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Sample records for cell scintillation proximity

  1. Scintillation proximity assay for human DNA topoisomerase I using recombinant biotinyl-fusion protein produced in baculovirus-infected insect cells.

    PubMed

    Lerner, C G; Saiki, A Y

    1996-09-05

    DNA topoisomerases are well-established targets of important therapeutic agents which include the antibacterial quinolones and anticancer camptothecins. Screens for new classes of topoisomerase inhibitors generally employ methods, such as gel electrophoresis, which are not readily amenable to a rapid high-throughput format. We describe here a high-throughput assay to screen for inhibitors of human DNA topoisomerase I based on the scintillation proximity assay. The assay employs recombinant biotinyl-topoisomerase I fusion protein, a hybrid protein which contains a domain that is biotinylated during in vivo expression. The hybrid topoisomerase I fusion protein is found to be biotinylated, active, and nuclear-localized when produced in insect cells using a baculovirus expression system. The biotinyl-topoisomerase I fusion protein can be captured from crude nuclear extracts by immobilization on streptavidin-coated scintillation proximity assay beads. The assay detects binding of 3H-labeled DNA to the bead-immobilized enzyme by scintillation counting. The method is also able to detect stabilization of covalent protein-DNA complexes by camptothecin, an inhibitor previously shown to stabilize covalent intermediates that form during catalysis.

  2. Scintillation Proximity Radioimmunoassay Utilizing 125I-Labeled Ligands

    NASA Astrophysics Data System (ADS)

    Udenfriend, Sidney; Diekmann Gerber, Louise; Brink, Larry; Spector, Sydney

    1985-12-01

    A unique type of radioimmunoassay is described that does not require centrifugation or separation. Microbeads containing a fluorophor are covalently linked to antibody. When an 125I-labeled antigen is added it binds to the beads and, by its proximity, the emitted short-range electrons of the 125I excite the fluorophor in the beads. The light emitted can be measured in a standard scintillation counter. Addition of unlabeled antigen from tissue extracts displaces the labeled ligand and diminishes the fluorescent signal. Application of scintillation proximity immunoassay to tissue enkephalins, serum thyroxin, and urinary morphine is described. Applications of the principle to study the kinetics of interaction between receptors and ligands are discussed.

  3. Scintillation proximity radioimmunoassay utilizing 125I-labeled ligands

    SciTech Connect

    Udenfriend, S.; Gerber, L.D.; Brink, L.; Spector, S.

    1985-12-01

    A unique type of radioimmunoassay is described that does not require centrifugation or separation. Microbeads containing a fluorophor are covalently linked to antibody. When an /sup 125/I-labeled antigen is added it binds to the beads and, by its proximity, the emitted short-range electrons of the /sup 125/I excite the fluorophor in the beads. The light emitted can be measured in a standard scintillation counter. Addition of unlabeled antigen from tissue extracts displaces the labeled ligand and diminishes the fluorescent signal. Application of scintillation proximity immunoassay to tissue enkephalins, serum thyroxin, and urinary morphine is described. Applications of the principle to study the kinetics of interaction between receptors and ligands are discussed.

  4. Development of a scintillation proximity binding assay for high-throughput screening of hematopoietic prostaglandin D2 synthase.

    PubMed

    Meleza, Cesar; Thomasson, Bobbie; Ramachandran, Chidambaram; O'Neill, Jason W; Michelsen, Klaus; Lo, Mei-Chu

    2016-10-15

    Prostaglandin D2 synthase (PGDS) catalyzes the isomerization of prostaglandin H2 (PGH2) to prostaglandin D2 (PGD2). PGD2 produced by hematopoietic prostaglandin D2 synthase (H-PGDS) in mast cells and Th2 cells is proposed to be a mediator of allergic and inflammatory responses. Consequently, inhibitors of H-PGDS represent potential therapeutic agents for the treatment of inflammatory diseases such as asthma. Due to the instability of the PGDS substrate PGH2, an in-vitro enzymatic assay is not feasible for large-scale screening of H-PGDS inhibitors. Herein, we report the development of a competition binding assay amenable to high-throughput screening (HTS) in a scintillation proximity assay (SPA) format. This assay was used to screen an in-house compound library of approximately 280,000 compounds for novel H-PGDS inhibitors. The hit rate of the H-PGDS primary screen was found to be 4%. This high hit rate suggests that the active site of H-PGDS can accommodate a large diversity of chemical scaffolds. For hit prioritization, these initial hits were rescreened at a lower concentration in SPA and tested in the LAD2 cell assay. 116 compounds were active in both assays with IC50s ranging from 6 to 807 nM in SPA and 82 nM to 10 μM in the LAD2 cell assay.

  5. High Throughput Scintillation Proximity Assay for the Identification of FKBP-12 Ligands.

    PubMed

    Graziani; Aldegheri; Terstappen

    1999-01-01

    A high throughput scintillation proximity assay (SPA) was developed to identify novel ligands of FKBP-12, an immunophilin with peptidyl prolyl isomerase (rotamase) activity. Recombinant histidine-tagged FKBP-12 was expressed in Escherichia coli, purified by metal ion affinity chromatography, and immobilized to SPA beads by an antibody that recognizes the histidine tag of the recombinant protein. Using 1 nM [3H] FK506, a well-known macrolid ligand of FKBP-12, specific binding was saturable and accounted for 95% of total binding. Analysis of saturation and homologous displacement isotherms indicated the existence of a single binding site with a Kd value of 1.6 nM. The specificity of [3H] FK506 binding was demonstrated in displacement experiments and showed that rapamycin, another macrolid, was as active as FK506 (IC50 of 3.5 and 3.2 nM, respectively), whereas GPI-1046, a prototype of small molecular compounds with neurotrophic properties and affinity for FKBP-type immunophilins, was more than 1000-fold less active. The high signal-to-noise ratio of 30, together with small standard deviations, makes this novel assay well suited for automated high throughput screening.

  6. Rapid colchicine competition-binding scintillation proximity assay using biotin-labeled tubulin.

    PubMed

    Tahir, S K; Kovar, P; Rosenberg, S H; Ng, S C

    2000-07-01

    We have developed a rapid [3H]colchicine competition-binding scintillation proximity assay (SPA) to evaluate antimitotic compounds that bind to the colchicine-binding site on tubulin. The premise of our assay is that compounds will compete with radiolabeled colchicine for the tubulin-binding domain. Biotin-labeled tubulin is incubated first with unlabeled compound and radiolabeled ligand. Streptavidin-labeled SPA beads are added, and the radiolabel associated with tubulin is directly counted with no separation steps. Under our experimental conditions, the dissociation constant of binding (Kd) for colchicine to tubulin was determined to be 1.4 microM, which was consistent with previously reported values. Assay validation was performed by competitively inhibiting [3H]colchicine binding to tubulin with known microtubule inhibitors and comparing their inhibition constants (Ki). Our SPA bead method is a powerful tool since it overcomes the disadvantage of traditional filtration techniques, as there are no separation steps. It is extremely easy to set up, multiple samples can be assayed and supply and labor costs are reduced because of the minimal volume and test reagents used.

  7. A scintillation proximity assay amenable for screening and characterization of DNA gyrase B subunit inhibitors.

    PubMed

    Gevi, Monica; Domenici, Enrico

    2002-01-01

    DNA gyrase is the target of coumarin and cyclothialidine antibacterials, which bind to the B subunit of the enzyme (GyrB). Currently available GyrB inhibitors have not been clinically successful, but their high in vitro potency against DNA gyrase has raised interest in the development of novel noncoumarin antibacterials acting at the same site. We report the development of a simple scintillation proximity assay (SPA) for the study of binding interactions between coumarin or noncoumarin antibacterials and GyrB, which prevents the needs of separation steps and can be run in microtiter plate formats. The assay is based on the detection of the binding of a radioligand, [3H]dihydronovobiocin, to a biotin-labeled 43-kDa fragment of GyrB (biotin-GyrB43), which is captured by streptavidin-coated SPA beads. The typical assay was conducted in 96-well microtiter plates, with final concentration of 10 nM for biotin-GyrB43, 20 nM for [3H]dihydronovobiocin, and 33 microg of SPA beads/well. From saturation experiments, an equilibrium dissociation constant (K(d)) for dihydronovobiocin of 8.10 nM was found. Displacement studies gave 50% inhibitory concentrations (IC(50)) of 42, 64, and 11 nM for novobiocin, dihydronovobiocin, and the cyclothialidine analogue GR122222X, respectively, consistent with previous findings. The assay was found to be robust to dimethyl sulfoxide up to 5% (v/v) and can be used for high-throughput screens of large chemical collections in the search of novel DNA gyrase inhibitors.

  8. Comparison of luminescence ADP production assay and radiometric scintillation proximity assay for Cdc7 kinase.

    PubMed

    Takagi, Toshimitsu; Shum, David; Parisi, Monika; Santos, Ruth E; Radu, Constantin; Calder, Paul; Rizvi, Zahra; Frattini, Mark G; Djaballah, Hakim

    2011-09-01

    Several assay technologies have been successfully adapted and used in HTS to screen for protein kinase inhibitors; however, emerging comparative analysis studies report very low hit overlap between the different technologies, which challenges the working assumption that hit identification is not dependent on the assay method of choice. To help address this issue, we performed two screens on the cancer target, Cdc7-Dbf4 heterodimeric protein kinase, using a direct assay detection method measuring [(33)P]-phosphate incorporation into the substrate and an indirect method measuring residual ADP production using luminescence. We conducted the two screens under similar conditions, where in one, we measured [(33)P]-phosphate incorporation using scintillation proximity assay (SPA), and in the other, we detected luminescence signal of the ATP-dependent luciferase after regenerating ATP from residual ADP (LUM). Surprisingly, little or no correlation were observed between the positives identified by the two methods; at a threshold of 30% inhibition, 25 positives were identified in the LUM screen whereas the SPA screen only identified two positives, Tannic acid and Gentian violet, with Tannic acid being common to both. We tested 20 out of the 25 positive compounds in secondary confirmatory study and confirmed 12 compounds including Tannic acid as Cdc7-Dbf4 kinase inhibitors. Gentian violet, which was only positive in the SPA screen, inhibited luminescence detection and categorized as a false positive. This report demonstrates the strong impact in detection format on the success of a screening campaign and the importance of carefully designed confirmatory assays to eliminate those compounds that target the detection part of the assay.

  9. Development of a Novel High-Density [3H]Hypoxanthine Scintillation Proximity Assay To Assess Plasmodium falciparum Growth

    PubMed Central

    Caballero, Iván; Colmenarejo, Gonzalo; Sanz, Laura M.; Álvarez-Ruiz, Emilio; Gamo, Francisco-Javier; Cid, Concepción

    2016-01-01

    The discovery and development of new antimalarial drugs are becoming imperative because of the spread of resistance to current clinical treatments. The lack of robustly validated antimalarial targets and the difficulties with the building in of whole-cell activity in screening hits are hampering target-based approaches. However, phenotypic screens of structurally diverse molecule libraries are offering new opportunities for the identification of novel antimalarials. Several methodologies can be used to determine the whole-cell in vitro potencies of antimalarial hits. The [3H]hypoxanthine incorporation assay is considered the “gold standard” assay for measurement of the activity of antimalarial compounds against intraerythrocytic forms of Plasmodium falciparum. However, the method has important limitations, as the assay is not amenable for high-throughput screening since it remains associated with the 96-well plate format. We have overcome this drawback by adapting the [3H]hypoxanthine incorporation method to a 384-well high-density format by coupling a homogeneous scintillation proximity assay (SPA) and thus eliminating the limiting filtration step. This SPA has been validated using a diverse set of 1,000 molecules, including both a representative set from the Tres Cantos Antimalarial Set (TCAMS) of compounds and molecules inactive against whole cells. The results were compared with those from the P. falciparum lactate dehydrogenase whole-cell assay, another method that is well established as a surrogate for parasite growth and is amenable for high-throughput screening. The results obtained demonstrate that the SPA-based [3H]hypoxanthine incorporation assay is a suitable design that is adaptable to high-throughput antimalarial drug screening and that maintains the features, robustness, and reliability of the standard filtration hypoxanthine incorporation method. PMID:27458216

  10. Functional Stability of the Human Kappa Opioid Receptor Reconstituted in Nanodiscs Revealed by a Time-Resolved Scintillation Proximity Assay

    PubMed Central

    Hansen, Randi Westh; Wang, Xiaole; Golab, Agnieszka; Bornert, Olivier; Oswald, Christine; Wagner, Renaud; Martinez, Karen Laurence

    2016-01-01

    Long-term functional stability of isolated membrane proteins is crucial for many in vitro applications used to elucidate molecular mechanisms, and used for drug screening platforms in modern pharmaceutical industry. Compared to soluble proteins, the understanding at the molecular level of membrane proteins remains a challenge. This is partly due to the difficulty to isolate and simultaneously maintain their structural and functional stability, because of their hydrophobic nature. Here we show, how scintillation proximity assay can be used to analyze time-resolved high-affinity ligand binding to membrane proteins solubilized in various environments. The assay was used to establish conditions that preserved the biological function of isolated human kappa opioid receptor. In detergent solution the receptor lost high-affinity ligand binding to a radiolabelled ligand within minutes at room temperature. After reconstitution in Nanodiscs made of phospholipid bilayer the half-life of high-affinity ligand binding to the majority of receptors increased 70-fold compared to detergent solubilized receptors—a level of stability that is appropriate for further downstream applications. Time-resolved scintillation proximity assay has the potential to screen numerous conditions in parallel to obtain high levels of stable and active membrane proteins, which are intrinsically unstable in detergent solution, and with minimum material consumption. PMID:27035823

  11. Effective Quenchers Are Required to Eliminate the Interference of Substrate: Cofactor Binding in the HAT Scintillation Proximity Assay

    PubMed Central

    Ngo, Liza; Wu, Jiang; Yang, Chao

    2015-01-01

    Abstract Histone acetyltransferases (HATs) mediate the transfer of an acetyl group from the cofactor, acetyl-CoA, to the side chain amino group of specific lysines in diverse protein substrates, most notably nuclear histones. The deregulation of HATs is connected to a number of disease states. Reliable and rapid biochemical assays for HATs are critical for understanding biological functions of protein acetylation, as well as for screening small-molecule inhibitors of HAT enzymes. In this report, we present a scintillation proximity assay (SPA) for the measurement of HAT enzymatic activities. The acetyl donor was [3H]Ac-CoA, and a biotin-modified histone peptide served as the HAT substrate. After the HAT reaction, streptavidin-coated beads were added to induce proximity of acetylated substrate to the scintillant molecules. However, we observed strong nonspecific binding between the cofactor and the histone peptide substrates, which adversely complicated the SPA performance. To prevent this problem, a set of chemical agents were evaluated to eliminate the cofactor–substrate interaction, thus providing reliable SPA readings. With optimization, the SPA showed consistent and robust performance for HAT activity measurement and HAT inhibitor evaluation. Overall, this mix-and-measure assay does not require any washing procedure, can be utilized in the microplate format, and is well suited for high-throughput screening of HAT chemical modulators. PMID:26065557

  12. Radon measurement of natural gas using alpha scintillation cells.

    PubMed

    Kitto, Michael E; Torres, Miguel A; Haines, Douglas K; Semkow, Thomas M

    2014-12-01

    Due to their sensitivity and ease of use, alpha-scintillation cells are being increasingly utilized for measurements of radon ((222)Rn) in natural gas. Laboratory studies showed an average increase of 7.3% in the measurement efficiency of alpha-scintillation cells when filled with less-dense natural gas rather than regular air. A theoretical calculation comparing the atomic weight and density of air to that of natural gas suggests a 6-7% increase in the detection efficiency when measuring radon in the cells. A correction is also applicable when the sampling location and measurement laboratory are at different elevations. These corrections to the measurement efficiency need to be considered in order to derive accurate concentrations of radon in natural gas.

  13. Novel method for the detection of receptors and membrane proteins by scintillation proximity radioassay

    SciTech Connect

    Nelson, N.

    1987-09-01

    A rapid and convenient binding assay for receptors and membrane proteins has been developed. It is based on the binding of /sup 125/I-labeled ligands to membrane proteins adsorbed to polyvinyltoluene plastic scintillation microspheres. Membranes or isolated membrane proteins adsorb to the beads upon mixing, and addition of /sup 125/I-labeled ligand induces photon emission which is proportional to the amount of added receptor or membrane protein. The interaction of acetylcholine receptor with /sup 125/I-labeled alpha-bungarotoxin and antigens with /sup 125/I-labeled antibodies or protein A were used as models to test the system. As little as 1 ng of acetylcholine receptor is detected by the assay and a linear relationship with receptor concentration is observed up to 50 ng of receptor per 250 microliter reaction medium. The effects of detergents, salts, soluble proteins, and neutral membranes were studied. Inclusion of bovine serum albumin up to 1 mg/ml, sodium chloride up to 0.5 M, and membranes up to 10 micrograms/ml cause little or no effect on the assay. Detergents at 10-fold below their critical micelle concentrations had little or no effect on the assay. The pharmacological effects of agonists such as acetylcholine were conveniently studied by following the displacement of the /sup 125/I-labeled ligand. Similarly, the amount of toxin in crude snake venom can be assayed by measuring competition with the labeled toxin. Only a few seconds are required to perform each binding assay.

  14. Modular design of long narrow scintillating cells for ILC detector

    SciTech Connect

    Beznosko, D.; Blazey, G.; Dyshkant, A.; Maloney, J.; Rykalin, V.; Schellpfeffer, J.; /Fermilab

    2005-09-01

    The experimental results for the narrow scintillating elements with effective area about 20 cm{sup 2} are reported. The elements were formed from the single piece of scintillator and were read out via wavelength shifting fibers with the MRS (Metal/Resistor/Semiconductor) photodiodes on both ends of each fiber. The formation of the cells from the piece of scintillator by using grooves is discussed. The cell performance was tested using the radioactive source by measuring the PMT current and a single rate after amplifier and discrimination with threshold at about three photo electrons in each channel and quad coincidences (double coincidences between sensors on each fiber and double coincidences between two neighboring fibers). This result is of high importance for large multi-channel systems, i.e. module may be used as an active element for calorimeter or muon system for the design of the future electron-positron linear collider detector because cell effective area can be smoothly enlarged or reduced (to 4 cm{sup 2} definitely).

  15. Development of a robust scintillation proximity assay for protein tyrosine phosphatase 1B using the catalytically inactive (C215S) mutant.

    PubMed

    Skorey, K I; Kennedy, B P; Friesen, R W; Ramachandran, C

    2001-04-15

    Protein tyrosine phosphatases are a class of enzymes that function to modulate tyrosine phosphorylation of cellular proteins and play an essential role in regulating cell function. PTP1B has been implicated in the negative regulation of the insulin signaling pathway by dephosphorylating the activated insulin receptor. Inhibiting this phosphatase and preventing the insulin-receptor downregulation has been suggested as a target for the treatment of Type II diabetes. A high-throughput screen for inhibitors of PTP1B was developed using a scintillation proximity assay (SPA) with GST-- or FLAG--PTP1B((1-320)) and a potent [(3)H]-tripeptide inhibitor. The problem of interference from extraneous oxidizing and alkylating agents which react with the cysteine active-site nucleophile was overcome by the use of the catalytically inactive C215S form of the native enzyme (GST--PTP1B(C215S)). The GST--PTP1B was linked to the protein A scintillation bead via GST antibody. The radiolabeled inhibitor when bound to the enzyme gave a radioactive signal that was competed away by the unknown competitive compounds. Further utility of PTP1B(C215S) was demonstrated by mixing in the same well both the catalytically inactive GST--PTP1B(C215S) and the catalytically active FLAG--CD45 with an inhibitor. Both a binding and kinetic assay was then performed in the same 96-well plate with the inhibition results determined for the PTP1B(C215S) (binding assay) and CD45 (activity assay). In this way inhibitors could be differentiated between the two phosphatases under identical assay conditions in one 96-well assay plate. The use of a mutant to reduce interference in a binding assay and compare with activity assays is also amenable for most cysteine active-site proteases.

  16. WGA-coated yttrium oxide beads enable an imaging-based adenosine 2a receptor binding scintillation proximity assay suitable for high throughput screening.

    PubMed

    Bryant, Robert; McGuinness, Debra; Turek-Etienne, Tammy; Guyer, Deborah; Yu, Liming; Howells, Leighton; Caravano, Joseph; Zhai, Ying; Lachowicz, Jean

    2004-06-01

    Adenosine receptors belong to the superfamily of G protein-coupled receptors and are involved in a variety of physiologic functions. Traditionally, binding assays to detect adenosine 2a (A2a) antagonists and agonists have used filtration methods that are cumbersome to run and not amenable to HTS. We developed scintillation proximity assays (SPA trade mark ) utilizing HEK293 RBHA2AM cell membranes, either wheat germ agglutinin (WGA)-coated yttrium silicate (YSi) or red-shifted yttrium oxide (YO) beads and the A2a-selective radioligand [(3)H]SCH 58261. Both beads gave windows (total binding/nonspecific binding) of >5 and K(d) values of 2-3 nM for the radioligand, in agreement with results obtained by filtration. In contrast, WGA-polyvinyltoluene as well as other bead types had windows of <3 and significant radioligand binding to the uncoated beads. A 384-well WGA-YO bead SPA was optimized utilizing a LEADseeker imaging system and an automated trituration process for dispensing the dense yttrium-based beads. Signals were stable after 4 h, and Z' values were 0.7-0.8. The LEADseeker imaging assay tolerated 2% dimethyl sulfoxide and generated IC(50) values of 3-5 nM for the A2a antagonist CGS 15943, comparable to that obtained by the filtration method. A number of adenosine and xanthine analogues were identified as hits in the Library of Pharmacologically Active Compounds (LOPAC). This imaging-based A2a SPA enables HTS and is a major improvement over the filtration method.

  17. Progenitor Cells in Proximal Airway Epithelial Development and Regeneration

    PubMed Central

    Lynch, Thomas J.; Engelhardt, John F.

    2015-01-01

    Multiple distinct epithelial domains are found throughout the airway that are distinguishable by location, structure, function, and cell-type composition. Several progenitor cell populations in the proximal airway have been identified to reside in confined microenvironmental niches including the submucosal glands (SMGs), which are embedded in the tracheal connective tissue between the surface epithelium and cartilage, and basal cells that reside within the surface airway epithelium (SAE). Current research suggests that regulatory pathways that coordinate development of the proximal airway and establishment of progenitor cell niches may overlap with pathways that control progenitor cell responses during airway regeneration following injury. SMGs have been shown to harbor epithelial progenitor cells, and this niche is dysregulated in diseases such as cystic fibrosis. However, mechanisms that regulate progenitor cell proliferation and maintenance within this glandular niche are not completely understood. Here we discuss glandular progenitor cells during development and regeneration of the proximal airway and compare properties of glandular progenitors to those of basal cell progenitors in the SAE. Further investigation into glandular progenitor cell control will provide a direction for interrogating therapeutic interventions to correct aberrant conditions affecting the SMGs in diseases such as cystic fibrosis, chronic bronchitis, and asthma. PMID:24818588

  18. Metastatic transitional cell carcinoma in proximal humerus of a dog

    PubMed Central

    Malek, Sarah; Murphy, Kimberly A.; Nykamp, Stephanie G.; Allavena, Rachel

    2011-01-01

    Transitional cell carcinoma (TCC) was diagnosed in the proximal humerus of a dog that was presented with persistent right forelimb lameness with no clinical signs of urinary tract involvement. A diagnosis of TCC was made from surgical biopsy of the humeral lesion with subsequent necropsy revealing the prostatic urethra as the primary site of the tumor. PMID:22379204

  19. Shifting scintillator neutron detector

    DOEpatents

    Clonts, Lloyd G; Cooper, Ronald G; Crow, Jr., Morris Lowell; Hannah, Bruce W; Hodges, Jason P; Richards, John D; Riedel, Richard A

    2014-03-04

    Provided are sensors and methods for detecting thermal neutrons. Provided is an apparatus having a scintillator for absorbing a neutron, the scintillator having a back side for discharging a scintillation light of a first wavelength in response to the absorbed neutron, an array of wavelength-shifting fibers proximate to the back side of the scintillator for shifting the scintillation light of the first wavelength to light of a second wavelength, the wavelength-shifting fibers being disposed in a two-dimensional pattern and defining a plurality of scattering plane pixels where the wavelength-shifting fibers overlap, a plurality of photomultiplier tubes, in coded optical communication with the wavelength-shifting fibers, for converting the light of the second wavelength to an electronic signal, and a processor for processing the electronic signal to identify one of the plurality of scattering plane pixels as indicative of a position within the scintillator where the neutron was absorbed.

  20. Human embryonic stem cells differentiate into functional renal proximal tubular-like cells.

    PubMed

    Narayanan, Karthikeyan; Schumacher, Karl M; Tasnim, Farah; Kandasamy, Karthikeyan; Schumacher, Annegret; Ni, Ming; Gao, Shujun; Gopalan, Began; Zink, Daniele; Ying, Jackie Y

    2013-04-01

    Renal cells are used in basic research, disease models, tissue engineering, drug screening, and in vitro toxicology. In order to provide a reliable source of human renal cells, we developed a protocol for the differentiation of human embryonic stem cells into renal epithelial cells. The differentiated stem cells expressed markers characteristic of renal proximal tubular cells and their precursors, whereas markers of other renal cell types were not expressed or expressed at low levels. Marker expression patterns of these differentiated stem cells and in vitro cultivated primary human renal proximal tubular cells were comparable. The differentiated stem cells showed morphological and functional characteristics of renal proximal tubular cells, and generated tubular structures in vitro and in vivo. In addition, the differentiated stem cells contributed in organ cultures for the formation of simple epithelia in the kidney cortex. Bioreactor experiments showed that these cells retained their functional characteristics under conditions as applied in bioartificial kidneys. Thus, our results show that human embryonic stem cells can differentiate into renal proximal tubular-like cells. Our approach would provide a source for human renal proximal tubular cells that are not affected by problems associated with immortalized cell lines or primary cells.

  1. Proximal Tubule Cell Hypothesis for Cardiorenal Syndrome in Diabetes

    PubMed Central

    Saito, Akihiko; Kaseda, Ryohei; Hosojima, Michihiro; Sato, Hiroyoshi

    2011-01-01

    Incidence of cardiovascular disease (CVD) is remarkably high among patients with chronic kidney disease (CKD), even in the early microalbuminuric stages with normal glomerular filtration rates. Proximal tubule cells (PTCs) mediate metabolism and urinary excretion of vasculotoxic substances via apical and basolateral receptors and transporters. These cells also retrieve vasculoprotective substances from circulation or synthesize them for release into the circulation. PTCs are also involved in the uptake of sodium and phosphate, which are critical for hemodynamic regulation and maintaining the mineral balance, respectively. Dysregulation of PTC functions in CKD is likely to be associated with the development of CVD and is linked to the progression to end-stage renal disease. In particular, PTC dysfunction occurs early in diabetic nephropathy, a leading cause of CKD. It is therefore important to elucidate the mechanisms of PTC dysfunction to develop therapeutic strategies for treating cardiorenal syndrome in diabetes. PMID:21197105

  2. Development of a high-throughput scintillation proximity assay for the identification of C-domain translational initiation factor 2 inhibitors.

