Investigation of Nanoparticles in High Resolution Scanning Electron Microscopy (SEM) and Low Voltage SEM by Digital Image-Analysis

Small particles (Cu, Ag, In, Sn, Au, also MgO and NaCl) were prepared in the diameter range from 1 nm to 100 nm on different conductive substrates by thermal evaporation in high-vacuum or in an inert gas atmosphere. Imaging of the particles was performed in a high resolution scanning electron microscope (HRSEM) that can also be operated at low beam voltages of a few hundred volts. This mode of operation is called low voltage SEM (LVSEM). Scanning electron micrographs were taken at different beam voltages VO (0.5-30 kV). The micrographs were digitally recorded and analyzed with an image processing system operated on-line to the HRSEM. Grey-value line profiles and densitometric quantities of single particles, as well as the contrast between particle and substrate, changed with VO. The results for tin-particles on a bulk carbon substrate are shown. In all cases considered, only positive contrasts, i.e., particles looking brighter than the substrate, were obtained. The main contrast producing mechanism is, therefore, assigned to effects that include the particle\u27s geometrical properties of size, shape and surface. Sn-, In-, and Ag-particles, imaged in the secondary electron (SE) mode showed significantly larger particle diameters, as did images simultaneously recorded with transmitted electrons; however, Au-particles did not show that difference. This effect may be qualitatively explained by SE resulting from decaying plasmons

Mechanical and optical degradation of flexible optical solar reflectors during simulated low earth orbit thermal cycling

Multilayer thin film systems on flexible polymer substrates are used as flexible optical solar reflectors or thermal insulation of satellites and spacecraft. During one year of operation, a satellite in low earth orbit typically encounters 6000 thermal cycles of ±100 °C. Due to the different coefficients of thermal expansion between the individual layers and the substrate it is important to investigate the thermo-mechanical stability of the multilayers as a function of the cyclic heat load. Scanning electron microscopy and focused ion beam cross-sectioning revealed that Inconel-Ag bilayers on fluorinated ethylene propylene (FEP) substrate severely degrade during thermal cycling of ±150 °C in a gaseous N2 atmosphere. After only 100 cycles through thickness cracks and subsurface voids in the Ag layer form as a result of equi-biaxial thermal stresses caused by the large difference in thermal expansion between film and substrate. Transmission Kikuchi Diffraction (TKD) before and after thermal cycling also revealed grain growth and twin widening in the Ag layer. Cracking and void formation are detrimental to application relevant material properties including corrosion protection (Inconel) and reflectivity (Ag). Reflectance measurements revealed that the amount of reflected energy as well as the reflection mode (specular vs. diffuse) significantly change during the first 100 cycles. Saturation of reflection characteristics was observed after 25 cycles, which correlates to a turning point in the evolution of Ag voids. Results of this study indicate that special focus should be directed towards thermal stress control (Δα) and tailoring of the metal-polymer interface to improve resistance of versatile metal-polymer systems against thermal cycling. © 2020 IA

CSF Protein Level of Neurotransmitter Secretion, Synaptic Plasticity, and Autophagy in PD and DLB

BACKGROUND: Molecular pathways associated with α-synuclein proteostasis have been detected in genetic studies and in cell models and include autophagy, ubiquitin-proteasome system, mitochondrial homeostasis, and synaptic plasticity. However, we lack biomarkers that are representative for these pathways in human biofluids. OBJECTIVE: The objective of this study was to evaluate CSF protein profiles of pathways related to α-synuclein proteostasis. METHODS: We assessed CSF protein profiles associated with neurotransmitter secretion, synapse plasticity, and autophagy in 2 monocentric cohorts with α-synucleinopathy (385 PD patients and 67 DLB patients). We included 80 PD patients and 17 DLB patients with variants in the glucocerebrosidase gene to serve as proxy for accelerated α-synuclein pathology with pronounced clinical trajectories. RESULTS: (1) Proteins associated with neurotransmitter secretion, synaptic plasticity, and endolysosomal autophagy were lower in PD and DLB patients compared with healthy controls. (2) These patterns were more pronounced in DLB than in PD patients, accentuated by GBA variant status in both entities. (3) CSF levels of these proteins were positively associated with CSF levels of total α-synuclein, with lower levels of proteostasis proteins related to lower levels of total α-synuclein. (4) These findings could be confirmed longitudinally. PD patients with low CSF profiles of proteostasis proteins showed lower CSF levels of α-synuclein longitudinally compared with PD patients with a normal proteostasis profile. CONCLUSION: CSF proteins associated with neurotransmitter secretion, synaptic plasticity, and endolysosomal autophagy might serve as biomarkers related to α-synuclein proteostasis in PD and DLB

