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At least 1,567 records · Page 87

Endothelial sirtuin 1 inactivation enhances capillary rarefaction and fibrosis following kidney injury through Notch activation

Peritubular capillary (PTC) rarefaction along with tissue fibrosis is a hallmark of chronic kidney disease (CKD). However, molecular mechanisms of PTC loss have been poorly understood. Previous studies have demonstrated that functional loss of endothelial sirtuin 1 (SIRT1) impairs angiogenesis during development and tissue damage. Here, we found that endothelial SIRT1 dysfunction causes activation of endothelial Notch1 signaling, which leads to PTC rarefaction and fibrosis following kidney injury. In mice lacking functional SIRT1 in the endothelium (Sirt1 mutant), kidney injury enhanced apoptosis and senescence of PTC endothelial cells with impaired endothelial proliferation and expanded myofibroblast population and collagen deposition. Compared to wild-type kidneys, Sirt1 mutant kidneys up-regulated expression of Delta-like 4 (DLL4, a potent Notch1 ligand), Hey1 and Hes1 (Notch target genes), and Notch intracellular domain-1 (NICD1, active form of Notch1) in microvascular endothelial cells (MVECs) post-injury. Sirt1 mutant primary kidney MVECs reduced motility and vascular assembly and enhanced senescence compared to wild-type kidney MVECs. This difference in the phenotype was negated with Notch inhibition. Concurrent stimulation of DLL4 and transforming growth factor (TGF)-β1 increased trans-differentiation of primary kidney pericytes into myofibroblast more than TGF-β1 treatment alone. Collectively, these results indicate that endothelial SIRT1 counteracts PTC rarefaction by repression of Notch1 signaling and antagonizes fibrosis via suppression of endothelial DLL4 expression. - Highlights: • SIRT1 represses Notch1 signaling in capillary endothelial cells in the kidney. • Endothelial SIRT1 is depleted in the kidney following injury. • Activation of endothelial Notch impairs angiogenesis in the kidney. • Increased expression of endothelial DLL4 enhances renal fibrosis.

60 APPLIED LIFE SCIENCES↗

Temporal Sampling of Enzymes from Live Cells by Localized Electroporation and Quantification of Activity by SAMDI Mass Spectrometry

Abstract Measuring changes in enzymatic activity over time from small numbers of cells remains a significant technical challenge. In this work, a method for sampling the cytoplasm of cells is introduced to extract enzymes and measure their activity at multiple time points. A microfluidic device, termed the live cell analysis device (LCAD), is designed, where cells are cultured in microwell arrays fabricated on polymer membranes containing nanochannels. Localized electroporation of the cells opens transient pores in the cell membrane at the interface with the nanochannels, enabling extraction of enzymes into nanoliter‐volume chambers. In the extraction chambers, the enzymes modify immobilized substrates, and their activity is quantified by self‐assembled monolayers for matrix‐assisted laser desorption/ionization (SAMDI) mass spectrometry. By employing the LCAD‐SAMDI platform, protein delivery into cells is demonstrated. Next, it is shown that enzymes can be extracted, and their activity measured without a loss in viability. Lastly, cells are sampled at multiple time points to study changes in phosphatase activity in response to oxidation by hydrogen peroxide. With this unique sampling device and label‐free assay format, the LCAD with SAMDI enables a powerful new method for monitoring the dynamics of cellular activity from small populations of cells.

