Journal articles on the topic 'Detector read-out techniques'

Precise measurements of the muon flux are important for different practical applications, both in environmental studies and for the estimation of the water equivalent depths of underground sites. A mobile detector for cosmic muon flux measurements has been set up at IFIN-HH, Romania. The device is used to measure the muon flux on different locations at the surface and underground. Its first configuration, not used in the present, has been composed of two 1 m2scintillator plates, each viewed by wave length shifters and read out by two Photomultiplier Tubes (PMTs). A more recent configuration, consists of two 1 m2detection layers, each one including four 1 · 0,25 m2large scintillator plates. The light output in each plate is collected by twelve optical fibers and then read out by one PMT. Comparative results were obtained with both configurations.

Elemental mapping of biological specimens by electron energy loss spectroscopy (EELS) can be carried out both in the scanning transmission electron microscope (STEM), and in the energy-filtering transmission electron microscope (EFTEM). Choosing between these two approaches is complicated by the variety of specimens that are encountered (e.g., cells or macromolecules; cryosections, plastic sections or thin films) and by the range of elemental concentrations that occur (from a few percent down to a few parts per million). Our aim here is to consider the strengths of each technique for determining elemental distributions in these different types of specimen.On one hand, it is desirable to collect a parallel EELS spectrum at each point in the specimen using the ‘spectrum-imaging’ technique in the STEM. This minimizes the electron dose and retains as much quantitative information as possible about the inelastic scattering processes in the specimen. On the other hand, collection times in the STEM are often limited by the detector read-out and by available probe current. For example, a 256 x 256 pixel image in the STEM takes at least 30 minutes to acquire with read-out time of 25 ms. The EFTEM is able to collect parallel image data using slow-scan CCD array detectors from as many as 1024 x 1024 pixels with integration times of a few seconds. Furthermore, the EFTEM has an available beam current in the µA range compared with just a few nA in the STEM. Indeed, for some applications this can result in a factor of ~100 shorter acquisition time for the EFTEM relative to the STEM. However, the EFTEM provides much less spectral information, so that the technique of choice ultimately depends on requirements for processing the spectrum at each pixel (viz., isolated edges vs. overlapping edges, uniform thickness vs. non-uniform thickness, molar vs. millimolar concentrations).

The design of a detector for tracking charged particles is presented together with the characterization techniques developed to extract the main design specifications. The goals for the final detector are to achieve real-time imaging performances, a large detection area, and a high spatial resolution, particularly suitable for medical imaging applications. This paper describes the prototype of the tracker plane, which has a 20 × 20 cm2sensitive area consisting of two crossed ribbons of 500 μm square scintillating fibers. The information about the hit position extracted real-time tracker in an innovative way, using a reduced number of the read-out channels to obtain a very large detection area but with moderate costs and complexity. The performances of the tracker have been investigated usingβsources, cosmic rays, and a 62 MeV proton beam.

Paleo-detectors are a proposed experimental technique to search for dark matter (DM). In lieu of the conventional approach of operating a tonne-scale real-time detector to search for DM-induced nuclear recoils, paleo-detectors take advantage of small samples of naturally occurring rocks on Earth that have been deep underground (≳5 km), accumulating nuclear damage tracks from recoiling nuclei for O(1)Gyr. Modern microscopy techniques promise the capability to read out nuclear damage tracks with nanometer resolution in macroscopic samples. Thanks to their O(1)Gyr integration times, paleo-detectors could constitute nuclear recoil detectors with keV recoil energy thresholds and 100 kilotonne-yr exposures. This combination would allow paleo-detectors to probe DM-nucleon cross sections orders of magnitude below existing upper limits from conventional direct detection experiments. In this article, we use improved background modeling and a new spectral analysis technique to update the sensitivity forecast for paleo-detectors. We demonstrate the robustness of the sensitivity forecast to the (lack of) ancillary measurements of the age of the samples and the parameters controlling the backgrounds, systematic mismodeling of the spectral shape of the backgrounds, and the radiopurity of the mineral samples. Specifically, we demonstrate that even if the uranium concentration in paleo-detector samples is 10−8 (per weight), many orders of magnitude larger than what we expect in the most radiopure samples obtained from ultra basic rock or marine evaporite deposits, paleo-detectors could still probe DM-nucleon cross sections below current limits. For DM masses ≲ 10 GeV/c2, the sensitivity of paleo-detectors could still reach down all the way to the conventional neutrino floor in a Xe-based direct detection experiment.

Residual stresses in welds are of major concern for the structure integrity assessment in industrial components. The stresses in the final weld can be determined relatively simple using well established destructive or non-destructive techniques. However, such measurements reflect only the final condition and it remains unclear how stresses built up during the welding process. In order to optimise the final residual stresses in the weld, it is important to monitor the formation of residual strain and stress during the welding process and as such to gain insight into the mechanisms of stress development. In this work non-destructive high energy dispersive synchrotron X-ray diffraction at a high count rate is applied in order to dissolve the welding process in-situ in time and temperature. However, the achievable time resolution at commonly used instruments is restricted by either a limited photon flux or the read out electronics of the detector system resulting in counting times usually much longer than 1sec. We present an energy dispersive detector and read-out-electronics setup realized at the high flux and energy beam line ID15A at the ESRF. The setup allowed for monitoring the strain evolution in two perpendicular directions simultaneously at a sampling rate of 5Hz, resulting in sufficient time and temperature resolution. The change in detector dead time is accounted for by a correction function, which was specifically determined for the detector setup as used for this in-situ experiment.

A liquid argon time projection chamber (LAr-TPC) working as an electronic bubblechamber, continually sensitive, self-triggering, able to provide 3-D imaging of any ionizing event together with a good calorimetric response was first proposed by Carlo Rubbia in 1977. In order to verify the feasibility of such a detector, the ICARUS collaboration started in 1985 an intensive R&D program aiming to solve the main technological problems. The satisfactory results obtained on small scale tests allowed us to start in 1989 the construction of a 3 ton prototype which is presently working at CERN under stable conditions without interruptions since May 1991, collecting events from cosmic rays and monochromatic gamma ray sources. The paper describes the working principles of such a detector, showing the results of the research program applied in the present prototype. Some emphasis is put into the description of the read-out apparatus, a custom designed VME-based multichannel waveform recorder. The huge number of channels and the high sampling rate essential to achieve the high resolution in the track detection make the data acquisition architecture a crucial point for event detection rate and self-triggering capabilities. At present a. complete signal analysis is performed to define those parameters needed to tune up algorithms and filtering methods that will become necessary in the future step of the ICARUS detector (hundred to thousands of tons of argon, i. e. tens of thousands of readout channels). A particular effort is made to optimize track localization algorithms, in order to achieve high efficiency data reduction by storing only those portions of signal which contain the track (thus lowering up to three orders of magnitude the event size) . From a human interface point of view, the data presentation relies on an event imaging very similar to the old fashioned bubble chambers. Besides from giving a direct feeling of what happens inside the detector, this kind of representations opens a way to the wide panorama of image processing applications: tasks as detail enhancement, pattern recognition or 3D reconstruction will helpfully inherit support from more general procedures.

Introduction Contemporary radiotherapy uses a number of highly specialized irradiation techniques dedicated to well-defined clinical diagnoses. Among these methods are techniques to irradiate the skin (TSEI), bone marrow (TMI) or the whole body of the patient (TBI). TBI has over the last century been used in the treatment of a variety of conditions, both benign and malignant. However, its importance has increased with the development of knowledge about the impact of ionizing radiation on the human body and the development of clinical dosimetry techniques. At present, however, this method is primarily used in the treatment of hyperplasia. Aim The general aim of the study is to compare dose distributions at selected points of the anthropomorphic phantom under full body radiation conditions for X: 6MV and 15MV radiation. Specific objectives are defined: comparison of percent depth and function of photon emission profiles: 6MV and 15MV measured with radiofrequency hydrophobic films; measurement of doses in selected cross sections of the anthropomorphic phantom. Material and method A number of measuring devices and materials used in daily work by staff of the Medical Physics Department of the Greater Poland Cancer Centre were used to carry out the study part, but Alderson's anthropomorphic phantom and the radiochromic films in the form of point detectors were essential. In addition to each step of the research part, a special measuring system was prepared to reproduce the conditions prevailing during the TBI session as closely as possible. The research was carried out in three stages: Calibration of radiochromic films; PDD and OCR measurement for: 6MV and 15MV photon beam under TBI conditions; Measurement of dose distribution in selected anthropomorphic phantom's cross sections using radiochromic films in the form of point detectors. Results For the lateral field irradiated with 6MV photon beam, the maximum compliance (less than 2%) was obtained for the elbows at the entrance and in the center of the phantom; abdomen for the detector positioned in the center of the phantom, the lungs at the entrance and the arms in the middle, and the neck at the position of the film at the entrance. In the case of the lateral field X 15MV, the highest correspondence occurred for the points: the head and the PC in the position of the film in the center of the phantom and the entrance neck. In the case of AP/PA fields for 6MV energy, the highest compatibility was obtained for the mediastinum in all positions of the film. A small difference was also obtained for the points: head in the middle and at the output of the beam; as well as PC on the output. For AP/PA X 15MV fields, the highest dose compliance not exceeding 1% was obtained for the location of the neck - at the beam entrance, and the lung and mediastinum at the detector position at the center of the phantom. Conclusions On the basis of measurements of dose distribution at selected points of the patient's body for radiation X: 6MV and 15MV in the TBI procedure, the following conclusions can be made: Gafchromic EBT (radiochromic type film) can be successfully used for dosimetric measurements, among others. Due to their properties, such as the ability to cut from the sheet of film spot detectors of any shape and size, flexibility, low sensitivity to daylight, resistance to humidity, etc.; Their main drawback is the high cost of buying films and the long time required to prepare the detectors and then read the measured doses. Because of the low popularity of point-based EBTs in point dosing, further research is needed to improve their response to ionizing radiation. There is a noticeable increase in the difference between the dose calculated and measured as the distance between the position of individual detectors increases from the center point. The difference between the dose measured and planned in any of the cases examined does not exceed 9%. The measurements show that the method used is fast, accurate, and can be successfully used as a validation tool not only for the TBI procedure but also for other methods of cancer radiotherapy.

A rapid nondestructive defect assessment and quantification method based on X-ray diffraction and three-dimensional reciprocal-space mapping has been established. A fast read-out two-dimensional detector with a high dynamic range of 20 bits, in combination with a powerful data analysis software package, is set up to provide fast feedback to crystal growers with the goal of supporting the development of reduced defect density GaN growth techniques. This would contribute strongly to the improvement of the crystal quality of epitaxial structures and therefore of optoelectronic properties. The method of normalized three-dimensional reciprocal-space mapping is found to be a reliable tool which shows clearly the influence of the parameters of the metal–organic vapour phase epitaxial and hydride vapour phase epitaxial (HVPE) growth methods on the extent of the diffuse scattering streak. This method enables determination of the basal stacking faults and an exploration of the presence of other types of defect such as partial dislocations and prismatic stacking faults. Three-dimensional reciprocal-space mapping is specifically used in the manuscript to determine basal stacking faults quantitatively and to discuss the presence of partial dislocations. This newly developed method has been applied to semipolar GaN structures grown on patterned sapphire substrates (PSSs). The fitting of the diffuse scattering intensity profiles along the stacking fault streaks with simulations based on a Monte Carlo approach has delivered an accurate determination of the basal plane stacking fault density. Three-dimensional reciprocal-space mapping is shown to be a method sensitive to the influence of crystallographic surface orientation on basal stacking fault densities during investigation of semipolar (11{\overline 2}2) GaN grown on anr-plane (1{\overline 1}02) PSS and semipolar (10{\overline 1}1) GaN grown on ann-plane (11{\overline 2}3) PSS. Moreover, the influence of HVPE overgrowth at reduced temperature on the quality of semipolar (11{\overline 2}2) GaN has been studied.

