Non-Ideal Program-Time Conservation in Charge Trap Flash for Deep Learning

Training deep neural networks (DNNs) is computationally intensive but arrays of non-volatile memories like Charge Trap Flash (CTF) can accelerate DNN operations using in-memory computing. Specifically, the Resistive Processing Unit (RPU) architecture uses the voltage-threshold program by stochastic encoded pulse trains and analog memory features to accelerate vector-vector outer product and weight update for the gradient descent algorithms. Although CTF, offering high precision, has been regarded as an excellent choice for implementing RPU, the accumulation of charge due to the applied stochastic pulse trains is ultimately of critical significance in determining the final weight update. In this paper, we report the non-ideal program-time conservation in CTF through pulsing input measurements. We experimentally measure the effect of pulse width and pulse gap, keeping the total ON-time of the input pulse train constant, and report three non-idealities: (1) Cumulative V_T shift reduces when total ON-time is fragmented into a larger number of shorter pulses, (2) Cumulative V_T shift drops abruptly for pulse widths < 2 {\mu}s, (3) Cumulative V_T shift depends on the gap between consecutive pulses and the V_T shift reduction gets recovered for smaller gaps. We present an explanation based on a transient tunneling field enhancement due to blocking oxide trap-charge dynamics to explain these non-idealities. Identifying and modeling the responsible mechanisms and predicting their system-level effects during learning is critical. This non-ideal accumulation is expected to affect algorithms and architectures relying on devices for implementing mathematically equivalent functions for in-memory computing-based acceleration

Autofluorescence of Model Polyethylene Terephthalate Nanoplastics for Cell Interaction Studies

This work contributes to fill one of the gaps regarding nanoplastic interactions with biological systems by producing polyethylene terephthalate (PET) model nanoplastics, similar to those found in the marine environment, by means of a fast top-down approach based on mechanical fragmentation. Their size distribution and morphology were characterized by laser diffraction and atomic force microscopy (AFM). Their autofluorescence was studied by spectrofluorimetry and fluorescence imaging, being a key property for the evaluation of their interaction with biota. The emission spectra of label-free nanoplastics were comparable with those of PET nanoplastics labeled with Nile red. Finally, the suitability of label-free nanoplastics for biological studies was assessed by in vitro exposure with Mytilus galloprovincialis hemolymphatic cells in a time interval up to 6 h. The nanoplastic internalization into these cells, known to be provided with phagocytic activity, was assessed by fluorescence microscopy. The obtained results underlined that the autofluorescence of the model PET nanoplastics produced in the laboratory was adequate for biological studies having the potential to overcome the disadvantages commonly associated with several fluorescent dyes, such as the tendency to also stain other organic materials different from plastics, to form aggregates due to intermolecular interactions at high concentrations with a consequent decrease in fluorescence intensity, and to dye desorption from nanoparticles. The results of the autofluorescence study provide an innovative approach for plastic risk assessment

Motility of Mytilus galloprovincialis hemocytes: Sensitivity to paracetamol in vitro exposure

Pharmaceuticals released into the environment (PiEs) represent an environmental problem of growing concern for the health of ecosystems and humans. An increasing number of studies show that PiEs pose a risk to aquatic organisms. The aim of the present work was to contribute to increasing the knowledge of the effects of PiE on marine biota focusing on the effect of paracetamol on the motility of hemocytes in Mytilus galloprovincialis, a bivalve mollusk species widely utilized as bioindicator organism. Hemocytes are the immunocompetent cells of bivalve mollusks. An early and key stage of mollusk immune response is represented by the recruitment and migration of these cells to the site of infection. Therefore, motility is an intrinsic characteristic of these cells. Here, we first characterized the spontaneous cell movement of M. galloprovincialis hemocytes when plated in a TC-treated polystyrene 96-well microplate. Two different cellular morphotypes were distinguished based on their appearance and motility behavior: spread cells and round-star-shaped cells. The two motility morphotypes were characterized by different velocities as well as movement directness, which were significantly lower in round-star-shaped cells with respect to spread cells. The sensitivity of the motility of M. galloprovincialis hemocytes to paracetamol at different concentrations (0.02, 0.2 and 2 mg/L) was investigated in vitro after 1h and 24h exposure. Paracetamol induced alterations in the motility behavior (both velocity and trajectories) of the hemocytes and the effects were cell-type specific. The study of hemocyte movements at the single cell level by cell tracking and velocimetric parameters analysis provides new sensitive tools for assessing the effects of emerging pollutants at the cellular levels in non-target organism