Mukherjee, Prithvijit↗

Single Crystal Microwires of p -DTS(FBTTh 2 ) 2 and Their Use in the Fabrication of Field-Effect Transistors and Photodetectors

Single crystal microwires of a well–studied organic semiconductor used in organic solar cells, namely p –DTS(FBTTh 2 ) 2 , are prepared via a self–assembly method in solution. The high level of intermolecular organization in the single crystals facilitates migration of charges, relative to solution–processed films, and provides insight into the intrinsic charge transport properties of p –DTS(FBTTh 2 ) 2 . Field–effect transistors based on the microwires can achieve hole mobilities on the order of ≈1.8 cm 2 V –1 s –1 . Furthermore, these microwires show photoresponsive electrical characteristics and can act as photoswitches, with switch ratios over 1000. Here, these experimental results are interpreted using theoretical simulations using an atomistic density functional theory approach. Based on the lattice organization, intermolecular couplings and reorganization energies are calculated, and hole mobilities for comparison with experimental measurements are further estimated. These results demonstrate a unique example of the optoelectronic applications of p –DTS(FBTTh 2 ) 2 microwires.

36 MATERIALS SCIENCE↗

Leptin promotes endothelial dysfunction in chronic kidney disease through AKT/GSK3β and β-catenin signals

Endothelial dysfunction (ED) is a well-recognized instigator of cardiovascular diseases and develops in chronic kidney disease (CKD) with high rate. Recent studies have implicated that leptin is associated with endothelial dysfunction. We investigated the relationship between leptin and markers of ED in CKD patients and how leptin contributed to endothelial damage. 140 CKD patients and 140 healthy subjects were studied. Serum leptin levels were significantly higher in CKD than in controls and displayed significantly positive association with the increase levels of sICAM-1 and sVCAM-1 but negative correlation with flow-mediated dilatation (FMD) reduction in patients. Our in vitro study demonstrated that leptin induced overexpression of ICAM-1 and VCAM-1, led to f-actin reorganization and vinculin assembly, increased endothelial monolayer permeability for FITC-dextran, and accelerated endothelial cell migration; these changes were markedly reversed when the cells were transfected with AKT or β-catenin shRNA vectors. Notably, high leptin resulted in hyper-phosphorylation of AKT and GSK3β, along with nuclear accumulation of β-catenin. In conclusion, serum leptin was elevated in CKD patients and it might contribute to endothelial dysfunction by disarrangement of f-actin cytoskeleton via a mechanism involving the AKT/GSK3β and β-catenin pathway. - Highlights: • Serum leptin was elevated in CKD patients and it was associated with endothelial dysfunction. • Leptin induced endothelial dysfunction by remodeling cytoskeleton in HUVECs. • Leptin promoted endothelial dysfunction via a mechanism involving the AKT/GSK3β and β-catenin signals.

60 APPLIED LIFE SCIENCES↗

LncRNA NEAT1 promotes autophagy in MPTP-induced Parkinson's disease through stabilizing PINK1 protein

Highlights: • LncRNA NEAT1 was first observed to significantly up-regulate in the midbrain of PD mice in present study. • NEAT1 stabilizes PINK1 protein through direct binding. • NEAT1 promotes the MPTP-induced autophagy in SH-SY5Y cells via stabilizing PINK1 protein. Long non-coding RNA nuclear paraspeckle assembly transcript 1 (lncRNA NEAT1) was found to be closely related to the pathological changes in brain and nervous system. However, the role of NEAT1 and its potential mechanism in Parkinson's disease (PD) largely remain uncharacterized.

60 APPLIED LIFE SCIENCES↗

2D and 3D Characterization of PtNi Nanowire Electrode Composition and Structure

Extended surface Pt or Pt-M (M = Ni or Co) catalysts are a viable alternative to supported nanoparticle catalysts for the oxygen reduction reaction (ORR) at the cathode in proton exchange membrane fuel cells (PEMFCs). The activity and durability of these catalysts in membrane electrode assemblies (MEAs) are greatly dependent on the surface and bulk properties of the catalyst material, integration with ionomer, and the three-dimensional structure of the electrode, necessitating extensive characterization of the catalyst in the electrode at multiple scales. In this work, extended surface PtNi nanowire-based electrocatalysts derived by spontaneous galvanic displacement, and post-treated to obtain high specific and mass activities, are characterized by X-ray spectroscopies to assess catalyst composition and to determine the extent of Pt-Ni alloying. Transmission X-ray microscopy (TXM) is used to generate two- and three-dimensional images of catalyst layers with varied compositions within the electrode to show the distribution of nanowires and to study Ni dissolution and redeposition within the electrode. These techniques are complemented by electron microscopy, which is used to confirm leaching of nickel from the nanowires and its redeposition within the electrode. This process is shown to have detrimental effect on the MEA performance, which can be at least in part recovered by soaking electrodes in acid. The combination of these techniques provides unprecedented detail of the evolution of the nanowire-based extended surface catalyst and electrode structure guiding the targeted design of high performance electrodes based on this class of materials.