The foreseen incremental luminosity for near-future high-energy physics experiments demands evolution for the read-out electronics in terms of event data-rate. However, the filtering necessary to reject noise and meet the signal-to-noise-ratio requirements imposes a restriction on the operational speed of the conventional read-out electronics. The stringent trade-off between signal-to-noise-ratio and the event data-rate originates from the time-invariant behavior of the conventional systems. In this paper, the cases of time-variant systems are addressed, studying a benchmark with the RC-CR shaping function used in time-over-threshold methods. It was demonstrated that the time-variant systems enable a higher data-rate for the given noise performance. Moreover, taking advantage of time-variant systems, the proposed rising-edge method enables further data-rate enhancement with respect to the traditional time-over-threshold technique by reading the data from the rising edge of the analog output waveform. A comparison between the conventional time-invariant time-over-threshold technique, its time-variant equivalent and rising-edge method confirms the better performance of the latter one in terms of data-rate enhancement for a target noise performance. Moreover, design challenges for time-variant systems are briefly discussed, considering the ATLAS Monitored Drift Tube detector as a design case.

A resonance detection scheme and some useful ideas for cavity-based searches of light cold dark matter particles (such as axions) are presented, as an effort to aid in the on-going endeavors in this direction as well as for future experiments, especially in possibly developing a table-top experiment. The scheme is based on our idea of a resonant detector, incorporating an integrated tunnel diode (TD) and GaAs HEMT/HFET (High-Electron Mobility Transistor/Heterogeneous FET) transistor amplifier, weakly coupled to a cavity in a strong transverse magnetic field. The TD-amplifier combination is suggested as a sensitive and simple technique to facilitate resonance detection within the cavity while maintaining excellent noise performance, whereas our proposed Halbach magnet array could serve as a low-noise and permanent solution replacing the conventional electromagnets scheme. We present some preliminary test results which demonstrate resonance detection from simulated test signals in a small optimal axion mass range with superior signal-to-noise ratios (SNR). Our suggested design also contains an overview of a simpler on-resonance dc signal read-out scheme replacing the complicated heterodyne read-out. We believe that all these factors and our propositions could possibly improve or at least simplify the resonance detection and read-out in cavity-based DM particle detection searches (and other spectroscopy applications) and reduce the complications (and associated costs), in addition to reducing the electromagnetic interference and background.

In this report we propose a sensor architecture and a corresponding read-out technique on silicon for detection of dynamic capacitance change that can be applied to rapid particle counting and single particle sensing in a fluidic system.

Despite an evermore complete plethora of complex domain-specific semiempirical models, no succinct recipe for large-scale carbon nanotube electromechanical systems design has been formulated. To combine the benefits of these highly sensitive miniaturized mechanical sensors with the vast functionalities available in electronics, we identify a reduced key parameter set of carbon nanotube properties, nanoelectromechanical system design, and operation that steers the sensor’s performance towards system applications, based on open- and closed-loop topologies. Suspended single-walled carbon nanotubes are reviewed in terms of their electromechanical properties with the objective of evaluating orders of magnitude of the electrical actuation and detection mechanisms. Open-loop time-averaging and 1ωor 2ωmixing methods are completed by a new 4ωactuation and detection technique. A discussion on their extension to closed-loop topologies and system applications concludes the analysis, covering signal-to-noise ratio, and the capability to spectrally isolate the motional information from parasitical feedthrough by contemporary electronic read-out techniques.

Though I concur with most of the authors’ comments and procedure, I must disagree with the sweeping conclusion “it is unlikely that a Cyprus‐type deposit could be detected using TEM alone.” I realize this quote is somewhat out of context, but others may also read the statement that way. It was not mentioned by the authors that these deposits may occur in different electrical environments than their study area and, thus, be readily observed using TEM techniques.

Tritium is released abundantly to the environment by nuclear power plants (NPP), as a product of neutron capture by hydrogen and deuterium. In normal running conditions, released cooling waters may contain levels of tritium close to or even larger than the maximum authorised limit for human consumption (drinking and irrigation). The European Council Directive 2013/51/Euratom requires a maximum level of tritium in water for human consumption lower than 100 Bq=L. Current monitoring of tritium activity in water by liquid scintillating method takes about two days and can only be carried out in a dedicated laboratory. This system is not appropriate for real time monitoring. At present, there exists no available detector device with enough sensitivity to monitor waters for human consumption with high enough sensitivity. The goal of the TRITIUM project is to build a tritium monitor capable to measure tritium activities with detection limit close to 100Bq=L, using instrumentation technique developed in recent years for Nuclear and Particle Physics, such as scintillating fibres and silicon photomultipliers (SiPM). In this paper the current status of the TRITIUM project is presented and he results of first prototypes are discussed. A detector system based on scintillating fibers read out either photomultiplier tubes (PMTs) or silicon photomultiplier (SiPM) arrays is under development and will be installed in the vicinity of Almaraz nuclear power plant (Cáceres, Spain) by the fourth term of 2019.

Bismuth iodide is a potentially active material for room temperature radiation detector, as it is well reported in the literature that it has both wide energy band gap and high atomic absorption coefficient. Crystalline films of high atomic number and high radiation absorption coefficient can absorb the X-rays and convert them directly into electrical charges which can be read by imaging devices. Therefore, it was proposed to grow thin films of Bismuth iodide on glass substrate using thermal evaporation technique in vacuum to avoid the inclusion of impurities in the films. The structural studies of the films were carried out using XRD and optical absorption measurement was carried out in the UV/VIS region using spectrophotometer. All Bismuth iodide films grown at room temperature are polycrystalline and show X-ray diffraction peaks at angles reported in research papers. The optical transmission spectra of BiI3 films show a high transmission of about 80% in visible region with a sharp fall near the fundamental absorption at 650 nm. Resistivity of the as-grown film was found to be around 1012 ohm-cm suitable value for X-ray detection application. Films were subjected to scanning electron microscopy to study the growth features of both as-grown and annealed films.

Neutron depth profiling (NDP) is a non-destructive technique used for identifying the concentration of impurity isotopes below the sample surface. NDP is carried out by detection of the emitted charged particles resulting from bombarding the sample with neutrons. NDP specifies the isotopic concentration versus the sample depth for a few micrometers below the surface. The sample is bombarded inside a research reactor using a thermal neutron beam. Charged particles like alpha particles or protons are produced from the neutron induced reactions in the sample. Each neutron isotopic interaction produces a certain Q, indicating a specific kinetic energy for the emitted charged particle. As the charged particle travels through the sample to eject the surface, it loses energy to atoms (electrons) on its path. The charged particle energy loss holds information regarding the number of atoms by which the emitted particle passed, thus indicating its original depth. The purpose of this work is to check the capability of Artificial Neural Networks (ANNs) in predicting the boron concentration profile across a boro-silicate sample of thickness 3.5 μm divided into 10 layers. Each layer included different boron concentration than the other. Also, the boron concentration had the values {0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1}. Training, validation, and test data were generated synthetically using MCNP6 in which the boron concentrations varied in the layer number from one sample to another. MCNP6 model consisted of a silicon barrier detector, boro-silicate sample, chamber body and an idealized thermal neutron source. The detector, sample, and the source were located in a voided chamber. The samples were irradiated with a 0.025 eV monoenergetic thermal neutron beam from a monodirectional disk source. To cover the whole area of the samples, the thermal neutron beam had a radius of 3 cm. The silicon detector active volume was modelled as a 100 μm thick and 3 cm radius facing the sample directly. The sample, beam, and the detector were placed on the same axis. Ten ANN regression models were developed, one for each layer boron concentration prediction where the input for each model was the alpha spectrum read by the detector, while the output was the boron concentration for each layer. Results showed regression values higher than 0.94 for all of the developed models. ANNs proved its capability of predicting the boron profile form the alpha spectrum read by the detector regarding neutron depth profiling in a boro-silicate samples.

One of the most important functions of the human visual system is automatic captioning. Caption generation is one of the more interesting and focused areas of AI, with numerous challenges to overcome. If there is an application that automatically captions the scenes in which a person is present and converts the caption into a clear message, people will benefit from it in a variety of ways. In this, we offer a deep learning model that detects things or features in images automatically, produces descriptions for the images, and transforms the descriptions to audio for louder readout. The model uses pre-trained CNN and LSTM models to perform the task of extracting objects or features to get the captions. In our model, first task is to detect objects within the image using pre trained Mobilenet model of CNN (Convolutional Neural Networks) and therefore the other is to caption the pictures based on the detected objects by using LSTM (Long Short Term Memory) and convert caption into speech to read out louder to the person by using SpeechSynthesisUtterance interface of the Web Speech API. The interface of the model is developed using NodeJS as a backend for the web page. Caption generation entails a number of complex steps, including selecting the dataset, training the model, validating the model, creating pre-trained models to check the images, detecting the images, and finally generating captions.

Mycotoxins contamination is a global public health concern. Therefore, highly sensitive and selective techniques are needed for their on-site monitoring. Several approaches are conceivable for mycotoxins analysis, among which colorimetric methods are the most attractive for commercialization purposes thanks to their visual read-out, easy operation, cost-effectiveness, and rapid response. This review covers the latest achievements in the last five years for the development of colorimetric methods specific to mycotoxins analysis, with a particular emphasis on their potential for large-scale applications in food industries. Gathering all types of (bio)receptors, main colorimetric methods are critically discussed, including enzyme-linked assays, lateral flow-assays, microfluidic devices, and homogenous in-solution strategies. This special focus on colorimetry as a versatile transduction method for mycotoxins analysis is comprehensively reviewed for the first time.

IntroductionEnergy loss spectroscopy is a well known technique allowing elemental analysis in thin films. When an energy loss spectrometer is coupled with a STEM it is possible in addition to perform chemical mapping. The U314 INSERM laboratory is equipped with a C.M. 30 PHILIPS STEM coupled with an EDAX 9900 and a GATAN 666 parallel detection electron spectrometer.We describe a complete system including an additional A.T. microcomputer enabling simultaneous energy loss analysis and chemical mapping .Material and methodAn interface fitted with the bus connecting the GATAN electronics and the EDAX multichannel analyzer (MCA) allows to read the data during a photodiode read-out, and to send them via a parallel port into the host A.T microcomputer. To do this, only 18 bus lines are used and the proprietary communication protocol between the photodiode interface and the MCA is not affected. All the functions provided by the EDAX PVEELP software are preserved.

The optimization of the Beetle readout ASIC and the performance of the software for the signal processing based on machine learning methods are presented. The Beetle readout chip was developed for the LHCb (Large Hadron Collider beauty) tracking detectors and was used in the VELO (Vertex Locator) during Run 1 and 2 of LHC data taking. The VELO, surrounding the LHC beam crossing region, was a leading part of the LHCb tracking system. The Beetle chip was used to read out the signal from silicon microstrips, integrating and amplifying it. The studies presented in this paper cover the optimization of its electronic configuration to achieve the lower power consumption footprint and the lower operational temperature of the sensors, while maintaining a good condition of the analogue response of the whole chip. The studies have shown that optimizing the operational temperature is possible and can be beneficial when the detector is highly irradiated. Even a single degree drop in silicon temperature can result in a significant reduction in the leakage current. Similar studies are being performed for the future silicon tracker, the Upstream Tracker (UT), which will start operating at LHC in 2021. It is expected that the inner part of the UT detector will suffer radiation damage similar to the most irradiated VELO sensors in Run 2. In the course of analysis we also developed a general approach for the pulse shape reconstruction using an ANN approach. This technique can be reused in case of any type of front-end readout chip.