The Carnian Humid Episode of the late Triassic: a review

From 1989 to 1994 a series of papers outlined evidence for a brief episode of climate change from arid to humid, and then back to arid, during the Carnian Stage of the late Triassic Epoch. This time of climate change was compared to marine and terrestrial biotic changes, mainly extinction and then radiation of flora and fauna. Subsequently termed, albeit incorrectly, the Carnian Pluvial Event (CPE) by successive authors, interest in this episode of climatic change has increased steadily, with new evidence being published as well as several challenges to the theory. The exact nature of this humid episode, whether reflecting widespread precipitation or more local effects, as well as its ultimate cause, remains equivocal. Bed-by-bed sampling of the Carnian in the Southern Alps (Dolomites) shows the episode began with a negative carbon isotope excursion that lasted for only part of one ammonoid zone (A. austriacum). However, that the Carnian Humid Episode represents a significantly longer period, both environmentally and biotically, is irrefutable. The evidence is strongest in the European, Middle Eastern, Himalayan, North American and Japanese successions, but not always so clear in South America, Antarctica and Australia. The eruption of the Wrangellia Large Igneous Province and global warming (causing increased evaporation in the Tethyan and Panthalassic oceans) are suggested as causes for the humid episode

Association between CSF alpha-synuclein seeding activity and genetic status in Parkinson’s disease and dementia with Lewy bodies

The clinicopathological heterogeneity in Lewy-body diseases (LBD) highlights the need for pathology-driven biomarkers in-vivo. Misfolded alpha-synuclein (α-Syn) is a lead candidate based on its crucial role in disease pathophysiology. Real-time quaking-induced conversion (RT-QuIC) analysis of CSF has recently shown high sensitivity and specificity for the detection of misfolded α-Syn in patients with Parkinson's disease (PD) and dementia with Lewy bodies (DLB). In this study we performed the CSF RT-QuIC assay in 236 PD and 49 DLB patients enriched for different genetic forms with mutations in GBA, parkin, PINK1, DJ1, and LRRK2. A subgroup of 100 PD patients was also analysed longitudinally. We correlated kinetic seeding parameters of RT-QuIC with genetic status and CSF protein levels of molecular pathways linked to α-Syn proteostasis. Overall, 85% of PD and 86% of DLB patients showed positive RT-QuIC α-Syn seeding activity. Seeding profiles were significantly associated with mutation status across the spectrum of genetic LBD. In PD patients, we detected positive α-Syn seeding in 93% of patients carrying severe GBA mutations, in 78% with LRRK2 mutations, in 59% carrying heterozygous mutations in recessive genes, and in none of those with bi-allelic mutations in recessive genes. Among PD patients, those with severe GBA mutations showed the highest seeding activity based on RT-QuIC kinetic parameters and the highest proportion of samples with 4 out of 4 positive replicates. In DLB patients, 100% with GBA mutations showed positive α-Syn seeding compared to 79% of wildtype DLB. Moreover, we found an association between α-Syn seeding activity and reduced CSF levels of proteins linked to α-Syn proteostasis, specifically lysosome-associated membrane glycoprotein 2 and neurosecretory protein VGF. These findings highlight the value of α-Syn seeding activity as an in-vivo marker of Lewy-body pathology and support its use for patient stratification in clinical trials targeting α-Syn

IL-10 transcription is negatively regulated by BAF180, a component of the SWI/SNF chromatin remodeling enzyme

<p>Abstract</p> <p>Background</p> <p>SWI/SNF chromatin remodeling enzymes play a critical role in the development of T helper lymphocytes, including Th2 cells, and directly program chromatin structure at Th2 cytokine genes. Different versions of SWI/SNF complexes, including BAF and PBAF, have been described based on unique subunit composition. However, the relative role of BAF and PBAF in Th cell function and cytokine expression has not been reported.</p> <p>Results</p> <p>Here we examine the role of the PBAF SWI/SNF complex in Th cell development and gene expression using mice deficient for a PBAF-specific component, BAF180. We find that T cell development in the thymus and lymphoid periphery is largely normal when the BAF180 gene is deleted late in thymic development. However, BAF180-deficient Th2 cells express high levels of the immunoregulatory cytokine IL-10. BAF180 binds directly to regulatory elements in the Il-10 locus but is replaced by BAF250 BAF complexes in the absence of BAF180, resulting in increased histone acetylation and CBP recruitment to the IL-10 locus.</p> <p>Conclusions</p> <p>These results demonstrate that BAF180 is a repressor of IL-10 transcription in Th2 cells and suggest that the differential recruitment of different SWI/SNF subtypes can have direct consequences on chromatin structure and gene transcription.</p

Green Fluorescent Protein (GFP) Color Reporter Gene Visualizes Parvovirus B19 Non-Structural Segment 1 (NS1) Transfected Endothelial Modification