36 MATERIALS SCIENCE↗

Electric Switching of Fluorescence Decay in Gold–Silica–Dye Nematic Nanocolloids Mediated by Surface Plasmons

Tunable composite materials with interesting physical behavior can be designed through integrating unique optical properties of solid nanostructures with facile responses of soft matter to weak external stimuli, but this approach remains challenged by their poorly controlled coassembly at the mesoscale. Using scalable wet chemical synthesis procedures, we fabricated anisotropic gold–silica-dye colloidal nanostructures and then organized them into the device-scale (demonstrated for square-inch cells) electrically tunable composites by simultaneously invoking molecular and colloidal self-assembly. We show that the ensuing ordered colloidal dispersions of shape-anisotropic nanostructures exhibit tunable fluorescence decay rates and intensity. Here in this paper, we characterize how these properties depend on low-voltage fields and polarization of both the excitation and emission light, demonstrating a great potential for the practical realization of an interesting breed of nanostructured composite materials.

42 ENGINEERING↗

Grape pomace compost harbors organohalide-respiring Dehalogenimonas species with novel reductive dehalogenase genes

Here, organohalide-respiring bacteria play key roles in the natural chlorine cycle; however, most of the current knowledge is based on cultures from contaminated environments. We demonstrate that grape pomace compost without prior exposure to chlorinated solvents harbors a Dehalogenimonas ( Dhgm ) species capable of using chlorinated ethenes, including the human carcinogen and common groundwater pollutant vinyl chloride (VC) as electron acceptors. Grape pomace microcosms and derived solid-free enrichment cultures were able to dechlorinate trichloroethene (TCE) to less chlorinated daughter products including ethene. 16S rRNA gene amplicon and qPCR analyses revealed the predominance of Dhgm sequences, but no Dehalococcoides mccartyi ( Dhc ) biomarker genes were detected. The enumeration of Dhgm 16S rRNA genes demonstrated VC-dependent growth, and 6.55 ± 0.64 x 10 8 cells were produced per µmole of chloride released. Metagenome sequencing enabled the assembly of a Dhgm draft genome, and 52 putative reductive dehalogenase (RDase) genes were identified. Proteomic workflows identified a putative VC RDase with 49% and 56.1% amino acid similarity to the known VC RDases VcrA and BvcA, respectively. A survey of 1,173 groundwater samples collected from 111 chlorinated solvent-contaminated sites revealed that Dhgm 16S rRNA genes were frequently detected and outnumbered Dhc in 65% of the samples. Dhgm may be more relevant contributors to chlorinated solvent reductive dechlorination in contaminated aquifers than is currently recognized, and non-polluted environments are a source of strictly organohalide-respiring bacteria with novel RDase genes.