Detection of mycotoxins, especially aflatoxin M1 (AFM1), in milk is crucial to be able to guarantee food quality and safety. In recent years, biosensors have been emerging as a fast, reliable and low-cost technique for the detection of this toxin. In this work, flexible biosensors were fabricated using dispense-printed electrodes, which were functionalized with single-walled carbon nanotubes (SWCNTs) and subsequently coated with specific antibodies to improve their sensitivity. Next, the immunosensor was tested for the detection of AFM1 in buffer solution and a spiked milk sample using a chronoamperometric technique. Results showed that the working range of the sensors was 0.01 µg/L at minimum and 1 µg/L at maximum in both buffer and spiked milk. The lower limit of detection of the SWCNT-functionalized sensor was 0.02 µg/L, which indicates an improved sensitivity compared to the sensors reported so far. The sensitivity and detection range were in accordance with the limitation values imposed by regulations on milk and its products. Therefore, considering the low fabrication cost, the ease of operation, and the rapid read-out, the use of this sensor could contribute to safeguarding consumers’ health.

The first experimental results from a new transmissive diagnostic instrument for synchrotron X-ray beamlines are presented. The instrument utilizes a single-crystal chemical-vapour-deposition diamond plate as the detector material, with graphitic wires embedded within the bulk diamond acting as electrodes. The resulting instrument is an all-carbon transmissive X-ray imaging detector. Within the instrument's transmissive aperture there is no surface metallization that could absorb X-rays, and no surface structures that could be damaged by exposure to synchrotron X-ray beams. The graphitic electrodes are fabricated in situ within the bulk diamond using a laser-writing technique. Two separate arrays of parallel graphitic wires are fabricated, running parallel to the diamond surface and perpendicular to each other, at two different depths within the diamond. One array of wires has a modulated bias voltage applied; the perpendicular array is a series of readout electrodes. X-rays passing through the detector generate charge carriers within the bulk diamond through photoionization, and these charge carriers travel to the nearest readout electrode under the influence of the modulated electrical bias. Each of the crossing points between perpendicular wires acts as an individual pixel. The simultaneous read-out of all pixels is achieved using a lock-in technique. The parallel wires within each array are separated by 50 µm, determining the pixel pitch. Readout is obtained at 100 Hz, and the resolution of the X-ray beam position measurement is 600 nm for a 180 µm size beam.

Metasurfaces provide opportunities for wavefront control, flat optics, and subwavelength light focusing. We developed an imaging-based nanophotonic method for detecting mid-infrared molecular fingerprints and implemented it for the chemical identification and compositional analysis of surface-bound analytes. Our technique features a two-dimensional pixelated dielectric metasurface with a range of ultrasharp resonances, each tuned to a discrete frequency; this enables molecular absorption signatures to be read out at multiple spectral points, and the resulting information is then translated into a barcode-like spatial absorption map for imaging. The signatures of biological, polymer, and pesticide molecules can be detected with high sensitivity, covering applications such as biosensing and environmental monitoring. Our chemically specific technique can resolve absorption fingerprints without the need for spectrometry, frequency scanning, or moving mechanical parts, thereby paving the way toward sensitive and versatile miniaturized mid-infrared spectroscopy devices.

AbstractWe present a nanospectroscopic device platform allowing simple and spatially resolved thermoelectric detection of molecular fingerprints of soft materials. Our technique makes use of a locally generated thermal gradient converted into a thermoelectric photocurrent that is read out in the underlying device. The thermal gradient is generated by an illuminated atomic force microscope tip that localizes power absorption onto the sample surface. The detection principle is illustrated using a concept device that contains a nanostructured strip of polymethyl methacrylate (PMMA) defined by electron beam lithography. The platform’s capabilities are demonstrated through a comparison between the spectrum obtained by on-chip thermoelectric nanospectroscopy with a nano-FTIR spectrum recorded by scattering-type scanning near-field optical microscopy at the same position. The subwavelength spatial resolution is demonstrated by a spectral line scan across the edge of the PMMA layer.

The multiwavelength anomalous dispersion (MAD) method of protein structure determination is becoming a routine technique in protein crystallography. The increased number of wavelength-tuneable synchrotron beamlines capable of performing challenging MAD experiments, coupled with the widespread availability of charge-coupled device (CCD) based X-ray detectors with fast read-out times have brought MAD structure determination to a new exciting level. Ultrafast MAD data collection is now possible and, with the widespread use of selenium in the form of selenomethionine for phase determination, the method is growing in popularity. Recent developments in crystallographic software are complementing the above advances, paving the way for rapid protein structure determination. An overview of a typical MAD experiment is described, with emphasis on the rates and quality of data acquisition now achievable at third-generation synchrotron sources.

To increase read-out speed, sensitivity or specificity, an often applied strategy in fluorescence-based biomolecular spectroscopy and imaging is to simultaneously record two or more of the fluorescence parameters: intensity, lifetime, polarization or wavelength. This review highlights how additional, to-date largely unexploited, information can be extracted by monitoring long-lived, photo-induced transient states of organic dyes and their dynamics. Two major approaches are presented, where the transient state information is obtained either from fluorescence fluctuation analysis or by recording the time-averaged fluorescence response to a time-modulated excitation. The two approaches combine the detection sensitivity of the fluorescence signal with the environmental sensitivity of the long-lived transient states. For both techniques, proof-of-principle experiments are reviewed, and advantages, limitations and possible applications for biomolecular cellular biology studies are discussed.

The 2m-grating radius, extreme grazing incidence (1.5°) Schwob-Fraenkel spectrograph was developed at the Racah Institute of Physics (under CEA contract) more than 10 years ago. The first results (using photographic plates) on the TFR tokamak permitted the indentification of the spectrum of highly ionised Mo ions in the 5-50 Å spectral region /1/. Subsequently, the system was modified by J.L. Schwob into a duochromator, using two channeltron electron multipliers independently movable along the Rowland circle. It was thus possible to obtain radial profiles of the emissivities of the strongest lines of the H-and He-like isoelectronic sequences of light impurities in the 18-42 Å spectral range /2/. Recently, the duochromator has been converted into a multichannel spectrometer by equipping it with a microchannelplate (MCP) detector again movable along the Rowland circle. The detector consists of a MgF2coated, funneled MCP, associated with a phosphor screen image intensifier and coupled by a flexible fiber optic conduit to a 1024 element photodiode array (controlled and read-out by a commercially available PAR-1461 EGG Princeton Applied Research optical multichannel analyser system). The first of this type of detector was developed at Princeton for the PLT and TFTR tokamaks and was described by Schwob et al /3/. An identical system has been installed on TFR, using a 20 /μm entrance slit and a 600 groove mm−1Jobin-Yvon holographic grating. This instrument has been routinely used during the last year of TFR operation to monitor spectra of both intrinsic impurities (C, 0, Cr, Fe, and Ni, with traces of Mn, Cl, and S) and purposely injected impurity elements in the 10-330 Å spectral range. The spectrometer has been used in both the spectrographic and the polychromator modes. In the former mode, spectra of highly-ionized, unstudied, heavy elements (injected either by the laser blow-off technique or as gaseous elements) have been obtained /4,5/. In the latter utilization (in which selected individual pixels are read-out as function of time) line radiance evolutions of several different Fe ions have been simultaneously obtained on a single discharge. This has allowed the impurity transport to be modelled /6/ even though the system was not absolutely calibrated, since different ionization degrees have different time evolutions.

Restriction site Associated DNA Sequencing (RAD-Seq) is a technique characterized by the sequencing of specific loci along the genome that is widely employed in the field of evolutionary biology since it allows to exploit variants (mainly Single Nucleotide Polymorphism—SNPs) information from entire populations at a reduced cost. Common RAD dedicated tools, such as STACKS or IPyRAD, are based on all-vs-all read alignments, which require consequent time and computing resources. We present an original method, DiscoSnp-RAD, that avoids this pitfall since variants are detected by exploiting specific parts of the assembly graph built from the reads, hence preventing all-vs-all read alignments. We tested the implementation on simulated datasets of increasing size, up to 1,000 samples, and on real RAD-Seq data from 259 specimens of Chiastocheta flies, morphologically assigned to seven species. All individuals were successfully assigned to their species using both STRUCTURE and Maximum Likelihood phylogenetic reconstruction. Moreover, identified variants succeeded to reveal a within-species genetic structure linked to the geographic distribution. Furthermore, our results show that DiscoSnp-RAD is significantly faster than state-of-the-art tools. The overall results show that DiscoSnp-RAD is suitable to identify variants from RAD-Seq data, it does not require time-consuming parameterization steps and it stands out from other tools due to its completely different principle, making it substantially faster, in particular on large datasets.

The Pencil Beam Scanning (PBS) technique in proton therapy uses fast magnets to scan the tumor volume rapidly. Changing the proton energy allows changing to layers in the third dimension, hence scanning the same volume several times. The PBS approach permits adapting the speed and/or current to modulate the delivered dose. We built a simple prototype that measures the dose distribution in a single step. The active detection material consists of a single layer of scintillating fibers (i.e., 1D) with an active length of 100 mm, a width of 18.25 mm, and an insignificant space (20 μm) between them. A commercial CMOS-based camera detects the scintillation light. Short exposure times allow running the camera at high frame rates, thus, monitoring the beam motion. A simple image processing method extracts the dose information from each fiber of the array. The prototype would allow scaling the concept to multiple layers read out by the same camera, such that the costs do not scale with the dimensions of the fiber array. Presented here are the characteristics of the prototype, studied under two modalities: spatial resolution, linearity, and energy dependence, characterized at the Center for Proton Therapy (Paul Scherrer Institute); the dose rate response, measured at an electron accelerator (Swiss Federal Institute of Metrology).

The Sun Coronal Ejection Tracker (SunCET) is an extreme ultraviolet imager and spectrograph instrument concept for tracking coronal mass ejections through the region where they experience the majority of their acceleration: the difficult-to-observe middle corona. It contains a wide field of view (0–4 R⊙) imager and a 1 Å spectral-resolution-irradiance spectrograph spanning 170–340 Å. It leverages new detector technology to read out different areas of the detector with different integration times, resulting in what we call “simultaneous high dynamic range”, as opposed to the traditional high dynamic range camera technique of subsequent full-frame images that are then combined in post-processing. This allows us to image the bright solar disk with short integration time, the middle corona with a long integration time, and the spectra with their own, independent integration time. Thus, SunCET does not require the use of an opaque or filtered occulter. SunCET is also compact – ~15 × 15 × 10 cm in volume – making it an ideal instrument for a CubeSat or a small, complementary addition to a larger mission. Indeed, SunCET is presently in a NASA-funded, competitive Phase A as a CubeSat and has also been proposed to NASA as an instrument onboard a 184 kg Mission of Opportunity.

The accurate knowledge of the neutron-induced fission cross-sections of actinides and other isotopes involved in the nuclear fuel cycle are essential for the design of advanced nuclear systems. These experimental data can also provide feedback for the adjustment of nuclear model parameters used in the evaluation process, resulting in further developments of nuclear fission models. In the present work, the 240Pu(n,f) cross-section was measured at CERN's n_TOF facility over a wide range of neutron energies, from a few meV to several MeV, using the time-of-flight technique and a set-up based on MicroMegas detectors. This measurement was the first experiment to be performed in n_TOF's new experimental area (EAR-2), which offers a significantly higher neutron flux compared to the existing experimental area. Preliminary results as well as the experimental procedure, including a brief description of the facility, the sample mounting, the read-out process and the data handling and analysis, are presented.

