from academia and industry. The extended abstracts and full papers of the contributions underwent a double - blind review process. The 24 accepted presentations are assigned, according to their subject, to one of the following sessions: ‘Direct Digital Manufacturing in the Context of Industry 4.0’, ‘Industrial Engineering and Lean Management’, ‘Management Techniques and Methodologies’, ‘Wood Processing Technologies and Furniture Production’ and ‘Innovation Techniques and Methodologies

Towards UWB microwave hyperthermia for brain cancer treatment

Despite numerous clinical trials demonstrating that microwave hyperthermia is a powerful adjuvant modality in the treatment of cancers, there have been few instances where this method has been applied to brain tumors. The reason is a combination of anatomical and physiological factors in this site that require an extra degree of accuracy and control in the thermal dose delivery which current systems are not able to provide. All clinical applicators available today are in fact based on a single-frequency technology. In terms of treatment planning options, the use of a single frequency is limiting as the size of the focal spot cannot be modified to accommodate the specific tumor volume and location. The introduction of UWB systems opens up an opportunity to overcome these limitations, as they convey the possibility to adapt the focal spot and to use multiple operating frequencies to reduce the power deposition in healthy tissues.In this thesis, we explore whether the current treatment planning methods can be meaningfully translated to the UWB setting and propose new solutions for UWB microwave hyperthermia. We analyze the most commonly used cost-functions for treatment planning optimization and discuss their suitability for use with UWB systems. Then, we propose a novel cost-function specifically tailored for UWB optimization (HCQ). To solve for the HCQ, we further describe a novel, time-reversal based, iterative scheme for the rapid and efficient optimization of UWB treatment plans. We show that the combined use of these techniques results in treatment plans that better exploit the benefits of UWB systems, yielding increased tumor coverage and lower peak temperatures outside the target. Next, we investigate the design possibilities of UWB applicators and introduce a fast E-field approximation scheme. The method can be used for the global optimization of the array parameters with respect to the multiple objectives and requirements of hyperthermia treatments. Together, the proposed solutions represent a step forward in the implementation of patient-specific hyperthermia treatments, increasing their accuracy and precision. The results suggest that UWB microwave hyperthermia for brain cancer treatment is feasible, and motivate the efforts for further development of UWB applicators and systems

Occupational health and safety issues in human-robot collaboration: State of the art and open challenges

Human-Robot Collaboration (HRC) refers to the interaction of workers and robots in a shared workspace. Owing to the integration of the industrial automation strengths with the inimitable cognitive capabilities of humans, HRC is paramount to move towards advanced and sustainable production systems. Although the overall safety of collaborative robotics has increased over time, further research efforts are needed to allow humans to operate alongside robots, with awareness and trust. Numerous safety concerns are open, and either new or enhanced technical, procedural and organizational measures have to be investigated to design and implement inherently safe and ergonomic automation solutions, aligning the systems performance and the human safety. Therefore, a bibliometric analysis and a literature review are carried out in the present paper to provide a comprehensive overview of Occupational Health and Safety (OHS) issues in HRC. As a result, the most researched topics and application areas, and the possible future lines of research are identified. Reviewed articles stress the central role played by humans during collaboration, underlining the need to integrate the human factor in the hazard analysis and risk assessment. Human-centered design and cognitive engineering principles also require further investigations to increase the worker acceptance and trust during collaboration. Deepened studies are compulsory in the healthcare sector, to investigate the social and ethical implications of HRC. Whatever the application context is, the implementation of more and more advanced technologies is fundamental to overcome the current HRC safety concerns, designing low-risk HRC systems while ensuring the system productivity

Modelling, Simulation and Data Analysis in Acoustical Problems

Modelling and simulation in acoustics is currently gaining importance. In fact, with the development and improvement of innovative computational techniques and with the growing need for predictive models, an impressive boost has been observed in several research and application areas, such as noise control, indoor acoustics, and industrial applications. This led us to the proposal of a special issue about “Modelling, Simulation and Data Analysis in Acoustical Problems”, as we believe in the importance of these topics in modern acoustics’ studies. In total, 81 papers were submitted and 33 of them were published, with an acceptance rate of 37.5%. According to the number of papers submitted, it can be affirmed that this is a trending topic in the scientific and academic community and this special issue will try to provide a future reference for the research that will be developed in coming years