Microporous Collagen Scaffolds for Regeneration of Peripheral Nerve

INTRODUCTION Peripheral nerve injuries often result in painful neuropathies owing to reduction in motor function and sensory perception. When large nerve gaps exist (20mm or longer in humans), sensory nerve autografts are conventionally used to treat neural defects. The main issues related to autografts are shortage of donor nerves, a mismatch of donor nerve size with the recipient site, and occurrences of neuroma formation. Recent advances in nanotechnology and tissue engineering have been found to cover a broad range of applications in regenerative medicine and offer the most effective strategy to repair neural defects. Prior work in this area has shown the utility of collagen-based scaffolds for the regeneration of nerve tissue. This work focuses on the fabrication of collagen scaffolds with two different pore sizes, with the aim of evaluating the effects of pore size on the migration of Schwann cell lines. EXPERIMENTAL METHODS Scaffold fabrication and crosslinking Porous cylindrical scaffolds (diameter=2mm, length=10mm) with aligned channels were fabricated by freeze-drying a 2wt% collagen suspension along a one-dimensional temperature gradient (along the length of the cylindrical scaffold). Scaffolds with two different pore sizes were fabricated by freezing the collagen suspension at two different final freezing temperatures (-20°C and -60°C). The scaffolds were then subjected to dehydrothermal (DHT) cross-linking, followed by a carbodiimide based chemical crosslinking. Qualitative characterization of the pore structure was performed by means of scanning electron microscopy (SEM). Cell culture and cytocompatibility A rat Schwann cell line, RSC96, was expanded in monolayer culture in a 96-well plate. The plate was then incubated at 37°C and 5% CO2 for 24 hours. After 24 hours, sterilized scaffolds were placed vertically to the wells of the 96-well plate and incubated again at 37°C and 5% CO2. At 1, 3, 7, and 10 days, the cell-seeded scaffolds were fixed in 10% formalin and processed for paraffin embedding. Schwann cells were quantified by embedding the cell-seeded scaffolds in paraffin blocks, sectioning, staining them with hematoxylin & eosin stain (H&E stain) and visualizing under a microscope. MTT assay was also performed at 1, 3, 7 and 10 days to evaluate the cell viability. RESULTS AND DISCUSSION SEM demonstrated that both freezing temperature and rate of freezing affect significantly the pore size. As shown in Fig.1, lower temperatures (-60°C) resulted in smaller pore sizes (~85µm), while higher temperatures (-20°C) resulted in much larger pores (~120µm). The longitudinal sections of the samples showed that the pores were in axial orientation disregard of the freezing temperature. MTT assay revealed that cell viability on the two different types of scaffolds increased gradually from first to the tenth day after seeding. Although there was not much difference between the two porous scaffolds on day 1, 3 and 7, on day 10 there was a slight increase in the cell number in the scaffolds with a larger pore size (-20°C). In spite of the different pore dimensions under investigation, the cell migration studies revealed that Schwann cells could migrate through the entire length of both types of scaffolds, by day 7. CONCLUSION Both types of scaffolds were found to support Schwann cell growth and migration, which is the key factor required for the regeneration of nerve tissue. Further studies are proposed regarding the addition of laminin and the evaluation of its effects on the cell growth and migration. REFERENCES 1. W. Daly et al., J. R. Soc. Interface 9:202-221, 2012

Scaffolds for bone regeneration made of hydroxyapatite microspheres in a collagen matrix