Abstract Abstract 3921 Background. Osteolytic disease is a major complication of multiple myeloma (MM) that may lead to devastating skeletal-related events (SREs). 70% of patients have osteolytic lesions at diagnosis, and up to 90% develop lytic lesions during the course of their disease. Furthermore, almost 15% of patients present with diffuse osteopenia at diagnosis. Various imaging techniques have been performed for the diagnosis and follow-up of bone disease in MM, however, conventional radiography (CR) remains the gold standard. CR has several limitations. One is that CR reveals lytic disease when more than 30% of the trabecular bone has been lost. Thus, some patients may have a suboptimal assessment of generalized osteopenia. Another limitation is that CR cannot be used for the assessment of response to therapy because the lytic bone lesions seldom show evidence of healing. On the other hand new vertebral fractures do not always indicate disease progression and may occur due to ongoing bone loss or reduction of tumor mass that supports the bony cortex. Other limitations include lack of accurate visualization of some areas, observer dependency, lengthy period on the examination table, and poor tolerance by patients with severe pain and extended lytic disease. The EOS System is a new 2D and 3D imaging system for musculo-skeletal physiology and pathology assessment with low radiation dose and standing position. We hypothesized that EOS would not be inferior to CR in terms of routine imaging and diagnosis of bone lytic lesions of patients with MM, but would improve on the quality of life during the procedure of the imaging for the frail patients with MM. Methods and Materials. Fifty six consecutive patients with symptomatic MM (at diagnostic and at first relapse) were included in this prospective study. Each patient provided informed consent. All patients underwent an EOS® examination (frontal and lateral views from skull to knees) and radiographs (axial skeleton: skull, spine, pelvis, femurs, humeri, ribs, as per International Myeloma Working Group guidelines) the same day, prior to start any treatment. Each imaging modality was read in random order by 2 reviewers independently for the detection of bone lesions (osteolytic lesions, vertebral collapses). Whole-body MRI was performed in case of disagreement between the 2 imaging modalities. Radiation dose and technical comfort were also assessed. The length of time of either exam was measured and the patients had to fill in a quality of life questionnaire aimed at comparing the roughness of the 2 techniques. Our study received prior approval from our Ethics Committee. Results. The median age was 62 (range, 32–90), gender ratio was 30 male / 26 female. CR and EOS® diagnosed 467 and 451 bone lytic lesions, respectively. There was no significant difference between the 2 imaging techniques, as 445 out of 473 bone lesions were detected by both the EOS®system and the CR. The median length of time to perform the CR exam was 6 to 8 times longer than EOS® technique. The average radiation dose with the EOS®system was 7.8 times less than with CR. The majority of the patients found the EOS®system examination to be more comfortable than the multiple radiographic incidences. The main limitation to EOS® technique was in patients with high BMI greater than 30, in whom CR remains the most sensitive technique. Furthermore, EOS® system was not able to differentiate old versus novel lytic lesions, as expected with standard radiographs. Finally, EOS presented with the same difficulty to count the number of lytic lesions per patient, as for the CR technique. Conclusion. EOS® system is a new low-dose radiation device which allows a quicker scan of the whole body. In this preliminary study performed in patients with MM, this technique allowed detection of bone lesions with better comfort for the patients as compared to CR. This is of paramount importance in patients with MM that often presented with altered health status and bone pain that hampered the ability to perform CR with optimal conditions. Disclosures: Facon: Celgene: Consultancy, Honoraria; Janssen-Cilag: Consultancy, Honoraria.

Background: Attention Deficit Hyperactivity Disorder (ADHD) is one of the most common mental disorders that manifest among children. Despite the fact that the teacher’s role is essential in the assessment and management of pupils with ADHD and the recommendation of participation of teachers for the success and efficiency of diagnoses and treatment, the vast majority of teachers have neither understanding nor knowledge of ADHD. Objectives: The current study explores the kindergarten and elementary school teachers’ knowledge regarding early detection and management of ADHD. Methods: Cross sectional survey using stratified random sampling technique was carried out in governmental and private elementary and Kindergarten schools in four cities of Qassim region, Saudi Arabia. Results: The study included 1095 teachers, 711 (59.3%) did not get information about ADHD during undergraduate studies. Teachers’ overall ADHD knowledge mean was 21.7+5.5 out of 38 marks. Teachers with high qualification degree and kindergarten specialty scored 56.4% and 60.2% respectively. Teachers who attended conferences related to ADHD scored 64.5%, while teachers who read about ADHD or have been ever asked to diagnose/teach an ADHD student had 59.4% and 62.1% respectively. The level of ADHD knowledge perception showed that 76% of teachers were knowledgeable in relation to overall perception about ADHD. Conclusion: Teachers who have higher qualifications or training in identifying ADHD children scored higher in our study.

Abstract Numerous recurrently mutated genes have been described in the last years for acute myeloid leukemia (AML). Nevertheless, only a few of them (NPM1, CEBPA, FLT3), and usually individually analyzed, have been systematically incorporated into clinical laboratories to stratify prognosis and guide risk-adapted therapy. So far, technology made impossible their simultaneous study, however, development of high throughput techniques as "next generation sequencing" (NGS) allows a parallel assay. Recently, our group designed a NGS panel (custom panel) interrogating the complete coding sequence of 87 genes potentially involved in AML. Interactome analysis revealed that 13 genes were highly implicated in AML pathogenesis. Although NGS has been broadly utilized in the investigational scope, its application to the routine diagnosis is still far from become a standard. The aim of this study is to validate the clinical applicability to routine laboratories of a hotspot NGS panel that includes these 13 genes and other potentially actionable targets. We included 62 "de novo" AML patients in which we tested the Ion Ampliseq AML community panel (Life Technologies) (IAAC panel) in the Ion PGM/Proton platforms. This panel includes hotspots of ASXL1 (exon 12), BRAF (V600E), CBL (exons 8-9), FLT3 (codons 676, 830-850), IDH1 (exon 4), IDH2 (exon 4), JAK 2 (exon 14), KIT (exons 8, 10, 11 and 17), KRAS (exons 2-4), NRAS (exons 2-4), PTPN11 (exons 3,7,8,13), RUNX1 (exons 3-8) y WT1 (exons 7 and 9), and the entire coding sequence of CEBPA, DNMT3A, GATA, TET2 and TP53. The design does not include the FLT3-ITD region. This panel requires only a total of 40 ng of DNA, far less than the custom panel (250 ng), which is very convenient in a clinical laboratory, where the input sample can be limited. The hotspot NGS panel detected 153 variants in 62 patients (2.47 mutations/patient). Mean read depth and uniformity were 1580 and 94.89%, respectively. The IAAC panel detected 25 NPM1 mutations, 20 TET2 and DNMT3A mutations, 14 in CEBPA, 12 in TP53, 11 in RUNX1, 8 in FLT3, 7 in IDH2, 6 in ASXL1, GATA2 and NRAS, 5 in PTPN11 and KRAS, 3 in WT1, 2 in CBL and 1 mutation in IDH1, KIT, and BRAF. Only 4 patients (6%) remained wild-type for these genes after the analysis with the IAAC panel. These samples had previously been analyzed by conventional molecular biology techniques (CMBT) for FLT3-D835, NPM1-T288, DNMT3A-R882 and CEBPA. The IAAC panel found 100% of these previously known mutations plus 5 extra mutations that were negative by CMBT. These mutations were reconfirmed by Sanger sequencing. Therefore, the IAAC panel detects more mutations than CMBT. To further assess the suitability of the hotspot panel, we selected a subset of 25 patients that had been also analyzed by the custom panel (Sure Desing Tool (Agilent) and an Illumina platform). This allowed us to analyze the complete coding sequence of the genes included in the IAAC panel and thus look for mutations outside the hotspots. A total of 53 variants were found with the custom panel. Fifty of these variants (95%) were also detected with the IAAC panel. The remaining 3 variants (5%) were located outside the hotspot regions. Additionally, the IAAC panel detected 13 variants in the overlapping regions that were not found with the custom panel. This could be explained because when focusing on recurrent regions of particular genes, it is possible to increase the mean read depth and therefore reach higher sensibility, which can be achieved with the IAAC panel but not with the custom panel. In conclusion, the Ion Ampliseq AML community panel detects mutations currently analyzed in most of clinical laboratories with validated prognostic relevance (NPM1 and CEBPA) in one assay, with low sample input requirement and with 100% sensibility compared with CMBT. In addition, this panel is able to find out alterations in these genes that are lost by CMBT. Moreover, other mutations with probable diagnostic or prognostic value and/or potential therapeutic targets are also studied and identified with high sensibility. Only a few changes are excluded of the covered regions. Therefore, IAAC panel is useful for routine diagnosis; however, detection of FLT3 internal tandem duplications is not possible, which limits its clinical utility. Disclosures No relevant conflicts of interest to declare.

BackgroundAttention deficit hyperactivity disorder (ADHD) is one of the most common mental disorders that manifest among children. Despite the fact that the teacher's role is essential in the assessment and management of pupils with ADHD and the recommendation of participation of teachers for the success and efficiency of diagnoses and treatment, the vast majority of teachers have neither understanding nor knowledge of ADHD.ObjectivesThe current study explores the kindergarten and elementary school teachers’ knowledge regarding early detection and management of ADHD.MethodsCross sectional survey using stratified random sampling technique was carried out in governmental and private elementary and kindergarten schools in four cities of Qassim region, Saudi Arabia.ResultsThe study included 1095 teachers, 711 (59.3%) did not get information about ADHD during undergraduate studies. Teachers’ overall ADHD knowledge mean was 21.7 + 5.5 out of 38 marks. Teachers with high qualification degree and kindergarten specialty scored 56.4% and 60.2%, respectively. Teachers who attended conferences related to ADHD scored 64.5%, while teachers who read about ADHD or have been ever asked to diagnose/teach an ADHD student had 59.4% and 62.1%, respectively. The sources for those who got their information through reading were statistically significant in overall knowledge, general knowledge and treatment dimensions. As scientific studies and books were the highest with percentage of 24.6% and 23.3%, respectively, 4% and 3.6%, respectively, 3% and 3%, respectively. The level of ADHD knowledge perception showed that 76% of teachers were knowledgeable in relation to overall ADHD perception.ConclusionTeachers who have higher qualifications or training in identifying ADHD children scored higher in our study.Disclosure of interestThe authors have not supplied their declaration of competing interest.

<b><i>Objective:</i></b> Based on the hypothesis that neonatal uterine bleedings (NUB), occurring mostly in the first week after birth, could represent a pathogenetic mechanism for early-onset endometriosis, this systematic review (SR) was undertaken to evaluate the prevalence and screening strategies used to assess and quantify NUB. <b><i>Design:</i></b> Both a SR and a sample literature search in PubMed and Embase were conducted to gather information on NUB prevalence and screening techniques. This was performed by an information specialist. Only full-text articles regarding the assessment of NUB in neonates in the first 2 weeks after birth were included. No limit on language or publication data was used. <b><i>Materials and Methods:</i></b> The SR was registered in PROSPERO (CRD42019138121). Data was first assessed for eligibility on title and abstract by 2 blinded review authors. Any disagreements were discussed with a third reviewer if necessary. Subsequently, full-text articles were read and assessed for quality using the Cochrane Collaboration Handbook. <b><i>Results:</i></b> Out of 1,988 articles in the systematic search, 10 relevant articles were selected, of which 8 were identified through the systematic search and 2 were found through other sources. The sample search of 4,445 articles did not bring up relevant articles. Results were not comparable due to the heterogeneity of screening techniques, although data showed consensus. The prevalence of visible bleeding ranged from 3.3 to 53.8% and the prevalence of occult bleeding from 25.4 to 96.7%. The occurrence was the highest between the 3rd and 7th day postpartum (PP) and the bleeding lasted for 3–4 days on average. Various screening techniques for detecting NUB were found in the literature, including the use of hemoglobin detection devices (such as Hemastix) in the vaginal vestibulum, comparison of diapers with stains of known volume, colposcopy, and ultrasonography. <b><i>Conclusion:</i></b> The reported prevalence of NUB varies considerably, with a consistent occurrence between the 3rd and the 7th day PP. Literature to assess NUB is dated. The techniques are poorly described and heterogeneous. Future research should focus on prospective cohort studies in order to attempt to correlate NUB cases to (early-onset) endometriosis.