    PubMed

    Delle Fratte, Sonia; Piubelli, Chiara; Domenici, Enrico

    2002-12-01

    Translational initiation factor 2 (IF2) is the largest of the 3 factors required for translation initiation in prokaryotes and has been shown to be essential in Escherichia coli. It stimulates the binding of fMet-tRNA(f)(Met) to the 30S ribosomal subunit in the presence of GTP. The selectivity is achieved through specific recognition of the tRNA(f)(Met) blocked alpha-amino group. IF2 is composed of 3 structural domains: N-domain, whose function is not known; G-domain, which contains the GTP/GDP binding site and the GTPase catalytic center; and C-domain, which recognizes and binds fMet-tRNA(f)(Met). Its activity is strictly bacteria specific and highly conserved among prokaryotes. So far, antibiotics targeting IF2 function are not known, and this makes it an ideal target for new drugs with mechanisms of resistance not yet developed. A few assays have been developed in the past, which allow the detection of IF2 activity either directly or indirectly. In both instances, the assays are based on radioactive detection and do not allow for high throughput because of the need for separation or solvent extraction steps. The authors describe a novel biochemical assay for IF2 that exploits the molecular recognition of fMet-tRNA(f)(Met) by the C-domain. The assay is based on the incubation of biotinyl-IF2 with fMet-tRNA(f)(Met) and the subsequent capture of the radiolabeled complex by streptavidin-coated beads, exploiting the scintillation proximity assay (SPA) technology. The assay has been designed in an automatable, homogeneous, miniaturized fashion suitable for high-throughput screening and is rapid, sensitive, and robust to dimethyl sulfoxide (DMSO) up to 10% v/v. The assay, used to screen a limited chemical collection of about 5000 compounds and a subset of compounds originated by a 2-D substructural search, has shown to be able to detect potential IF2 inhibitors.

  3. Cell volume regulation in the proximal convoluted tubule.

    PubMed

    Gagnon, J; Ouimet, D; Nguyen, H; Laprade, R; Le Grimellec, C; Carrière, S; Cardinal, J

    1982-10-01

    To evaluate the effect of hyper- and hypotonicity on proximal convoluted tubule (PCT) cell volume, nonperfused PCT were studied in vitro with hypertonic solutions containing sodium chloride, urea, or mannitol (450 mosmol/kg H2O) and with hypotonic low sodium chloride solutions (160 mosmol/kg H2O). When the tubules were subjected to hypertonic peritubular solutions containing NaCl, cell volume immediately decreased by 15.5% and remained constant throughout the experimental period (60 min). With mannitol, the initial decrease was identical to that with NaCl (17.7%), but the PCT volume increased slightly during the experimental period. With urea, the decrease in cell volume was smaller (7%) and transient. In hypotonicity, the PCT swelled rapidly, but this swelling was followed by a rapid regulatory phase in which PCT volume nearly returned to control values after less than 10 min. With a potassium-free peritubular medium or 10(-3) M ouabain, the regulatory phase of hypotonicity completely disappeared, whereas the cells did not maintain their reduced volume in NaCl-induced hypertonicity. These results suggest that Na-K-ATPase plays an important role in the maintenance of a reduced cellular volume in hypertonicity and in the regulatory phase of hypotonicity, probably by an active extrusion of sodium and water from the cell.

  4. Sex Differences in Renal Proximal Tubular Cell Homeostasis.

    PubMed

    Seppi, Thomas; Prajczer, Sinikka; Dörler, Maria-Magdalena; Eiter, Oliver; Hekl, Daniel; Nevinny-Stickel, Meinhard; Skvortsova, Iraida; Gstraunthaler, Gerhard; Lukas, Peter; Lechner, Judith

    2016-10-01

    Studies in human patients and animals have revealed sex-specific differences in susceptibility to renal diseases. Because actions of female sex hormones on normal renal tissue might protect against damage, we searched for potential influences of the female hormone cycle on basic renal functions by studying excretion of urinary marker proteins in healthy human probands. We collected second morning spot urine samples of unmedicated naturally ovulating women, postmenopausal women, and men daily and determined urinary excretion of the renal tubular enzymes fructose-1,6-bisphosphatase and glutathione-S-transferase-α Additionally, we quantified urinary excretion of blood plasma proteins α1-microglobulin, albumin, and IgG. Naturally cycling women showed prominent peaks in the temporal pattern of urinary fructose-1,6-bisphosphatase and glutathione-S-transferase-α release exclusively within 7 days after ovulation or onset of menses. In contrast, postmenopausal women and men showed consistently low levels of urinary fructose-1,6-bisphosphatase excretion over comparable periods. We did not detect changes in urinary α1-microglobulin, albumin, or IgG excretion. Results of this study indicate that proximal tubular tissue architecture, representing a nonreproductive organ-derived epithelium, undergoes periodical adaptations phased by the female reproductive hormone cycle. The temporally delimited higher rate of enzymuria in ovulating women might be a sign of recurring increases of tubular cell turnover that potentially provide enhanced repair capacity and thus, higher resistance to renal damage.

  5. Mitigation of Memory Effects in Beta Scintillation Cells for Radioactive Gas Detection

    SciTech Connect

    Seifert, Carolyn E; McIntyre, Justin I; Antolick, Kathryn C; Carman, April J; Cooper, Matthew W; Hayes, James C; Heimbigner, Tom R; Hubbard, C W; Litke, Kevin E; Ripplinger, Mike D; Suarez, Reynold

    2005-08-31

    The Automated Radioxenon Sampler/Analyzer (ARSA) developed at PNNL measures the relative concentrations of xenon isotopes using a coincidence system. Previous tests of the ARSA system have shown that latent radioactivity remains in the plastic cells after evacuation of the gases, leading to a “memory effect” in which the background count rate is dependent on the sample history. The increased background results in lower detection sensitivity. Two possible solutions to the memory effect are explored in this work: depositing a thin layer of metal on the plastic cell (“metallization”), and using an inorganic scintillating cell composed of yttrium aluminum perovskite (YAP). In both cases, the presence of inorganic material at the surface is intended to inhibit the diffusion of gases into the cell walls.

  6. Light collection and pulse-shape discrimination in elongated scintillator cells for the PROSPECT reactor antineutrino experiment

    SciTech Connect

    Ashenfelter, J.; Jaffe, D.; Diwan, M. V.; Dolph, J.; Qian, X.; Sharma, R.; Viren, B.; Zhang, C.

    2015-11-06

    A meter-long, 23-liter EJ-309 liquid scintillator detector has been constructed to study the light collection and pulse-shape discrimination performance of elongated scintillator cells for the PROSPECT reactor antineutrino experiment. The magnitude and uniformity of light collection and neutron-gamma discrimination power in the energy range of antineutrino inverse beta decay products have been studied using gamma and spontaneous fission calibration sources deployed along the cell axis. We also study neutron-gamma discrimination and light collection abilities for differing PMT and reflector configurations. As a result, key design features for optimizing MeV-scale response and background rejection capabilities are identified.

  7. Light collection and pulse-shape discrimination in elongated scintillator cells for the PROSPECT reactor antineutrino experiment

    NASA Astrophysics Data System (ADS)

    Ashenfelter, J.; Balantekin, B.; Band, H. R.; Barclay, G.; Bass, C. D.; Berish, D.; Bowden, N. S.; Bowes, A.; Brodsky, J. P.; Bryan, C. D.; Cherwinka, J. J.; Chu, R.; Classen, T.; Commeford, K.; Davee, D.; Dean, D.; Deichert, G.; Diwan, M. V.; Dolinski, M. J.; Dolph, J.; Dwyer, D. A.; Gaison, J. K.; Galindo-Uribarri, A.; Gilje, K.; Glenn, A.; Goddard, B. W.; Green, M.; Han, K.; Hans, S.; Heeger, K. M.; Heffron, B.; Jaffe, D. E.; Langford, T. J.; Littlejohn, B. R.; Martinez Caicedo, D. A.; McKeown, R. D.; Mendenhall, M. P.; Mueller, P.; Mumm, H. P.; Napolitano, J.; Neilson, R.; Norcini, D.; Pushin, D.; Qian, X.; Romero, E.; Rosero, R.; Saldana, L.; Seilhan, B. S.; Sharma, R.; Sheets, S.; Stemen, N. T.; Surukuchi, P. T.; Varner, R. L.; Viren, B.; Wang, W.; White, B.; White, C.; Wilhelmi, J.; Williams, C.; Wise, T.; Yao, H.; Yeh, M.; Yen, Y. R.; Zangakis, G.; Zhang, C.; Zhang, X.

    2015-11-01

    A meter-long, 23-liter EJ-309 liquid scintillator detector has been constructed to study the light collection and pulse-shape discrimination performance of elongated scintillator cells for the PROSPECT reactor antineutrino experiment. The magnitude and uniformity of light collection and neutron-gamma discrimination power in the energy range of antineutrino inverse beta decay products have been studied using gamma and spontaneous fission calibration sources deployed along the cell axis. We also study neutron-gamma discrimination and light collection abilities for differing PMT and reflector configurations. Key design features for optimizing MeV-scale response and background rejection capabilities are identified.

  8. Registration of Charged Particles by Scintillating Fibers Coupled with μ-CELL SI APDG

    NASA Astrophysics Data System (ADS)

    Basharuli, N.; Bondarenko, G.; Bekenov, B.; Golovin, V.; Petrov, V.; Ponomarev, N.; Grigoriev, E.

    2002-11-01

    Silicon μ-cell Avalanche Photodiode operating in Geiger mode (APDG) was used to detect light produced in scintillating fibers of 1 mm diameter by electrons from a 90Sr-source and by α-particles from a 238Pu-source. This recently developed in mesa-technology square 1 mm2 APDG, consisting of 1370 μ-cells, has enhanced inter-cell optical isolation and individual quenching resistors. It showed at room temperature and low biasing voltages (45-47 V) very high gain (up to 106), low dark counting rates (below 3 × 105sec-1) and high detection efficiency for photons of green light (> 35%). Basic characteristics - internal gain, dark counting rate and average number of detected photoelectrons as a function of bias voltage were measured.

  9. Errors in measurements of 222Rn in methane and carbon dioxide using scintillation cells calibrated for 222Rn in air.

    PubMed

    Jenkins, Phillip H; Burkhart, James F; Camley, Robert E

    2014-03-01

    Scintillation cells are used typically for measuring the concentration of (222)Rn in air and are calibrated for that purpose. However, scintillation cells are sometimes used for measuring (222)Rn in natural gas or carbon dioxide. The counting efficiencies of scintillation cells for measurements of (222)Rn in these gases should be different from those for measuring (222)Rn in air because the ranges of alpha particles emitted by (222)Rn and its progeny are greater in methane and smaller in carbon dioxide than in air. If these effects are not taken into consideration, measurements of (222)Rn in natural gas will be biased high and in carbon dioxide will be biased low. The authors previously investigated the effects of barometric pressure on measurements of (222)Rn in air using scintillation cells. A modeling technique was used in a previous study to calculate theoretical errors that would result if atmospheric pressure were not considered. In the current study, the same modeling technique was used to calculate theoretical errors that would be made for measurements of (222)Rn in methane and carbon dioxide if the calibration for (222)Rn in air were used. Results are presented for four types of scintillation cells of varying geometries and for barometric pressures representative of four elevations ranging from sea level to 1,963 m (6,440 feet). These results indicate that the errors introduced by the ranges of the alpha particles in gases different from air can be significant. Depending on the type of cell and the local pressure, a measurement of (222)Rn in methane may be biased high by 2-7%, while a measurement of (222)Rn in CO2 may be biased low by 15-20% if the calibration for (222)Rn in air is used.

  10. A pathway-centric view of spatial proximity in the 3D nucleome across cell lines

    PubMed Central

    Karathia, Hiren; Kingsford, Carl; Girvan, Michelle; Hannenhalli, Sridhar

    2016-01-01

    In various contexts, spatially proximal genes have been shown to be functionally related. However, the extent to which spatial proximity of genes in a pathway contributes to the pathway’s context-specific activity is not known. Leveraging Hi-C data in six human cell-lines, we show that spatial proximity of genes in a pathway is highly correlated with the pathway’s context-specific expression and function. Furthermore, spatial proximity of pathway genes correlates with interactions of their protein products, and the specific pathway genes that are proximal to one another tend to occupy higher levels in the regulatory hierarchy. In addition to intra-pathway proximity, related pathways are spatially proximal to one another and housekeeping-genes tend to be proximal to several other pathways suggesting their coordinating role. Substantially extending previous works, our study reveals a pathway-centric organization of 3D-nucleome, whereby, functionally related interacting driver genes tend to be in spatial-proximity in a context-specific manner. PMID:27976707

  11. Comparative testing of various flow-cell detectors fabricated using CaF{sub 2} solid scintillator

    SciTech Connect

    Kawano, T.; Ohashi, H.; Hamada, Y.; Jamsranjav, E.

    2015-03-15

    A monitoring system based on a flow-cell detector was developed for measuring the tritium concentration in water. The flow-cell detector was fabricated using a granular CaF{sub 2} solid scintillator. This system does not use a liquid scintillation counting system and does not generate radioactive organic liquid waste. Moreover, continuous real-time measurements are possible, in contrast to a liquid scintillation counting system, which requires batch measurements. For further development of the system, four flow-cell detectors were fabricated. They included a single 3-mm-diameter cell, three 3-mm-diameter cells in series, a single 5-mm-diameter cell, and three 5-mm-diameter cells in series. Continuously flowing water containing tritium at various concentrations was passed through the flow cells, and tritium count were measured for 600 and 10000 s. Investigating the relation between the count rate and concentration, the three 5-mm-diameter cells were most sensitive, with a linear relation maintained down to approximately 2 Bq/ml and 10 Bq/ml for 10000- and 600-s measurements, respectively. (authors)

  12. Scintillator material

    DOEpatents

    Anderson, D.F.; Kross, B.J.

    1992-07-28

    An improved scintillator material comprising cerium fluoride is disclosed. Cerium fluoride has been found to provide a balance of good stopping power, high light yield and short decay constant that is superior to known scintillator materials such as thallium-doped sodium iodide, barium fluoride and bismuth germanate. As a result, cerium fluoride is favorably suited for use as a scintillator material in positron emission tomography. 4 figs.

  13. Scintillator material

    DOEpatents

    Anderson, David F.; Kross, Brian J.

    1994-01-01

    An improved scintillator material comprising cerium fluoride is disclosed. Cerium fluoride has been found to provide a balance of good stopping power, high light yield and short decay constant that is superior to known scintillator materials such as thallium-doped sodium iodide, barium fluoride and bismuth germanate. As a result, cerium fluoride is favorably suited for use as a scintillator material in positron emission tomography.

  14. Scintillator material

    DOEpatents

    Anderson, David F.; Kross, Brian J.

    1992-01-01

    An improved scintillator material comprising cerium fluoride is disclosed. Cerium fluoride has been found to provide a balance of good stopping power, high light yield and short decay constant that is superior to known scintillator materials such as thallium-doped sodium iodide, barium fluoride and bismuth germanate. As a result, cerium fluoride is favorably suited for use as a scintillator material in positron emission tomography.

  15. Scintillator material

    DOEpatents

    Anderson, D.F.; Kross, B.J.

    1994-06-07

    An improved scintillator material comprising cerium fluoride is disclosed. Cerium fluoride has been found to provide a balance of good stopping power, high light yield and short decay constant that is superior to known scintillator materials such as thallium-doped sodium iodide, barium fluoride and bismuth germanate. As a result, cerium fluoride is favorably suited for use as a scintillator material in positron emission tomography. 4 figs.

  16. Scintillation detector for carbon-14

    NASA Technical Reports Server (NTRS)

    Knoll, G. F.; Rogers, W. L.

    1971-01-01

    Detector consists of plastic, cylindrical double-wall scintillation cell, which is filled with gas to be analyzed. Thin, inner cell wall is isolated optically from outer (guard) scintillator wall by evaporated-aluminum coating. Bonding technique provides mechanical support to cell wall when device is exposed to high temperatures.

  17. Scintillation Counters

    NASA Astrophysics Data System (ADS)

    Bell, Zane W.

    Scintillators find wide use in radiation detection as the detecting medium for gamma/X-rays, and charged and neutral particles. Since the first notice in 1895 by Roentgen of the production of light by X-rays on a barium platinocyanide screen, and Thomas Edison's work over the following 2 years resulting in the discovery of calcium tungstate as a superior fluoroscopy screen, much research and experimentation have been undertaken to discover and elucidate the properties of new scintillators. Scintillators with high density and high atomic number are prized for the detection of gamma rays above 1 MeV; lower atomic number, lower-density materials find use for detecting beta particles and heavy charged particles; hydrogenous scintillators find use in fast-neutron detection; and boron-, lithium-, and gadolinium-containing scintillators are used for slow-neutron detection. This chapter provides the practitioner with an overview of the general characteristics of scintillators, including the variation of probability of interaction with density and atomic number, the characteristics of the light pulse, a list and characteristics of commonly available scintillators and their approximate cost, and recommendations regarding the choice of material for a few specific applications. This chapter does not pretend to present an exhaustive list of scintillators and applications.

  18. Responses of proximal tubular cells to injury in congenital renal disease: fight or flight.

    PubMed

    Chevalier, Robert L; Forbes, Michael S; Galarreta, Carolina I; Thornhill, Barbara A

    2014-04-01

    Most chronic kidney disease in children results from congenital or inherited disorders, which can be studied in mouse models. Following 2 weeks of unilateral ureteral obstruction (UUO) in the adult mouse, nephron loss is due to proximal tubular mitochondrial injury and cell death. In neonatal mice, proximal tubular cell death is delayed beyond 2 weeks of complete UUO, and release of partial UUO allows remodeling of remaining nephrons. Progressive cyst expansion develops in polycystic kidney disease (PKD), a common inherited renal disorder. The polycystic kidney and fibrosis (pcy)-mutant mouse (which develops late-onset PKD) develops thinning of the glomerulotubular junction in parallel with growth of cysts in adulthood. Renal insufficiency in nephropathic cystinosis, a rare inherited renal disorder, results from progressive tubular cystine accumulation. In the Ctns knockout mouse (a model of cystinosis), proximal tubular cells become flattened, with loss of mitochondria and thickening of tubular basement membrane. In each model, persistent obstructive or metabolic stress leads ultimately to the formation of atubular glomeruli. The initial "fight" response (proximal tubular survival) switches to a "flight" response (proximal tubular cell death) with ongoing oxidative injury and mitochondrial damage. Therapies should be directed at reducing proximal tubular mitochondrial oxidative injury to enhance repair and regeneration.

  19. Residential Proximity to Major Roadways is Associated with Increased Levels of AC133+ Circulating Angiogenic Cells

    PubMed Central

    DeJarnett, Natasha; Yeager, Ray; Conklin, Daniel J.; Lee, Jongmin; O'Toole, Timothy E.; McCracken, James; Abplanalp, Wes; Srivastava, Sanjay; Riggs, Daniel W.; Hamzeh, Ihab; Wagner, Stephen; Chugh, Atul; DeFilippis, Andrew; Ciszewski, Tiffany; Wyatt, Brad; Becher, Carrie; Higdon, Deirdre; Ramos, Kenneth S.; Tollerud, David J.; Myers, John A.; Rai, Shesh N.; Shah, Jasmit; Zafar, Nagma; Krishnasamy, Sathya S.; Prabhu, Sumanth D.; Bhatnagar, Aruni

    2016-01-01

    Objective Previous studies have shown that residential proximity to a roadway is associated with increased cardiovascular disease (CVD) risk. Yet the nature of this association remains unclear, and its impact on individual CVD risk factors has not been assessed. The objective of this study was to determine whether residential proximity to roadways influences systemic inflammation and the levels of circulating angiogenic cells. Approach and Results In a cross-sectional study, CVD risk factors, blood levels of C-reactive protein (CRP), and 15 antigenically-defined circulating angiogenic cell populations were measured in participants (n=316) with moderate to high CVD risk. Attributes of roadways surrounding residential locations were assessed using Geographic Information Systems. Associations between road proximity and cardiovascular indices were analyzed using Generalized Linear Models. Close proximity (<50m) to a major roadway was associated with lower income and higher rates of smoking, but not CRP levels. After adjustment for potential confounders, levels of circulating angiogenic cell in peripheral blood were significantly elevated in people living in close proximity to a major roadway (CD31+/AC133+, AC133+, CD34+/AC133+, and CD34+/45dim/AC133+ cells); and positively associated with road segment distance (CD31+/AC133+, AC133+, and CD34+/AC133+ cells), traffic intensity (CD31+/AC133+ and AC133+ cells), and distance-weighted traffic intensity (CD31+/34+/45+/AC133+ cells). Conclusions Living close to a major roadway is associated with elevated levels of circulating cells positive for the “early” stem marker, AC133+. This may reflect an increased need for vascular repair. Levels of these cells in peripheral blood may be a sensitive index of cardiovascular injury due to residential proximity to roadways. PMID:26293462

  20. p-Cresol mediates autophagic cell death in renal proximal tubular cells.

    PubMed

    Lin, Hsin-Hung; Huang, Chiu-Ching; Lin, Tze-Yi; Lin, Ching-Yuang

    2015-04-02

    Higher serum level of p-cresol (PC) in chronic kidney disease (CKD) patients has been linked with CKD progression. The toxic effect of PC on diverse cells has been reported by prior studies, except for renal tubular cells. Both autophagy and apoptosis contribute to renal tubular cell death, yet evidence of its response to PC is limited and their crosstalk is still unclear. Autophagy is an important cellular process involved in toxin-induced cell death. Renal tubular cell death in tubular injury is thought to be one of the key events causing the progression of CKD. Thus, we treated rat (NRK-52E) and human (HRPTEC) renal proximal tubular cells (RPTC) with PC and found the cell proliferation was significantly decreased. Cell apoptosis was significantly increased and accompanied with the activation of autophagy as evidenced by increases in LC3-II, beclin 1 and Atg 4. We also found an increase of p62 by c-Jun activation. p62 accumulation could mediate the activation of caspase 8-dependent cell apoptosis. Conversely, knockdown of p62 by siRNA of p62 had the opposite effect by arresting LC3-II accumulation and promoting increasing cell viability. We conclude that PC triggered autophagic RPTC death via JNK-mediated p62 accumulation and then activated caspase 8-dependent cell death pathway. PC can be considered as one of the key events causing progression of CKD, which might affect drug disposition in CKD cases.

  1. Exosomal GAPDH from Proximal Tubule Cells Regulate ENaC Activity

    PubMed Central

    Jella, Kishore Kumar; Yu, Ling; Yue, Qiang; Friedman, Daniel; Duke, Billie J.; Alli, Abdel A.

    2016-01-01

    Exosomes are nanometer-scale, cell-derived vesicles that contain various molecules including nucleic acids, proteins, and lipids. These vesicles can release their cargo into adjacent or distant cells and mediate intercellular communication and cellular function. Here we examined the regulation of epithelial sodium channels in mpkCCD cells and distal tubule Xenopus 2F3 cells by exosomes isolated from proximal tubule LLC-PK1 cells. Cultured mpkCCD cells were stained with CTX coupled to a green fluorophore in order to label the cell membranes and freshly isolated exosomes from LLC-PK1 cells were labeled with the red lipophilic dye PKH26 in order to visualize uptake of exosomes into the cells. Single-channel patch clamp recordings showed the open probability of ENaC in Xenopus 2F3 cells and in freshly isolated split-open tubules decreased in response to exogenous application of exosomes derived from LLC-PK1 proximal tubule cells. Active GAPDH was identified within exosomes derived from proximal tubule LLC-PK1 cells. The effect on ENaC activity in Xenopus 2F3 cells was blunted after application of exosomes transfected with the GAPDH inhibitor heptelidic acid. Also, we show GAPDH and ENaC subunits associate in mpkCCD cells. These studies examine a potential role for exosomes in the regulation of ENaC activity and examine a possible mechanism for communication from proximal tubule cells to distal tubule and collecting duct cells. PMID:27802315

  2. Giant Cell Tumour of Proximal Phalanx of Ring Finger: Case Report and Review of Literature

    PubMed Central

    Soni, Rishit; Shah, Malkesh; Patel, Amit; Golwala, Paresh

    2016-01-01

    Giant cell tumour (GCT) of bone arising from a phalanx of a finger is extremely rare. Only two percent of all reported GCTs are found in the hand, which show a higher rate of recurrence as compared to those occurring at a more proximal location. Here we report a rare case of giant cell tumour of proximal phalanx of the ring finger in a 20-year-old male, which was treated with extended curettage and bone grafting. After two years of follow-up, the patient was asymptomatic with complete functional recovery and no signs of recurrence. PMID:27900230

  3. Giant Cell Tumor within the Proximal Tibia after ACL Reconstruction.

    PubMed

    Takahashi, Takashi; MacCormick, Lauren; Ellermann, Jutta; Clohisy, Denis; Marette, Shelly

    2016-01-01

    26-year-old female with prior anterior cruciate ligament reconstruction developed an enlarging lytic bone lesion around the tibial screw with sequential imaging over the course of one year demonstrating progression of this finding, which was confirmed histologically to be a giant cell tumor of bone. The lesion originated around the postoperative bed, making the diagnosis challenging during the early course of the presentation. The case demonstrates giant cell tumor which originated in the metaphysis and subsequently grew to involve the epiphysis; therefore, early course of the disease not involving the epiphysis should not exclude this diagnosis.

  4. Giant Cell Tumor within the Proximal Tibia after ACL Reconstruction

    PubMed Central

    Takahashi, Takashi; MacCormick, Lauren; Ellermann, Jutta; Clohisy, Denis; Marette, Shelly

    2016-01-01

    26-year-old female with prior anterior cruciate ligament reconstruction developed an enlarging lytic bone lesion around the tibial screw with sequential imaging over the course of one year demonstrating progression of this finding, which was confirmed histologically to be a giant cell tumor of bone. The lesion originated around the postoperative bed, making the diagnosis challenging during the early course of the presentation. The case demonstrates giant cell tumor which originated in the metaphysis and subsequently grew to involve the epiphysis; therefore, early course of the disease not involving the epiphysis should not exclude this diagnosis. PMID:26981302

  5. Hybrid scintillators for neutron discrimination

    DOEpatents

    Feng, Patrick L; Cordaro, Joseph G; Anstey, Mitchell R; Morales, Alfredo M

    2015-05-12

    A composition capable of producing a unique scintillation response to neutrons and gamma rays, comprising (i) at least one surfactant; (ii) a polar hydrogen-bonding solvent; and (iii) at least one luminophore. A method including combining at least one surfactant, a polar hydrogen-bonding solvent and at least one luminophore in a scintillation cell under vacuum or an inert atmosphere.

  6. Are endothelial cell bioeffects from acoustic droplet vaporization proximity dependent?

    NASA Astrophysics Data System (ADS)

    Seda, Robinson; Li, David; Fowlkes, J. Brian; Bull, Joseph

    2013-11-01

    Acoustic droplet vaporization (ADV) produces gas microbubbles that provide a means of selective occlusion in gas embolotherapy. Vaporization and subsequent occlusion occur inside blood vessels supplying the targeted tissue, such as tumors. Theoretical and computational studies showed that ADV within a vessel can impart high fluid mechanical stresses on the vessel wall. Previous in vitro studies have demonstrated that vaporization at an endothelial layer may affect cell attachment and viability. The current study is aimed at investigating the role of vaporization distance away from the endothelial layer. HUVECs were cultured in OptiCell™ chambers until reaching confluence. Dodecafluoropentane microdroplets were added, attaining a 10:1 droplet to cell ratio. A single ultrasound pulse (7.5 MHz) consisting of 16 cycles (~ 2 μs) and a 5 MPa peak rarefactional pressure was used to produce ADV while varying the vaporization distance from the endothelial layer (0 μm, 500 μm, 1000 μm). Results indicated that cell attachment and viability was significantly different if the distance was 0 μm (at the endothelial layer). Other distances were not significantly different from the control. ADV will significantly affect the endothelium if droplets are in direct contact with the cells. Droplet concentration and flow conditions inside blood vessels may play an important role. This work was supported by NIH grant R01EB006476.