Biomimetic scaffolds with a structural and chemical composition similar to native bone tissue may be promising for bone tissue regeneration. In the present work hydroxyapatite mesoporous microspheres (mHA) were incorporated into collagen scaffolds containing an ordered interconnected macroporosity. The mHA were obtained by spray drying of a nano hydroxyapatite slurry prepared by the precipitation technique. X-ray diffraction (XRD) analysis revealed that the microspheres were composed only of hydroxyapatite (HA) phase, and energy-dispersive x-ray spectroscopy (EDS) analysis revealed the Ca/P ratio to be 1.69 which is near the value for pure HA. The obtained microspheres had an average diameter of 6 μm, a specific surface area of 40 m(2)/g as measured by Brunauer-Emmett-Teller (BET) analysis, and Barrett-Joyner-Halenda (BJH) analysis showed a mesoporous structure with an average pore diameter of 16 nm. Collagen/HA-microsphere (Col/mHA) composite scaffolds were prepared by freeze-drying followed by dehydrothermal crosslinking. SEM observations of Col/mHA scaffolds revealed HA microspheres embedded within a porous collagen matrix with a pore size ranging from a few microns up to 200 μm, which was also confirmed by histological staining of sections of paraffin embedded scaffolds. The compressive modulus of the composite scaffold at low and high strain values was 1.7 and 2.8 times, respectively, that of pure collagen scaffolds. Cell proliferation measured by the MTT assay showed more than a 3-fold increase in cell number within the scaffolds after 15 days of culture for both pure collagen scaffolds and Col/mHA composite scaffolds. Attractive properties of this composite scaffold include the potential to load the microspheres for drug delivery and the controllability of the pore structure at various length scales

Apoptotic volume decrease (AVD) in A549 cells exposed to water-soluble fraction of particulate matter (PM10)

Exposure to atmospheric particulate matter (PM) is recognized as a human health risk factor of great concern. The present work aimed to study the cellular mechanisms underlying cytotoxic effects of airborne particulate matter <10 mu m in size (PM10), sampled in an urban background site from January to May 2020, on A549 cells. In particular, the study addressed if PM10 exposure can be a main factor in the induction of the Apoptotic Volume Decrease (AVD), which is one of the first events of apoptosis, and if the generation of intracellular oxidative stress can be involved in the PM10 induction of apoptosis in A549 cells. The cytotoxicity of PM10 samples was measured by MTT test on cells exposed for 24 h to the PM10 aqueous extracts, cell volume changes were monitored by morphometric analysis of the cells, apoptosis appearance was detected by annexin V and the induction of intracellular oxidative stress was evaluated by the ROS sensitive CM-H2DCFDA fluorescent probe. The results showed cytotoxic effects ascribable to apoptotic death in A549 cells exposed for 24 h to aqueous extracts of airborne winter PM10 samples characterized by high PM10 value and organic carbon content. The detected reduced cell viability in winter samples ranged from 55% to 100%. Normotonic cell volume reduction (ranging from about 60% to 30% cell volume decrease) after PM10 exposure was already detectable after the first 30 min clearly indicating the ability of PM10, mainly arising from biomass burning, to induce Apoptotic Volume Decrease (AVD) in A549 cells. AVD was prevented by the pre-treatment with 0.5 mM SITS indicating the activation of Clefflux presumably through the activation of VRAC channels. The exposure of A549 cells to PM10 aqueous extracts was able to induce intracellular oxidative stress detected by using the ROS-sensitive probe CM-H2DCFDA. The PM10induced oxidative stress was statistically significantly correlated with cell viability inhibition and with apoptotic cell shrinkage. It was already evident after 15 min exposure representing one of the first cellular effects caused by PM exposure. This result suggests the role of oxidative stress in the PM10 induction of AVD as one of the first steps in cytotoxicity

Intracellular Antioxidant Activity of Biocompatible Citrate-Capped Palladium Nanozymes

A method for the aqueous synthesis of stable and biocompatible citrate-coated palladium nanoparticles (PdNPs) in the size range comparable to natural enzymes (4&ndash;8 nm) has been developed. The toxicological profile of PdNPs was assessed by different assays on several cell lines demonstrating their safety in vitro also at high particle concentrations. To elucidate their cellular fate upon uptake, the localization of PdNPs was analyzed by Transmission Electron Microscopy (TEM). Moreover, crucial information about their intracellular stability and oxidation state was obtained by Sputtering-Enabled Intracellular X-ray Photoelectron Spectroscopy (SEI-XPS). TEM/XPS results showed significant stability of PdNPs in the cellular environment, an important feature for their biocompatibility and potential for biomedical applications. On the catalytic side, these PdNPs exhibited strong and broad antioxidant activities, being able to mimic the three main antioxidant cellular enzymes, i.e., peroxidase, catalase, and superoxide dismutase. Remarkably, using an experimental model of a human oxidative stress-related disease, we demonstrated the effectiveness of PdNPs as antioxidant nanozymes within the cellular environment, showing that they are able to completely re-establish the physiological Reactive Oxygen Species (ROS) levels in highly compromised intracellular redox conditions