Purpose of research. Nowadays optical character recognition systems have a high level of dependence on the specific type of marking that is to be recognized, and therefore, the creation of a universal solution is an important and difficult task. The paper considers the issue of creating a system for recognizing symbolic information that can be used at various stages of production to automate processes in control systems, in particular, to analyze the labeling of circuit breakers.Methods. Binarization, filtering, and boundary detection are digital image processing techniques. Line search method, baseline search method, word splitting algorithms, image enhancement methods by segmentation, damaged characters recognition method, an algorithm for increasing the final recognition quality are character recognition methods.Results. The analysis of algorithms used for preprocessing and subsequent recognition of images containing marking of circuit breakers is carried out. The mathematical model of image processing for subsequent recognition has been created. We have described methods used to define marking symbols. Illustrative examples of the operation of the algorithms on which the system is built are given. The obtained solution was tested. The ways of system development are described here, they can lead to improved results, for particular use cases.Conclusion. It is proposed a solution that recognizes symbolic information on the labeling of circuit breakers, which can be the basis for the development and description of systems serving for the automation of production, by transferring information read from the product during the production process. This system, by its example, describes the components of character recognition systems, and for direct use, it needs to be refined in accordance with the technical requirements and the specifics of the conditions in which it will be used.

Abstract Abstract 2487 Genome wide analyses of regulatory elements have identified successive waves of coherent transcriptional programs guiding through and controlling cell cycle progression. Each of these modules induces its successor and, at the same time, inactivates the key kinases active in the antecedent phase of the cycle. In an initial signaling module, the RAF-MEK-ERK pathway is central for phosphorylation of the tumor suppressor Rb (retinoblastoma), release of transcription factors of the E2F family and subsequent cell cycle entry. CDK2 is the driving force and guardian of the consecutive phase where DNA replication occurs. After having initiated CDK2 activity, ERK is inactive and non-inducible (Pouyssegur 2003). The pRB/CDK2 node thus is a bistable switch that converts mitogenic information into an irreversible cellular commitment to duplicate, and uncouples the cell's growth and division program from environmental or internal control. Comparing primary leukemic blasts with healthy CD34+ hematopoietic progenitor cells, we here show that simultaneous rather than serial activation of the canonical ERK and CDK2 pathways is a recurrent aberrant signaling motif indicative for malignant transformation. This phenomenon was observed in leukemic blasts and also in cancer cells of epithelial and mesenchymal provenience, but not in healthy or regenerating tissues using flow cytometry and confocal microscopy techniques and phospho-epitope specific antibodies. To further visualize this “forbidden ERK/CDK2 signal combination”, we developed a biosensor comprising both, the optimal peptide substrates for ERK and CDK kinases attached to a fluorescein tag and a membrane-penetrating nona-arginine (9R) moiety. This compound enables detection of simultaneous ERK and CDK2 activity on single cell level by means of electrophoretic mobility shift. Site directed mutagenesis of the two phospho sites confirmed mutual exclusive substrate recognition thus ascertaining the specificity of the read out system. In healthy CD34+ cells, single but never dual phosphorylation of the biosensor was detected even after stimulation with PMA/Ionomycin, the latter establishing a non-physiological, proliferative compartment. These findings confirm the serial and mutually exclusive piloting of ERK and CDK2 signaling pathways in non-transformed cells. In stark contrast, dual phosphorylation of the biosensor peptide was observed upon analysis of leukemic blasts. Thus, simultaneous and therefore aberrant recruitment of ERK and Rb/E2F/CDK2 cell cycle elements in transformed but not healthy cells discloses leukemia. Our data do not add to the increasing complexity of genetic insults associated with malignant transformation. Rather, we demonstrate that violations of the orderly recruitment of common signaling pathways are indicative for a malignant phenotype which can be detected on the basis of aberrant spatiotemporal organization of essential cell signaling nodes. Our findings do not only anticipate a novel way to diagnose malignancy. Based on the structural information provided by our template peptide, we further envisage a new class of cancer therapeutics where a nontoxic prodrug is converted into a tumoricidal substance exclusively in malignant cells after simultaneous double-phosphorylation by aberrantly active kinases. Disclosures: No relevant conflicts of interest to declare.

ObjectiveThe mechanisms underlying the association between diabetes and inner ear dysfunction are not known yet. The aim of the present study is to evaluate the impact of obesity/insulin resistance on inner ear fluid homeostasis in vivo, and to investigate whether the organ of Corti could be a target tissue for insulin signaling using auditory House Ear Institute-Organ of Corti 1 (HEI-OC1) cells as an in vitro model.MethodsHigh fat diet (HFD) fed C57BL/6J mice were used as a model to study the impact of insulin resistance on the inner ear. In one study, 12 C57BL/6J mice were fed either control diet or HFD and the size of the inner ear endolymphatic fluid compartment (EFC) was measured after 30 days using MRI and gadolinium contrast as a read-out. In another study, the size of the inner ear EFC was evaluated in eight C57BL/6J mice both before and after HFD feeding, with the same techniques. HEI-OC1 auditory cells were used as a model to investigate insulin signaling in organ of Corti cells.ResultsHFD feeding induced an expansion of the EFC in C57BL/6J mice, a hallmark of inner ear dysfunction. Insulin also induced phosphorylation of protein kinase B (PKB/Akt) at Ser473, in a PI3-kinase-dependent manner. The phosphorylation of PKB was inhibited by isoproterenol and IBMX, a general phosphodiesterase (PDE) inhibitor. PDE1B, PDE4D and the insulin-sensitive PDE3B were found expressed and catalytically active in HEI-OC1 cells. Insulin decreased and AICAR, an activator of AMP-activated protein kinase, increased the phosphorylation at the inhibitory Ser79 of acetyl-CoA carboxylase, the rate-limiting enzyme in de novo lipogenesis. Furthermore, the activity of hormone-sensitive lipase, the rate-limiting enzyme in lipolysis, was detected in HEI-OC1 cells.ConclusionsThe organ of Corti could be a target tissue for insulin action, and inner ear insulin resistance might contribute to the association between diabetes and inner ear dysfunction.

ABL1 Kinase Domain (ABL1-KD) mutations are a common resistance mechanism to tyrosine-kinase inhibitors (TKIs) in Chronic Myeloid Leukemia (CML) and Philadelphia Positive Acute Lymphoblastic Leukemia (ALL). Different ABL1-KD mutations induce different degrees of resistance to different TKIs. The early detection of these resistant mutations helps to adjust patient's treatment. Here we present an Ultra-Deep Sequencing approach to detect and quantify acquired ABL1-KD mutations in genomic DNA (gDNA), aiming to define a robust test to detect such alterations in TKIs exposed Philadelphia-Positive Leukemia Patients with a resolution below 1E-4. Firstly, we defined an ABL1 specific next-generation sequencing (NGS) panel designed to cover all coding regions of ABL1 exons 4-10. The 9 amplicons were designed to cover full exons where possible to detect co-occurring mutations (Figure 1A). A panel was then applied to 3 biological replicates of 3 Healthy control donors (9 NGS data points each with 220ng of gDNA). The average coverage per amplicon in all samples was at least 500,000x. The NGS data was then analyzed applying the NGS-MRD algorithm described elsewhere (Onecha, E et al. Haematologica 2019) to 25 known ABL1-KD hotspots. After applying our error correcting algorithm, we obtained an average of 135,000 (22,000-503,000) refined reads for the 25 hotspots. The limit of detection (LOD) was calculated for every position in the DNA as the mean noise (Variant Read Frequency; VRF) per position in the controls ± 3SD (standard deviation); the limit of quantification (LOQ) being defined as mean ± 10SD. For all the hotspots analyzed, the LOD was below 1E-4 and the LOQ below 3E-4 (Figure 1B), except for p.F311L (c.931T&gt;C; LOD=2.7E-3). The high level of noise in this position, constant in the different control samples sequenced in different sequencing runs, is most likely related to the high number of homopolymers in the region. Ten Philadelphia-Positive Leukemia patients were then screened after TKI treatment (8 CML and 2 ALL). The median BCR-ABL1 defined by quantitative PCR (ratio BCR-ABL1 vs ABL1) in these follow-up samples was 0.6% (0.034% - 95%). All patients were screened in triplicates (220ng gDNA each) and the data-points ± 1SD from the mean were considered outliers (NGS false positives) and excluded from further analysis. Five patients presented a signal above the LOD for p.T315I (c.944C&gt;T). This position is covered by 2 different amplicons in our panel. By bioinformatically demultiplexing the signal, the detection of those five mutations in both amplicons was confirmed (Amp_4; LOD=3E-5, Amp_5; LOD=4E-5). Moreover, aiming to validate this new approach, we applied to paired RNA samples an in-house BCR-ABL1/ABL1 nested PCR + NGS approach designed to quantify those alterations in cDNA. This approach confirmed the presence of 4 out of 5 gDNA detected mutations, with a Pearson correlation of 0.92 (Pval&lt;0.001) (Figure 1C). The only mutation not confirmed by nested PCR presented the lowest BCR-ABL1 quantification (0.034%). Here we show an Ultra-Deep NGS based test which allows the early detection of TKI resistant emerging clones in genomic DNA samples with a resolution of 1E-4. Despite the facts that in Phi-positive leukemia patients' other techniques such as the nested PCR are available, for most of heme- dyscrasias it is not easy to detect acquired mutations below 1% VRF. Our test can reduce this limit by at least 2 logarithms. The clinical impact of this approach is illustrated by the two LLA patients included, both under dasatinib therapy when the p.T315I mutations were detected. Those 2 patients were changed to ponatinib, reducing BCR-ABL1 levels. An extension of the cohort and the validation of our test at clinical level will be presented at the meeting. Figure Disclosures Heredia: Altum sequencing: Current Employment. Carrillo:Altum sequencing: Current Employment. Rufian:Altum sequencing: Current Employment. Wang:Hosea Precision Medical Technology Co., Ltd: Current Employment. Ribera:Pfizer, Amgen: Research Funding; Pfizer, Amgen, Ariad, Novartis: Consultancy, Speakers Bureau. Martinez-López:Janssen, BMS, Sanofi, Novartis, Incyte, F. Hoffmann-La Roche and Amgen: Honoraria, Other: Advisory boards; Hosea and Altum: Membership on an entity's Board of Directors or advisory committees; Janssen, Novartis, BMS, Incyte: Consultancy.

Abstract Introduction Immunoglobulin (Ig)/ T-cell receptor (TCR) gene rearrangements are the most widely used clonal marker to detect residual leukemic cells in patients with Acute Lymphoblastic Leukemia (ALL). Ig/TCR gene rearrangements based molecular minimum residual disease (MRD) monitoring has become one of the most powerful prognostic indicators for patients with ALL. Although the standard method of real-time quantitative PCR (RQ-PCR) provides very good sensitivity in MRD measurement, the workflow is very complicated and time-consuming, requiring expert technique and much work for operation, which limits the number of patients to be examined for MRD monitoring and the number of testable markers per patient. Material and methods To reveal clonal architecture and detect appropriate MRD marker, we designed capture probes covering the coding and recognition signal sequences of V, D, J genes of the Ig/TCR loci. We performed high-throughput target-capture sequencing in 208 pediatric cases with BCP-ALL and 35 pediatric cases with T-cell ALL, including 20 relapsed cases and 14 MRD marker negative cases. Extracted DNA samples were enriched with about 420 capture probes (Agilent Technology) and sequenced by HiSeq2500 platform (Illumina) in order to obtain enough sequence coverage (> 500 mean depth). Sequenced data were analyzed with Ig/TCR recombination analysis tool Vidjil (Giraud et al, 2014) and V(D)J recombination clones were listed according to a number of detected read for each clone. Results Total 2379 clonal Ig/TCR gene rearrangements (median 9 per patient, range 0-82) were detected by capture sequencing among 236 (97%) cases. A clonal IGH sequence with V(D)J recombination was identified in 91% of BCP-ALL cases, followed by TRG (68 %), IGK (67 %), TRA+D (66%), TRD (59 %), TRB (49%), and IGL (15 %), respectively. On the other hand, clonal TRG V(D)J recombination was detected in 74% of T-ALL cases, followed by TRB (69%), TRD (57%), IGH (26%), and TRA+D (6%), respectively. About half of BCP-ALL cases were identified two independent IGH rearrangements. These frequencies agree with previous reports obtained by PCR based experiments. In the cases in this study with well-characterized clonal Ig/TCR gene rearrangements by PCR and Sanger sequencing, our capture sequencing was able to detect all rearrangements used in MRD measurements. Although 8 BCP-ALL cases in this study were marker-negative in standard PCR-MRD diagnostics, clonal Ig/TCR gene rearrangements were identified for 5 out of 8 cases by capture sequencing. Some of the hidden clonal rearrangements showed specific and good quantitative amplification by RQ-PCR and can be used as sensitive PCR-MRD targets. On the other hand, all the MRD marker negative 6 T-ALL cases were not detected clonal Ig/TCR gene rearrangements. Finally, we compared the clonal architecture based on Ig/TCR gene rearrangements between diagnosis and relapse in relapsed B-ALL patients. Changes in the clonal architecture were associated with remission duration. In very early relapse cases, detected Ig/TCR rearrangements and their proportion at relapse are very similar to those at diagnosis. In early to late relapse cases, some major Ig/TCR gene rearrangements were lost at relapse and other minor rearrangements expanded at relapse. Most of the identified Ig/TCR gene rearrangements were different between at diagnosis and at relapse in a case relapsed after more than 10 years. Loss of rearrangements were commonly seen in TRA, TRB, and IgL, while most of the IgK and TRD rearrangements were steady during disease course. Conclusion Introducing target capture sequencing enables to high throughput sample preparation and automated data analysis. Capture sequencing is a useful method for comprehensive detection of Ig/TCR gene rearrangements and contributes to better understanding clonal architecture and detecting appropriate MRD markers in ALL patients. Disclosures No relevant conflicts of interest to declare.