  7. The Integrin β1 Subunit Regulates Paracellular Permeability of Kidney Proximal Tubule Cells*

    PubMed Central

    Elias, Bertha C.; Mathew, Sijo; Srichai, Manakan B.; Palamuttam, Riya; Bulus, Nada; Mernaugh, Glenda; Singh, Amar B.; Sanders, Charles R.; Harris, Raymond C.; Pozzi, Ambra; Zent, Roy

    2014-01-01

    Epithelial cells lining the gastrointestinal tract and kidney have different abilities to facilitate paracellular and transcellular transport of water and solutes. In the kidney, the proximal tubule allows both transcellular and paracellular transport, while the collecting duct primarily facilitates transcellular transport. The claudins and E-cadherin are major structural and functional components regulating paracellular transport. In this study we present the novel finding that the transmembrane matrix receptors, integrins, play a role in regulating paracellular transport of renal proximal tubule cells. Deleting the integrin β1 subunit in these cells converts them from a “loose” epithelium, characterized by low expression of E-cadherin and claudin-7 and high expression of claudin-2, to a “tight” epithelium with increased E-cadherin and claudin-7 expression and decreased claudin-2 expression. This effect is mediated by the integrin β1 cytoplasmic tail and does not entail β1 heterodimerization with an α-subunit or its localization to the cell surface. In addition, we demonstrate that deleting the β1 subunit in the proximal tubule of the kidney results in a major urine-concentrating defect. Thus, the integrin β1 tail plays a key role in regulating the composition and function of tight and adherens junctions that define paracellular transport properties of terminally differentiated renal proximal tubule epithelial cells. PMID:24509849

  8. IgE epitope proximity determines immune complex shape and effector cell activation capacity

    PubMed Central

    Gieras, Anna; Linhart, Birgit; Roux, Kenneth H.; Dutta, Moumita; Khodoun, Marat; Zafred, Domen; Cabauatan, Clarissa R.; Lupinek, Christian; Weber, Milena; Focke-Tejkl, Margarete; Keller, Walter; Finkelman, Fred D.; Valenta, Rudolf

    2016-01-01

    Background IgE-allergen complexes induce mast cell and basophil activation and thus immediate allergic inflammation. They are also important for IgE-facilitated allergen presentation to T cells by antigen-presenting cells. Objective To investigate whether the proximity of IgE binding sites on an allergen affects immune complex shape and subsequent effector cell activation in vitro and in vivo. Methods We constructed artificial allergens by grafting IgE epitopes in different numbers and proximity onto a scaffold protein. The shape of immune complexes formed between artificial allergens and the corresponding IgE was studied by negative-stain electron microscopy. Allergenic activity was determined using basophil activation assays. Mice were primed with IgE, followed by injection of artificial allergens to evaluate their in vivo allergenic activity. Severity of systemic anaphylaxis was measured by changes in body temperature. Results We could demonstrate simultaneous binding of 4 IgE antibodies in close vicinity to each other. The proximity of IgE binding sites on allergens influenced the shape of the resulting immune complexes and the magnitude of effector cell activation and in vivo inflammation. Conclusions Our results demonstrate that the proximity of IgE epitopes on an allergen affects its allergenic activity. We thus identified a novel mechanism by which IgE-allergen complexes regulate allergic inflammation. This mechanism should be important for allergy and other immune complex–mediated diseases. PMID:26684291

  9. Demonstration of neutron detection utilizing open cell foam and noble gas scintillation

    SciTech Connect

    Lavelle, C. M. Miller, E. C.; Coplan, M.; Thompson, Alan K.; Vest, Robert E.; Yue, A. T.; Kowler, A. L.; Koeth, T.; Al-Sheikhly, M.; Clark, Charles W.

    2015-03-02

    We present results demonstrating neutron detection via a closely spaced converter structure coupled to low pressure noble gas scintillation instrumented by a single photo-multiplier tube (PMT). The converter is dispersed throughout the gas volume using a reticulated vitreous carbon foam coated with boron carbide (B{sub 4}C). A calibrated cold neutron beam is used to measure the neutron detection properties, using a thin film of enriched {sup 10}B as a reference standard. Monte Carlo computations of the ion energy deposition are discussed, including treatment of the foam random network. Results from this study indicate that the foam shadows a significant portion of the scintillation light from the PMT. The high scintillation yield of Xe appears to overcome the light loss, facilitating neutron detection and presenting interesting opportunities for neutron detector design.

  10. Development and Application of Human Renal Proximal Tubule Epithelial Cells for Assessment of Compound Toxicity

    PubMed Central

    Li, Shuaizhang; Zhao, Jinghua; Huang, Ruili; Steiner, Toni; Bourner, Maureen; Mitchell, Michael; Thompson, David C.; Zhao, Bin; Xia, Menghang

    2017-01-01

    Kidney toxicity is a major problem both in drug development and clinical settings. It is difficult to predict nephrotoxicity in part because of the lack of appropriate in vitro cell models, limited endpoints, and the observation that the activity of membrane transporters which plays important roles in nephrotoxicity by affecting the pharmacokinetic profile of drugs is often not taken into account. We developed a new cell model using pseudo-immortalized human primary renal proximal tubule epithelial cells. This cell line (SA7K) was characterized by the presence of proximal tubule cell markers as well as several functional properties, including transporter activity and response to a few well-characterized nephrotoxicants. We subsequently evaluated a group of potential nephrotoxic compounds in SA7K cells and compared them to a commonly used human immortalized kidney cell line (HK-2). Cells were treated with test compounds and three endpoints were analyzed, including cell viability, apoptosis and mitochondrial membrane potential. The results showed that most of the known nephrotoxic compounds could be detected in one or more of these endpoints. There were sensitivity differences in response to several of the chemicals between HK-2 and SA7K cells, which may relate to differences in expressions of key transporters or other components of nephrotoxicity pathways. Our data suggest that SA7K cells appear as promising for the early detection of renal toxicants.

  11. Autocrine inhibition of Na+/K(+)-ATPase by nitric oxide in mouse proximal tubule epithelial cells.

    PubMed Central

    Guzman, N J; Fang, M Z; Tang, S S; Ingelfinger, J R; Garg, L C

    1995-01-01

    An inducible nitric oxide synthase has recently been described in proximal tubule epithelium. To investigate the effects of proximal tubule NO on Na+/K(+)-ATPase, we induced NO production in mouse proximal tubule epithelial cells by treatment with lipopolysaccharide (LPS) and interferon-gamma (IFN gamma) followed by determinations of ouabain-sensitive ATPase activity. Na+/K(+)-ATPase activity decreased after 4 h of LPS/IFN gamma treatment, reaching maximal inhibition after 24 h (34% reduction in activity). The inhibition of Na+/K(+)-ATPase activity by LPS/IFN gamma was prevented by simultaneous incubation with N omega-nitro L-arginine and markedly blunted by removal of L-arginine from the medium. The NO donors sodium nitroprusside and SIN-1 also inhibited Na+/K(+)-ATPase activity to a similar extent than LPS/IFN gamma. However, treatment with 8-pCPT-cGMP only modestly reduced Na+/K(+)-ATPase activity. Interestingly, superoxide dismutase prevented the inhibitory effects of NO on Na+/K(+)-ATPase activity, suggesting a role for peroxynitrite in this inhibition. We conclude that NO generated by mouse proximal tubule epithelial cell iNOS inhibits Na/K ATPase activity in an autocrine fashion and that this inhibition is accompanied by a reduction in Na-dependent solute transport. PMID:7537754

  12. PKB and megalin determine the survival or death of renal proximal tubule cells.

    PubMed

    Caruso-Neves, Celso; Pinheiro, Ana Acacia S; Cai, Hui; Souza-Menezes, Jackson; Guggino, William B

    2006-12-05

    Renal proximal tubule cells have a remarkable ability to reabsorb large quantities of albumin through megalin-mediated endocytosis. This is an essential process for overall body homeostasis. Overstressing this endocytic system with a prolonged excess of albumin is injurious to proximal tubule cells. How these cells function and protect themselves from injury is unknown. Here, we show that megalin is the sensor that determines whether cells will be protected or injured by albumin. Megalin, through a novel mechanism, binds PKB in a D-3-phosphorylated phospholipid-insensitive manner, anchoring PKB in the luminal plasma membrane. Whereas low doses of albumin are protective, an overload of albumin decreases megalin expression followed by a reduction of plasma membrane PKB, PKB activity, and Bad phosphorylation induced by PKB. The result is albumin-induced apoptosis. These results reveal a model for PKB distribution in the plasma membrane and elucidate mechanisms involved in both the protective and toxic effects of albumin on proximal tubule cells. In addition, our findings suggest a mechanism for the progression of chronic kidney disease to end-stage renal disease.

  13. Ankle arthrodesis fusion rates for mesenchymal stem cell bone allograft versus proximal tibia autograft.

    PubMed

    Anderson, John J; Boone, Joshua J; Hansen, Myron; Brady, Chad; Gough, Adam; Swayzee, Zflan

    2014-01-01

    Ankle arthrodesis is commonly used in the treatment of ankle arthritis. The present study compared mesenchymal stem cell (MSC) bone allografts and proximal tibia autografts as adjuncts in performing ankle arthrodesis. A total of 109 consecutive ankle fusions performed from 2002 to 2008 were evaluated retrospectively. Of the 109 fusions, 24 were excluded from the present study, leaving 85 patients who had undergone ankle arthrodesis. Of the 85 patients, 41 had received a proximal tibia autograft and 44, an MSC bone allograft. These 2 groups were reviewed and compared retrospectively at least 2 years postoperatively for the overall fusion rate, interval to radiographic fusion, and interval to clinical fusion. A modified and adjusted American College of Foot and Ankle Surgeons ankle scale was used to measure patient satisfaction. The overall fusion rate was 84.1% in the MSC bone allograft group and 95.1% in the proximal tibia autograft group (p = .158). The corresponding mean intervals to radiographic fusion were 13.0 ± 2.5 weeks and 11.3 ± 2.8 weeks (p ≤ .001). The interval to clinical fusion was 13.1 ± 2.1 weeks and 11.0 ± 1.5 weeks (p ≤ .001) in the MSC bone allograft and proximal tibia autograft group, respectively. No statistically significant difference was found in the fusion rates between the MSC bone allograft and proximal tibia autograft groups. Also, no statistically significant difference was found between the preoperative and postoperative scores using a modified and adjusted American College of Foot and Ankle Surgeons ankle scale between the 2 groups (p = .41 and p = .44, respectively). A statistically significant delay to radiographic and clinical fusion was present in the MSC bone allograft group compared with the proximal tibia autograft group; however, no difference was found in patient satisfaction.

  14. A promiscuous biotin ligase fusion protein identifies proximal and interacting proteins in mammalian cells

    PubMed Central

    Kim, Dae In; Raida, Manfred; Burke, Brian

    2012-01-01

    We have developed a new technique for proximity-dependent labeling of proteins in eukaryotic cells. Named BioID for proximity-dependent biotin identification, this approach is based on fusion of a promiscuous Escherichia coli biotin protein ligase to a targeting protein. BioID features proximity-dependent biotinylation of proteins that are near-neighbors of the fusion protein. Biotinylated proteins may be isolated by affinity capture and identified by mass spectrometry. We apply BioID to lamin-A (LaA), a well-characterized intermediate filament protein that is a constituent of the nuclear lamina, an important structural element of the nuclear envelope (NE). We identify multiple proteins that associate with and/or are proximate to LaA in vivo. The most abundant of these include known interactors of LaA that are localized to the NE, as well as a new NE-associated protein named SLAP75. Our results suggest BioID is a useful and generally applicable method to screen for both interacting and neighboring proteins in their native cellular environment. PMID:22412018

  15. The COOH terminus of megalin regulates gene expression in opossum kidney proximal tubule cells

    PubMed Central

    Li, Yuanli; Cong, Rong; Biemesderfer, Daniel

    2008-01-01

    We recently reported that megalin is subjected to regulated intramembrane proteolysis (RIP) and includes 1) protein kinase C (PKC)-regulated, metalloprotease-mediated ectodomain shedding producing a membrane-bound megalin COOH-terminal fragment (MCTF) and 2) γ-secretase-mediated cleavage of the MCTF producing a soluble megalin intracellular domain (MICD). Based on studies of RIP of other receptors, the MICD is predicted to target to the nucleus and regulate gene expression. To determine whether RIP of megalin regulates proximal tubule gene expression, we stably expressed the transfected MCTF (tMCTF) or transfected MICD (tMICD) in opossum kidney proximal tubule (OKP) cells and examined the resulting phenotype. Immunoblotting and immunocytochemical analysis of tMCTF cells showed the tMCTF was expressed and constitutively processed by γ-secretase. Analysis of specific protein expression in tMCTF- and tMICD-transfected cells using Western blot showed endogenous megalin and Na+/H+ exchanger 3 (NHE3) protein expression to be dramatically lower than that of control cells. Expression of other proteins including myosin VI, β-adaptin, and the Na-K-ATPase appeared unchanged. Analysis of specific mRNA expression using quantitative real-time PCR showed megalin and NHE3 mRNA levels were significantly lower in tMCTF- and tMICD-transfected cells compared with controls. Inhibition of γ-secretase activity in tMCTF cells resulted in an 8- to 10-fold recovery of megalin mRNA within 4 h. These data show that the COOH-terminal domain of megalin regulates expression of specific proteins in OKP cells and provides the first evidence that RIP of megalin may be part of a signaling pathway linking protein absorption and gene expression in proximal tubule. PMID:18495814

  16. SCINTILLATION SPECTROMETER

    DOEpatents

    Bell, P.R.; Francis, J.E.

    1960-06-21

    A portable scintillation spectrometer is described which is especially useful in radio-biological studies for determining the uptake and distribution of gamma -emitting substances in tissue. The spectrometer includes a collimator having a plurality of apertures that are hexagonal in cross section. Two crystals are provided: one is activated to respond to incident rays from the collimator; the other is not activated and shields the first from external radiation.

  17. Plastic scintillation dosimetry: Optimal selection of scintillating fibers and scintillators

    SciTech Connect

    Archambault, Louis; Arsenault, Jean; Gingras, Luc; Sam Beddar, A.; Roy, Rene; Beaulieu, Luc

    2005-07-15

    Scintillation dosimetry is a promising avenue for evaluating dose patterns delivered by intensity-modulated radiation therapy plans or for the small fields involved in stereotactic radiosurgery. However, the increase in signal has been the goal for many authors. In this paper, a comparison is made between plastic scintillating fibers and plastic scintillator. The collection of scintillation light was measured experimentally for four commercial models of scintillating fibers (BCF-12, BCF-60, SCSF-78, SCSF-3HF) and two models of plastic scintillators (BC-400, BC-408). The emission spectra of all six scintillators were obtained by using an optical spectrum analyzer and they were compared with theoretical behavior. For scintillation in the blue region, the signal intensity of a singly clad scintillating fiber (BCF-12) was 120% of that of the plastic scintillator (BC-400). For the multiclad fiber (SCSF-78), the signal reached 144% of that of the plastic scintillator. The intensity of the green scintillating fibers was lower than that of the plastic scintillator: 47% for the singly clad fiber (BCF-60) and 77% for the multiclad fiber (SCSF-3HF). The collected light was studied as a function of the scintillator length and radius for a cylindrical probe. We found that symmetric detectors with nearly the same spatial resolution in each direction (2 mm in diameter by 3 mm in length) could be made with a signal equivalent to those of the more commonly used asymmetric scintillators. With augmentation of the signal-to-noise ratio in consideration, this paper presents a series of comparisons that should provide insight into selection of a scintillator type and volume for development of a medical dosimeter.

  18. Characterisation of human tubular cell monolayers as a model of proximal tubular xenobiotic handling

    SciTech Connect

    Brown, Colin D.A. Sayer, Rachel; Windass, Amy S.; Haslam, Iain S.; Broe, Marc E. de; D'Haese, Patrick C.; Verhulst, Anja

    2008-12-15

    The aim of this study was to determine whether primary human tubular cell monolayers could provide a powerful tool with which to investigate the renal proximal tubular handling of xenobiotics. Human proximal and distal tubule/collecting duct cells were grown as monolayers on permeable filter supports. After 10 days in culture, proximal tubule cells remained differentiated and expressed a wide palette of transporters at the mRNA level including NaPi-IIa, SGLT1, SGLT2, OCT2, OCTN2, OAT1, OAT3, OAT4, MDR1, MRP2 and BCRP. At the protein level, the expression of a subset of transporters including NaPi-IIa, OAT1 and OAT3 was demonstrated using immunohistochemistry. Analysis of the expression of the ATP binding cassette efflux pumps MDR1, MRP2 and BCRP confirmed their apical membrane localisation. At the functional level, tubule cell monolayers retain the necessary machinery to mediate the net secretion of the prototypic substrates; PAH and creatinine. PAH secretion across the monolayer consisted of the uptake of PAH across the basolateral membrane by OAT1 and OAT3 and the apical exit of PAH by a probenecid and MK571-sensitive route consistent with actions of MRP2 or MRP4. Creatinine secretion was by OCT2-mediated uptake at the basolateral membrane and via MDR1 at the apical membrane. Functional expression of MDR1 and BCRP at the apical membrane was also demonstrated using a Hoechst 33342 dye. Similarly, measurement of calcein efflux demonstrated the functional expression of MRP2 at the apical membrane of cell monolayers. In conclusion, human tubular cell monolayers provide a powerful tool to investigate renal xenobiotic handling.

  19. Endo-Lysosomal Dysfunction in Human Proximal Tubular Epithelial Cells Deficient for Lysosomal Cystine Transporter Cystinosin

    PubMed Central

    Van Den Heuvel, Lambertus; Pastore, Anna; Dijkman, Henry; De Matteis, Maria Antonietta; Levtchenko, Elena N.

    2015-01-01

    Nephropathic cystinosis is a lysosomal storage disorder caused by mutations in the CTNS gene encoding cystine transporter cystinosin that results in accumulation of amino acid cystine in the lysosomes throughout the body and especially affects kidneys. Early manifestations of the disease include renal Fanconi syndrome, a generalized proximal tubular dysfunction. Current therapy of cystinosis is based on cystine-lowering drug cysteamine that postpones the disease progression but offers no cure for the Fanconi syndrome. We studied the mechanisms of impaired reabsorption in human proximal tubular epithelial cells (PTEC) deficient for cystinosin and investigated the endo-lysosomal compartments of cystinosin-deficient PTEC by means of light and electron microscopy. We demonstrate that cystinosin-deficient cells had abnormal shape and distribution of the endo-lysosomal compartments and impaired endocytosis, with decreased surface expression of multiligand receptors and delayed lysosomal cargo processing. Treatment with cysteamine improved surface expression and lysosomal cargo processing but did not lead to a complete restoration and had no effect on the abnormal morphology of endo-lysosomal compartments. The obtained results improve our understanding of the mechanism of proximal tubular dysfunction in cystinosis and indicate that impaired protein reabsorption can, at least partially, be explained by abnormal trafficking of endosomal vesicles. PMID:25811383

  20. Influence of uranium speciation on normal rat kidney (NRK-52E) proximal cell cytotoxicity.

    PubMed

    Carrière, M; Avoscan, L; Collins, R; Carrot, F; Khodja, H; Ansoborlo, E; Gouget, B

    2004-03-01

    Uranium is a naturally occurring heavy metal. Its extensive use in the nuclear cycle and for military applications has focused attention on its potential health effects. Acute exposures to uranium are toxic to the kidneys where they mainly cause damage to proximal tubular epithelium. The purpose of this study was to investigate the biological consequences of acute in vitro uranyl exposure and the influence of uranyl speciation on its cytotoxicity. NRK-52E cells, representative of rat kidney proximal epithelium, were exposed to uranyl-carbonate and -citrate complexes, which are the major complexes transiting through renal tubules after acute in vivo contamination. Before NRK-52E cell exposure, these complexes were diluted in classical or modified cell culture media, which can possibly modify uranyl speciation. In these conditions, uranium cytotoxicity appears after 16 h of exposure. The CI50 cytotoxicity index, the uranium concentration leading to 50% dead cells after 24 h of exposure, is 500 microM (+/-100 microM) and strongly depends on uranyl counterion and cell culture medium composition. Computer modeling of uranyl speciation is reported, enabling one to draw a parallel between uranyl speciation and its cytotoxicity.

  1. Tim-3 enhances FcεRI-proximal signaling to modulate mast cell activation.

    PubMed

    Phong, Binh L; Avery, Lyndsay; Sumpter, Tina L; Gorman, Jacob V; Watkins, Simon C; Colgan, John D; Kane, Lawrence P

    2015-12-14

    T cell (or transmembrane) immunoglobulin and mucin domain protein 3 (Tim-3) has attracted significant attention as a novel immune checkpoint receptor (ICR) on chronically stimulated, often dysfunctional, T cells. Antibodies to Tim-3 can enhance antiviral and antitumor immune responses. Tim-3 is also constitutively expressed by mast cells, NK cells and specific subsets of macrophages and dendritic cells. There is ample evidence for a positive role for Tim-3 in these latter cell types, which is at odds with the model of Tim-3 as an inhibitory molecule on T cells. At this point, little is known about the molecular mechanisms by which Tim-3 regulates the function of T cells or other cell types. We have focused on defining the effects of Tim-3 ligation on mast cell activation, as these cells constitutively express Tim-3 and are activated through an ITAM-containing receptor for IgE (FcεRI), using signaling pathways analogous to those in T cells. Using a variety of gain- and loss-of-function approaches, we find that Tim-3 acts at a receptor-proximal point to enhance Lyn kinase-dependent signaling pathways that modulate both immediate-phase degranulation and late-phase cytokine production downstream of FcεRI ligation.

  2. Tim-3 enhances FcεRI-proximal signaling to modulate mast cell activation

    PubMed Central

    Phong, Binh L.; Avery, Lyndsay; Sumpter, Tina L.; Gorman, Jacob V.; Watkins, Simon C.; Colgan, John D.

    2015-01-01

    T cell (or transmembrane) immunoglobulin and mucin domain protein 3 (Tim-3) has attracted significant attention as a novel immune checkpoint receptor (ICR) on chronically stimulated, often dysfunctional, T cells. Antibodies to Tim-3 can enhance antiviral and antitumor immune responses. Tim-3 is also constitutively expressed by mast cells, NK cells and specific subsets of macrophages and dendritic cells. There is ample evidence for a positive role for Tim-3 in these latter cell types, which is at odds with the model of Tim-3 as an inhibitory molecule on T cells. At this point, little is known about the molecular mechanisms by which Tim-3 regulates the function of T cells or other cell types. We have focused on defining the effects of Tim-3 ligation on mast cell activation, as these cells constitutively express Tim-3 and are activated through an ITAM-containing receptor for IgE (FcεRI), using signaling pathways analogous to those in T cells. Using a variety of gain- and loss-of-function approaches, we find that Tim-3 acts at a receptor-proximal point to enhance Lyn kinase-dependent signaling pathways that modulate both immediate-phase degranulation and late-phase cytokine production downstream of FcεRI ligation. PMID:26598760

  3. Localization of the calcium-regulated citrate transport process in proximal tubule cells.

    PubMed

    Hering-Smith, Kathleen S; Mao, Weibo; Schiro, Faith R; Coleman-Barnett, Joycelynn; Pajor, Ana M; Hamm, L Lee

    2014-06-01

    Urinary citrate is an important inhibitor of calcium-stone formation. Most of the citrate reabsorption in the proximal tubule is thought to occur via a dicarboxylate transporter NaDC1 located in the apical membrane. OK cells, an established opossum kidney proximal tubule cell line, transport citrate but the characteristics change with extracellular calcium such that low calcium solutions stimulate total citrate transport as well as increase the apparent affinity for transport. The present studies address several fundamental properties of this novel process: the polarity of the transport process, the location of the calcium-sensitivity and whether NaDC1 is present in OK cells. OK cells grown on permeable supports exhibited apical >basolateral citrate transport. Apical transport of both citrate and succinate was sensitive to extracellular calcium whereas basolateral transport was not. Apical calcium, rather than basolateral, was the predominant determinant of changes in transport. Also 2,3-dimethylsuccinate, previously identified as an inhibitor of basolateral dicarboxylate transport, inhibited apical citrate uptake. Although the calcium-sensitive transport process in OK cells is functionally not typical NaDC1, NaDC1 is present in OK cells by Western blot and PCR. By immunolocalization studies, NaDC1 was predominantly located in discrete apical membrane or subapical areas. However, by biotinylation, apical NaDC1 decreases in the apical membrane with lowering calcium. In sum, OK cells express a calcium-sensitive/regulated dicarboxylate process at the apical membrane which responds to variations in apical calcium. Despite the functional differences of this process compared to NaDC1, NaDC1 is present in these cells, but predominantly in subapical vesicles.

  4. Diabetes increases susceptibility of primary cultures of rat proximal tubular cells to chemically induced injury

    SciTech Connect

    Zhong Qing; Terlecky, Stanley R.; Lash, Lawrence H.

    2009-11-15

    Diabetic nephropathy is characterized by increased oxidative stress and mitochondrial dysfunction. In the present study, we prepared primary cultures of proximal tubular (PT) cells from diabetic rats 30 days after an ip injection of streptozotocin and compared their susceptibility to oxidants (tert-butyl hydroperoxide, methyl vinyl ketone) and a mitochondrial toxicant (antimycin A) with that of PT cells isolated from age-matched control rats, to test the hypothesis that PT cells from diabetic rats exhibit more cellular and mitochondrial injury than those from control rats when exposed to these toxicants. PT cells from diabetic rats exhibited higher basal levels of reactive oxygen species (ROS) and higher mitochondrial membrane potential, demonstrating that the PT cells maintain the diabetic phenotype in primary culture. Incubation with either the oxidants or mitochondrial toxicant resulted in greater necrotic and apoptotic cell death, greater evidence of morphological damage, greater increases in ROS, and greater decreases in mitochondrial membrane potential in PT cells from diabetic rats than in those from control rats. Pretreatment with either the antioxidant N-acetyl-L-cysteine or a catalase mimetic provided equivalent protection of PT cells from both diabetic and control rats. Despite the greater susceptibility to oxidative and mitochondrial injury, both cytoplasmic and mitochondrial glutathione concentrations were markedly higher in PT cells from diabetic rats, suggesting an upregulation of antioxidant processes in diabetic kidney. These results support the hypothesis that primary cultures of PT cells from diabetic rats are a valid model in which to study renal cellular function in the diabetic state.

  5. Cadmium activates extracellular signal-regulated kinase 5 in HK-2 human renal proximal tubular cells

    SciTech Connect

    Kondo, Mio; Inamura, Hisako; Matsumura, Ken-ichi; Matsuoka, Masato

    2012-05-11

    Highlights: Black-Right-Pointing-Pointer Cadmium exposure induces ERK5 phosphorylation in HK-2 renal proximal tubular cells. Black-Right-Pointing-Pointer BIX02189 treatment suppresses cadmium-induced ERK5 but not ERK1/2 phosphorylation. Black-Right-Pointing-Pointer BIX02189 treatment suppresses cadmium-induced CREB and c-Fos phosphorylation. Black-Right-Pointing-Pointer ERK5 activation by cadmium exposure may play an anti-apoptotic role in HK-2 cells. -- Abstract: We examined the effects of cadmium chloride (CdCl{sub 2}) exposure on the phosphorylation and functionality of extracellular signal-regulated kinase 5 (ERK5), a recently identified member of the mitogen-activated protein kinase (MAPK) family, in HK-2 human renal proximal tubular cells. Following exposure to CdCl{sub 2}, ERK5 phosphorylation increased markedly, but the level of total ERK5 was unchanged. ERK5 phosphorylation following CdCl{sub 2} exposure was rapid and transient, similar to the time course of ERK1/2 phosphorylation. Treatment of HK-2 cells with the MAPK/ERK kinase 5 inhibitor, BIX02189, suppressed CdCl{sub 2}-induced ERK5 but not ERK1/2 phosphorylation. The CdCl{sub 2}-induced increase of phosphorylated cAMP response element-binding protein (CREB) and activating transcription factor-1 (ATF-1), as well as the accumulation of mobility-shifted c-Fos protein, were suppressed by BIX02189 treatment. Furthermore, BIX02189 treatment enhanced cleavage of poly(ADP-ribose) polymerase and increased the level of cytoplasmic nucleosomes in HK-2 cells exposed to CdCl{sub 2}. These findings suggest that ERK5 pathway activation by CdCl{sub 2} exposure might induce the phosphorylation of cell survival-transcription factors, such as CREB, ATF-1, and c-Fos, and may exert a partial anti-apoptotic role in HK-2 cells.