54 ENVIRONMENTAL SCIENCES↗

High p CO 2 -induced exopolysaccharide-rich ballasted aggregates of planktonic cyanobacteria could explain Paleoproterozoic carbon burial

Here, the contribution of planktonic cyanobacteria to burial of organic carbon in deep-sea sediments before the emergence of eukaryotic predators ~1.5 Ga has been considered negligible owing to the slow sinking speed of their small cells. However, global, highly positive excursion in carbon isotope values of inorganic carbonates ~2.22–2.06 Ga implies massive organic matter burial that had to be linked to oceanic cyanobacteria. Here to elucidate that link, we experiment with unicellular planktonic cyanobacteria acclimated to high partial CO 2 pressure ( p CO 2 ) representative of the early Paleoproterozoic. We find that high p CO 2 boosts generation of acidic extracellular polysaccharides (EPS) that adsorb Ca and Mg cations, support mineralization, and aggregate cells to form ballasted particles. The down flux of such self-assembled cyanobacterial aggregates would decouple the oxygenic photosynthesis from oxidative respiration at the ocean scale, drive export of organic matter from surface to deep ocean and sustain oxygenation of the planetary surface.

59 BASIC BIOLOGICAL SCIENCES↗

Emergence of distinct electronic states in epitaxially-fused PbSe quantum dot superlattices

Abstract Quantum coupling in arrayed nanostructures can produce novel mesoscale properties such as electronic minibands to improve the performance of optoelectronic devices, including ultra-efficient solar cells and infrared photodetectors. Colloidal PbSe quantum dots (QDs) that self-assemble into epitaxially-fused superlattices (epi-SLs) are predicted to exhibit such collective phenomena. Here, we show the emergence of distinct local electronic states induced by crystalline necks that connect individual PbSe QDs and modulate the bandgap energy across the epi-SL. Multi-probe scanning tunneling spectroscopy shows bandgap modulation from 0.7 eV in the QDs to 1.1 eV at their necks. Complementary monochromated electron energy-loss spectroscopy demonstrates bandgap modulation in spectral mapping, confirming the presence of these distinct energy states from necking. The results show the modification of the electronic structure of a precision-made nanoscale superlattice, which may be leveraged in new optoelectronic applications.

36 MATERIALS SCIENCE↗

Asymmetric electrode ionomer for low relative humidity operation of anion exchange membrane fuel cells

The operation of fuel cells under low relative humidity (RH) conditions gives substantial cost and performance benefits. Nonetheless, it is not currently feasible to operate anion exchange membrane fuel cells (AEMFCs) at low RH conditions because current materials for membrane electrode assembly cannot provide sufficient water for the oxygen reduction reaction. In this work, we synthesized polyfluorene ionomers with different ammonium concentrations for anode and cathode to control water management. We designed several asymmetric electrodes that enable high performance under low RH conditions via not only conventional backward water diffusion (anode to cathode) but also forward diffusion (cathode to anode). The AEMFCs using optimized asymmetric electrodes exhibited high H 2 /CO 2 -free air performance (rated power density of circa 540 mW cm -2 at 90 °C under 75% (anode) and 50% RH (cathode) conditions), which is comparable to those of state-of-the-art AEMFCs under nearly water-saturated conditions. The durability of the AEMFCs is excellent, generating 0.6 A cm -2 for >900 h at 80 °C under 50% RH (cathode) conditions. This study demonstrates that high-performance and durable AEMFCs under low RH and high current generating conditions are possible.

25 ENERGY STORAGE↗

The Era GTPase recognizes the GAUCACCUCC sequence and binds helix 45 near the 3; end of 16S rRNA