Abstract In Philadelphia-positive (Ph+) Acute Lymphoblastic Leukemia (ALL) patients (pts), resistance to tyrosine kinase inhibitors (TKIs) is frequently associated with the selection of one or more mutations in the BCR-ABL1 kinase domain (KD). The swift emergence of mutant clones as early as during induction therapy supports the hypothesis that, at least in some cases, mutations may already be present at diagnosis. Next Generaton Sequencing (NGS) has been proposed as an alternative to Sanger sequencing (seq) for BCR-ABL1 KD mutation screening because of its greater sensitivity and accuracy, but no studies have so far evaluated its prospective use in Ph+ ALL. Between 2015 and 2018, we have used NGS in parallel to Sanger seq to analyze a consecutive series of 126 Ph+ ALL pts who were newly diagnosed (n=39) or who had relapsed/refractory disease (n=87) on TKI therapy. In 22 cases, both bone marrow and peripheral blood were analyzed and compared. NGS of ≈400bp amplicons generated by nested RT-PCR was performed on a Roche GS Junior (until April 2017) or on an Illumina MiSeq (from May 2017 on). Read alignment and variant calling (with a lower limit set to 3%) were done with the AmpSuite software (SmartSeq srl). When multiple mutations mapped within the same sequence reads, assessment of cis vs trans configuration was done correcting for the probability of PCR recombination. Three out of 39 (7.7%) de novo Ph+ ALL pts had low burden point mutations detectable by NGS: one had a V289A (variant frequency, 3.4%); one had a D276G (4.0%) and a F359V (3.5%); one had an E255K mutation (3.3%). The first pt was enrolled in the GIMEMA LAL1811 study of frontline ponatinib; the second and the third pts were enrolled in the GIMEMA D-ALBA study of frontline sequential treatment with dasatinib and blinatumomab. All pts achieved molecular remission, consistently with the mutations being sensitive to the TKIs received. The 35INS insertion/truncation mutant was detected in 27 (69%) pts, who all have so far achieved molecular remission. This is in line with the report by O'Hare et al (Blood 2011) suggesting that the 35INS variant is kinase-inactive and does not contribute to TKI resistance. For this reason, the 35INS was excluded from subsequent analyses. Relapsed/refractory pts positive for mutations by Sanger seq were 57 (65%); those positive for mutations by NGS were 69 (79%). Fifty-six out of 87 (49%) pts had >1 mutation (up to 13) detected by NGS. NGS identified low burden mutations (i.e., mutations present in a proportion of transcripts between 3 and 20%) in 12 pts who were negative for mutations by Sanger seq. Most importantly, NGS provided a more accurate picture of BCR-ABL1 mutations status in 40 (46%) pts who turned out to have one or more low burden mutations in addition to the dominant mutation(s) detectable by Sanger seq. In all cases, each low burden mutation detected by NGS could be recognized as poorly sensitive either to the TKI the pt was receiving at the time of testing, or to the previous TKI. The clonal nature of NGS-based analysis further proved its utility i) in 4 pts where Sanger seq had shown 2 base substitutions in the same codon so that the actual amino-acid change(s) were impossible to infer (a ponatinib-resistant pt with a T315M mutation, 2 dasatinib-resistant pts with various combinations of F317I, F317C and/or F1317L, a dasatinib-resistant pt with 2 different nucleotide substitutions both leading to the V299L), and ii) in 48/56 pts who had ≥2 mutations whose clonal configuration could not be resolved. Twenty-eight out of these 48 pts were found to carry one or more (up to 3) compound mutants. Compound mutants were more common in pts who had failed ≥2 lines of therapy, whereas polyclonality was more common in pts who had failed first line therapy. The most frequent compound mutants were T315I+E255K and T315I+E255V. Interestingly, the latter was associated with poor or no response to ponatinib. Our results in a relatively large series of Ph+ ALL pts suggest that an NGS-based approach provides a more accurate characterization of the complexity of BCR-ABL1 KD mutation status, including compound mutants some of whom may be poorly sensitive even to ponatinib. Mutations may already be detected at the time of diagnosis. It remains to be assessed whether more sensitive techniques like digital PCR may identify a greater number of pts with pre-therapy mutations and whether the detection of pre-therapy mutations may be used to guide 1st-line treatment selection. Disclosures Soverini: Incyte Biosciences: Consultancy; Bristol Myers Squibb: Consultancy; Novartis: Consultancy. Pagano:Gilead: Speakers Bureau; Basilea: Speakers Bureau; Merck: Speakers Bureau; Janssen: Speakers Bureau; Pfizer: Speakers Bureau. Abruzzese:Ariad: Consultancy; BMS: Consultancy; Novartis: Consultancy; Pfizer: Consultancy. Martinelli:Roche: Consultancy; Celgene: Consultancy, Speakers Bureau; Jazz Pharmaceuticals: Consultancy; Pfizer: Consultancy, Speakers Bureau; Novartis: Speakers Bureau; Abbvie: Consultancy; Janssen: Consultancy; Ariad/Incyte: Consultancy; Amgen: Consultancy. Cavo:Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Bristol-Myers Squibb: Honoraria, Membership on an entity's Board of Directors or advisory committees; Janssen: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; AbbVie: Honoraria, Membership on an entity's Board of Directors or advisory committees; GlaxoSmithKline: Honoraria, Membership on an entity's Board of Directors or advisory committees; Amgen: Honoraria, Membership on an entity's Board of Directors or advisory committees; Takeda: Honoraria, Membership on an entity's Board of Directors or advisory committees; Adaptive Biotechnologies: Honoraria, Membership on an entity's Board of Directors or advisory committees.

Abstract Introduction: Dyskeratosis Congenita (DKC) is caused by defective telomere maintenance, mostly due to mutations impacting on the functional activity of telomerase. Classical DKC is characterized by mucocutaneous features and bone marrow failure diagnosed during childhood. In contrast, the cryptic form of DKC often first manifests itself in adulthood and presents clinically with less characteristic manifestations. Correct identification of DKC patients is of utmost importance because of significant implications for the affected patients (treatment, toxicities) and her/his family (donor selection, genetic counselling). Accelerated shortening of telomere length (TL) in peripheral blood leucocytes (PBL) represents the functional read-out and consequently, clinically useful screening parameter of altered telomere function. Consequently, in cases without distinct genetics or clinical presentation, significantly shortened TL is frequently used as a disease-defining marker. TL in PBL of pediatric DKC patients typically is below the first percentile (<1%). However, the difference between the normal range and critically short telomeres decreases with age, thus complicating the diagnosis of DKC in adult patients. Consequently, the TL threshold below 10% of age-adjusted healthy controls is frequently recommended in adults. Currently, fluorescence in situ hybridization and flow cytometry (Flow-FISH), especially of the lymphocyte subpopulation, or monochrome multiplex quantitative polymerase chain reaction (MM-qPCR) are preferred techniques for TL assessment. In our study, we compared MM-qPCR to Flow-FISH regarding their ability to properly identify DKC patients. Methods and Patients: 105 patients enrolled in the Aachen Telomeropathy Registry from 2015 to 2017 were analysed. Patient inclusion criteria were based on the clinical suspicion of cryptic DKC by the treating physician. Patient`s median age was 47.5 [range 2-88] years. Initial diagnoses were aplastic anemia/paroxysmal nocturnal hemoglobinuria (n=50), unexplained cytopenia (n=20), genetically confirmed DKC mutation/polymorphism or screening of family members (n=14) as well as other disorders (e.g. lung fibrosis, n=21). All patients underwent customized amplicon-based next-generation sequencing analysis (TERT, TERC, DKC1, NOP10, NHP2, USB1, CTC1, RTEL1, TIN2, TCAB1) on a Miseq sequencer. 16/105 patients were identified with a known DKC-related mutation (TERC n=6, TERT n=6, RTEL1 n=3, DKC1 n=1). Median age of the DKC patient cohort was 44.5 (range 21-77) years. Age adaption was performed by using healthy control and TL assessment of PB granulocytes and lymphocytes was carried out in parallel by Flow-FISH and MM-qPCR with all experimental samples assayed in triplicates Results: We observed that TL measured by MM-qPCR correlates with lymphocyte and granulocyte TL measured by flow-FISH (R²=0.33 and R²=0.25, respectively; both p≤0.001). Assessment of lymphocyte TL by Flow-FISH led to values below the 1% or 10% threshold of normal individuals in 24% (n=26) or 49% (n=51) of the patients, respectively leading to a detection rate of 75% (n=12) or 88% (n=14) of the known DKC patients. In comparison, TL assessment by MM-qPCR resulted in 29% (n=31) or 74% (n=78) of the patients below the 1% or 10% percentile, respectively leading to a detection rate of known DKC cases of 68% (n=11) or 94% (n=15). Thus, the sensitivity of both techniques basically did not differ. When we used the 1% cut off, the specificity of flow-FISH (84%, n=75/89) and MM-qPCR (78%, n=69/89) again did not differ significantly (p=0.74). However, using the 10% threshold, specificity of MM-qPCR (29%, n=26/89) was significantly lower compared to flow-FISH (58%, n=52/89, p=0.02). Finally, the rate of false positive patients using flow-FISH was 13% (n=14) and 19% (n=20) for MM-qPCR for the 1% threshold. Using the 10% threshold, we observed a significantly higher rate of false positive patients using MM-qPCR with 60% (n=63/105) compared to Flow-FISH with 35% (n=37/105, p=0.04). Conclusions: Depending on the applied threshold, the use of MM-qPCR as a screening and confirmatory method for telomere maintenance disorders was inferior in specificity and showed a significant higher rate of false positive results compared to flow-FISH. Our results suggest the use of flow FISH as the gold standard method of screening especially in adults with suspected cryptic DKC. Disclosures Kirschner: Basilea Pharmaceutica: Other: travel support; BMS: Consultancy; Roche: Consultancy; Bayer: Consultancy. Wilop:Medizinwelten-Services GmbH: Honoraria; Amgen: Consultancy; Celgene: Consultancy, Honoraria, Other: Travel grant; Bristol-Myers Squibb: Honoraria. Brümmendorf:Novartis: Consultancy, Research Funding; Merck: Consultancy; Janssen: Consultancy; Takeda: Consultancy; Pfizer: Consultancy, Research Funding. Beier:Celgene: Other: travel support; Gilead: Other: travel support.