  6. Importance of adenosine triphosphate in phospholipase A2-induced rabbit renal proximal tubule cell injury.

    PubMed Central

    Nguyen, V D; Cieslinski, D A; Humes, H D

    1988-01-01

    The pathogenesis of ischemic renal tubular cell injury involves a complex interaction of different processes, including membrane phospholipid alterations and depletion of high-energy phosphate stores. To assess the role of membrane phospholipid changes due to activation of phospholipases in renal tubule cell injury, suspensions enriched in rabbit renal proximal tubule segments were incubated with exogenous phospholipase A2 (PLA2). Exogenous PLA2 did not produce any significant change in various metabolic parameters reflective of cell injury in control nonhypoxic preparations despite a significant decrease in phosphatidylethanolamine (PE) and moderate increases in lysophosphatidylcholine (LPC) and lysophosphatidylethanolamine (LPE). In contrast, exogenous PLA2 treatment of hypoxic tubules resulted in a severe degree of cell injury, as demonstrated by marked declines in tubule K+ and ATP contents and significant decreases in tubule uncoupled respiratory rates, and was associated with significant phospholipid alterations, including marked declines in phosphatidylcholine (PC) and PE and significant rises in LPC, LPE, and free fatty acids (FFA). The injurious metabolic effects of exogenous PLA2 on hypoxic tubules were reversed by addition of ATP-MgCl2 to the tubules. The protective effect of ATP-MgCl2 was associated with increases in tubule PC and PE contents and declines in LPC, LPE, and FFA contents. These experiments thus indicate that an increase in exogenous PLA2 activity produces renal proximal tubule cell injury when cell ATP levels decline, at which point phospholipid resynthesis cannot keep pace with phospholipid degradation with resulting depletion of phospholipids and accumulation of lipid by-products. High-energy phosphate store depletion appears to be an important condition for exogenous PLA2 activity to induce renal tubule cell injury. PMID:3417866

  7. PKCθ links proximal T cell and Notch signaling through localized regulation of the actin cytoskeleton

    PubMed Central

    Britton, Graham J; Ambler, Rachel; Clark, Danielle J; Hill, Elaine V; Tunbridge, Helen M; McNally, Kerrie E; Burton, Bronwen R; Butterweck, Philomena; Sabatos-Peyton, Catherine; Hampton-O’Neil, Lea A; Verkade, Paul; Wuelfing, Christoph; Wraith, David Cameron

    2017-01-01

    Notch is a critical regulator of T cell differentiation and is activated through proteolytic cleavage in response to ligand engagement. Using murine myelin-reactive CD4 T cells, we demonstrate that proximal T cell signaling modulates Notch activation by a spatiotemporally constrained mechanism. The protein kinase PKCθ is a critical mediator of signaling by the T cell antigen receptor and the principal costimulatory receptor CD28. PKCθ selectively inactivates the negative regulator of F-actin generation, Coronin 1A, at the center of the T cell interface with the antigen presenting cell (APC). This allows for effective generation of the large actin-based lamellum required for recruitment of the Notch-processing membrane metalloproteinase ADAM10. Such enhancement of Notch activation is critical for efficient T cell proliferation and Th17 differentiation. We reveal a novel mechanism that, through modulation of the cytoskeleton, controls Notch activation at the T cell:APC interface thereby linking T cell receptor and Notch signaling pathways. DOI: http://dx.doi.org/10.7554/eLife.20003.001 PMID:28112644

  8. Proliferation and intracellular pH in cultured proximal tubular cells

    SciTech Connect

    Larsson, S.H.; Fukuda, Y.; Koelare, S.A.; Aperia, A. )

    1990-03-01

    Renal proximal tubule (PT) cells from adult rats will maintain much of their functional characteristics in short-term primary culture. This study examines the growth regulation of these highly differentiated cells with particular reference to cell density, intracellular pH (pHi), and the expression of the Na(+)-H+ exchanger. PT cells were obtained from young adult rats and studied after 48 h in culture. The mitotic rate was determined as the labeling index (LI) after (3H)thymidine autoradiography, and pHi was determined by 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein quantitative fluorescence microscopy in single cells. Cells were grown either continuously in serum (S) or were serum deprived after 24 h (D). The cells were nonconfluent and grew in colonies. We defined the two peripheral layers of cells in a colony as peripheral (P) cells and the remaining cells as central (C). In C cells LI/h and pHi were in the range of what has been observed under in vivo conditions. In S condition LI/h was 2.2 +/- 0.3% and in D condition was 0.3 +/- 0.1%. LI was significantly higher in P than in C cells both under S (2.5 +/- 0.4-fold) and D conditions (5.6 +/- 0.8-fold). The rapidly growing P cells had a significantly lower pHi than the growth-retarded C cells both under S (7.25 +/- 0.02 vs. 7.30 +/- 0.01, P less than 0.05) and D conditions (7.21 +/- 0.02 vs. 7.28 +/- 0.01, P less than 0.05).

  9. Directed Induction of Functional Multi-ciliated Cells in Proximal Airway Epithelial Spheroids from Human Pluripotent Stem Cells

    PubMed Central

    Konishi, Satoshi; Gotoh, Shimpei; Tateishi, Kazuhiro; Yamamoto, Yuki; Korogi, Yohei; Nagasaki, Tadao; Matsumoto, Hisako; Muro, Shigeo; Hirai, Toyohiro; Ito, Isao; Tsukita, Sachiko; Mishima, Michiaki

    2015-01-01

    Summary Multi-ciliated airway cells (MCACs) play a role in mucociliary clearance of the lung. However, the efficient induction of functional MCACs from human pluripotent stem cells has not yet been reported. Using carboxypeptidase M (CPM) as a surface marker of NKX2-1+-ventralized anterior foregut endoderm cells (VAFECs), we report a three-dimensional differentiation protocol for generating proximal airway epithelial progenitor cell spheroids from CPM+ VAFECs. These spheroids could be induced to generate MCACs and other airway lineage cells without alveolar epithelial cells. Furthermore, the directed induction of MCACs and of pulmonary neuroendocrine lineage cells was promoted by adding DAPT, a Notch pathway inhibitor. The induced MCACs demonstrated motile cilia with a “9 + 2” microtubule arrangement and dynein arms capable of beating and generating flow for mucociliary transport. This method is expected to be useful for future studies on human airway disease modeling and regenerative medicine. PMID:26724905

  10. IL-6 Augments Angiotensinogen in Primary Cultured Renal Proximal Tubular Cells

    PubMed Central

    Satou, Ryousuke; Gonzalez-Villalobos, Romer A.; Miyata, Kayoko; Ohashi, Naro; Urushihara, Maki; Acres, Omar W.; Navar, L. Gabriel; Kobori, Hiroyuki

    2009-01-01

    In human kidneys, the mechanisms underlying angiotensinogen (AGT) augmentation by interleukin 6 (IL-6) are poorly understood and the only information available is in HK-2, immortalized human renal proximal tubular epithelial cells. Therefore, the present study was performed to elucidate the effects of IL-6 on AGT expression in primary cultured human renal proximal tubular epithelial cells (RPTEC) after characterization of HK-2 and RPTEC. RPTEC showed low basal AGT mRNA (11±1%) and protein (7.0±0.9%) expression, high IL-6 receptor (IL-6R) expression (282±17%), and low basal NF-κB (43±7%) and STAT3 (43±7%) activities compared to those in HK-2. In RPTEC, AGT mRNA and protein expressions were enhanced by IL-6 (172±31% and 378±39%, respectively). This AGT augmentation was attenuated by an IL-6R antibody. STAT3 phosphorylation (366±55% at 30 min) and translocation were enhanced by IL-6. The AGT augmentation was attenuated by a STAT3 inhibitor. These data indicate that IL-6 increases AGT expression via STAT3 pathway in RPTEC. PMID:19583994

  11. Dopamine and angiotensin type 2 receptors cooperatively inhibit sodium transport in human renal proximal tubule cells.

    PubMed

    Gildea, John J; Wang, Xiaoli; Shah, Neema; Tran, Hanh; Spinosa, Michael; Van Sciver, Robert; Sasaki, Midori; Yatabe, Junichi; Carey, Robert M; Jose, Pedro A; Felder, Robin A

    2012-08-01

    Little is known regarding how the kidney shifts from a sodium and water reclaiming state (antinatriuresis) to a state where sodium and water are eliminated (natriuresis). In human renal proximal tubule cells, sodium reabsorption is decreased by the dopamine D(1)-like receptors (D(1)R/D(5)R) and the angiotensin type 2 receptor (AT(2)R), whereas the angiotensin type 1 receptor increases sodium reabsorption. Aberrant control of these opposing systems is thought to lead to sodium retention and, subsequently, hypertension. We show that D(1)R/D(5)R stimulation increased plasma membrane AT(2)R 4-fold via a D(1)R-mediated, cAMP-coupled, and protein phosphatase 2A-dependent specific signaling pathway. D(1)R/D(5)R stimulation also reduced the ability of angiotensin II to stimulate phospho-extracellular signal-regulated kinase, an effect that was partially reversed by an AT(2)R antagonist. Fenoldopam did not increase AT(2)R recruitment in renal proximal tubule cells with D(1)Rs uncoupled from adenylyl cyclase, suggesting a role of cAMP in mediating these events. D(1)Rs and AT(2)Rs heterodimerized and cooperatively increased cAMP and cGMP production, protein phosphatase 2A activation, sodium-potassium-ATPase internalization, and sodium transport inhibition. These studies shed new light on the regulation of renal sodium transport by the dopaminergic and angiotensin systems and potential new therapeutic targets for selectively treating hypertension.

  12. Na sup + -H sup + exchanger in proximal cells isolated from rabbit kidney. I. Functional characteristics

    SciTech Connect

    Bidet, M.; Tauc, M.; Merot, J.; Vandewalle, A.; Poujeol, P. )

    1987-11-01

    The purpose of this study was to investigate the characteristics of the Na{sup +}-H{sup +} exchange in isolated proximal cells from rabbit kidney cortex. The cells were prepared by mechanical dissociation and sequential passages through nylon meshes. The intracellular pH (pH{sub i}) was measured in a bicarbonate-free medium using the fluorescent dye 2,7-biscarboxyethyl-5(6)-carboxyfluorescein (BCECF). Resting pH{sub i} was 7.13 {+-} 0.04. Cells were acid loaded with nigericin in choline solution and H{sup +} efflux, induced by extracellular Na{sup +} (Na{sub e}), was calculated using a buffering power of 23.6 {+-} 0.6 mmol {center dot} l{sup {minus}1} {center dot} pH unit{sup {minus}1} estimated by NH{sub 4}Cl exposure. The intracellular H{sup +} concentration dependence did not follow simple Michaelis-Menten kinetics. Of the different cations tested on pH{sub i} recovery, such as Li{sup +}, choline{sup +}, K{sup +}, and tetramethylammonium, only Li{sup +} induced an alkalinization of acidified cells similar to that of Na{sup +}. {sup 22}Na influx measurements indicated that cellular depletion of Na{sup +} stimulated Na{sup +}-H{sup +} exchange. The results permit the conclusion that the isolation procedures did not impair the main features of the Na{sup +}-H{sup +} antiporter, at least as compared with those previously described in renal brush-border membrane vesicles or in other cellular systems. The integrity of the transporter in isolated proximal cells would permit the direct study of its hormonal and metabolic control.

  13. Deferasirox-induced iron depletion promotes BclxL downregulation and death of proximal tubular cells.

    PubMed

    Martin-Sanchez, Diego; Gallegos-Villalobos, Angel; Fontecha-Barriuso, Miguel; Carrasco, Susana; Sanchez-Niño, Maria Dolores; Lopez-Hernandez, Francisco J; Ruiz-Ortega, Marta; Egido, Jesus; Ortiz, Alberto; Sanz, Ana Belén

    2017-01-31

    Iron deficiency has been associated with kidney injury. Deferasirox is an oral iron chelator used to treat blood transfusion-related iron overload. Nephrotoxicity is the most serious and common adverse effect of deferasirox and may present as an acute or chronic kidney disease. However, scarce data are available on the molecular mechanisms of nephrotoxicity. We explored the therapeutic modulation of deferasirox-induced proximal tubular cell death in culture. Deferasirox induced dose-dependent tubular cell death and AnexxinV/7AAD staining showed features of apoptosis and necrosis. However, despite inhibiting caspase-3 activation, the pan-caspase inhibitor zVAD-fmk failed to prevent deferasirox-induced cell death. Moreover, zVAD increased deferasirox-induced cell death, a feature sometimes found in necroptosis. Electron microscopy identified mitochondrial injury and features of necrosis. However, neither necrostatin-1 nor RIP3 knockdown prevented deferasirox-induced cell death. Deferasirox caused BclxL depletion and BclxL overexpression was protective. Preventing iron depletion protected from BclxL downregulation and deferasirox cytotoxicity. In conclusion, deferasirox promoted iron depletion-dependent cell death characterized by BclxL downregulation. BclxL overexpression was protective, suggesting a role for BclxL downregulation in iron depletion-induced cell death. This information may be used to develop novel nephroprotective strategies. Furthermore, it supports the concept that monitoring kidney tissue iron depletion may decrease the risk of deferasirox nephrotoxicity.

  14. Mitogenic action of lysophosphatidic acid in proximal tubular epithelial cells obtained from voided human urine.

    PubMed

    Kumagai, N; Inoue, C N; Kondo, Y; Iinuma, K

    2000-12-01

    Focal tubular cell multiplication at sites on an injured nephron is a critical event in the recovery phase following acute tubular necrosis. During this process, numerous viable tubular cells exfoliate and are shed into the urine. Lysophosphatidic acid (LPA) is generated in the plasma membrane of injured cells and acts as an intercellular mediator of various biological processes, including inflammation, proliferation and repair. In the present study, exfoliated proximal tubule (PT) cells were isolated from human urine and the mitogenic effects of LPA were investigated as a model of repair and proliferation following renal injury. LPA stimulated a 23. 5% increase in DNA synthesis, a 29.4% increase in cell number and an 86.6% decrease in cAMP content. All of these responses were pertussis toxin sensitive, indicating the involvement of G(i)-type G-proteins in LPA signalling. Conversely, the LPA-induced DNA synthesis and the decrease in intracellular cAMP content were insensitive to wortmannin, an inhibitor of phosphatidylinositol 3-kinase (PI3K), suggesting a mitogenic response via PI3K-independent mechanisms. Furthermore, we detected specific mRNA transcripts for the recently cloned human LPA-receptors, endothelial differentiation gene (Edg)-2 and Edg-4 (Edg-2>Edg-4) by reverse transcription-PCR in PT cells. Our data suggest that LPA may behave as a local growth factor in PT cells following tubular injury.

  15. Deferasirox-induced iron depletion promotes BclxL downregulation and death of proximal tubular cells

    PubMed Central

    Martin-Sanchez, Diego; Gallegos-Villalobos, Angel; Fontecha-Barriuso, Miguel; Carrasco, Susana; Sanchez-Niño, Maria Dolores; Lopez-Hernandez, Francisco J; Ruiz-Ortega, Marta; Egido, Jesus; Ortiz, Alberto; Sanz, Ana Belén

    2017-01-01

    Iron deficiency has been associated with kidney injury. Deferasirox is an oral iron chelator used to treat blood transfusion-related iron overload. Nephrotoxicity is the most serious and common adverse effect of deferasirox and may present as an acute or chronic kidney disease. However, scarce data are available on the molecular mechanisms of nephrotoxicity. We explored the therapeutic modulation of deferasirox-induced proximal tubular cell death in culture. Deferasirox induced dose-dependent tubular cell death and AnexxinV/7AAD staining showed features of apoptosis and necrosis. However, despite inhibiting caspase-3 activation, the pan-caspase inhibitor zVAD-fmk failed to prevent deferasirox-induced cell death. Moreover, zVAD increased deferasirox-induced cell death, a feature sometimes found in necroptosis. Electron microscopy identified mitochondrial injury and features of necrosis. However, neither necrostatin-1 nor RIP3 knockdown prevented deferasirox-induced cell death. Deferasirox caused BclxL depletion and BclxL overexpression was protective. Preventing iron depletion protected from BclxL downregulation and deferasirox cytotoxicity. In conclusion, deferasirox promoted iron depletion-dependent cell death characterized by BclxL downregulation. BclxL overexpression was protective, suggesting a role for BclxL downregulation in iron depletion-induced cell death. This information may be used to develop novel nephroprotective strategies. Furthermore, it supports the concept that monitoring kidney tissue iron depletion may decrease the risk of deferasirox nephrotoxicity. PMID:28139717

  16. Effects of cumene hydroperoxide on cellular cation composition in frog kidney proximal tubular cells.

    PubMed

    Petrovic, S; Cemerikic, D

    2000-06-01

    Effects of cumene hydroperoxide were studied on the peritubular membrane potential and cellular cation composition in frog kidney proximal tubular cells. After perfusion of isolated frog kidneys for 30 min with 1.3x10(-4) mol l(-1) cumene hydroperoxide Ringer solution, the peritubular membrane potential gradually declined. The ouabain-like effects were demonstrated on cell Na and K activities after 1 h of perfusion with cumene hydroperoxide. The peritubular apparent transference number for potassium was decreased. Intracellular pH was not altered in the presence of cumene hydroperoxide. Intracellular free Ca(2+) concentration increased slowly and moderately. The concentration of the malondialdehyde in the kidney homogenates, measured as an index of lipid peroxidation, was increased. A previously observable effect of cumene hydroperoxide on the peritubular membrane potential was prevented by oxygen radical scavengers.

  17. Phosphoinositide binding differentially regulates NHE1 Na+/H+ exchanger-dependent proximal tubule cell survival.

    PubMed

    Abu Jawdeh, Bassam G; Khan, Shenaz; Deschênes, Isabelle; Hoshi, Malcolm; Goel, Monu; Lock, Jeffrey T; Shinlapawittayatorn, Krekwit; Babcock, Gerald; Lakhe-Reddy, Sujata; DeCaro, Garren; Yadav, Satya P; Mohan, Maradumane L; Naga Prasad, Sathyamangla V; Schilling, William P; Ficker, Eckhard; Schelling, Jeffrey R

    2011-12-09

    Tubular atrophy predicts chronic kidney disease progression, and is caused by proximal tubular epithelial cellcaused by proximal tubular epithelial cell (PTC) apoptosis. The normally quiescent Na(+)/H(+) exchanger-1 (NHE1) defends against PTC apoptosis, and is regulated by PI(4,5)P(2) binding. Because of the vast array of plasma membrane lipids, we hypothesized that NHE1-mediated cell survival is dynamically regulated by multiple anionic inner leaflet phospholipids. In membrane overlay and surface plasmon resonance assays, the NHE1 C terminus bound phospholipids with low affinity and according to valence (PIP(3) > PIP(2) > PIP = PA > PS). NHE1-phosphoinositide binding was enhanced by acidic pH, and abolished by NHE1 Arg/Lys to Ala mutations within two juxtamembrane domains, consistent with electrostatic interactions. PI(4,5)P(2)-incorporated vesicles were distributed to apical and lateral PTC domains, increased NHE1-regulated Na(+)/H(+) exchange, and blunted apoptosis, whereas NHE1 activity was decreased in cells enriched with PI(3,4,5)P(3), which localized to basolateral membranes. Divergent PI(4,5)P(2) and PI(3,4,5)P(3) effects on NHE1-dependent Na(+)/H(+) exchange and apoptosis were confirmed by selective phosphoinositide sequestration with pleckstrin homology domain-containing phospholipase Cδ and Akt peptides, PI 3-kinase, and Akt inhibition in wild-type and NHE1-null PTCs. The results reveal an on-off switch model, whereby NHE1 toggles between weak interactions with PI(4,5)P(2) and PI(3,4,5)P(3). In response to apoptotic stress, NHE1 is stimulated by PI(4,5)P(2), which leads to PI 3-kinase activation, and PI(4,5)P(2) phosphorylation. The resulting PI(3,4,5)P(3) dually stimulates sustained, downstream Akt survival signaling, and dampens NHE1 activity through competitive inhibition and depletion of PI(4,5)P(2).

  18. Etoposide induced cytotoxicity mediated by ROS and ERK in human kidney proximal tubule cells

    PubMed Central

    Shin, Hyeon-Jun; Kwon, Hyuk-Kwon; Lee, Jae-Hyeok; Anwar, Muhammad Ayaz; Choi, Sangdun

    2016-01-01

    Etoposide (ETO) is a commonly used chemotherapeutic drug that inhibits topoisomerase II activity, thereby leading to genotoxicity and cytotoxicity. However, ETO has limited application due to its side effects on normal organs, especially the kidney. Here, we report the mechanism of ETO-induced cytotoxicity progression in human kidney proximal tubule (HK-2) cells. Our results show that ETO perpetuates DNA damage, activates mitogen-activated protein kinase (MAPK), and triggers morphological changes, such as cell and nuclear swelling. When NAC, a well-known reactive oxygen species (ROS) scavenger, is co-treated with ETO, it inhibits an ETO-induced increase in mitochondrial mass, mitochondrial DNA (ND1 and ND4) copy number, intracellular ATP level, and mitochondrial biogenesis activators (TFAM, PGC-1α and PGC-1β). Moreover, co-treatment with ETO and NAC inhibits ETO-induced necrosis and cell swelling, but not apoptosis. Studies using MAPK inhibitors reveal that inhibition of extracellular signal regulated kinase (ERK) protects ETO-induced cytotoxicity by inhibiting DNA damage and caspase 3/7 activity. Eventually, ERK inhibitor treated cells are protected from ETO-induced nuclear envelope (NE) rupture and DNA leakage through inhibition of caspase activity. Taken together, these data suggest that ETO mediates cytotoxicity in HK-2 cells through ROS and ERK pathways, which highlight the preventive avenues in ETO-induced cytotoxicity in kidney. PMID:27666530

  19. Cell swelling, co-transport activation and potassium conductance in isolated perfused rabbit kidney proximal tubules.

    PubMed Central

    Beck, J S; Potts, D J

    1990-01-01

    1. Isolated, perfused rabbit proximal tubules were used to study the effects of activation of the apical membrane sodium co-transporters, and of the effects of osmotically induced cell swelling, upon cell volume, basolateral membrane potential and apparent partial conductance of potassium. 2. Activation of electrogenic apical sodium co-transport caused a depolarization of the basolateral membrane and a reduction of the basolateral apparent potassium transference number. This was followed by a spontaneous partial recovery of potential and increase in apparent potassium transference number. 3. Stimulation of apical sodium co-transport led to a sustained increase in cell volume. 4. A sustained increase in cell volume (of similar magnitude to that seen after activation of apical membrane sodium co-transporters) was also caused by reduction of bath and perfusate osmolality by removal of 89 mmol l-1 mannitol from both lumen and bath solutions. 5. This reduction in bath and perfusate osmolality also led to a basolateral membrane hyperpolarization and an increase in basolateral apparent potassium transference number. 6. These observations support the possibility that some of the partial recovery of basolateral membrane potential (Vb1) during apical sodium co-transport stimulation is due to a cell volume sensitive change in basolateral potassium conductance. PMID:2213582

  20. Short term exposure to elevated levels of leptin reduces proximal tubule cell metabolic activity.

    PubMed

    Briffa, Jessica F; Grinfeld, Esther; McAinch, Andrew J; Poronnik, Philip; Hryciw, Deanne H

    2014-01-25

    Leptin plays a pathophysiological role in the kidney, however, its acute effects on the proximal tubule cells (PTCs) are unknown. In opossum kidney (OK) cells in vitro, Western blot analysis identified that exposure to leptin increases the phosphorylation of the mitogen-activated protein kinase (MAPK) p44/42 and the mammalian target of rapamycin (mTOR). Importantly leptin (0.05, 0.10, 0.25 and 0.50 μg/ml) significantly reduced the metabolic activity of PTCs, and significantly decreased protein content per cell. Investigation of the role of p44/42 and mTOR on metabolic activity and protein content per cell, demonstrated that in the presence of MAPK inhibitor U0126 and mTOR inhibitor Ku-63794, that the mTOR pathway is responsible for the reduction in PTC metabolic activity in response to leptin. However, p44/42 and mTOR play no role the reduced protein content per cell in OKs exposed to leptin. Therefore, leptin modulates metabolic activity in PTCs via an mTOR regulated pathway.

  1. A critical synopsis: Continuous growth of proximal tubular kidney epithelial cells in hormone-supplemented serum-free medium

    NASA Technical Reports Server (NTRS)

    Chuman, L. M.; FINE; COHEN; Saier, M. H.

    1985-01-01

    The kidney forms urine and reabsorbs electrolytes and water. Kidney cell lines and hormone supplemented serum free medium were used for growth. The hormones were insulin, transferrin, vasopressin, cholesterol, prostaglandins, hydrocortisone, and triidothyronine. Epithelial cell lines are polar and form hemicysts. The Madin-Darby canine kidney(MDCK) cell line used is distal tubulelike. LLC-PK sub 1 cells are derived from pig kidneys and have the properties of different kidney segments. The LLC-PK sub 1 cells with proximal tubule properties were maintained in hormone-supplemented serum free medium. Seven factors (the aforementioned homrones and selenium) were needed for growth. Hormone-defined medium supported LLC-PK sub 1 cell growth, allowed transport (as seen by hemicyst formation), and influenced cell morphology. Vasopressin (used for growth and morphology) could be partially replaced by isobutylmethylxanthine or dibutyryl cAMP. The defined medium was used to isolate rabbit proximal tubule kidney epithelial cells free of fibroblasts.

  2. Wnt signaling induces transcription, spatial proximity, and translocation of fusion gene partners in human hematopoietic cells.

    PubMed

    Ugarte, Giorgia D; Vargas, Macarena F; Medina, Matías A; León, Pablo; Necuñir, David; Elorza, Alvaro A; Gutiérrez, Soraya E; Moon, Randall T; Loyola, Alejandra; De Ferrari, Giancarlo V

    2015-10-08

    Chromosomal translocations are frequently associated with a wide variety of cancers, particularly hematologic malignancies. A recurrent chromosomal abnormality in acute myeloid leukemia is the reciprocal translocation t(8;21) that fuses RUNX1 and ETO genes. We report here that Wnt/β-catenin signaling increases the expression of ETO and RUNX1 genes in human hematopoietic progenitors. We found that β-catenin is rapidly recruited into RNA polymerase II transcription factories (RNAPII-Ser5) and that ETO and RUNX1 genes are brought into close spatial proximity upon Wnt3a induction. Notably, long-term treatment of cells with Wnt3a induces the generation a frequent RUNX1-ETO translocation event. Thus, Wnt/β-catenin signaling induces transcription and translocation of RUNX1 and ETO fusion gene partners, opening a novel window to understand the onset/development of leukemia.