Background: Human Parvovirus B19 (PVB19) has been associated with myocarditis putative due to endothelial infection. Whether PVB19 infects endothelial cells and causes a modification of endothelial function and inflammation and, thus, disturbance of microcirculation has not been elucidated and could not be visualized so far. Methods and Findings: To examine the PVB19-induced endothelial modification, we used green fluorescent protein (GFP) color reporter gene in the non-structural segment 1 (NS1) of PVB19. NS1-GFP-PVB19 or GFP plasmid as control were transfected in an endothelial-like cell line (ECV304). The endothelial surface expression of intercellular-adhesion molecule-1 (CD54/ICAM-1) and extracellular matrix metalloproteinase inducer (EMMPRIN/CD147) were evaluated by flow cytometry after NS-1-GFP or control-GFP transfection. To evaluate platelet adhesion on NS-1 transfected ECs, we performed a dynamic adhesion assay (flow chamber). NS-1 transfection causes endothelial activation and enhanced expression of ICAM-1 (CD54: mean±standard deviation: NS1-GFP vs. control-GFP: 85.3±11.2 vs. 61.6±8.1; P<0.05) and induces endothelial expression of EMMPRIN/CD147 (CD147: mean±SEM: NS1-GFP vs. control-GFP: 114±15.3 vs. 80±0.91; P<0.05) compared to control-GFP transfected cells. Dynamic adhesion assays showed that adhesion of platelets is significantly enhanced on NS1 transfected ECs when compared to control-GFP (P<0.05). The transfection of ECs was verified simultaneously through flow cytometry, immunofluorescence microscopy and polymerase chain reaction (PCR) analysis. Conclusions: GFP color reporter gene shows transfection of ECs and may help to visualize NS1-PVB19 induced endothelial activation and platelet adhesion as well as an enhanced monocyte adhesion directly, providing in vitro evidence of possible microcirculatory dysfunction in PVB19-induced myocarditis and, thus, myocardial tissue damage

Magnetically-Regulated Fragmentation Induced by Nonlinear Flows and Ambipolar Diffusion

We present a parameter study of simulations of fragmentation regulated by gravity, magnetic fields, ambipolar diffusion, and nonlinear flows. The thin-sheet approximation is employed with periodic lateral boundary conditions, and the nonlinear flow field ("turbulence") is allowed to freely decay. In agreement with previous results in the literature, our results show that the onset of runaway collapse (formation of the first star) in subcritical clouds is significantly accelerated by nonlinear flows in which a large-scale wave mode dominates the power spectrum. In addition, we find that a power spectrum with equal energy on all scales also accelerates collapse, but by a lesser amount. For a highly super-Alfvenic initial velocity field with most power on the largest scales, the collapse occurs promptly during the initial compression wave. However, for trans-Alfvenic perturbations, a subcritical magnetic field causes a rebound from the initial compression, and the system undergoes several oscillations before runaway collapse occurs. Models that undergo prompt collapse have highly supersonic infall motions at the core boundaries. Cores in magnetically subcritical models with trans-Alfvenic initial perturbations also pick up significant systematic speeds by inheriting motions associated with magnetically-driven oscillations. Core mass distributions are much broader than in models with small-amplitude initial perturbations, although the disturbed structure of cores that form due to nonlinear flows does not guarantee subsequent monolithic collapse. Our simulations also demonstrate that significant power can (if present initially) be maintained with negligible dissipation in large-scale compressive modes of a magnetic thin sheet, in the limit of perfect flux freezing.Comment: v2, 26 pages, 12 figures, to appear in New Astronomy, animations at http://www.astro.uwo.ca/~basu/pb.ht

Parkinson’s disease: evolution of cognitive impairment and CSF Aβ₁−₄₂ profiles in a prospective longitudinal study

OBJECTIVE: To evaluate the evolution of cognitive impairment in relation to cerebrospinal fluid (CSF) profiles of amyloid-β (Aβ), total-Tau and phosphorylated-Tau in Parkinson’s disease (PD). METHODS: Prospective, longitudinal, observational study up to 10 years with follow-up every 2  years. We assessed CSF profiles in 415 patients with sporadic PD (median age 66; 63% men) and 142 healthy controls (median age 62; 43% men). RESULTS: Patients with PD with low CSF Aβ₁−₄₂ levels at baseline were more often cognitively impaired than patients with intermediate and high Aβ₁−₄₂ levels. Sixty-seven per cent of the patients with low Aβ₁−₄₂ levels at baseline and normal cognition developed cognitive impairment during follow-up, compared with 41% and 37% of patients having intermediate and high CSF Aβ₁−₄₂ levels. Kaplan-Meier survival curves and Cox regression revealed that patients with low CSF Aβ₁−₄₂ levels at baseline developed cognitive impairment more frequently and earlier during follow-up. CONCLUSION: We conclude that in patients with sporadic PD, low levels of Aβ₁−₄₂ are associated with a higher risk of developing cognitive impairment earlier in the disease process at least in a subgroup of patients