Background Acute leukemia is the most common malignancy of childhood. Most of pediatric acute leukemias are derived from B-cell precursor, and about 15% are from T-cell origin and lead to T-cell acute lymphoblastic leukemia (T-ALL). Despite recent progress in treatment, the prognosis of T-ALL patients is worse, and ~30% of cases relapse (Pui CH, Seminars in hematology 2009). Despite the need for targeting molecules to treat resistant and high risk pediatric T-ALLs, in vitro drug testing may not be predictive of efficacy in vivo due to the complexity of the signalling network and to the cell heterogeneity within the individual's sample. High-resolution analysis of signalling profile at single-cell level may represent an innovative tool in the comprehension of signalling network and its role in the response to drugs (Bodenmiller B et al, Science signaling 2010). Currently mass cytometry (CyTOF) technique allows the measurement of more than 30 parameter per single cell (Bendall SC et al, Science 2011), rendering this approach an ideal method to investigate the complex biology of T-ALL (Girardi T et al, Blood. 2017) and to assay drug candidates for cellular targeting. Methods Using single-cell mass cytometry we developed panel of 39 metal-labeled monoclonal antibodies (moAbs) identifying T-ALL blasts and non-malignant T-cells. Using phospho-specific moAbs and moAbs targeting proliferation and apoptosis we detected signal transduction upon in vitro treatment of 17 diagnostic and 5 relapse T-ALL samples with IL-7 (Jak/STAT5 pathway activator) , BEZ-235 (inhibitor of PI3K and mTOR), and Pervanadate (inhibitor of tyrosine phosphatases). We have used sample barcoding by anti-CD45 antibodies to unify sample preparation and acquisition for all treatment conditions and we resolved the cells of interest by manual gating. We evaluated both, the individual change sin p-kinases and the overall changes using dimensionality reduction approach. Results T-ALL cells showed constitutive activation of various signaling pathways as well as proliferation markers compared to residual bone marrow T-cells and T-cells isolated from healthy donors. Up-regulated activity of PI3K-mTOR pathway (p4E-BP1, pAkt, pS6), proliferation rate (pRb, Ki-67), MAPK pathway (p-p38, pErk1/2), translation (pCREB) in T-ALL cells was detected, whereas lower levels of anti-apoptotic protein Bcl2 in T-ALL cells were detected. Interleukin 7 (IL-7) activates three main signalling pathways such as STAT5, PI3K/Akt/mTOR and MEK/Erk, leading to the promotion of leukemia cell viability, cell cycle progression and growth. Thus we interrogated T-ALLs in their ability to respond to IL -7 in vitro . We used an hierarchical clustering analysis (with Euclidean distance metrics and an average linkage) and we were able to divide T-ALL samples in IL-7 responder (6 out of 17) and IL-7 non responder (11 out of 17). Of note, no significant differences in IL7Ra (CD127) expression was observed between the two groups. Interestingly IL-7 responders had higher levels of pRb and Ki-67 proliferation markers as compared to both IL-7 non-responders and non-malignant T-cells. Moreover IL-7 non-response correlate with poor response in vivo to prednisone and higher level of minimal residual disease (MRD) at day15 of remission induction treatment. Finally T-ALL cells were treated ex vivo with PI3K/AkT/mTOR dual inhibitor BEZ-235. Of the 17 T-ALL patients tested, 9 responded to BEZ-235 but not to IL-7 , by contrast 4 did not respond to BEZ-235 being IL-7 responders, two patients responded both to IL-7 activation and BEZ-235 inhibition. One single patient did not respond neither to IL-7 nor to BEZ-235 Conclusions In summary we characterized pediatric T-ALL samples demonstrating the feasibility of CyTOF-based single-cell profiling of signal transduction pathways in this setting. We detected constitutively active pathways in T-ALL blasts as compared to residual non-malignant T-cells. Importantly we identified by functional read outs distinct clusters of IL-7 and BEZ-235 T-ALL responders patients, supporting the notion of a mutual exclusivity between JAK-STAT (or Ras) pathway genomic alterations and PI3K-AKT pathway alterations (Liu et al. Nat Genet. 2017). Our observation can contribute to the better understanding of the complex signalling network governing T-ALL behaviour and response to therapy. Disclosures No relevant conflicts of interest to declare.

<p><strong>Abstract.</strong> Flow maps, where counts of people or things are shown travelling between origin-destination pairs with paths or arrows, are difficult to draw well, mostly because they rapidly get cluttered as more and more paths are added. Researchers in cartographic and information visualization fields have sought to mitigate these problems in various ways, including cluster detection and edge bundling (Buchin, Speckmann, &amp; Verbeek, 2011; Guo, 2009), force-directed path placement (Holten &amp; van Wijk, 2009; Jenny et al., 2017), use of network topological space instead of planimetric space (Xiao &amp; Chun, 2009), interactive spatial filtering of flows (Vrotsou, Fuchs, Andrienko, &amp; Andrienko, 2017), and by changing the drawing medium to accommodate flows that arc through 3D space (Raposo, 2017; Yang et al., 2019; Zhang, Zhang, Li, &amp; Li, 2018). Other geovisualization techniques recently developed involve and use of matrixes or grids to store and visualize flows; Wood, Dykes, &amp; Slingsby (2010) use spatial tessellation and a set of small multiples of the same tessellation representing origin and destination spaces, respectively, creating a sort of travel heat map when tiles are colored as a choropleth. A comparable approach is taken by Yang et al. (2017) in a system that combines an abstract matrix of flow records with inflow and outflow maps.</p><p>This early-stage project builds on previous matrix-based flow visualization methods while leveraging the power of Discrete Global Grid Systems (DGGS), which present a naturally Earth-oriented hierarchical tessellation upon which a matrix of origins and destinations can be stored and visualized. DGGS (Sahr, White, &amp; Kimerling, 2003; Raposo, Robinson, &amp; Brown, in press) partition the round Earth into nested tiles in a manner similar to quad-trees, and have the desirable properties of covering the whole spherical globe in equal-area (or near equal-area) tiles, and of being hierarchical to an arbitrary number of levels. The hierarchical quality allows flows in a DGGS to be naturally bundled by origin and destination points as a function of grid size; greater or lesser detail (i.e., more or less bundling) in the overall pattern of flows is effected by descending or ascending DGGS levels. As DGGS are becoming increasingly popular for geospatial data computation and collection in movement analysis (Brodsky, 2018), the present work provides a natural bridge to the visualization of such data when it pertains to movement phenomena.</p><p>The present research will develop a suite of flow visualization techniques upon an existing open-source DGGS rendering platform (Raposo, Robinson, &amp; Brown, in press): 1) grid facet coloring in a choropleth scheme to indicate magnitudes of flow into or out of a particular facet; 2) vertical-space cubic spline flow arcs, and 3) tabular representation of the flow data. All three visualization techniques will function across multiple spatial resolutions, corresponding to each of the hierarchical levels of our chosen DGGS, so that users can interact both with each of the three visualizations of the data as well as the spatial granularity to which the data are displayed. All views will be interactive and linked.</p><p>We use Dutton’s (1999) quaternary triangular mesh (QTM), being a DGGS based on recursively subdividing the faces of an octahedron over the globe (Figure 1). Our method begins by finding, at many nested QTM levels, the facets of the QTM with which each origin and destination point intersects; in practice, we use up to 16 levels, as the triangles at that level in the QTM are small enough to address typical individual buildings. Importantly, point-in-polygon intersection is calculated geodetically, and not in 2D projected space, since failing to do this can cause topological errors (Raposo, Robinson, Brown, in press). At each QTM facet and at each level, we build two lists: one of all the other facets at that level that contain an origin point for a flow that arrives at this facet, and another of those that contain a destination point for flows originating in this facet. These lists are stored to each facet as polygon attributes. A flow path curving through vertical space via a cubic spline is also derived for each origin-destination pair and stored as a renderable 3D solid.</p><p>The QTM is then plotted in our software on a virtual globe using NASA’s World Wind application programming interface. As users select any one QTM facet, its attributes are read to find all the other facets either contributing or receiving flows to or from the selected facet; these are then colored according to a choropleth scheme to visualize magnitudes of flow (Figure 2). Coloration is controlled for various descriptive statistics on the flows (e.g., total, mean, maximum, etc.). Linear paths curving up and around the globe (Figure 3) will also be available to be toggled on or off, giving users multiple redundant symbolizations of travel paths.</p>