  3. GIANT CELL TUMOR IN THE PROXIMAL PHALANX WITH PULMONARY METASTASIS: CASE REPORT AND LITERATURE REVIEW

    PubMed Central

    de Medeiros, Frederico Carvalho; de Medeiros, Fernando Carvalho; de Campos Carvalho Lopes, Izabella; de Medeiros, Guilherme Carvalho; de Medeiros, Eduardo Carvalho

    2015-01-01

    This is a case report on a giant cell tumor (GCT) in the proximal phalanx of the third finger of the left hand, with pulmonary metastasis. The patient presented pain in the finger without any previous history of trauma. Clinical examination, radiographic imaging and magnetic resonance imaging were carried out. A histological evaluation was carried out from an incisional biopsy, taking the hypothesis of GCT. The patient underwent amputation of the finger and the diagnosis was confirmed by means of microscopy on the specimen. The patient was followed up because of the risk of lung metastasis, which was shown by radiographic examination and computed tomography on the chest, and thoracotomy was performed. Since then, there has been an improvement in the symptoms that had been reported preoperatively, and no local recurrence or new metastasis has been found. PMID:27027012

  4. Intracellular trafficking pathway of BK virus in human renal proximal tubular epithelial cells

    PubMed Central

    Moriyama, Takahito; Sorokin, Andrey

    2009-01-01

    Intracellular trafficking of BK Virus (BKV) in human renal proximal tubular epithelial cells (HRPTEC) is critical for BKV nephritis. However, the major trafficking components utilized by BKV remain unknown. Co-incubation of HRPTEC with BKV and microtubule disrupting agents prevented BKV infection as detected by immunofluorescence and western blot analysis with antibodies which recognize BKV large T antigen. However, inhibition of a dynein, cellular motor protein, did not interfere with BKV infection in HRPTEC. A colocalization study of BKV with the markers of the endoplasmic reticulum (ER) and the Golgi apparatus (GA), indicated that BKV reached the ER from 6 to 10 hours, while bypassing the GA or passing through the GA too transiently to be detected. This study contributes to the understanding of mechanisms of intracellular trafficking used by BKV in the infection of HRPTEC. PMID:17976677

  5. Akt Links Insulin Signaling to Albumin Endocytosis in Proximal Tubule Epithelial Cells.

    PubMed

    Coffey, Sam; Costacou, Tina; Orchard, Trevor; Erkan, Elif

    2015-01-01

    Diabetes mellitus (DM) has become an epidemic, causing a significant decline in quality of life of individuals due to its multisystem involvement. Kidney is an important target organ in DM accounting for the majority of patients requiring renal replacement therapy at dialysis units. Microalbuminuria (MA) has been a valuable tool to predict end-organ damage in DM but its low sensitivity has driven research efforts to seek other alternatives. Albumin is taken up by albumin receptors, megalin and cubilin in the proximal tubule epithelial cells. We demonstrated that insulin at physiological concentrations induce albumin endocytosis through activation of protein kinase B (Akt) in proximal tubule epithelial cells. Inhibition of Akt by a phosphorylation deficient construct abrogated insulin induced albumin endocytosis suggesting a role for Akt in insulin-induced albumin endocytosis. Furthermore we demonstrated a novel interaction between Akt substrate 160kDa (AS160) and cytoplasmic tail of megalin. Mice with type 1 DM (T1D) displayed decreased Akt, megalin, cubilin and AS160 expression in their kidneys in association with urinary cubilin shedding preceding significant MA. Patients with T1D who have developed MA in the EDC (The Pittsburgh Epidemiology of Diabetes Complications) study demonstrated urinary cubilin shedding prior to development of MA. We hypothesize that perturbed insulin-Akt cascade in DM leads to alterations in trafficking of megalin and cubilin, which results in urinary cubilin shedding as a prelude to MA in early diabetic nephropathy. We propose that utilization of urinary cubilin shedding, as a urinary biomarker, will allow us to detect and intervene in diabetic nephropathy (DN) at an earlier stage.

  6. Label retaining cells (LRCs) with myoepithelial characteristic from the proximal acinar region define stem cells in the sweat gland.

    PubMed

    Leung, Yvonne; Kandyba, Eve; Chen, Yi-Bu; Ruffins, Seth; Kobielak, Krzysztof

    2013-01-01

    Slow cycling is a common feature shared among several stem cells (SCs) identified in adult tissues including hair follicle and cornea. Recently, existence of unipotent SCs in basal and lumenal layers of sweat gland (SG) has been described and label retaining cells (LRCs) have also been localized in SGs; however, whether these LRCs possess SCs characteristic has not been investigated further. Here, we used a H2BGFP LRCs system for in vivo detection of infrequently dividing cells. This system allowed us to specifically localize and isolate SCs with label-retention and myoepithelial characteristics restricted to the SG proximal acinar region. Using an alternative genetic approach, we demonstrated that SG LRCs expressed keratin 15 (K15) in the acinar region and lineage tracing determined that K15 labeled cells contributed long term to the SG structure but not to epidermal homeostasis. Surprisingly, wound healing experiments did not activate proximal acinar SG cells to participate in epidermal healing. Instead, predominantly non-LRCs in the SG duct actively divided, whereas the majority of SG LRCs remained quiescent. However, when we further challenged the system under more favorable isolated wound healing conditions, we were able to trigger normally quiescent acinar LRCs to trans-differentiate into the epidermis and adopt its long term fate. In addition, dissociated SG cells were able to regenerate SGs and, surprisingly, hair follicles demonstrating their in vivo plasticity. By determining the gene expression profile of isolated SG LRCs and non-LRCs in vivo, we identified several Bone Morphogenetic Protein (BMP) pathway genes to be up-regulated and confirmed a functional requirement for BMP receptor 1A (BMPR1A)-mediated signaling in SG formation. Our data highlight the existence of SG stem cells (SGSCs) and their primary importance in SG homeostasis. It also emphasizes SGSCs as an alternative source of cells in wound healing and their plasticity for regenerating

  7. Characterization of hyaluronan cable structure and function in renal proximal tubular epithelial cells.

    PubMed

    Selbi, W; de la Motte, C A; Hascall, V C; Day, A J; Bowen, T; Phillips, A O

    2006-10-01

    Alteration in the glycosaminoglycan hyaluronan (HA) has been demonstrated in numerous renal diseases. We have demonstrated that renal proximal tubular epithelial cells (PTCs) surround themselves in vitro with HA in an organized pericellular matrix or 'coat', which is associated with cell migration, and also form pericellular HA cable-like structures which modulate PTC-mononuclear leukocytes interactions. The aim of this study was to characterize potential regulatory mechanism in the assembly of PTC-HA into pericellular cables. HA cables are generated by PTCs in the absence of serum. Immunohistochemical analysis demonstrates the incorporation of components of the inter-alpha-inhibitor (IalphaI) family of proteins and versican into HA cables. Addition of an antibody to IalphaI/PalphaI (pre-alpha-inhibitor) inhibits cable formation. In contrast, inhibition of tumor necrosis factor-alpha-stimulated gene 6 (TSG-6) has no effect on cable formation, suggesting that their generation is independent of the known heavy-chain transfer activity of TSG-6. Overexpression of HAS3 is associated with induction of HA cable formation, and also increased incorporation of HA into pericellular coats. Functionally, this resulted in enhanced HA-dependent monocyte binding and cell migration, respectively. Cell surface expression of CD44 and trypsin-released cell-associated HA were increased in HAS3-overexpressing cells. In addition, hyaluronidase (hyal1 and hyal2) and bikunin mRNA expression were increased, whereas PalphaI HC3 mRNA expression was unchanged in the transfected cells. The data demonstrate the importance of IalphaI/PalphaI in cable formation and suggest that expression of HAS3 may be critical for HA cable assembly.

  8. Interleukin 2 mediates stimulation of complement C3 biosynthesis in human proximal tubular epithelial cells.

    PubMed Central

    Brooimans, R A; Stegmann, A P; van Dorp, W T; van der Ark, A A; van der Woude, F J; van Es, L A; Daha, M R

    1991-01-01

    Previous reports have suggested the production of complement components C4, C2, and factor B by renal tissue. However, the cells involved in production of complement have not been identified. In this study metabolic labeling experiments demonstrated that human proximal tubular epithelial cells (PTEC) synthesize a 180-kD precursor of C3 that is secreted after proteolytic cleavage into a disulphide linked two-chain molecule as found in plasma. C3 present in culture supernatants of PTEC was functionally active, however, during the culture period there was a partial inactivation of the C3 molecule as assessed by hemolytic titration. Recombinant IL-2 enhances the rate of C3 synthesis in a dose-dependent manner reaching maximal stimulation at doses of 200-400 U/ml IL-2. Northern blot analysis demonstrated a 5.2-kb C3 mRNA species present in PTEC that was increased within 24 h of IL-2 treatment. IL-2-induced enhancement of C3 production by PTEC could be neutralized with specific antibodies to IL-2. This study demonstrates that C3 synthesis in PTEC is upregulated by IL-2, the major cytokine produced by activated T cells. Images PMID:1864952

  9. Human proximal tubule epithelial cells cultured on hollow fibers: living membranes that actively transport organic cations.

    PubMed

    Jansen, J; De Napoli, I E; Fedecostante, M; Schophuizen, C M S; Chevtchik, N V; Wilmer, M J; van Asbeck, A H; Croes, H J; Pertijs, J C; Wetzels, J F M; Hilbrands, L B; van den Heuvel, L P; Hoenderop, J G; Stamatialis, D; Masereeuw, R

    2015-11-16

    The bioartificial kidney (BAK) aims at improving dialysis by developing 'living membranes' for cells-aided removal of uremic metabolites. Here, unique human conditionally immortalized proximal tubule epithelial cell (ciPTEC) monolayers were cultured on biofunctionalized MicroPES (polyethersulfone) hollow fiber membranes (HFM) and functionally tested using microfluidics. Tight monolayer formation was demonstrated by abundant zonula occludens-1 (ZO-1) protein expression along the tight junctions of matured ciPTEC on HFM. A clear barrier function of the monolayer was confirmed by limited diffusion of FITC-inulin. The activity of the organic cation transporter 2 (OCT2) in ciPTEC was evaluated in real-time using a perfusion system by confocal microscopy using 4-(4-(dimethylamino)styryl)-N-methylpyridinium iodide (ASP(+)) as a fluorescent substrate. Initial ASP(+) uptake was inhibited by a cationic uremic metabolites mixture and by the histamine H2-receptor antagonist, cimetidine. In conclusion, a 'living membrane' of renal epithelial cells on MicroPES HFM with demonstrated active organic cation transport was successfully established as a first step in BAK engineering.

  10. Human proximal tubule epithelial cells cultured on hollow fibers: living membranes that actively transport organic cations

    PubMed Central

    Jansen, J.; De Napoli, I. E; Fedecostante, M.; Schophuizen, C. M. S.; Chevtchik, N. V.; Wilmer, M. J.; van Asbeck, A. H.; Croes, H. J.; Pertijs, J. C.; Wetzels, J. F. M.; Hilbrands, L. B.; van den Heuvel, L. P.; Hoenderop, J. G.; Stamatialis, D.; Masereeuw, R.

    2015-01-01

    The bioartificial kidney (BAK) aims at improving dialysis by developing ‘living membranes’ for cells-aided removal of uremic metabolites. Here, unique human conditionally immortalized proximal tubule epithelial cell (ciPTEC) monolayers were cultured on biofunctionalized MicroPES (polyethersulfone) hollow fiber membranes (HFM) and functionally tested using microfluidics. Tight monolayer formation was demonstrated by abundant zonula occludens-1 (ZO-1) protein expression along the tight junctions of matured ciPTEC on HFM. A clear barrier function of the monolayer was confirmed by limited diffusion of FITC-inulin. The activity of the organic cation transporter 2 (OCT2) in ciPTEC was evaluated in real-time using a perfusion system by confocal microscopy using 4-(4-(dimethylamino)styryl)-N-methylpyridinium iodide (ASP+) as a fluorescent substrate. Initial ASP+ uptake was inhibited by a cationic uremic metabolites mixture and by the histamine H2-receptor antagonist, cimetidine. In conclusion, a ‘living membrane’ of renal epithelial cells on MicroPES HFM with demonstrated active organic cation transport was successfully established as a first step in BAK engineering. PMID:26567716

  11. Iron repletion relocalizes hephaestin to a proximal basolateral compartment in polarized MDCK and Caco2 cells

    SciTech Connect

    Lee, Seung-Min; Attieh, Zouhair K.; Son, Hee Sook; Chen, Huijun; Bacouri-Haidar, Mhenia; Vulpe, Chris D.

    2012-05-11

    Highlights: Black-Right-Pointing-Pointer Hephaestin localizes in the perinuclear space in non-polarized cells. Black-Right-Pointing-Pointer Hephaestin localizes in the perinuclear space in iron deficient and polarized cells. Black-Right-Pointing-Pointer Hephaestin with apical iron moves near to basolateral membrane of polarized cells. Black-Right-Pointing-Pointer Peri-basolateral location of hephaestin is accessible to the extracellular space. Black-Right-Pointing-Pointer Hephaestin is involved in iron mobilization from the intestine to circulation. -- Abstract: While intestinal cellular iron entry in vertebrates employs multiple routes including heme and non-heme routes, iron egress from these cells is exclusively channeled through the only known transporter, ferroportin. Reduced intestinal iron export in sex-linked anemia mice implicates hephaestin, a ferroxidase, in this process. Polarized cells are exposed to two distinct environments. Enterocytes contact the gut lumen via the apical surface of the cell, and through the basolateral surface, to the body. Previous studies indicate both local and systemic control of iron uptake. We hypothesized that differences in iron availability at the apical and/or basolateral surface may modulate iron uptake via cellular localization of hephaestin. We therefore characterized the localization of hephaestin in two models of polarized epithelial cell lines, MDCK and Caco2, with varying iron availability at the apical and basolateral surfaces. Our results indicate that hephaestin is expressed in a supra-nuclear compartment in non-polarized cells regardless of the iron status of the cells and in iron deficient and polarized cells. In polarized cells, we found that both apical (as FeSO{sub 4}) and basolateral iron (as the ratio of apo-transferrin to holo-transferrin) affect mobilization of hephaestin from the supra-nuclear compartment. We find that the presence of apical iron is essential for relocalization of hephaestin to a

  12. Scintillators and applications thereof

    DOEpatents

    Williams, Richard T.

    2015-09-01

    Scintillators of various constructions and methods of making and using the same are provided. In some embodiments, a scintillator comprises at least one radiation absorption region and at least one spatially discrete radiative exciton recombination region.

  13. Scintillators and applications thereof

    DOEpatents

    Williams, Richard T.

    2014-07-15

    Scintillators of various constructions and methods of making and using the same are provided. In some embodiments, a scintillator comprises at least one radiation absorption region and at least one spatially discrete radiative exciton recombination region.

  14. Effect of gentamicin on phospholipid metabolism in cultured rabbit proximal tubular cells

    SciTech Connect

    Ramsammy, L.S.; Josepovitz, C.; Lane, B.; Kaloyanides, G.J.

    1989-01-01

    We examined the hypothesis that the accumulation of phospholipid in cells exposed to gentamicin is due to impaired degradation. Experiments were performed in rabbit proximal tubular cells grown in primary culture. Cells exposed to 10(-3) M gentamicin manifested myeloid body formation and a progressive increase in total phospholipid that by day 6 was 44% higher than that of control cells and reflected increases of phosphatidylinositol of 235%, phosphatidylcholine of 60%, phosphatidylethanolamine of 90%, and phosphatidylserine of 55% above control values. Gentamicin impaired the degradation of these phospholipids. The t1/2 of the phospholipid pool labeled with (3H)myoinositol increased 146% from 1.17 (control) to 2.88 days (gentamicin); the t1/2 of the (3H)choline pool increased 34% from 1.77 to 2.38 days; the t1/2 of the (3H)ethanolamine pool increased 57% from 3.14 to 4.93 days; the t1/2 of the (3H) serine pool increased 37% from 6.30 to 8.63 days. Exposure of cells to gentamicin for 2 days also stimulated increased incorporation of (3H)myoinositol (68%) and (3H)ethanolamine (59%) into phospholipid. The data are consistent with the hypothesis that gentamicin inhibits the activity of lysosomal phospholipases that results in the accumulation of phospholipid within the lysosome in the form of myeloid bodies. Increased phospholipid synthesis may represent a compensatory response to the impaired lysosomal degradation of phospholipid. We postulate that the preferential increase of phosphatidylinositol reflects the capacity of the polycationic gentamicin to interact electrostatically with the anionic phosphoinositides and inhibit their turnover.

  15. MDR1 transporter protects against paraquat-induced toxicity in human and mouse proximal tubule cells.

    PubMed

    Wen, Xia; Gibson, Christopher J; Yang, Ill; Buckley, Brian; Goedken, Michael J; Richardson, Jason R; Aleksunes, Lauren M

    2014-10-01

    Paraquat is a herbicide that is highly toxic to the lungs and kidneys following acute exposures. Prior studies have demonstrated that the organic cation transporter 2 and multidrug and toxin extrusion protein 1 contribute to the urinary secretion of paraquat in the kidneys. The purpose of this study was to determine whether the multidrug resistance protein 1 (MDR1/Mdr1, ABCB1, or P-glycoprotein) also participates in the removal of paraquat from the kidneys and protects against renal injury. Paraquat transport and toxicity were quantified in human renal proximal tubule epithelial cells (RPTEC) that endogenously express MDR1, HEK293 cells overexpressing MDR1, and Mdr1a/1b knockout mice. In RPTEC cells, reduction of MDR1 activity using the antagonist PSC833 or siRNA transfection increased the cellular accumulation of paraquat by 50%. Reduced efflux of paraquat corresponded with enhanced cytotoxicity in PSC833-treated cells. Likewise, stable overexpression of the human MDR1 gene in HEK293 cells reduced intracellular levels of paraquat by 50%. In vivo studies assessed the renal accumulation and subsequent nephrotoxicity of paraquat (10 or 30 mg/kg ip) in wild-type and Mdr1a/1b knockout mice. At 4 h after paraquat treatment, renal concentrations of paraquat in the kidneys of Mdr1a/1b knockout mice were 750% higher than wild-type mice. By 72 h, paraquat-treated Mdr1a/1b knockout mice had more extensive tubular degeneration and significantly greater mRNA expression of kidney injury-responsive genes, including kidney injury molecule-1, lipocalin-2, and NAD(P)H quinone oxidoreductase 1, compared with wild-type mice. In conclusion, MDR1/Mdr1 participates in the elimination of paraquat from the kidneys and protects against subsequent toxicity.

  16. Scintillator materials for calorimetry

    SciTech Connect

    Weber, M.J.

    1994-09-01

    Requirements for fast, dense scintillator materials for calorimetry in high energy physics and approaches to satisfying these requirements are reviewed with respect to possible hosts and luminescent species. Special attention is given to cerium-activated crystals, core-valence luminescence, and glass scintillators. The present state of the art, limitations, and suggestions for possible new scintillator materials are presented.

  17. Scintillator manufacture at Fermilab

    SciTech Connect

    Mellott, K.; Bross, A.; Pla-Dalmau, A.

    1998-08-01

    A decade of research into plastic scintillation materials at Fermilab is reviewed. Early work with plastic optical fiber fabrication is revisited and recent experiments with large-scale commercial methods for production of bulk scintillator are discussed. Costs for various forms of scintillator are examined and new development goals including cost reduction methods and quality improvement techniques are suggested.

  18. Lead carbonate scintillator materials

    DOEpatents

    Derenzo, Stephen E.; Moses, William W.

    1991-01-01

    Improved radiation detectors containing lead carbonate or basic lead carbonate as the scintillator element are disclosed. Both of these scintillators have been found to provide a balance of good stopping power, high light yield and short decay constant that is superior to other known scintillator materials. The radiation detectors disclosed are favorably suited for use in general purpose detection and in medical uses.

  19. Lack of induction of single-strand breaks in mammalian cells by sodium azide and its proximal mutagen

    SciTech Connect

    Arenaz, P.; Nilan, R.A.; Kleinhofs, A.

    1981-01-01

    The mutagenicity of sodium azide in both higher plants and bacteria is well documented. However, in mammalian cells, research on the effects of azide on gene mutations has produced conflicting results. Furthermore, no research has been conducted on the effects of azide and its proximal mutagen (mutagenic metabolite) on single-strand breaks. The experiments herein were designed to overcome this lack of information on azide mutagenicity and to evaluate the potential hazard of azide exposure to man. Chinese hamster V/sub 79/ cells were treated with either azide or its proximal mutagen(s) for 2 h or 6 h respectively and analyzed by alkaline elution for single-strand breaks. The data showed that there was no effect of either azide or the proximal mutagen(s) on single-strand DNA breaks nor was there any indication that azide or the proximal mutagen(s) induced DNA-protein crosslinks. The data suggest that neither azide nor its proximal mutagen(s) interact directly with DNA and suggest that the methods may be applied to any specified population and dose scenario.

  20. BAG3 regulates ECM accumulation in renal proximal tubular cells induced by TGF-β1.

    PubMed

    Du, Feng; Li, Si; Wang, Tian; Zhang, Hai-Yan; Li, De-Tian; Du, Zhen-Xian; Wang, Hua-Qin; Wang, Yan-Qiu

    2015-01-01

    Previously we have demonstrated that Bcl-2-associated athanogene 3 (BAG3) is increased in renal fibrosis using a rat unilateral ureteral obstruction model. The current study investigated the role of BAG3 in renal fibrosis using transforming growth factor (TGF)-β1-treated human proximal tubular epithelial (HK-2) cells. An upregulation of BAG3 in vitro models was observed, which correlated with the increased synthesis of extracellular matrix (ECM) proteins and expression of tissue-type plasminogen activator inhibitor (PAI)-1. Blockade of BAG3 induction by shorting hairpin RNA suppressed the expression of ECM proteins but had no effect on PAI-1 expression induced by TGF-β1. Forced overexpression of BAG3 selectively increased collagens. TGF-β1-induced BAG3 expression in HK-2 cells was attenuated by ERK1/2 and JNK MAPK inhibitors. In addition, forced BAG3 overexpression blocked attenuation of collagens expression by ERK1/2 and JNK inhibitors. These data suggest that ERK1/2 and JNK signaling events are involved in modulating the expression of BAG3, which would ultimately contribute to renal fibrosis by enhancing the synthesis and deposition of ECM proteins.

  1. T-cell functional regions of the human IL-3 proximal promoter.

    PubMed

    Ryan, G R; Vadas, M A; Shannon, M F

    1994-10-01

    The human interleukin-3 (IL-3) gene is expressed almost exclusively in activated T cells. Its expression is regulated at both the transcriptional and post-transcriptional level. We have previously shown that treatment of Jurkat T cells with phytohemaglutinin (PHA) and the phorbol ester, PMA, activated transcription initiation from the IL-3 gene. To define the regions of the gene required for transcription activation, we generated a series of reporter constructs containing different regions of the IL-3 gene 5' and 3' flanking sequences. Both positive and negative regulatory elements were identified in the proximal 5' flanking region of the IL-3 gene. The promoter region between -173 and -60 contained the strongest activating elements. The transcription factor AP-1 could bind to this positive activator region of the promoter. We also examined the function of the IL-3 CK-1/CK-2 elements that are present in many cytokine genes and found that they acted as a repressor of basal level expression when cloned upstream of a heterologous promoter but were also inducible by PMA/PHA.

  2. Acute leptin exposure reduces megalin expression and upregulates TGFβ1 in cultured renal proximal tubule cells.

    PubMed

    Briffa, Jessica F; Grinfeld, Esther; Mathai, Michael L; Poronnik, Phillip; McAinch, Andrew J; Hryciw, Deanne H

    2015-02-05

    Increased leptin concentrations observed in obesity can lead to proteinuria, suggesting that leptin may play a role in obesity-related kidney disease. Obesity reduces activation of AMP-activated protein kinase (AMPK) and increases transforming growth factor-β1 (TGF-β1) expression in the kidney, leading to albuminuria. Thus we investigated if elevated leptin altered AMPK and TGF-β1 signaling in proximal tubule cells (PTCs). In opossum kidney (OK) PTCs Western blot analysis demonstrated that leptin upregulates TGF-β1 secretion (0.50 µg/ml) and phosphorylated AMPKα (at 0.25, and 0.50 µg/ml), and downregulates megalin expression at all concentrations (0.05-0.50 µg/ml). Using the AMPK inhibitor, Compound C, leptin exposure regulated TGF-β1 expression and secretion in PTCs via an AMPK mediated pathway. In addition, elevated leptin exposure (0.50 µg/ml) reduced albumin handling in OK cells independently of megalin expression. This study demonstrates that leptin upregulates TGF-β1, reduces megalin, and reduces albumin handling in PTCs by an AMPK mediated pathway.

  3. The Replisomes Remain Spatially Proximal throughout the Cell Cycle in Bacteria

    PubMed Central

    Mangiameli, Sarah M.; Veit, Brian T.; Wiggins, Paul A.

    2017-01-01

    The positioning of the DNA replication machinery (replisome) has been the subject of several studies. Two conflicting models for replisome localization have been proposed: In the Factory Model, sister replisomes remain spatially co-localized as the replicating DNA is translocated through a stationary replication factory. In the Track Model, sister replisomes translocate independently along a stationary DNA track and the replisomes are spatially separated for the majority of the cell cycle. Here, we used time-lapse imaging to observe and quantify the position of fluorescently labeled processivity-clamp (DnaN) complexes throughout the cell cycle in two highly-divergent bacterial model organisms: Bacillus subtilis and Escherichia coli. Because DnaN is a core component of the replication machinery, its localization patterns should be an appropriate proxy for replisome positioning in general. We present automated statistical analysis of DnaN positioning in large populations, which is essential due to the high degree of cell-to-cell variation. We find that both bacteria show remarkably similar DnaN positioning, where any potential separation of the two replication forks remains below the diffraction limit throughout the majority of the replication cycle. Additionally, the localization pattern of several other core replisome components is consistent with that of DnaN. These data altogether indicate that the two replication forks remain spatially co-localized and mostly function in close proximity throughout the replication cycle. The conservation of the observed localization patterns in these highly divergent species suggests that the subcellular positioning of the replisome is a functionally critical feature of DNA replication. PMID:28114307

  4. The Replisomes Remain Spatially Proximal throughout the Cell Cycle in Bacteria.

    PubMed

    Mangiameli, Sarah M; Veit, Brian T; Merrikh, Houra; Wiggins, Paul A

    2017-01-01

    The positioning of the DNA replication machinery (replisome) has been the subject of several studies. Two conflicting models for replisome localization have been proposed: In the Factory Model, sister replisomes remain spatially co-localized as the replicating DNA is translocated through a stationary replication factory. In the Track Model, sister replisomes translocate independently along a stationary DNA track and the replisomes are spatially separated for the majority of the cell cycle. Here, we used time-lapse imaging to observe and quantify the position of fluorescently labeled processivity-clamp (DnaN) complexes throughout the cell cycle in two highly-divergent bacterial model organisms: Bacillus subtilis and Escherichia coli. Because DnaN is a core component of the replication machinery, its localization patterns should be an appropriate proxy for replisome positioning in general. We present automated statistical analysis of DnaN positioning in large populations, which is essential due to the high degree of cell-to-cell variation. We find that both bacteria show remarkably similar DnaN positioning, where any potential separation of the two replication forks remains below the diffraction limit throughout the majority of the replication cycle. Additionally, the localization pattern of several other core replisome components is consistent with that of DnaN. These data altogether indicate that the two replication forks remain spatially co-localized and mostly function in close proximity throughout the replication cycle. The conservation of the observed localization patterns in these highly divergent species suggests that the subcellular positioning of the replisome is a functionally critical feature of DNA replication.

  5. Autophagy protects kidney proximal tubule epithelial cells from mitochondrial metabolic stress.

    PubMed

    Kimura, Tomonori; Takahashi, Atsushi; Takabatake, Yoshitsugu; Namba, Tomoko; Yamamoto, Takeshi; Kaimori, Jun-Ya; Matsui, Isao; Kitamura, Harumi; Niimura, Fumio; Matsusaka, Taiji; Soga, Tomoyoshi; Rakugi, Hiromi; Isaka, Yoshitaka

    2013-11-01

    Chronic metabolic stress is related to diseases, whereas autophagy supplies nutrients by recycling the degradative products. Cyclosporin A (CsA), a frequently used immunosuppressant, induces metabolic stress via effects on mitochondrial respiration, and thereby, its chronic usage is often limited. Here we show that autophagy plays a protective role against CsA-induced metabolic stress in kidney proximal tubule epithelial cells. Autophagy deficiency leads to decreased mitochondrial membrane potential, which coincides with metabolic abnormalities as characterized by decreased levels of amino acids, increased tricarboxylic acid (TCA) ratio (the levels of intermediates of the latter part of the TCA cycle, over levels of intermediates in the earlier part), and decreased products of oxidative phosphorylation (ATP). In addition to the altered profile of amino acids, CsA decreased the hyperpolarization of mitochondria with the disturbance of mitochondrial energy metabolism in autophagy-competent cells, i.e., increased TCA ratio and worsening of the NAD(+)/NADH ratio, coupled with decreased energy status, which suggests that adaptation to CsA employs autophagy to supply electron donors from amino acids via intermediates of the latter part of the TCA cycle. The TCA ratio of autophagy-deficient cells was further worsened with decreased levels of amino acids in response to CsA, and, as a result, the deficiency of autophagy failed to adapt to the CsA-induced metabolic stress. Deterioration of the TCA ratio further worsened energy status. The CsA-induced metabolic stress also activated regulatory genes of metabolism and apoptotic signals, whose expressions were accelerated in autophagy-deficient cells. These data provide new perspectives on autophagy in conditions of chronic metabolic stress in disease.