Era, composed of a GTPase domain and a K homology domain, is essential for bacterial cell viability. It is required for the maturation of 16S rRNA and assembly of the 30S ribosomal subunit. We showed previously that the protein recognizes nine nucleotides (1531{sup AUCACCUCC}1539) near the 3{prime} end of 16S rRNA, and that this recognition stimulates GTP-hydrolyzing activity of Era. In all three kingdoms of life, the 1530{sup GAUCA}1534 sequence and helix 45 (h45) (nucleotides 1506-1529) are highly conserved. It has been shown that the 1530{sup GA}1531 to 1530{sup AG}1531 double mutation severely affects the viability of bacteria. However, whether Era interacts with G1530 and/or h45 and whether such interactions (if any) contribute to the stimulation of Era's GTPase activity were not known. Here, we report two RNA structures that contain nucleotides 1506-1542 (RNA301), one in complex with Era and GDPNP (GNP), a nonhydrolysable GTP-analogue, and the other in complex with Era, GNP, and the KsgA methyltransferase. The structures show that Era recognizes 10 nucleotides, including G1530, and that Era also binds h45. Moreover, GTPase assay experiments show that G1530 does not stimulate Era's GTPase activity. Rather, A1531 and A1534 are most important for stimulation and h45 further contributes to the stimulation. Although G1530 does not contribute to the intrinsic GTPase activity of Era, its interaction with Era is important for binding and is essential for the protein to function, leading to the discovery of a new cold-sensitive phenotype of Era.

59 BASIC BIOLOGICAL SCIENCES↗

Direct visualization of degradation microcompartments at the ER membrane

To promote the biochemical reactions of life, cells can compartmentalize molecular interaction partners together within separated non–membrane-bound regions. It is unknown whether this strategy is used to facilitate protein degradation at specific locations within the cell. Leveraging in situ cryo-electron tomography to image the native molecular landscape of the unicellular alga Chlamydomonas reinhardtii , we discovered that the cytosolic protein degradation machinery is concentrated within ∼200-nm foci that contact specialized patches of endoplasmic reticulum (ER) membrane away from the ER–Golgi interface. These non–membrane-bound microcompartments exclude ribosomes and consist of a core of densely clustered 26S proteasomes surrounded by a loose cloud of Cdc48. Active proteasomes in the microcompartments directly engage with putative substrate at the ER membrane, a function canonically assigned to Cdc48. Live-cell fluorescence microscopy revealed that the proteasome clusters are dynamic, with frequent assembly and fusion events. We propose that the microcompartments perform ER-associated degradation, colocalizing the degradation machinery at specific ER hot spots to enable efficient protein quality control.

Science & Technology - Other Topics↗

Pectin methylesterase selectively softens the onion epidermal wall yet reduces acid-induced creep

De-esterification of homogalacturonan (HG) is thought to stiffen pectin gels and primary cell walls by increasing calcium cross-linking between HG chains. Contrary to this idea, recent studies found that HG de-esterification correlated with reduced stiffness of living tissues, measured by surface indentation. The physical basis of such apparent wall softening is unclear, but possibly involves complex biological responses to HG modification. To assess the direct physical consequences of HG de-esterification on wall mechanics without such complications, we treated isolated onion (Allium cepa) epidermal walls with pectin methylesterase (PME) and assessed wall biomechanics with indentation and tensile tests. In nanoindentation assays, PME action softened the wall (reduced the indentation modulus). In tensile force/extension assays, PME increased plasticity, but not elasticity. These softening effects are attributed, at least in part, to increased electrostatic repulsion and swelling of the wall after PME treatment. Despite softening and swelling upon HG de-esterification, PME treatment alone failed to induce cell wall creep. Instead, acid-induced creep, mediated by endogenous α-expansin, was reduced. We conclude that HG de-esterification physically softens the onion wall, yet reduces expansin-mediated wall extensibility.

59 BASIC BIOLOGICAL SCIENCES↗

Experimental Studies of Graphene-Coated Polymer Electrolyte Membranes for Direct Methanol Fuel Cells