Internet of Things (IoT) is regarded as a next-generation wave of Information Technology (IT) after the widespread emergence of the Internet and mobile communication technologies. IoT supports information exchange and networked interaction of appliances, vehicles and other objects, making sensing and actuation possible in a low-cost and smart manner. On the other hand, cyber-physical systems (CPS) are described as the engineered systems which are built upon the tight integration of the cyber entities (e.g., computation, communication, and control) and the physical things (natural and man-made systems governed by the laws of physics). The IoT and CPS are not isolated technologies. Rather it can be said that IoT is the base or enabling technology for CPS and CPS is considered as the grownup development of IoT, completing the IoT notion and vision. Both are merged into closed-loop, providing mechanisms for conceptualizing, and realizing all aspects of the networked composed systems that are monitored and controlled by computing algorithms and are tightly coupled among users and the Internet. That is, the hardware and the software entities are intertwined, and they typically function on different time and location-based scales. In fact, the linking between the cyber and the physical world is enabled by IoT (through sensors and actuators). CPS that includes traditional embedded and control systems are supposed to be transformed by the evolving and innovative methodologies and engineering of IoT. Several applications areas of IoT and CPS are smart building, smart transport, automated vehicles, smart cities, smart grid, smart manufacturing, smart agriculture, smart healthcare, smart supply chain and logistics, etc. Though CPS and IoT have significant overlaps, they differ in terms of engineering aspects. Engineering IoT systems revolves around the uniquely identifiable and internet-connected devices and embedded systems; whereas engineering CPS requires a strong emphasis on the relationship between computation aspects (complex software) and the physical entities (hardware). Engineering CPS is challenging because there is no defined and fixed boundary and relationship between the cyber and physical worlds. In CPS, diverse constituent parts are composed and collaborated together to create unified systems with global behaviour. These systems need to be ensured in terms of dependability, safety, security, efficiency, and adherence to real‐time constraints. Hence, designing CPS requires knowledge of multidisciplinary areas such as sensing technologies, distributed systems, pervasive and ubiquitous computing, real-time computing, computer networking, control theory, signal processing, embedded systems, etc. CPS, along with the continuous evolving IoT, has posed several challenges. For example, the enormous amount of data collected from the physical things makes it difficult for Big Data management and analytics that includes data normalization, data aggregation, data mining, pattern extraction and information visualization. Similarly, the future IoT and CPS need standardized abstraction and architecture that will allow modular designing and engineering of IoT and CPS in global and synergetic applications. Another challenging concern of IoT and CPS is the security and reliability of the components and systems. Although IoT and CPS have attracted the attention of the research communities and several ideas and solutions are proposed, there are still huge possibilities for innovative propositions to make IoT and CPS vision successful. The major challenges and research scopes include system design and implementation, computing and communication, system architecture and integration, application-based implementations, fault tolerance, designing efficient algorithms and protocols, availability and reliability, security and privacy, energy-efficiency and sustainability, etc. It is our great privilege to present Volume 21, Issue 3 of Scalable Computing: Practice and Experience. We had received 30 research papers and out of which 14 papers are selected for publication. The objective of this special issue is to explore and report recent advances and disseminate state-of-the-art research related to IoT, CPS and the enabling and associated technologies. The special issue will present new dimensions of research to researchers and industry professionals with regard to IoT and CPS. Vivek Kumar Prasad and Madhuri D Bhavsar in the paper titled "Monitoring and Prediction of SLA for IoT based Cloud described the mechanisms for monitoring by using the concept of reinforcement learning and prediction of the cloud resources, which forms the critical parts of cloud expertise in support of controlling and evolution of the IT resources and has been implemented using LSTM. The proper utilization of the resources will generate revenues to the provider and also increases the trust factor of the provider of cloud services. For experimental analysis, four parameters have been used i.e. CPU utilization, disk read/write throughput and memory utilization. Kasture et al. in the paper titled "Comparative Study of Speaker Recognition Techniques in IoT Devices for Text Independent Negative Recognition" compared the performance of features which are used in state of art speaker recognition models and analyse variants of Mel frequency cepstrum coefficients (MFCC) predominantly used in feature extraction which can be further incorporated and used in various smart devices. Mahesh Kumar Singh and Om Prakash Rishi in the paper titled "Event Driven Recommendation System for E-Commerce using Knowledge based Collaborative Filtering Technique" proposed a novel system that uses a knowledge base generated from knowledge graph to identify the domain knowledge of users, items, and relationships among these, knowledge graph is a labelled multidimensional directed graph that represents the relationship among the users and the items. The proposed approach uses about 100 percent of users' participation in the form of activities during navigation of the web site. Thus, the system expects under the users' interest that is beneficial for both seller and buyer. The proposed system is compared with baseline methods in area of recommendation system using three parameters: precision, recall and NDGA through online and offline evaluation studies with user data and it is observed that proposed system is better as compared to other baseline systems. Benbrahim et al. in the paper titled "Deep Convolutional Neural Network with TensorFlow and Keras to Classify Skin Cancer" proposed a novel classification model to classify skin tumours in images using Deep Learning methodology and the proposed system was tested on HAM10000 dataset comprising of 10,015 dermatoscopic images and the results observed that the proposed system is accurate in order of 94.06\% in validation set and 93.93\% in the test set. Devi B et al. in the paper titled "Deadlock Free Resource Management Technique for IoT-Based Post Disaster Recovery Systems" proposed a new class of techniques that do not perform stringent testing before allocating the resources but still ensure that the system is deadlock-free and the overhead is also minimal. The proposed technique suggests reserving a portion of the resources to ensure no deadlock would occur. The correctness of the technique is proved in the form of theorems. The average turnaround time is approximately 18\% lower for the proposed technique over Banker's algorithm and also an optimal overhead of O(m). Deep et al. in the paper titled "Access Management of User and Cyber-Physical Device in DBAAS According to Indian IT Laws Using Blockchain" proposed a novel blockchain solution to track the activities of employees managing cloud. Employee authentication and authorization are managed through the blockchain server. User authentication related data is stored in blockchain. The proposed work assists cloud companies to have better control over their employee's activities, thus help in preventing insider attack on User and Cyber-Physical Devices. Sumit Kumar and Jaspreet Singh in paper titled "Internet of Vehicles (IoV) over VANETS: Smart and Secure Communication using IoT" highlighted a detailed description of Internet of Vehicles (IoV) with current applications, architectures, communication technologies, routing protocols and different issues. The researchers also elaborated research challenges and trade-off between security and privacy in area of IoV. Deore et al. in the paper titled "A New Approach for Navigation and Traffic Signs Indication Using Map Integrated Augmented Reality for Self-Driving Cars" proposed a new approach to supplement the technology used in self-driving cards for perception. The proposed approach uses Augmented Reality to create and augment artificial objects of navigational signs and traffic signals based on vehicles location to reality. This approach help navigate the vehicle even if the road infrastructure does not have very good sign indications and marking. The approach was tested locally by creating a local navigational system and a smartphone based augmented reality app. The approach performed better than the conventional method as the objects were clearer in the frame which made it each for the object detection to detect them. Bhardwaj et al. in the paper titled "A Framework to Systematically Analyse the Trustworthiness of Nodes for Securing IoV Interactions" performed literature on IoV and Trust and proposed a Hybrid Trust model that seperates the malicious and trusted nodes to secure the interaction of vehicle in IoV. To test the model, simulation was conducted on varied threshold values. And results observed that PDR of trusted node is 0.63 which is higher as compared to PDR of malicious node which is 0.15. And on the basis of PDR, number of available hops and Trust Dynamics the malicious nodes are identified and discarded. Saniya Zahoor and Roohie Naaz Mir in the paper titled "A Parallelization Based Data Management Framework for Pervasive IoT Applications" highlighted the recent studies and related information in data management for pervasive IoT applications having limited resources. The paper also proposes a parallelization-based data management framework for resource-constrained pervasive applications of IoT. The comparison of the proposed framework is done with the sequential approach through simulations and empirical data analysis. The results show an improvement in energy, processing, and storage requirements for the processing of data on the IoT device in the proposed framework as compared to the sequential approach. Patel et al. in the paper titled "Performance Analysis of Video ON-Demand and Live Video Streaming Using Cloud Based Services" presented a review of video analysis over the LVS \& VoDS video application. The researchers compared different messaging brokers which helps to deliver each frame in a distributed pipeline to analyze the impact on two message brokers for video analysis to achieve LVS & VoS using AWS elemental services. In addition, the researchers also analysed the Kafka configuration parameter for reliability on full-service-mode. Saniya Zahoor and Roohie Naaz Mir in the paper titled "Design and Modeling of Resource-Constrained IoT Based Body Area Networks" presented the design and modeling of a resource-constrained BAN System and also discussed the various scenarios of BAN in context of resource constraints. The Researchers also proposed an Advanced Edge Clustering (AEC) approach to manage the resources such as energy, storage, and processing of BAN devices while performing real-time data capture of critical health parameters and detection of abnormal patterns. The comparison of the AEC approach is done with the Stable Election Protocol (SEP) through simulations and empirical data analysis. The results show an improvement in energy, processing time and storage requirements for the processing of data on BAN devices in AEC as compared to SEP. Neelam Saleem Khan and Mohammad Ahsan Chishti in the paper titled "Security Challenges in Fog and IoT, Blockchain Technology and Cell Tree Solutions: A Review" outlined major authentication issues in IoT, map their existing solutions and further tabulate Fog and IoT security loopholes. Furthermore, this paper presents Blockchain, a decentralized distributed technology as one of the solutions for authentication issues in IoT. In addition, the researchers discussed the strength of Blockchain technology, work done in this field, its adoption in COVID-19 fight and tabulate various challenges in Blockchain technology. The researchers also proposed Cell Tree architecture as another solution to address some of the security issues in IoT, outlined its advantages over Blockchain technology and tabulated some future course to stir some attempts in this area. Bhadwal et al. in the paper titled "A Machine Translation System from Hindi to Sanskrit Language Using Rule Based Approach" proposed a rule-based machine translation system to bridge the language barrier between Hindi and Sanskrit Language by converting any test in Hindi to Sanskrit. The results are produced in the form of two confusion matrices wherein a total of 50 random sentences and 100 tokens (Hindi words or phrases) were taken for system evaluation. The semantic evaluation of 100 tokens produce an accuracy of 94\% while the pragmatic analysis of 50 sentences produce an accuracy of around 86\%. Hence, the proposed system can be used to understand the whole translation process and can further be employed as a tool for learning as well as teaching. Further, this application can be embedded in local communication based assisting Internet of Things (IoT) devices like Alexa or Google Assistant. Anshu Kumar Dwivedi and A.K. Sharma in the paper titled "NEEF: A Novel Energy Efficient Fuzzy Logic Based Clustering Protocol for Wireless Sensor Network" proposed a a deterministic novel energy efficient fuzzy logic-based clustering protocol (NEEF) which considers primary and secondary factors in fuzzy logic system while selecting cluster heads. After selection of cluster heads, non-cluster head nodes use fuzzy logic for prudent selection of their cluster head for cluster formation. NEEF is simulated and compared with two recent state of the art protocols, namely SCHFTL and DFCR under two scenarios. Simulation results unveil better performance by balancing the load and improvement in terms of stability period, packets forwarded to the base station, improved average energy and extended lifetime.

Methods based on X-ray area detectors are presented for acquiring and analysing single-crystal X-ray diffraction data from quasicrystals and approximants. These methods are complementary to high-resolution diffractometry and fully exploit the possibilities of modern two-dimensional X-ray detector systems (imaging plates and charge coupled devices) such as the on-line read-out capability and the possibility to collect large volumes of reciprocal space in a quantitative and rapid way. In contrast to a classical point detector, area detectors allow the measurement of a very large number of Bragg reflections in a reasonable time scale, as is shown with a large unit-cell periodic approximant in the Al-Co-Ni alloy system. For large approximants exhibiting strong pseudosymmetry, direct methods for structure solution usually fail. Recently implemented Patterson deconvolution techniques are shown to succeed. The extension of techniques for area detectors – beyond the standard application of collecting integrated Bragg intensities – to the quantitative acquisition of continuous and diffuse scattering data is presented with particular emphasis on quasicrystals. One of the key techniques is the quantitative reconstruction of undistorted layers and volumes in reciprocal space. This is crucial for developing and comparing possible models for structural disorder in quasicrystals. Examples for decagonal Al-Co-Ni and icosahedral Al-Mn-Pd are given in respect to possibilities and limitations of X-ray area detectors.

Shifted excitation Raman difference spectroscopy (SERDS) can provide effective, chemically specific information on fluorescent samples. However, the restricted ability for fast alternating detection (usually < 10 Hz) of spectra excited at two shifted laser wavelengths can limit its effectiveness when rapidly varying emission backgrounds are present. This paper presents a novel charge-shifting lock-in approach permitting fast SERDS operation (exemplarily demonstrated at 1000 Hz) using a specialized dual-wavelength diode laser (emitting at 829.40 nm and 828.85 nm) and a custom-built charge-coupled device (CCD) enabling charge retention and shifting back and forth on the CCD chip. For six selected mineral samples (moved irregularly during spectral acquisition), results demonstrate superior reproducibility of the fast charge-shifting read-out over the conventional read-out (operated at 5.4 Hz). Partial least squares discriminant analysis revealed improved classification performance of charge-shifting (four latent variables, sensitivity: 99%, specificity: 94%) versus conventional read-out (six latent variables, sensitivity: 90%, specificity: 92%). The charge-shifting concept was also successfully translated to sub-surface analysis using spatially offset Raman spectroscopy (SORS). Charge-shifting SERDS-SORS spectra recorded from a polytetrafluoroethylene layer, concealed behind a 0.25 mm thick, opaque, heterogeneous layer, matched reference spectra much more closely and exhibited a signal-to-background-noise (S/NB) ratio two times higher than that achieved with conventional CCD read-out SERDS-SORS. The novel approach overcomes fundamental limitations of conventional CCDs. In conjunction with the inherent capability of the charge-shifting lock-in technique to suppress rapidly varying ambient light interference demonstrated by us earlier it is expected to be particularly beneficial with heterogeneous fluorescent samples in field applications.

New free electron lasers, such as SLAC’s LCLS-II, will provide unique scientific imaging opportunities. In order to fully utilize these facilities, we need to develop detectors with shallow entrance windows that will enable detection of soft x-rays from 250 eV to 1.5 KeV. Achieving adequately shallow entrance windows is challenging because the high temperature anneal needed to activate the dopant also drives the dopant profile deeper, growing the region that is insensitive to soft x-rays. A new microwave annealing technology provides an efficient way to achieve shallow entrance windows in fully depleted high-resistivity silicon sensors. The microwave anneal technique can activate dopants at low substrate temperature, with minimal dopant diffusion, and can be used to fabricate both n-type and p-type entrance windows. SRP and SIMS measurements were used to verify dopant activation with negligible dopant diffusion. We then applied the microwave anneal process to a planar sensor wafer, using the new process to create the backside diode contact. Electrical test of the resulting sensors shows good reverse bias characteristics. The sensors have been bump-bonded to a read-out ASIC and used successfully to measure an Fe-55 x-ray spectrum. Process and device simulations were performed to characterize the quantum efficiency of the entrance window for soft x-rays. This technique is useful for other sensor applications requiring a shallow entrance window, including detectors for UV photons, low energy ions and low energy electrons.