  6. Cysteamine restores glutathione redox status in cultured cystinotic proximal tubular epithelial cells.

    PubMed

    Wilmer, Martijn J; Kluijtmans, Leo A J; van der Velden, Thea J; Willems, Peter H; Scheffer, Peter G; Masereeuw, Rosalinde; Monnens, Leo A; van den Heuvel, Lambertus P; Levtchenko, Elena N

    2011-06-01

    Recent evidence implies that impaired metabolism of glutathione has a role in the pathogenesis of nephropathic cystinosis. This recessive inherited disorder is characterized by lysosomal cystine accumulation and results in renal Fanconi syndrome progressing to end stage renal disease in the majority of patients. The most common treatment involves intracellular cystine depletion by cysteamine, delaying the development of end stage renal disease by a yet elusive mechanism. However, cystine depletion does not arrest the disease nor cures Fanconi syndrome in patients, indicating involvement of other yet unknown pathologic pathways. Using a newly developed proximal tubular epithelial cell model from cystinotic patients, we investigate the effect of cystine accumulation and cysteamine on both glutathione and ATP metabolism. In addition to the expected increase in cystine and defective sodium-dependent phosphate reabsorption, we observed less negative glutathione redox status and decreased intracellular ATP levels. No differences between control and cystinosis cell lines were observed with respect to protein turnover, albumin uptake, cytosolic and mitochondrial ATP production, total glutathione levels, protein oxidation and lipid peroxidation. Cysteamine treatment increased total glutathione in both control and cystinotic cells and normalized cystine levels and glutathione redox status in cystinotic cells. However, cysteamine did not improve decreased sodium-dependent phosphate uptake. Our data implicate that cysteamine increases total glutathione and restores glutathione redox status in cystinosis, which is a positive side-effect of this agent next to cystine depletion. This beneficial effect points to a potential role of cysteamine as anti-oxidant for other renal disorders associated with enhanced oxidative stress.

  7. Scintillator reflective layer coextrusion

    DOEpatents

    Yun, Jae-Chul; Para, Adam

    2001-01-01

    A polymeric scintillator has a reflective layer adhered to the exterior surface thereof. The reflective layer comprises a reflective pigment and an adhesive binder. The adhesive binder includes polymeric material from which the scintillator is formed. A method of forming the polymeric scintillator having a reflective layer adhered to the exterior surface thereof is also provided. The method includes the steps of (a) extruding an inner core member from a first amount of polymeric scintillator material, and (b) coextruding an outer reflective layer on the exterior surface of the inner core member. The outer reflective layer comprises a reflective pigment and a second amount of the polymeric scintillator material.

  8. Dead cell and side leakage correction for a lead-scintillating fiber electromagnetic calorimeter

    NASA Astrophysics Data System (ADS)

    Zhang, Cheng; Li, Zu-Hao; Tang, Zhi-Cheng; Basegmez du Pree, Suzan; Zhang, Shao-Wen; Wang, Xue-Qiang; Yang, Min; Chen, Guo-Ming; Chen, He-Sheng

    2016-09-01

    The electromagnetic calorimeter (ECAL) of the Alpha Magnetic Spectrometer (AMS-02) is one of the key detectors for dark matter searches. It measures the energies of electrons, positrons and photons and seperates them from hadrons. Currently, there are 5 dead cells in the ECAL, which affect the reconstructed energy of 4.2% of total events in the ECAL acceptance. When an electromagnetic shower axis is close to the ECAL border, due to the side leakage, the reconstructed energy is affected as well. In this paper, methods for dead cells and side leakage corrections for the ECAL energy reconstruction are presented. For events with the shower axis crossing dead cells, applying dead cell correction improves the difference in the reconstructed energy from 12% to 1%, while for events near the ECAL border, with side leakage correction it is improved from 4% to 1%. Supported by National Natural Science Foundation of China(11220101004)

  9. Electrical and freeze-fracture analysis of the effects of ionic cadmium on cell membranes of human proximal tubule cells.

    PubMed Central

    Hazen-Martin, D J; Todd, J H; Sens, M A; Khan, W; Bylander, J E; Smyth, B J; Sens, D A

    1993-01-01

    We previously reported that cell cultures of human proximal tubule (HPT) cells respond to ionic cadmium in a manner consistent with well-defined Cd(2+)-elicited responses reported for in vivo systems. However, one unique finding was that the transepithelial electrical resistance and tight junction sealing strands were altered as a result of Cd2+ exposure at micromolar concentrations. These alterations are reexamined in detail in the present report to determine whether the Cd(2+)-induced alterations are specific alterations in the tight junction structure or reflect a general alteration in the cell membrane. Exhaustive analysis of tight junction sealing strands demonstrated no significant alterations due to Cd2+ exposure, even at the concentration that elicited a significant reduction in transepithelial resistance. Further analysis of intramembrane particle distribution demonstrated a significant increase in apical intramembrane particles, indicating that Cd2+ exposure altered the characteristics of the apical cell membrane. Overall, the results were consistent with evidence of Cd(2+)-induced alteration in the apical cell membrane of the HPT cell. Images Figure 1. Figure 2. Figure 3. a Figure 3. b Figure 3. c Figure 3. d Figure 4. Figure 5. PMID:8137780

  10. Effects of SGLT2 inhibition in human kidney proximal tubular cells--renoprotection in diabetic nephropathy?

    PubMed

    Panchapakesan, Usha; Pegg, Kate; Gross, Simon; Komala, Muralikrishna Gangadharan; Mudaliar, Harshini; Forbes, Josephine; Pollock, Carol; Mather, Amanda

    2013-01-01

    Sodium/glucose cotransporter 2 (SGLT2) inhibitors are oral hypoglycemic agents used to treat patients with diabetes mellitus. SGLT2 inhibitors block reabsorption of filtered glucose by inhibiting SGLT2, the primary glucose transporter in the proximal tubular cell (PTC), leading to glycosuria and lowering of serum glucose. We examined the renoprotective effects of the SGLT2 inhibitor empagliflozin to determine whether blocking glucose entry into the kidney PTCs reduced the inflammatory and fibrotic responses of the cell to high glucose. We used an in vitro model of human PTCs. HK2 cells (human kidney PTC line) were exposed to control 5 mM, high glucose (HG) 30 mM or the profibrotic cytokine transforming growth factor beta (TGFβ1; 0.5 ng/ml) in the presence and absence of empagliflozin for up to 72 h. SGLT1 and 2 expression and various inflammatory/fibrotic markers were assessed. A chromatin immunoprecipitation assay was used to determine the binding of phosphorylated smad3 to the promoter region of the SGLT2 gene. Our data showed that TGFβ1 but not HG increased SGLT2 expression and this occurred via phosphorylated smad3. HG induced expression of Toll-like receptor-4, increased nuclear deoxyribonucleic acid binding for nuclear factor kappa B (NF-κB) and activator protein 1, induced collagen IV expression as well as interleukin-6 secretion all of which were attenuated with empagliflozin. Empagliflozin did not reduce high mobility group box protein 1 induced NF-κB suggesting that its effect is specifically related to a reduction in glycotoxicity. SGLT1 and GLUT2 expression was not significantly altered with HG or empagliflozin. In conclusion, empagliflozin reduces HG induced inflammatory and fibrotic markers by blocking glucose transport and did not induce a compensatory increase in SGLT1/GLUT2 expression. Although HG itself does not regulate SGLT2 expression in our model, TGFβ increases SGLT2 expression through phosphorylated smad3.

  11. Hormone-specific regulation of the kidney androgen-regulated gene promoter in cultured mouse renal proximal-tubule cells.

    PubMed Central

    Soler, Montse; Tornavaca, Olga; Solé, Esther; Menoyo, Anna; Hardy, Dianne; Catterall, James F; Vandewalle, Alain; Meseguer, Anna

    2002-01-01

    The kidney androgen-regulated protein (KAP) is specifically expressed and differentially regulated by androgens and tri-iodothyronine (T(3)) in intact mouse early (PCT) and late (PR) proximal-tubule cells. Until now, detailed characterization of the molecular elements mediating androgen-responsive gene expression in the kidney has been hampered by the lack of appropriate cultured cell systems suitable for DNA transfection studies. In the present study we have analysed the hormone-dependent transactivation of the KAP gene promoter in immortalized differentiated PCT and PR proximal-tubule cells derived from L-PK/Tag1 transgenic mice. Transient transfection studies with different KAP promoter constructs indicated that a 224 bp-truncated fragment was sufficient to mediate cell-specific expression of the KAP promoter. Dihydrotestosterone (DHT) stimulated in an androgen-dependent manner the transactivation of KAP in PCT and PR cells, while mutation of a putative androgen-response element (ARE) sequence located at -39 bp from the transcription initiation site abolished the transactivation induced by DHT. Furthermore, insulin-like growth factor 1 (IGF-1), but not T(3), enhanced the androgen-dependent transactivation of KAP in cultured PCT cells. These results demonstrate that the short 224 bp fragment of the KAP promoter is sufficient to drive the proximal-tubule androgen-specific regulated expression of KAP and reveal synergistic interactions between IGF-1 and androgens for KAP regulation in PCT cells. PMID:12030848

  12. The renoprotective role of autophagy activation in proximal tubular epithelial cells in diabetic nephropathy.

    PubMed

    Xu, Ying; Liu, Lei; Xin, Wei; Zhao, Xu; Chen, Liyong; Zhen, Junhui; Wan, Qiang

    2015-01-01

    With intensive investigations recently, autophagy is hoped to be a potential therapeutic target to prevent or alleviate diabetic nephropathy (DN). Our previous study revealed that lipotoxicity participated in epithelial-to-mesenchymal transition (EMT) of proximal tubular epithelial cells (PTECs) under diabetic conditions. Based on evidences that autophagy and lipid metabolism are closely related, we investigated autophagy under diabetic conditions and how it contributed in the lipotoxicity and EMT. In high-glucose-cultured PTECs, we found that Beclin1 and LC3-II were elevated, while p62 was decreased. These results indicate that autophagy activity was elevated under diabetic conditions. Autophagy deficiency induced by autophagy inhibitors, chloroquine diphosphate (CQ) and 3-Methyladenine (3-MA), and by Atg5 siRNA transfection exacerbated lipid accumulation and EMT. This supports that the elevated autophagy activity acts as a renoprotective response under diabetic conditions. Treatment of rapamycin, which is a mammalian target of rapamycin (mTOR) receptor-specific inhibitor and a known autophagy activator, attenuated high-glucose-induced lipid accumulation and EMT. The Atg5 silence counteracted the protective effect of rapamycin. The present study deepens our understanding of the role of autophagy in DN, suggesting a complex interplay of autophagy, metabolic pathways, lipotoxicity and EMT.

  13. The Endocytic Receptor Megalin and its Associated Proteins in Proximal Tubule Epithelial Cells

    PubMed Central

    De, Shankhajit; Kuwahara, Shoji; Saito, Akihiko

    2014-01-01

    Receptor-mediated endocytosis in renal proximal tubule epithelial cells (PTECs) is important for the reabsorption and metabolization of proteins and other substances, including carrier-bound vitamins and trace elements, in glomerular filtrates. Impairment of this endocytic process results in the loss of such substances and development of proteinuria, which is an important clinical indicator of kidney diseases and is also a risk marker for cardiovascular disease. Megalin, a member of the low-density lipoprotein receptor gene family, is a multiligand receptor expressed in the apical membrane of PTECs and plays a central role in the endocytic process. Megalin interacts with various intracellular adaptor proteins for intracellular trafficking and cooperatively functions with other membrane molecules, including the cubilin-amnionless complex. Evidence suggests that megalin and the cubilin-amnionless complex are involved in the uptake of toxic substances into PTECs, which leads to the development of kidney disease. Studies of megalin and its associated molecules will be useful for future development of novel strategies for the diagnosis and treatment of kidney diseases. PMID:25019425

  14. In vitro safety assessment of food ingredients in canine renal proximal tubule cells.

    PubMed

    Koči, J; Jeffery, B; Riviere, J E; Monteiro-Riviere, N A

    2015-03-01

    In vitro models are useful tools to initially assess the toxicological safety hazards of food ingredients. Toxicities of cinnamaldehyde (CINA), cinnamon bark oil, lemongrass oil (LGO), thymol, thyme oil (TO), clove leaf oil, eugenol, ginger root extract (GRE), citric acid, guanosine monophosphate, inosine monophosphate and sorbose (SORB) were assessed in canine renal proximal tubule cells (CPTC) using viability assay and renal injury markers. At LC50, CINA was the most toxic (0.012mg/ml), while SORB the least toxic (>100mg/ml). Toxicities (LC50) of positive controls were as follows: 4-aminophenol (0.15mg/ml in CPTC and 0.083mg/ml in human PTC), neomycin (28.6mg/ml in CPTC and 27.1mg/ml in human PTC). XYL displayed lowest cytotoxic potency (LC50=82.7mg/ml in CPTC). In vivo renal injury markers in CPTC were not significantly different from controls. The LGO toxicity mechanism was analyzed using qPCR and electron microscopy. Out of 370 genes, 57 genes (15.4%) were significantly up (34, 9.1%) or down (23, 6.2%) regulated, with the most upregulated gene gsta3 (∼200-fold) and the most affected pathway being oxidative stress. LGO induced damage of mitochondria, phospholipid accumulation and lack of a brush border. Viability assays along with mechanistic studies in the CPTC model may serve as a valuable in vitro toxicity screening tool.

  15. Calcium dependence of integrity of the actin cytoskeleton of proximal tubule cell microvilli.

    PubMed

    Sogabe, K; Roeser, N F; Davis, J A; Nurko, S; Venkatachalam, M A; Weinberg, J M

    1996-08-01

    To better define the role of Ca2+ in pathophysiological alterations of the proximal tubule microvillus actin cytoskeleton, we studied freshly isolated tubules in which intracellular free Ca2+ was equilibrated with highly buffered, precisely defined medium Ca2+ levels using a combination of the metabolic inhibitor, antimycin, and the ionophore, ionomycin, in the presence of glycine, to prevent lethal membrane damage and resulting nonspecific changes. Increases of Ca2+ to > or = 10 microM were sufficient to initiate concurrent actin depolymerization, fragmentation of F-actin into forms requiring high-speed centrifugation for recovery, redistribution of villin to sedimentable fractions, and structural microvillar damage consisting of severe swelling and fragmentation of actin cores. These observations implicate Ca(2+)-dependent, villin-mediated actin cytoskeletal disruption in tubule cell microvillar damage under conditions conceivably present during pathophysiological states. However, despite prior evidence for cytosolic free Ca2+ increases of the same order of magnitude and similar structural microvillar alterations, Ca(2+)- and villin-mediated events did not appear to account for the initial microvillar damage that occurs during ATP depletion induced by antimycin alone or hypoxia.

  16. The CpG Island in the Murine Foxl2 Proximal Promoter Is Differentially Methylated in Primary and Immortalized Cells

    PubMed Central

    Tran, Stella; Wang, Ying; Lamba, Pankaj; Zhou, Xiang; Boehm, Ulrich; Bernard, Daniel J.

    2013-01-01

    Forkhead box L2 (Foxl2), a member of the forkhead transcription factor family, plays important roles in pituitary follicle-stimulating hormone synthesis and in ovarian maintenance and function. Mutations in the human FOXL2 gene cause eyelid malformations and premature ovarian failure. FOXL2/Foxl2 is expressed in pituitary gonadotrope and thyrotrope cells, the perioptic mesenchyme of the developing eyelid, and ovarian granulosa cells. The mechanisms governing this cell-restricted expression have not been described. We mapped the Foxl2 transcriptional start site in immortalized murine gonadotrope-like cells, LβT2, by 5’ rapid amplification of cDNA ends and then PCR amplified approximately 1 kb of 5’ flanking sequence from murine genomic DNA. When ligated into a reporter plasmid, the proximal promoter conferred luciferase activity in both homologous (LβT2) and, unexpectedly, heterologous (NIH3T3) cells. In silico analyses identified a CpG island in the proximal promoter and 5’ untranslated region, suggesting that Foxl2 transcription might be regulated epigenetically. Indeed, pyrosequencing and quantitative analysis of DNA methylation using real-time PCR revealed Foxl2 proximal promoter hypomethylation in homologous compared to some, though not all, heterologous cell lines. The promoter was also hypomethylated in purified murine gonadotropes. In vitro promoter methylation completely silenced reporter activity in heterologous and homologous cells. Collectively, the data suggest that differential proximal promoter DNA methylation may contribute to cell-specific Foxl2 expression in some cellular contexts. However, gonadotrope-specific expression of the gene cannot be explained by promoter hypomethylation alone. PMID:24098544

  17. Nitro-Arachidonic Acid Prevents Angiotensin II-Induced Mitochondrial Dysfunction in a Cell Line of Kidney Proximal Tubular Cells.

    PubMed

    Sánchez-Calvo, Beatriz; Cassina, Adriana; Rios, Natalia; Peluffo, Gonzalo; Boggia, José; Radi, Rafael; Rubbo, Homero; Trostchansky, Andres

    2016-01-01

    Nitro-arachidonic acid (NO2-AA) is a cell signaling nitroalkene that exerts anti-inflammatory activities during macrophage activation. While angiotensin II (ANG II) produces an increase in reactive oxygen species (ROS) production and mitochondrial dysfunction in renal tubular cells, little is known regarding the potential protective effects of NO2-AA in ANG II-mediated kidney injury. As such, this study examines the impact of NO2-AA on ANG II-induced mitochondrial dysfunction in an immortalized renal proximal tubule cell line (HK-2 cells). Treatment of HK-2 cells with ANG II increases the production of superoxide (O2●-), nitric oxide (●NO), inducible nitric oxide synthase (NOS2) expression, peroxynitrite (ONOO-) and mitochondrial dysfunction. Using high-resolution respirometry, it was observed that the presence of NO2-AA prevented ANG II-mediated mitochondrial dysfunction. Attempting to address mechanism, we treated isolated rat kidney mitochondria with ONOO-, a key mediator of ANG II-induced mitochondrial damage, in the presence or absence of NO2-AA. Whereas the activity of succinate dehydrogenase (SDH) and ATP synthase (ATPase) were diminished upon exposure to ONOO-, they were restored by pre-incubating the mitochondria with NO2-AA. Moreover, NO2-AA prevents oxidation and nitration of mitochondrial proteins. Combined, these data demonstrate that ANG II-mediated oxidative damage and mitochondrial dysfunction is abrogated by NO2-AA, identifying this compound as a promising pharmacological tool to prevent ANG II-induced renal disease.

  18. Protein Kinase C-δ Mediates Shedding of Angiotensin-Converting Enzyme 2 from Proximal Tubular Cells

    PubMed Central

    Xiao, Fengxia; Zimpelmann, Joseph; Burger, Dylan; Kennedy, Christopher; Hébert, Richard L.; Burns, Kevin D.

    2016-01-01

    Angiotensin-converting enzyme 2 (ACE2) degrades angiotensin (Ang) II to Ang-(1–7), and protects against diabetic renal injury. Soluble ACE2 fragments are shed from the proximal tubule, and appear at high levels in the urine with diabetes. High glucose-induced shedding of ACE2 from proximal tubular cells is mediated by the enzyme “a disintegrin and metalloproteinase-17″ (ADAM17). Here, we investigated the mechanism for constitutive shedding of ACE2. Mouse proximal tubular cells were cultured and ACE2 shedding into the media was assessed by enzyme activity assay and immunoblot analysis. Cells were incubated with pharmacologic inhibitors, or transfected with silencing (si) RNA. Incubation of proximal tubular cells with increasing concentrations of D-glucose stimulated ACE2 shedding, which peaked at 16 mM, while L-glucose (osmotic control) had no effect on shedding. In cells maintained in 7.8 mM D-glucose, ACE2 shedding was significantly inhibited by the pan-protein kinase C (PKC) competitive inhibitor sotrastaurin, but not by an inhibitor of ADAM17. Incubation of cells with the PKC-α and -β1-specific inhibitor Go6976, the PKC β1 and β2-specific inhibitor ruboxistaurin, inhibitors of matrix metalloproteinases-2,-8, and -9, or an inhibitor of ADAM10 (GI250423X) had no effect on basal ACE2 shedding. By contrast, the PKC-δ inhibitor rottlerin significantly inhibited both constitutive and high glucose-induced ACE2 shedding. Transfection of cells with siRNA directed against PKC-δ reduced ACE2 shedding by 20%, while knockdown of PKC-ε was without effect. These results indicate that constitutive shedding of ACE2 from proximal tubular cells is mediated by PKC-δ, which is also linked to high glucose-induced shedding. Targeting PKC-δ may preserve membrane-bound ACE2 in proximal tubule in disease states and diminish Ang II-stimulated adverse signaling. PMID:27313531

  19. Packed Red Blood Cells Are an Abundant and Proximate Potential Source of Nitric Oxide Synthase Inhibition

    PubMed Central

    Zwemer, Charles F.; Davenport, Robertson D.; Gomez-Espina, Juan; Blanco-Gonzalez, Elisa; Whitesall, Steven E.; D'Alecy, Louis G.

    2015-01-01

    Objective We determined, for packed red blood cells (PRBC) and fresh frozen plasma, the maximum content, and ability to release the endogenous nitric oxide synthase (NOS) inhibitors asymmetric dimethylarginine (ADMA) and monomethylarginine (LNMMA). Background ADMA and LNMMA are near equipotent NOS inhibitors forming blood’s total NOS inhibitory content. The balance between removal from, and addition to plasma determines their free concentrations. Removal from plasma is by well-characterized specific hydrolases while formation is restricted to posttranslational protein methylation. When released into plasma they can readily enter endothelial cells and inhibit NOS. Fresh rat and human whole blood contain substantial protein incorporated ADMA however; the maximum content of ADMA and LNMMA in PRBC and fresh frozen plasma has not been determined. Methods We measured total (free and protein incorporated) ADMA and LNMMA content in PRBCs and fresh frozen plasma, as well as their incubation induced release, using HPLC with fluorescence detection. We tested the hypothesis that PRBC and fresh frozen plasma contain substantial inhibitory methylarginines that can be released chemically by complete in vitro acid hydrolysis or physiologically at 37°C by enzymatic blood proteolysis. Results In vitro strong-acid-hydrolysis revealed a large PRBC reservoir of ADMA (54.5 ± 9.7 µM) and LNMMA (58.9 ± 28.9 μM) that persisted over 42-d at 6° or -80°C. In vitro 5h incubation at 37°C nearly doubled free ADMA and LNMMNA concentration from PRBCs while no change was detected in fresh frozen plasma. Conclusion The compelling physiological ramifications are that regardless of storage age, 1) PRBCs can rapidly release pathologically relevant quantities of ADMA and LNMMA when incubated and 2) PRBCs have a protein-incorporated inhibitory methylarginines reservoir 100 times that of normal free inhibitory methylarginines in blood and thus could represent a clinically relevant and proximate

  20. Specific estrogen-induced cell proliferation of cultured Syrian hamster renal proximal tubular cells in serum-free chemically defined media

    SciTech Connect

    Oberley, T.D.; Lauchner, L.J.; Pugh, T.D.; Gonzalez, A.; Goldfarb, S. ); Li, S.A.; Li, J.J. )

    1989-03-01

    It has long been recognized that the renal proximal tubular epithelium of the hamster is a bona fide estrogen target tissue. The effect of estrogens on the growth of proximal tubule cell explants and dissociated single cells derived from these explant outgrowths has been studied in culture. Renal tubular cells were grown on a PF-HR-9 basement membrane under serum-free chemically defined culture conditions. At 7-14 days in culture, cell number was enhanced 3-fold in the presence of either 17{beta}-estradiol or diethylstilbestrol. A similar 3-fold increase in cell number was also seen at 1 nM 17{beta}-estradiol in subcultured dissociated single tubular cells derived from hamster renal tubular explant outgrowths at 21 days in culture. Concomitant exposure of tamoxifen at 3-fold molar excess in culture completely abolished the increase in cell number seen with 17{beta}-estradiol. The proliferation effect of estrogens on proximal tubular cell growth appears to be species specific since 17{beta}-estradiol did not alter the growth of either rat or guinea pig proximal tubules in culture. In addition, at 7-10 days in culture in the presence of 17{beta}-estradiol, ({sup 3}H)thymidine labeling of hamster tubular cells was enhanced 3-fold. These results clearly indicate that estrogens can directly induce primary epithelial cell proliferation at physiologic concentrations and provide strong additional evidence for an important hormonal role in the neoplastic transformation of the hamster kidney.

  1. Intestinal Neuronal Dysplasia-Like Submucosal Ganglion Cell Hyperplasia at the Proximal Margins of Hirschsprung Disease Resections.

    PubMed

    Swaminathan, Maya; Oron, Assaf P; Chatterjee, Sumantra; Piper, Hannah; Cope-Yokoyama, Sandy; Chakravarti, Aravinda; Kapur, Raj P

    2015-01-01

    Intestinal neuronal dysplasia type B (IND) denotes an increased proportion of hyperplastic submucosal ganglia, as resolved histochemically in 15-μm-thick frozen sections. IND has been reported proximal to the aganglionic segment in patients with Hirschsprung disease (HSCR) and is putatively associated with a higher rate of postsurgical dysmotility. We developed and validated histological criteria to diagnose IND-like submucosal ganglion cell hyperplasia (IND-SH) in paraffin sections and used the approach to study the incidence and clinical and/or genetic associations of IND-SH at the proximal margins of HSCR pull-through resection specimens. Full-circumference paraffin sections from the proximal margins of 64 HSCR colonic pull-through specimens and 24 autopsy controls were immunostained for neuron-specific Hu antigen, and nucleated ganglion cells in each submucosal ganglion were counted. In controls, an age-related decline in the relative abundance of "giant" ganglia (≥7 nucleated Hu-positive [Hu+] ganglion cells) was observed. A conservative diagnostic threshold for IND-SH (control mean ± 3× standard deviation) was derived from 15 controls less than 25 weeks of age. No control exceeded this threshold, whereas in the same age range, IND-SH was observed at the proximal margins in 15% (7 of 46) of HSCR resections, up to 15 cm proximal to the aganglionic segment. No significant correlation was observed between IND-SH and length of or distance from the aganglionic segment, sex, trisomy 21, RET or SEMA3C/D polymorphisms, or clinical outcome, but analysis of more patients, with better long-term follow-up will be required to clarify the significance of this histological phenotype.

  2. Isolation and Characterization of a Primary Proximal Tubular Epithelial Cell Model from Human Kidney by CD10/CD13 Double Labeling

    PubMed Central

    Gnemmi, Viviane; Glowacki, François; Pottier, Nicolas; Bouillez, Audrey; Maboudou, Patrice; Zini, Laurent; Leroy, Xavier; Cauffiez, Christelle; Perrais, Michaël; Aubert, Sébastien

    2013-01-01

    Renal proximal tubular epithelial cells play a central role in renal physiology and are among the cell types most sensitive to ischemia and xenobiotic nephrotoxicity. In order to investigate the molecular and cellular mechanisms underlying the pathophysiology of kidney injuries, a stable and well-characterized primary culture model of proximal tubular cells is required. An existing model of proximal tubular cells is hampered by the cellular heterogeneity of kidney; a method based on cell sorting for specific markers must therefore be developed. In this study, we present a primary culture model based on the mechanical and enzymatic dissociation of healthy tissue obtained from nephrectomy specimens. Renal epithelial cells were sorted using co-labeling for CD10 and CD13, two renal proximal tubular epithelial markers, by flow cytometry. Their purity, phenotypic stability and functional properties were evaluated over several passages. Our results demonstrate that CD10/CD13 double-positive cells constitute a pure, functional and stable proximal tubular epithelial cell population that displays proximal tubule markers and epithelial characteristics over the long term, whereas cells positive for either CD10 or CD13 alone appear to be heterogeneous. In conclusion, this study describes a method for establishing a robust renal proximal tubular epithelial cell model suitable for further experimentation. PMID:23799132

  3. Measurement of radionuclides using ion chromatography and flow-cell scintillation counting with pulse shape discrimination

    SciTech Connect

    DeVol, T.A.; Fjeld, R.A.