The two main technical limitations of direct methanol fuel cells (DMFCs) are the slow kinetic reactions of the methanol oxidation reaction (MOR) in the anode and the crossing over of unreacted methanol through the proton exchange membrane (PEM). It is a common practice to use Nafion membranes as PEMs, which have high ion exchange capacity. However, Nafion-based membranes also have high fuel permeability, decreasing fuel utilization, and reducing the potential power density. This article focuses on using graphene-coated (Gr-coated) PEMs to reduce fuel crossover. Protons can permeate across graphene, and thus, it can be employed in various devices as a proton conductive membrane. Here, we report the efficiency of Gr-coated Nafions. We tested performance and crossover at three different temperatures with four different fuel concentrations and compared it to a Nafion PEM that underwent the same test conditions. We found that the adhesion of Gr on to PEMs is insufficient for prolonging fuel cell operation, resulting in Gr delamination at high temperatures and higher fuel crossover values compared to lower temperature testing. Furthermore, the results for 7.5M methanol fuel show a reduction of up to 25% in methanol crossover, translating to a peak power density that increases from 3.9 to 9.5 mW/cm 2 when using a Gr-coated PEM compared to a Nafion PEM at 30 °C.

25 ENERGY STORAGE↗

Characterization of local hydride re-orientation in high burn-up PWR fuel rods induced by high pressure at high temperatures

Hydride re-orientation in high burn-up PWR fuel rods was induced by high pressure at high temperatures. The high-burnup specimens were sectioned from PWR rods taken from a 15×15 assembly of the H.B. Robinson (HBR) Unit 2 reactor. Out-of cell benchmark tests performed on unirradiated hydrided Zircaloy-4 specimens were conducted to determine the appropriate temperature, pressure, cooling rate, and number of cooling cycles for the reorientation of the irradiated in-cell specimens. The in-cell hydride reorientation tests were performed using high-burnup fuel specimens under a hoop stress ≈145 MPa at 400°C. The specimens were heated to the target temperature of 400°C, held for 3 hours, cooled at 1°C/min to 170°C,and then heated at 1°C/min to the target temperature again for five cycles. Post test metallographic examinations showed that a significant amount of radial hydrides were induced in the HBR fuel rods. The length of radial hydride was up to 60 μm. For unirradiated materials, the ductility of the radial hydride treated specimens is significantly reduced as compared to the as-hydrided specimens having the same hydrogen concentration (≈300 wppm in this work). The mechanical testing on irradiated fueled samples with hydride reorientation experiments have been performed, and will be reported separately in the near future.

73 NUCLEAR PHYSICS AND RADIATION PHYSICS↗

RETRACTED: Proteasome storage granules protect proteasomes from autophagic degradation upon carbon starvation

26S proteasome abundance is tightly regulated at multiple levels, including the elimination of excess or inactive particles by autophagy. In yeast, this proteaphagy occurs upon nitrogen starvation but not carbon starvation, which instead stimulates the rapid sequestration of proteasomes into cytoplasmic puncta termed proteasome storage granules (PSGs). Here, we show that PSGs help protect proteasomes from autophagic degradation. Both the core protease and regulatory particle sub-complexes are sequestered separately into PSGs via pathways dependent on the accessory proteins Blm10 and Spg5, respectively. Modulating PSG formation, either by perturbing cellular energy status or pH, or by genetically eliminating factors required for granule assembly, not only influences the rate of proteasome degradation, but also impacts cell viability upon recovery from carbon starvation. PSG formation and concomitant protection against proteaphagy also occurs in Arabidopsis , suggesting that PSGs represent an evolutionarily conserved cache of proteasomes that can be rapidly re-mobilized based on energy availability.

59 BASIC BIOLOGICAL SCIENCES↗

Status of Rockwell-ERC high efficiency solar cell programs

Programs aimed at developing large area, high efficiency GaAs heteroface cells for low concentration space applications and high concentration terrestrial applications as well as other programs aimed at developing high efficiency multicolor devices for use in similar applications are described. An additional program aimed at achieving improved power to weight ratio by parting thin film solar cells from their growth substrates prior to their incorporation into an array assembly is also described. There is potential for multiple reuse of the substrates which could lead to reduced costs for such devices. Highlights of these programs and their interrelated contributions toward the goals of reducing specific weight, volume and cost of photovoltaic space power systems are discussed. Overall goals are summarized and current programs and their funding sources are listed.

Zehr, S. W.↗