    1995-10-01

    The use of ion chromatography (IC) for radiochemical separations is a well established technique. IC is commonly used in routine environmental monitoring applications as well as in specialized research applications. Typical usage involves the separation of a single radionuclide from the non-radioactive constituents. During the past decade, a limited amount of research has been conducted using automated IC systems in actinide separation applications (e.g.). More recently, separation procedures for common non-gamma emitting activation and fission products were developed utilizing a high performance liquid chromatography (HPLC) system. In addition, a separation procedure for six common actinides has been developed using a HPLC system. These latter systems used on-line flow-cell detectors for quantification of the radioactive constituents of the effluent stream.

  4. Study of equatorial scintillations

    NASA Technical Reports Server (NTRS)

    Pomalaza, J.; Woodman, R.; Tisnado, G.; Nakasone, E.

    1972-01-01

    Observations of the amplitude scintillations produced by the F-region in equatorial areas are presented. The equipment used for conducting the observations is described. The use of transmissions from the ATS-1, ATS-3, and ATS-5 for obtaining data is described. The two principal subjects discussed are: (1) correlation between satellite and incoherent radar observations of scintillations and (2) simultaneous observations of scintillations at 136 MHz and 1550 MHz.

  5. Lead carbonate scintillator materials

    DOEpatents

    Derenzo, S.E.; Moses, W.W.

    1991-05-14

    Improved radiation detectors containing lead carbonate or basic lead carbonate as the scintillator element are disclosed. Both of these scintillators have been found to provide a balance of good stopping power, high light yield and short decay constant that is superior to other known scintillator materials. The radiation detectors disclosed are favorably suited for use in general purpose detection and in medical uses. 3 figures.

  6. NKT cell modulates NAFLD potentiation of metabolic oxidative stress-induced mesangial cell activation and proximal tubular toxicity

    PubMed Central

    Alhasson, Firas; Dattaroy, Diptadip; Das, Suvarthi; Chandrashekaran, Varun; Seth, Ratanesh Kumar; Schnellmann, Rick G.

    2015-01-01

    Obesity and nonalcoholic fatty liver disease (NAFLD) are associated with the development and progression of chronic kidney disease. We recently showed that NAFLD induces liver-specific cytochrome P-450 (CYP)2E1-mediated metabolic oxidative stress after administration of the CYP2E1 substrate bromodichloromethane (BDCM) (Seth RK, Das S, Kumar A, Chanda A, Kadiiska MB, Michelotti G, Manautou J, Diehl AM, Chatterjee S. Toxicol Appl Pharmacol 274: 42–54, 2014; Seth RK, Kumar A, Das S, Kadiiska MB, Michelotti G, Diehl AM, Chatterjee S. Toxicol Sci 134:291–303, 2013). The present study examined the effects of CYP2E1-mediated oxidative stress in NAFLD leading to kidney toxicity. Mice were fed a high-fat diet for 12 wk to induce NAFLD. NAFLD mice were exposed to BDCM, a CYP2E1 substrate, for 4 wk. NAFLD + BDCM increased CYP2E1-mediated lipid peroxidation in proximal tubular cells compared with mice with NAFLD alone or BDCM-treated lean mice, thus ruling out the exclusive role of BDCM. Lipid peroxidation increased IL-1β, TNF-α, and interferon-γ. In parallel, mesangial cell activation was observed by increased α-smooth muscle actin and transforming growth factor-β, which was blocked by the CYP2E1 inhibitor diallyl sulphide both in vivo and in vitro. Mice lacking natural killer T cells (CD1d knockout mice) showed elevated (>4-fold) proinflammatory mediator release, increased Toll-like receptor (TLR)4 and PDGF2 mRNA, and mesangial cell activation in the kidney. Finally, NAFLD CD1D knockout mice treated with BDCM exhibited increased high mobility group box 1 and Fas ligand levels and TUNEL-positive nuclei, indicating that higher cell death was attenuated in TLR4 knockout mice. Tubular cells showed increased cell death and cytokine release when incubated with activated mesangial cells. In summary, an underlying condition of progressive NAFLD causes renal immunotoxicity and aberrant glomerular function possibly through high mobility group box 1-dependent TLR4 signaling

  7. Ginsenosides protect apical transporters of cultured proximal tubule cells from dysfunctions induced by h(2)o(2).

    PubMed

    Han, Ho Jae; Yoon, Byung Cheol; Park, Soo Hyun; Park, Ji Yeong; Oh, Young Joon; Lee, Yun Jung; Park, Kwon Moo

    2002-01-01

    Oxidative stress has been implicated as a primary cause of renal failure in certain renal diseases. Indeed, renal proximal tubule is a very sensitive site to oxidative stress and retains functionally fully characterized transporters. It has been reported that ginsenosides have a beneficial effect on diverse diseases including oxidative stress. However, the protective effect of ginsenosides on oxidative stress has not been elucidated in renal proximal tubule cells. Thus, we examined the effect of ginsenosides on oxidative stress-induced alteration of apical transporters and its related mechanism in renal proximal tubule cells. In the present study, hydrogen peroxide (H(2)O(2)) (>10(-5) M) inhibited alpha-methyl-D-glucopyranoside uptake in a dose-dependent manner (p < 0.05). It also inhibited Pi and Na(+) uptake. At a concentration of 20 microg/ml, total ginsenosides significantly reduced H(2)O(2)-induced inhibition of apical transporters. In contrast, protopanaxadiol (PD) and protopanaxatriol (PT) saponins exhibited a less preventive effect than total ginsenosides (p < 0.05). Furthermore, we examined its action mechanism. H(2)O(2) increased lipid peroxide formation, arachidonic acid (AA) release, and Ca(2+) uptake. These effects on H(2)O(2) were significantly prevented by total ginsenosides and PD or PT sanponins. However, total ginsenosides appear to be more protective than PD and PT saponins (p < 0.05). In conclusion, ginsenosides prevented H(2)O(2)-induced inhibition of apical transporters via a decrease in oxidative stress, AA release, and Ca(2+) uptake in primary cultured renal proximal tubule cells.

  8. Proximal Nephron

    PubMed Central

    Zhuo, Jia L.; Li, Xiao C.

    2013-01-01

    The kidney plays a fundamental role in maintaining body salt and fluid balance and blood pressure homeostasis through the actions of its proximal and distal tubular segments of nephrons. However, proximal tubules are well recognized to exert a more prominent role than distal counterparts. Proximal tubules are responsible for reabsorbing approximately 65% of filtered load and most, if not all, of filtered amino acids, glucose, solutes, and low molecular weight proteins. Proximal tubules also play a key role in regulating acid-base balance by reabsorbing approximately 80% of filtered bicarbonate. The purpose of this review article is to provide a comprehensive overview of new insights and perspectives into current understanding of proximal tubules of nephrons, with an emphasis on the ultrastructure, molecular biology, cellular and integrative physiology, and the underlying signaling transduction mechanisms. The review is divided into three closely related sections. The first section focuses on the classification of nephrons and recent perspectives on the potential role of nephron numbers in human health and diseases. The second section reviews recent research on the structural and biochemical basis of proximal tubular function. The final section provides a comprehensive overview of new insights and perspectives in the physiological regulation of proximal tubular transport by vasoactive hormones. In the latter section, attention is particularly paid to new insights and perspectives learnt from recent cloning of transporters, development of transgenic animals with knockout or knockin of a particular gene of interest, and mapping of signaling pathways using microarrays and/or physiological proteomic approaches. PMID:23897681

  9. Silencing megalin and cubilin genes inhibits myeloma light chain endocytosis and ameliorates toxicity in human renal proximal tubule epithelial cells.

    PubMed

    Li, Min; Balamuthusamy, Saravanan; Simon, Eric E; Batuman, Vecihi

    2008-07-01

    Using target-specific short interfering (si) RNAs, we silenced the tandem endocytic receptors megalin and cubilin genes in cultured human renal proximal tubule epithelial cells. Transfection by siRNA resulted in up to 90% suppression of both megalin and cubilin protein and mRNA expression. In HK-2 cells exposed to kappa-light chain for up to 24 h, light chain endocytosis was reduced in either megalin- or cubilin-silenced cells markedly but incompletely. Simultaneous silencing of both the cubilin and megalin genes, however, resulted in near-complete inhibition of light chain endocytosis, as determined by measuring kappa-light chain protein concentration in cell cytoplasm and by flow cytometry using FITC-labeled kappa-light chain. In these cells, light chain-induced cytokine responses (interleukin-6 and monocyte chemoattractant protein-1) and epithelial-to-mesenchymal transition as well as the associated cellular and morphological alterations were also markedly suppressed. The results demonstrate that light chain endocytosis is predominantly mediated by the megalin-cubilin tandem endocytic receptor and identify endocytosis as a key step in light chain cytotoxicity. Blocking light chain endocytosis prevents its nephrotoxic effects on human kidney proximal tubule cells.

  10. Differential roles of hydrogen peroxide and hydroxyl radical in cisplatin-induced cell death in renal proximal tubular epithelial cells.

    PubMed

    Baek, Su Mi; Kwon, Chae Hwa; Kim, Jae Ho; Woo, Jae Suk; Jung, Jin Sup; Kim, Yong Keun

    2003-09-01

    Reactive oxygen species (ROS) have been suggested as important mediators of cisplatin-induced acute renal failure in vivo. However, our previous studies have shown that cisplatin-induced cell death in vitro could not be prevented by scavengers of hydrogen peroxide and hydroxyl radical in rabbit renal cortical slices. This discrepancy may be attributed to differential roles of ROS in necrotic and apoptotic cell death. We therefore examined, in this study, the roles of ROS in necrosis and apoptosis induced by cisplatin in primary cultured rabbit proximal tubule. Cisplatin induced necrosis at high concentrations over a few hours and apoptosis at much lower concentrations over longer periods. Necrosis induced by high concentration of cisplatin was prevented by a cell-permeable superoxide scavenger (tiron), hydrogen peroxide scavengers (catalase and pyruvate), and antioxidants (Trolox and deferoxamine), whereas hydroxyl radical scavengers (dimethythiourea and thiourea) did not affect the cisplatin-induced necrosis. However, apoptosis induced by lower concentration of cisplatin was partially prevented by tiron and hydroxyl radical scavengers but not by hydrogen peroxide scavengers and antioxidants. Cisplatin-induced apoptosis was mediated by the signaling pathway that is associated with cytochrome c release from mitochondria and caspase-3 activation. These effects were prevented by tiron and dimethylthiourea but not by catalase. Dimethylthiourea produced a significant protection against cisplatin-induced acute renal failure, and the effect was associated with an inhibition of apoptosis. These results suggest that hydrogen peroxide is involved in the cisplatin-induced necrosis, whereas hydroxyl radical is responsible for the cisplatin-induced apoptosis. The protective effects of hydroxyl radical scavengers are associated with an inhibition of cytochrome c release and caspase activation.

  11. Stearoyl-CoA Desaturase-1 Protects Cells against Lipotoxicity-Mediated Apoptosis in Proximal Tubular Cells

    PubMed Central

    Iwai, Tamaki; Kume, Shinji; Chin-Kanasaki, Masami; Kuwagata, Shogo; Araki, Hisazumi; Takeda, Naoko; Sugaya, Takeshi; Uzu, Takashi; Maegawa, Hiroshi; Araki, Shin-ichi

    2016-01-01

    Saturated fatty acid (SFA)-related lipotoxicity is a pathogenesis of diabetes-related renal proximal tubular epithelial cell (PTEC) damage, closely associated with a progressive decline in renal function. This study was designed to identify a free fatty acid (FFA) metabolism-related enzyme that can protect PTECs from SFA-related lipotoxicity. Among several enzymes involved in FFA metabolism, we identified stearoyl-CoA desaturase-1 (SCD1), whose expression level significantly decreased in the kidneys of high-fat diet (HFD)-induced diabetic mice, compared with non-diabetic mice. SCD1 is an enzyme that desaturates SFAs, converting them to monounsaturated fatty acids (MUFAs), leading to the formation of neutral lipid droplets. In culture, retrovirus-mediated overexpression of SCD1 or MUFA treatment significantly ameliorated SFA-induced apoptosis in PTECs by enhancing intracellular lipid droplet formation. In contrast, siRNA against SCD1 exacerbated the apoptosis. Both overexpression of SCD1 and MUFA treatment reduced SFA-induced apoptosis via reducing endoplasmic reticulum stress in cultured PTECs. Thus, HFD-induced decrease in renal SCD1 expression may play a pathogenic role in lipotoxicity-induced renal injury, and enhancing SCD1-mediated desaturation of SFA and subsequent formation of neutral lipid droplets may become a promising therapeutic target to reduce SFA-induced lipotoxicity. The present study provides a novel insight into lipotoxicity in the pathogenesis of diabetic nephropathy. PMID:27834856

  12. Na sup + -H sup + exchanger in proximal cells isolated from kidney. II. Short-term regulation by glucocorticoids

    SciTech Connect

    Bidet, M.; Merot, J.; Tauc, M.; Poujeol, P. )

    1987-11-01

    The purpose of this study was to investigate the acute regulation by glucocorticoid of the Na{sup +}-H{sup +} exchanger in isolated renal proximal cells of the rabbit. The changes of intracellular pH (pH{sub i}) were determined in a bicarbonate-free buffer by the use of a fluorescent pH probe that may be trapped intracellularly, 2,7-biscarboxyethyl- 5(6)- carboxyfluorescein (BCECF). The activity of the Na{sup +}-H{sup +} exchanger was estimated by measuring the Na{sup +}-induced H{sup +} efflux in BCECF-loaded cells acid loaded with nigericin in choline medium. The uptake of 1.5 mM {sup 22}Na was also studied in Na{sup +}-depleted cells. Acute application of dexamethasone increased the activity of the Na{sup +}-H{sup +} exchanger. The effect on the kinetics of amiloride-sensitive Na{sup +}-H{sup +} exchange indicated that dexamethasone (DEX) increased the activity by increasing the V{sub max} of the carrier for external sodium and for external H{sup +}. The apparent affinity was not modified either for Na{sup +} or for H{sup +}. The glucocorticoid action was undetectable after pretreatment of cells with actinomycin D or cycloheximide. Acute glucocorticoid activation of the Na{sup +}-H{sup +} exchanger in isolated proximal cells required RNA and protein synthesis and was consistent with an increase in the number of carriers in the membrane.

  13. Receptor-mediated endocytosis of albumin by kidney proximal tubule cells is regulated by phosphatidylinositide 3-kinase.

    PubMed Central

    Brunskill, N J; Stuart, J; Tobin, A B; Walls, J; Nahorski, S

    1998-01-01

    Receptor-mediated endocytosis of albumin is an important function of the kidney proximal tubule epithelium. We have measured endocytosis of [125I]-albumin in opossum kidney cells and examined the regulation of this process by phosphatidylinositide 3-kinase (PI 3-kinase). Albumin endocytosis was inhibited by both wortmannin (IC50 6.9 nM) and LY294002 (IC50 6.5 microM) at concentrations that suggested the involvement of PI 3-kinase in its regulation. Recycling rates were unaffected. We transfected OK cells with either a wild-type p85 subunit of PI 3-kinase, or a dominant negative form of the p85 subunit (Deltap85) using the LacSwitch expression system. Transfects were screened by immunoblotting with anti-PI 3-kinase antibodies. Under basal conditions, transfects demonstrated no expression of p85 or Deltap85, but expression was briskly induced by treatment of the cells with IPTG (EC50 13.7 microM). Inhibition of PI 3-kinase activity by Deltap85 was confirmed by in vitro kinase assay of anti-phosphotyrosine immunoprecipitates from transfected cells stimulated with insulin. Expression of Deltap85 resulted in marked inhibition of albumin endocytosis, predominantly as a result of reduction of the Vmax of the transport process. Expression of p85 had no significant effect on albumin uptake. The results demonstrate that PI 3-kinase regulates an early step in the receptor-mediated endocytosis of albumin by kidney proximal tubular cells. PMID:9593770

  14. ZAP-70, CTLA-4 and proximal T cell receptor signaling in cows infected with Mycobacterium avium subsp. paratuberculosis.

    PubMed

    Leite, Fernando L; Eslabão, Livia B; Pesch, Bruce; Bannantine, John P; Reinhardt, Timothy A; Stabel, Judith R

    2015-09-15

    Paratuberculosis is a chronic intestinal disease of ruminant animals caused by Mycobacterium avium subsp. paratuberculosis (MAP). A hallmark of paratuberculosis is a transition from a cell-mediated Th1 type response to a humoral Th2 response with the progression of disease from a subclinical to clinical state. The objective of this study was to investigate the expression of two crucial molecules in T cell function, ZAP-70 (zeta-chain-associated protein of 70 kDa) and CTLA-4 (cytotoxic T-lymphocyte antigen-4), in cows naturally infected with MAP. Peripheral blood mononuclear cells (PBMCs) isolated from control non-infected cows (n=5), and cows in subclinical (n=6) and clinical stages of paratuberculosis (n=6) were cultured alone (medium only), and with concanavalin A, and a whole cell sonicate of MAP for 24, 72 and 144 h to measure the dynamic changes of ZAP-70 and CTLA-4 expression on CD4, CD8, and gamma delta (γδ) T cells. Flow cytometry was also performed to measure ZAP-70 phosphorylation to examine proximal T cell receptor signaling in animals of different disease status. The surface expression of CTLA-4 was increased in animals in subclinical stage of infection while levels of ZAP-70 were decreased in CD4+ T cells of both subclinical and clinical animals, indicating a change in T cell phenotype with disease state. Interestingly, proximal T cell receptor signaling was not altered in infected animals. This study demonstrated changes in crucial signaling molecules in animals infected with MAP, thereby elucidating T cell alterations associated with disease progression.

  15. Mitochondrial Fission Increases Apoptosis and Decreases Autophagy in Renal Proximal Tubular Epithelial Cells Treated with High Glucose.

    PubMed

    Lee, Wen-Chin; Chiu, Chien-Hua; Chen, Jin-Bor; Chen, Chiu-Hua; Chang, Hsueh-Wei

    2016-11-01

    The aim of this study was to examine the effect of mitochondrial morphogenesis changes on apoptosis and autophagy of high-glucose-treated proximal tubular epithelial cells (HK2). Cell viability, apoptosis, and mitochondrial morphogenesis were examined using crystal violet, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), and mitotracker staining, respectively. High glucose inhibited cell viability and induced mitochondrial fission in HK2 cells. After depleting mitofusin 1 (MFN1), the MFN1(-) HK2 cells (fission type) became more susceptible to high-glucose-induced apoptosis and mitochondrial fragmentation observed by TUNEL and mitotracker assays. In siMFN2 HK2 cells (fission type), mitochondria were highly fragmented (>80% fission rate) with or without high-glucose treatment; however, siFIS1 (mitochondrial fission protein 1) HK2 cells (fusion type) exhibited little fragmentation (<13%). High-glucose treatment induced autophagy, characterized by the formation of autophagosome and microtubule-associated protein light chain 3 (LC3) B-II, as observed by transmission electron microscopy and western blotting, respectively. LC3B-II levels decreased in both MFN1(-) and siMFN2 HK2 cells, but increased in siFIS1 HK2 cells. Moreover, autophagy displays a protective role against high-glucose-induced cell death based on cotreatment with autophagy inhibitors (3-methyladenine and chloroquine). Mitochondrial fission may increase apoptosis and decrease autophagy of high-glucose-treated HK2 cells.

  16. Renal expression of the C3a receptor and functional responses of primary human proximal tubular epithelial cells.

    PubMed

    Braun, Michael C; Reins, Rose Y; Li, Tong-Bin; Hollmann, Travis J; Dutta, Ranjan; Rick, Wetsel A; Teng, Ba-Bie; Ke, Baozhen

    2004-09-15

    Although complement activation and deposition have been associated with a variety of glomerulopathies, the pathogenic mechanisms by which complement directly mediates renal injury remain to be fully elucidated. Renal parenchymal tissues express a limited repertoire of receptors that directly bind activated complement proteins. We report the renal expression of the receptor for the C3 cleavage product C3a, a member of the anaphylatoxin family. C3aR is highly expressed in normal human and murine kidney, as demonstrated by immunohistochemistry and in situ hybridization. Its distribution is limited to epithelial cells only, as glomerular endothelial and mesangial cells showed no evidence of C3aR expression. The C3aR is also expressed by primary renal proximal tubular epithelial cells in vitro as demonstrated by FACS, Western blot, and RT-PCR. In vitro C3aR is functional in terms of its capacity to bind 125I-labeled C3a and generate inositol triphosphate. Finally, using microarray analysis, four novel genes were identified and confirmed as transcriptionally regulated by C3aR activation in proximal tubular cells. These studies define a new pathway by which complement activation may directly modulate the renal response to immunologic injury.

  17. Differential cytoprotection by glycine against oxidant damage to proximal tubule cells.

    PubMed

    Sogabe, K; Roeser, N F; Venkatachalam, M A; Weinberg, J M

    1996-09-01

    Tert-butyl hydroperoxide (tBHP) injured freshly isolated proximal tubules in an Fe-dependent fashion that was ameliorated by a lipophilic antioxidant, diphenyl-p-phenylenediamine (DPPD), but was only minimally affected by glycine. Menadione-induced injury was Fe-independent and was unaffected by DPPD, but was strongly blocked by glycine. Fe was highly toxic when intracellular loading was facilitated by concomitant treatment with hydroxyquinoline (HQ). This toxicity was blocked by DPPD or chelating the Fe, but not by glycine. All of the lesions were characterized by severe depletion of glutathione and other soluble thiols. Menadione induced large increases in protein associated with the Triton-insoluble cytoskeleton and decreases in protein thiol content, consistent with extensive cross linking, but did not increase thiobarbituric acid reactive substances (TBARS). tBHP and HQ + Fe had either no effect or only moderate, delayed effects on cytoskeletal proteins, but induced substantial increases of TBARS. Glycine did not the alter changes in cytoskeletal proteins, thiols, or TBARS produced by any of the agents. Protection against tBHP toxicity by deferoxamine and DPPD was accompanied by substantial suppression of TBARS accumulation. Superimposition of hypoxia during tBHP exposure reduced TBARS accumulation and restored cytoprotective activity to glycine. Thus, in contrast to its consistently strong cytoprotection against a number of other insults, glycine is only variably cytoprotective against oxidant lesions in freshly isolated proximal tubules. Extensive oxidative crosslinking of proteins is compatible with maintenance of glycine cytoprotection against lethal membrane damage. Fe-induced injury to proximal tubules associated with lipid peroxidation as manifested by TBARS formation is a relatively glycine-insensitive insult.

  18. Distribution of CD105 and CD166 positive cells in the proximal epiphysis of developing rat humerus.

    PubMed

    Ozbey, Ozlem; Sahin, Zeliha; Acar, Nuray; Ustunel, Ismail

    2010-11-01

    The expression of cell surface receptors, CD105 and CD166, are characteristic of mesenchymal stem cells in cartilage. However, there is limited data regarding their immunolocalization in the cartilage of developing rat epiphysis. The purpose of this study was to determine the presence of CD105 and CD 166 positive cells in the proximal epiphysis of developing rat humerus and specify their zonal distribution with age. The tissues of rat humerus were taken on embryonic day 15 (E15), embryonic day 19 (E19), postnatal day 10 (PN10), postnatal day 20 (PN20) and adult rats and studied for the immunolocalization of CD105 and CD166. Our results showed that CD105 and CD166 positive cells were scattered in early stages of development of humerus epiphysis. For E15, only the hypertrophic zone was positive, whereas for E19 almost all zones of the epiphysis were positively stained for these markers. For PN10 and PN20, the CD105 and CD166 positive cells were mainly localized on the surface of the articular cartilage. In adult articular cartilage the CD105 and CD166 positive cells were localized in the superficial and transitional zones and in the upper regions of the deep zone. Our study provides evidence that in the developing cartilage tissue the localization of CD105 and CD166 positive cells is both dynamic and stage dependent, which may imply the existence of stem cell-like cells in cartilage from an early age to adult.

  19. Carbohydrate-binding modules promote the enzymatic deconstruction of intact plant cell walls by targeting and proximity effects.

    PubMed

    Hervé, Cécile; Rogowski, Artur; Blake, Anthony W; Marcus, Susan E; Gilbert, Harry J; Knox, J Paul

    2010-08-24

    Cell wall degrading enzymes have a complex molecular architecture consisting of catalytic modules and noncatalytic carbohydrate-binding modules (CBMs). The function of CBMs in cell wall degrading processes is poorly understood. Here, we have evaluated the potential enzyme-targeting function of CBMs in the context of intact primary and secondary cell wall deconstruction. The capacity of a pectate lyase to degrade pectic homogalacturonan in primary cell walls was potentiated by cellulose-directed CBMs but not by xylan-directed CBMs. Conversely, the arabinofuranosidase-mediated removal of side chains from arabinoxylan in xylan-rich and cellulose-poor wheat grain endosperm cell walls was enhanced by a xylan-binding CBM but less so by a crystalline cellulose-specific module. The capacity of xylanases to degrade xylan in secondary cell walls was potentiated by both xylan- and cellulose-directed CBMs. These studies demonstrate that CBMs can potentiate the action of a cognate catalytic module toward polysaccharides in intact cell walls through the recognition of nonsubstrate polysaccharides. The targeting actions of CBMs therefore have strong proximity effects within cell wall structures, explaining why cellulose-directed CBMs are appended to many noncellulase cell wall hydrolases.

  20. Cytotoxicity of Portuguese Propolis: The Proximity of the In Vitro Doses for Tumor and Normal Cell Lines

    PubMed Central

    Falcão, Soraia; Queiroz, Maria João R. P.; Ferreira, Isabel C. F. R.

    2014-01-01

    With a complex chemical composition rich in phenolic compounds, propolis (resinous substance collected by Apis mellifera from various tree buds) exhibits a broad spectrum of biological activities. Recently, in vitro and in vivo data suggest that propolis has anticancer properties, but is the cytoxicity of propolis specific for tumor cells? To answer this question, the cytotoxicity of phenolic extracts from Portuguese propolis of different origins was evaluated using human tumor cell lines (MCF7—breast adenocarcinoma, NCI-H460—non-small cell lung carcinoma, HCT15—colon carcinoma, HeLa—cervical carcinoma, and HepG2—hepatocellular carcinoma), and non-tumor primary cells (PLP2). The studied propolis presented high cytotoxic potential for human tumor cell lines, mostly for HCT15. Nevertheless, excluding HCT15 cell line, the extracts at the GI50 obtained for tumor cell lines showed, in general, cytotoxicity for normal cells (PLP2). Propolis phenolic extracts comprise phytochemicals that should be further studied for their bioactive properties against human colon carcinoma. In the other cases, the proximity of the in vitro cytotoxic doses for tumor and normal cell lines should be confirmed by in vivo tests and may highlight the need for selection of specific compounds within the propolis extract. PMID:24982911

  1. Calcium Dobesilate Prevents Diabetic Kidney Disease by Decreasing Bim and Inhibiting Apoptosis of Renal Proximal Tubular Epithelial Cells.

    PubMed

    Cai, Tian; Wu, Xiao-Yun; Zhang, Xiao-Qian; Shang, Hong-Xia; Zhang, Zhong-Wen; Liao, Lin; Dong, Jian-Jun

    2017-04-01

    Apoptosis of renal proximal tubular epithelial cells (PTECs) plays a vital role in the pathogenesis and progression of diabetic kidney disease (DKD). Calcium dobesilate is a vascular protective compound used for treatment of diabetic retinopathy and chronic venous insufficiency. The aim of this study was to determine whether calcium dobesilate can protect PTECs from glucose-induced apoptosis and the potential mechanism of this effect. It is indicated that high glucose promoted abnormal apoptosis of HK2 cells, which was inhibited by treatment of calcium dobesilate, while Bim expression decreased in response to calcium dobesilate in high-glucose-treated HK2 cells. These findings confirmed the therapeutic effects of calcium dobesilate on DKD and emphasized the importance of it as a potentially crucial drug in treatment of DKD.

  2. Scintillating pad detectors

    SciTech Connect

    Adams, D.; Baumbaugh, B.; Borcherding, F.

    1996-12-31

    We have been investigating the performance of scintillating pad detectors, individual small tiles of scintillator that are read out with wavelength-shifting fibers and visible light photon counters, for application in high luminosity colliding beam experiments such as the D0 Upgrade. Such structures could provide {open_quotes}pixel{close_quotes} type readout over large fiducial volumes for tracking, preshower detection and triggering.

  3. Uremic Toxins Induce ET-1 Release by Human Proximal Tubule Cells, which Regulates Organic Cation Uptake Time-Dependently.

    PubMed

    Schophuizen, Carolien M S; Hoenderop, Joost G J; Masereeuw, Rosalinde; Heuvel, Lambert P van den

    2015-06-26

    In renal failure, the systemic accumulation of uremic waste products is strongly associated with the development of a chronic inflammatory state. Here, the effect of cationic uremic toxins on the release of inflammatory cytokines and endothelin-1 (ET-1) was investigated in conditionally immortalized proximal tubule epithelial cells (ciPTEC). Additionally, we examined the effects of ET-1 on the cellular uptake mediated by organic cation transporters (OCTs). Exposure of ciPTEC to cationic uremic toxins initiated production of the inflammatory cytokines IL-6 (117 ± 3%, p < 0.001), IL-8 (122 ± 3%, p < 0.001), and ET-1 (134 ± 5%, p < 0.001). This was accompanied by a down-regulation of OCT mediated 4-(4-(dimethylamino)styryl)-N-methylpyridinium-iodide (ASP+) uptake in ciPTEC at 30 min (23 ± 4%, p < 0.001), which restored within 60 min of incubation. Exposure to ET-1 for 24 h increased the ASP+ uptake significantly (20 ± 5%, p < 0.001). These effects could be blocked by BQ-788, indicating activation of an ET-B-receptor-mediated signaling pathway. Downstream the receptor, iNOS inhibition by (N(G)-monomethyl-l-arginine) l-NMMA acetate or aminoguanidine, as well as protein kinase C activation, ameliorated the short-term effects. These results indicate that uremia results in the release of cytokines and ET-1 from human proximal tubule cells, in vitro. Furthermore, ET-1 exposure was found to regulate proximal tubular OCT transport activity in a differential, time-dependent, fashion.

  4. Angiotensin II counteracts the effects of cAMP/PKA on NHE3 activity and phosphorylation in proximal tubule cells.

    PubMed

    Crajoinas, Renato O; Polidoro, Juliano Z; Carneiro de Morais, Carla P A; Castelo-Branco, Regiane C; Girardi, Adriana C C

    2016-11-01

    Binding of angiotensin II (ANG II) to the AT1 receptor (AT1R) in the proximal tubule stimulates Na(+)/H(+) exchanger isoform 3 (NHE3) activity through multiple signaling pathways. However, the effects of ANG II/AT1R-induced inihibitory G protein (Gi) activation and subsequent decrease in cAMP accumulation on NHE3 regulation are not well established. We therefore tested the hypothesis that ANG II reduces cAMP/PKA-mediated phosphorylation of NHE3 on serine 552 and, in doing so, stimulates NHE3 activity. Under basal conditions, ANG II stimulated NHE3 activity but did not affect PKA-mediated NHE3 phosphorylation at serine 552 in opossum kidney (OKP) cells. However, in the presence of the cAMP-elevating agent forskolin (FSK), ANG II blocked FSK-induced NHE3 inhibition, reduced intracellular cAMP concentrations, lowered PKA activity, and prevented the FSK-mediated increase in NHE3 serine 552 phosphorylation. All effects of ANG II were blocked by pretreating OKP cells with the AT1R antagonist losartan, highlighting the contribution of the AT1R/Gi pathway in ANG II-mediated NHE3 upregulation under cAMP-elevating conditions. Accordingly, Gi inhibition by pertussis toxin treatment decreased NHE3 activity both in vitro and in vivo and, more importantly, prevented the stimulatory effect of ANG II on NHE3 activity in rat proximal tubules. Collectively, our results suggest that ANG II counteracts the effects of cAMP/PKA on NHE3 phosphorylation and inhibition by activating the AT1R/Gi pathway. Moreover, these findings support the notion that NHE3 dephosphorylation at serine 552 may represent a key event in the regulation of renal proximal tubule sodium handling by ANG II in the presence of natriuretic hormones that promote cAMP accumulation and transporter phosphorylation.

  5. Proximal and distal regulation of the HYAL1 gene cluster by the estrogen receptor α in breast cancer cells

    PubMed Central

    Edjekouane, Lydia; Benhadjeba, Samira; Jangal, Maïka; Fleury, Hubert; Gévry, Nicolas; Carmona, Euridice; Tremblay, André

    2016-01-01

    Chromosomal and genome abnormalities at the 3p21.3 locus are frequent events linked to epithelial cancers, including ovarian and breast cancers. Genes encoded in the 3p21.3 cluster include HYAL1, HYAL2 and HYAL3 members of hyaluronidases involved in the breakdown of hyaluronan, an abundant component of the vertebrate extracellular matrix. However, the transcriptional regulation of HYAL genes is poorly defined. Here, we identified the estrogen receptor ERα as a negative regulator of HYAL1 expression in breast cancer cells. Integrative data mining using METABRIC dataset revealed a significant inverse correlation between ERα and HYAL1 gene expression in human breast tumors. ChIP-Seq analysis identified several ERα binding sites within the 3p21.3 locus, supporting the role of estrogen as an upstream signal that diversely regulates the expression of 3p21.3 genes at both proximal and distal locations. Of these, HYAL1 was repressed by estrogen through ERα binding to a consensus estrogen response element (ERE) located in the proximal promoter of HYAL1 and flanked by an Sp1 binding site, required to achieve optimal estrogen repression. The repressive chromatin mark H3K27me3 was increased at the proximal HYAL1 ERE but not at other EREs contained in the cluster, providing a mechanism to selectively downregulate HYAL1. The HYAL1 repression was also specific to ERα and not to ERβ, whose expression did not correlate with HYAL1 in human breast tumors. This study identifies HYAL1 as an ERα target gene and provides a functional framework for the direct effect of estrogen on 3p21.3 genes in breast cancer cells. PMID:27764788

  6. Uremic Toxins Induce ET-1 Release by Human Proximal Tubule Cells, which Regulates Organic Cation Uptake Time-Dependently

    PubMed Central

    Schophuizen, Carolien M. S.; Hoenderop, Joost G. J.; Masereeuw, Rosalinde; van den Heuvel, Lambert P.

    2015-01-01

    In renal failure, the systemic accumulation of uremic waste products is strongly associated with the development of a chronic inflammatory state. Here, the effect of cationic uremic toxins on the release of inflammatory cytokines and endothelin-1 (ET-1) was investigated in conditionally immortalized proximal tubule epithelial cells (ciPTEC). Additionally, we examined the effects of ET-1 on the cellular uptake mediated by organic cation transporters (OCTs). Exposure of ciPTEC to cationic uremic toxins initiated production of the inflammatory cytokines IL-6 (117 ± 3%, p < 0.001), IL-8 (122 ± 3%, p < 0.001), and ET-1 (134 ± 5%, p < 0.001). This was accompanied by a down-regulation of OCT mediated 4-(4-(dimethylamino)styryl)-N-methylpyridinium-iodide (ASP+) uptake in ciPTEC at 30 min (23 ± 4%, p < 0.001), which restored within 60 min of incubation. Exposure to ET-1 for 24 h increased the ASP+ uptake significantly (20 ± 5%, p < 0.001). These effects could be blocked by BQ-788, indicating activation of an ET-B-receptor-mediated signaling pathway. Downstream the receptor, iNOS inhibition by (N(G)‐monomethyl‐l‐arginine) l-NMMA acetate or aminoguanidine, as well as protein kinase C activation, ameliorated the short-term effects. These results indicate that uremia results in the release of cytokines and ET-1 from human proximal tubule cells, in vitro. Furthermore, ET-1 exposure was found to regulate proximal tubular OCT transport activity in a differential, time-dependent, fashion. PMID:26132391

  7. Expression of VHL Causes Three-Dimensional Morphological Changes in Renal Cells Indicative of Proximal Tubule Differentiation

    PubMed Central

    Chiatar, Shivannah S; Eze, Ogechukwu P; Schoenfeld, Alan R

    2013-01-01

    Mutations in the von Hippel-Lindau (VHL) tumor suppressor gene are responsible for the VHL hereditary cancer syndrome, and are associated with the majority of clear cell renal cell carcinomas. In this study, scanning electron microscopy of VHL-negative renal carcinoma cells was utilized to examine the effects of VHL re-expression on the morphology of these cells. Significant differences were observed between the morphology of VHL-negative control cells and those with reintroduced VHL, with VHL expression mediating an apical surface that mounded upward, as opposed to the flat surfaces seen with VHL-negative cells. In long term cultures, rounded VHL-expressing cells grew in clusters on top the monolayer, and microvilli were observed on the apical face of these cells, in a manner suggestive of proximal tubule differentiation. In contrast, VHL-negative cells remained flat and did not develop microvilli in long-term cultures. Since VHL is a key member of an ubiquitin E3 ligase complex whose best known target is hypoxia-inducible factor alpha (HIF-α), we looked at the effects of HIF-α expression on cell morphology. Knockdown of HIF-2α in cells that only express this isoform had no effect on the morphology of the cells. These results indicate that VHL expression directs three dimensional morphological changes in renal cells indicative of differentiation, and while dysregulation of HIF-α may be necessary for tumorigenesis following VHL loss, it is not the major determinant of these VHL-mediated morphological changes. PMID:24308012

  8. Rab11a-positive compartments in proximal tubule cells sort fluid-phase and membrane cargo

    PubMed Central

    Mattila, Polly E.; Raghavan, Venkatesan; Rbaibi, Youssef; Baty, Catherine J.

    2013-01-01

    The proximal tubule (PT) reabsorbs the majority of sodium, bicarbonate, and chloride ions, phosphate, glucose, water, and plasma proteins from the glomerular filtrate. Despite the critical importance of endocytosis for PT cell (PTC) function, the organization of the endocytic pathway in these cells remains poorly understood. We have used immunofluorescence and live-cell imaging to dissect the itinerary of apically internalized fluid and membrane cargo in polarized primary cultures of PTCs isolated from mouse kidney cortex. Cells from the S1 segment could be distinguished from those from more distal PT segments by their robust uptake of albumin and comparatively low expression of γ-glutamyltranspeptidase. Rab11a in these cells is localized to variously sized spherical compartments that resemble the apical vacuoles observed by electron microscopy analysis of PTCs in vivo. These Rab11a-positive structures are highly dynamic and receive membrane and fluid-phase cargo. In contrast, fluid-phase cargoes are largely excluded from Rab11a-positive compartments in immortalized kidney cell lines. The unusual morphology and sorting capacity of Rab11a compartments in primary PTCs may reflect a unique specialization of these cells to accommodate the functional demands of handling a high endocytic load. PMID:24153428

  9. Surface preparation and coupling in plastic scintillator dosimetry.

    PubMed

    Ayotte, Guylaine; Archambault, Louis; Gingras, Luc; Lacroix, Frédéric; Beddar, A Sam; Beaulieu, Luc

    2006-09-01

    One way to improve the performance of scintillation dosimeters is to increase the light-collection efficiency at the coupling interfaces of the detector system. We performed a detailed study of surface preparation of scintillating fibers and their coupling with clear optical fibers to minimize light loss and increase the amount of light collected. We analyzed fiber-surface polishing with aluminum oxide sheets, coating fibers with magnesium oxide, and the use of eight different coupling agents (air, three optical gels, an optical curing agent, ultraviolet light, cyanoacrylate glue, and acetone). We prepared 10 scintillating fiber and clear optical fiber light guide samples to test different coupling methods. To test the coupling, we first cut both the scintillating fiber and the clear optical fiber. Then, we cleaned and polished both ends of both fibers. Finally, we coupled the scintillating fiber with the clear optical fiber in either a polyethylene jacket or a V-grooved support depending on the coupling agent used. To produce more light, we used an ultraviolet lamp to stimulate scintillation. A typical series of similar couplings showed a standard deviation in light-collection efficiency of 10%. This can be explained by differences in the surface preparation quality and alignment of the scintillating fiber with the clear optical fiber. Absence of surface polishing reduced the light collection by approximately 40%, and application of magnesium oxide on the proximal end of the scintillating fiber increased the amount of light collected from the optical fiber by approximately 39%. Of the coupling agents, we obtained the best results using one of the optical gels. Because a large amount of the light produced inside a scintillator is usually lost, better light-collection efficiency will result in improved sensitivity.

  10. Surface preparation and coupling in plastic scintillator dosimetry

    SciTech Connect

    Ayotte, Guylaine; Archambault, Louis; Gingras, Luc; Lacroix, Frederic; Beddar, A. Sam; Beaulieu, Luc

    2006-09-15

    One way to improve the performance of scintillation dosimeters is to increase the light-collection efficiency at the coupling interfaces of the detector system. We performed a detailed study of surface preparation of scintillating fibers and their coupling with clear optical fibers to minimize light loss and increase the amount of light collected. We analyzed fiber-surface polishing with aluminum oxide sheets, coating fibers with magnesium oxide, and the use of eight different coupling agents (air, three optical gels, an optical curing agent, ultraviolet light, cyanoacrylate glue, and acetone). We prepared 10 scintillating fiber and clear optical fiber light guide samples to test different coupling methods. To test the coupling, we first cut both the scintillating fiber and the clear optical fiber. Then, we cleaned and polished both ends of both fibers. Finally, we coupled the scintillating fiber with the clear optical fiber in either a polyethylene jacket or a V-grooved support depending on the coupling agent used. To produce more light, we used an ultraviolet lamp to stimulate scintillation. A typical series of similar couplings showed a standard deviation in light-collection efficiency of 10%. This can be explained by differences in the surface preparation quality and alignment of the scintillating fiber with the clear optical fiber. Absence of surface polishing reduced the light collection by approximately 40%, and application of magnesium oxide on the proximal end of the scintillating fiber increased the amount of light collected from the optical fiber by approximately 39%. Of the coupling agents, we obtained the best results using one of the optical gels. Because a large amount of the light produced inside a scintillator is usually lost, better light-collection efficiency will result in improved sensitivity.

  11. RhoA and MAPK signal transduction pathways regulate NHE1-dependent proximal tubule cell apoptosis after mechanical stretch.

    PubMed

    Bocanegra, Victoria; Gil Lorenzo, Andrea Fernanda; Cacciamani, Valeria; Benardón, María Eugenia; Costantino, Valeria Victoria; Vallés, Patricia G

    2014-10-01

    Mechanical deformation after congenital ureteral obstruction is traduced into biochemical signals leading to tubular atrophy due to epithelial cell apoptosis. We investigated whether Na(+)/H(+) exchanger 1 (NHE1) could be responsible for HK-2 cell apoptosis induction in response to mechanical stretch through its ability to function as a control point of RhoA and MAPK signaling pathways. When mechanical stretch was applied to HK-2 cells, cell apoptosis was associated with diminished NHE1 expression and RhoA activation. The RhoA signaling pathway was confirmed to be upstream from the MAPK cascade when HK-2 cells were transfected with the active RhoA-V14 mutant, showing higher ERK1/2 expression and decreased p38 activation associated with NHE1 downregulation. NHE1 participation in apoptosis induction was confirmed by specific small interfering RNA NHE1 showing caspase-3 activation and decreased Bcl-2 expression. The decreased NHE1 expression was correlated with abnormal NHE1 activity addressed by intracellular pH measurements. These results demonstrate that mitochondrial proximal tubule cell apoptosis in response to mechanical stretch is orchestrated by signaling pathways initiated by the small GTPase RhoA and followed by the opposing effects of ERK1/2 and p38 MAPK phosphorylation, regulating NHE1 decreased expression and activity.

  12. Haptoglobin attenuates hemoglobin-induced heme oxygenase-1 in renal proximal tubule cells and kidneys of a mouse model of sickle cell disease.

    PubMed

    Chintagari, Narendranath Reddy; Nguyen, Julia; Belcher, John D; Vercellotti, Gregory M; Alayash, Abdu I

    2015-03-01

    Sickle cell disease (SCD), a hereditary hemolytic disorder is characterized by chronic hemolysis, oxidative stress, vaso-occlusion and end-organ damage. Hemolysis releases toxic cell-free hemoglobin (Hb) into circulation. Under physiologic conditions, plasma Hb binds to haptoglobin (Hp) and forms Hb-Hp dimers. The dimers bind to CD163 receptors on macrophages for further internalization and degradation. However, in SCD patients plasma Hp is depleted and free Hb is cleared primarily by proximal tubules of kidneys. Excess free Hb in plasma predisposes patients to renal damage. We hypothesized that administration of exogenous Hp reduces Hb-mediated renal damage. To test this hypothesis, human renal proximal tubular cells (HK-2) were exposed to HbA (50μM heme) for 24h. HbA increased the expression of heme oxygenase-1 (HO-1), an enzyme which degrades heme, reduces heme-mediated oxidative toxicity, and confers cytoprotection. Similarly, infusion of HbA (32μM heme/kg) induced HO-1 expression in kidneys of SCD mice. Immunohistochemistry confirmed the increased HO-1 expression in the proximal tubules of the kidney. Exogenous Hp attenuated the HbA-induced HO-1 expression in vitro and in SCD mice. Our results suggest that Hb-mediated oxidative toxicity may contribute to renal damage in SCD and that Hp treatment reduces heme/iron toxicity in the kidneys following hemolysis.

  13. Exposure of cultured human proximal tubular cells to cadmium, mercury, zinc and bismuth: toxicity and metallothionein induction.

    PubMed

    Rodilla, V; Miles, A T; Jenner, W; Hawksworth, G M

    1998-08-14

    The kidney, in particular the proximal convoluted tubule, is a major target site for the toxic effects of various metals. However, little is known about the early effects of these metals after acute exposure in man. In the present study we have evaluated the toxicity of several inorganic metal compounds (CdCl2, HgCl2, ZnCl2, and Bi(NO3)3) and the induction of metallothionein by these compounds in cultured human proximal tubular (HPT) cells for up to 4 days. The results showed that bismuth was not toxic even at the highest dose (100 microM) used, while zinc, cadmium and mercury exhibited varying degrees of toxicity, zinc being the least toxic and mercury the most potent. A significant degree of interindividual variation between the different isolates used in these experiments was also observed. All metals used in the present study induced MT, as revealed by immunocytochemistry. All metals showed maximal induction between 1 and 3 days after treatment. Although a certain amount of constitutive MT was present in the cultures, the intensity of the staining varied with time in culture and between the different isolates studied. No correlation could be made between the intensity of the staining in control cultures (indicating total amount of constitutive MT) and the susceptibility of a given isolate to metal toxicity. Furthermore, no correlation could be made between metal-induced MT and the susceptibility of a given isolate to that particular metal.

  14. Establishment of renal proximal tubule cell lines by targeted oncogenesis in transgenic mice using the L-pyruvate kinase-SV40 (T) antigen hybrid gene.

    PubMed

    Cartier, N; Lacave, R; Vallet, V; Hagege, J; Hellio, R; Robine, S; Pringault, E; Cluzeaud, F; Briand, P; Kahn, A

    1993-03-01

    Targeted oncogenesis allowed us to obtain two cell lines which have been derived from the proximal tubule of kidney from transgenic mice harbouring the simian virus (SV40) large T and small t antigens placed under the control of the 5' regulatory sequence from the rat L-type pyruvate kinase (L-PK) gene. The cell lines (PKSV-PCT and PKSV-PR cells) were derived from early (PCT) and late (Pars Recta, PR) microdissected proximal tubules grown in D-glucose-enriched medium. In such conditions of culture, both cell lines exhibited L-PK transcripts, a stable expression of SV40-encoded nuclear large T antigen, a prolonged life span but failed to induce tumors when injected sub-cutaneously into athymic (nu-nu) mice. Confluent cells, grown on plastic support or porous filters, were organized as monolayers of polarized cuboid cells with well developed apical microvilli and formed domes. Both cell lines exhibited morphological features of proximal tubule cells with villin located in the apical brush-border and substantial amounts of hydrolase activity. By immunofluorescence studies using specific antibodies, aminopeptidase N appeared restricted to the apical microvillar domain, whereas the H2 histocompatibility antigen was distributed in the cytoplasm and lateral membranes. These results demonstrate that the proximal morphological phenotype has been fully preserved in these cultured cells derived from tissue-specific targeted oncogenesis in transgenic mice.

  15. Scintillator Measurements for SNO+

    NASA Astrophysics Data System (ADS)

    Kaptanoglu, Tanner; SNO+ Collaboration

    2016-03-01

    SNO+ is a neutrino detector located 2km underground in the SNOLAB facility with the primary goal of searching for neutrinoless double beta decay. The detector will be filled with a liquid scintillator target primarily composed of linear alkyl benzene (LAB). As charged particles travel through the detector the LAB produces scintillation light which is detected by almost ten thousand PMTs. The LAB is loaded with Te130, an isotope known to undergo double beta decay. Additionally, the LAB is mixed with an additional fluor and wavelength shifter to improve the light output and shift the light to a wavelength regime in which the PMTs are maximally efficient. The precise scintillator optics drastically affect the ultimate sensitivity of SNO+. I will present work being done to measure the optical properties of the SNO+ scintillator cocktail. The measured properties are used as input to a scintillation model that allows us to extrapolate to the SNO+ scale and ultimately predict the sensitivity of the experiment. Additionally, I will present measurements done to characterize the R5912 PMT, a candidate PMT for the second phase of SNO+ that provides better light collection, improved charge resolution, and a narrower spread in timing.

  16. Toxicological significance of renal Bcrp: Another potential transporter in the elimination of mercuric ions from proximal tubular cells

    SciTech Connect

    Bridges, Christy C. Zalups, Rudolfs K.; Joshee, Lucy

    2015-06-01

    Secretion of inorganic mercury (Hg{sup 2+}) from proximal tubular cells into the tubular lumen has been shown to involve the multidrug resistance-associated protein 2 (Mrp2). Considering similarities in localization and substrate specificity between Mrp2 and the breast cancer resistance protein (Bcrp), we hypothesize that Bcrp may also play a role in the proximal tubular secretion of mercuric species. In order to test this hypothesis, the uptake of Hg{sup 2+} was examined initially using inside-out membrane vesicles containing Bcrp. The results of these studies suggest that Bcrp may be capable of transporting certain conjugates of Hg{sup 2+}. To further characterize the role of Bcrp in the handling of mercuric ions and in the induction of Hg{sup 2+}-induced nephropathy, Sprague–Dawley and Bcrp knockout (bcrp{sup −/−}) rats were exposed intravenously to a non-nephrotoxic (0.5 μmol·kg{sup −1}), a moderately nephrotoxic (1.5 μmol·kg{sup −1}) or a significantly nephrotoxic (2.0 μmol·kg{sup −1}) dose of HgCl{sub 2}. In general, the accumulation of Hg{sup 2+} was greater in organs of bcrp{sup −/−} rats than in Sprague–Dawley rats, suggesting that Bcrp may play a role in the export of Hg{sup 2+} from target cells. Within the kidney, cellular injury and necrosis was more severe in bcrp{sup −/−} rats than in controls. The pattern of necrosis, which was localized in the inner cortex and the outer stripe of the outer medulla, was significantly different from that observed in Mrp2-deficient animals. These findings suggest that Bcrp may be involved in the cellular export of select mercuric species and that its role in this export may differ from that of Mrp2. - Highlights: • Bcrp may mediate transport of mercury out of proximal tubular cells. • Hg-induced nephropathy was more severe in Bcrp knockout rats. • Bcrp and Mrp2 may differ in their ability to transport Hg.

  17. Scintillator plate calorimetry

    SciTech Connect

    Price, L.E.

    1990-01-01

    Calorimetry using scintillator plates or tiles alternated with sheets of (usually heavy) passive absorber has been proven over multiple generations of collider detectors. Recent detectors including UA1, CDF, and ZEUS have shown good results from such calorimeters. The advantages offered by scintillator calorimetry for the SSC environment, in particular, are speed (<10 nsec), excellent energy resolution, low noise, and ease of achieving compensation and hence linearity. On the negative side of the ledger can be placed the historical sensitivity of plastic scintillators to radiation damage, the possibility of nonuniform response because of light attenuation, and the presence of cracks for light collection via wavelength shifting plastic (traditionally in sheet form). This approach to calorimetry is being investigated for SSC use by a collaboration of Ames Laboratory/Iowa State University, Argonne National Laboratory, Bicron Corporation, Florida State University, Louisiana State University, University of Mississippi, Oak Ridge National Laboratory, Virginia Polytechnic Institute and State University, Westinghouse Electric Corporation, and University of Wisconsin.

  18. Bright Lu2O3:Eu thin-film scintillators for high-resolution radioluminescence microscopy

    PubMed Central

    Sengupta, Debanti; Miller, Stuart; Marton, Zsolt; Chin, Frederick; Nagarkar, Vivek

    2015-01-01

    We investigate the performance of a new thin-film Lu2O3:Eu scintillator for single-cell radionuclide imaging. Imaging the metabolic properties of heterogeneous cell populations in real time is an important challenge with clinical implications. We have developed an innovative technique called radioluminescence microscopy, to quantitatively and sensitively measure radionuclide uptake in single cells. The most important component of this technique is the scintillator, which converts the energy released during radioactive decay into luminescent signals. The sensitivity and spatial resolution of the imaging system depend critically on the characteristics of the scintillator, i.e. the material used and its geometrical configuration. Scintillators fabricated using conventional methods are relatively thick, and therefore do not provide optimal spatial resolution. We compare a thin-film Lu2O3:Eu scintillator to a conventional 500 μm thick CdWO4 scintillator for radioluminescence imaging. Despite its thinness, the unique scintillation properties of the Lu2O3:Eu scintillator allow us to capture single positron decays with over fourfold higher sensitivity, a significant achievement. The thin-film Lu2O3:Eu scintillators also yield radioluminescence images where individual cells appear smaller and better resolved on average than with the CdWO4 scintillators. Coupled with the thin-film scintillator technology, radioluminescence microscopy can yield valuable and clinically relevant data on the metabolism of single cells. PMID:26183115

  19. Quantitative Proteomic Analysis of Enriched Nuclear Fractions from BK Polyomavirus-infected Primary Renal Proximal Tubule Epithelial Cells

    PubMed Central

    Justice, Joshua L.; Verhalen, Brandy; Kumar, Ranjit; Lefkowitz, Elliot J.; Imperiale, Michael J.; Jiang, Mengxi

    2016-01-01

    Polyomaviruses are a family of small DNA viruses that are associated with a number of severe human diseases, particularly in immunocompromised individuals. The detailed virus-host interactions during lytic polyomavirus infection are not fully understood. Here we report the first nuclear proteomic study with BK polyomavirus (BKPyV) in a primary renal proximal tubule epithelial cell culture system using stable isotope labeling by amino acids in cell culture (SILAC) proteomic profiling coupled with LC-MS/MS. We demonstrated the feasibility of SILAC labeling in these primary cells and subsequently performed reciprocal labeling-infection experiments to identify proteins that are altered by BKPyV infection. Our analyses revealed specific proteins that are significantly up- or down-regulated in the infected nuclear proteome. The genes encoding many of these proteins were not identified in a previous microarray study, suggesting that differential regulation of these proteins may be independent of transcriptional control. Western blotting experiments verified the SILAC proteomic findings. Finally, pathway and network analyses indicated that the host cell DNA damage response signaling and DNA repair pathways are among the cellular processes most affected at the protein level during polyomavirus infection. Our study provides a comprehensive view of the host nuclear proteomic changes during polyomavirus lytic infection and suggests potential novel host factors required for a productive polyomavirus infection. PMID:26354146

  20. Hypoxia response and VEGF-A expression in human proximal tubular epithelial cells in stable and progressive renal disease.

    PubMed

    Rudnicki, Michael; Perco, Paul; Enrich, Julia; Eder, Susanne; Heininger, Dorothea; Bernthaler, Andreas; Wiesinger, Martin; Sarközi, Rita; Noppert, Susie-Jane; Schramek, Herbert; Mayer, Bernd; Oberbauer, Rainer; Mayer, Gert

    2009-03-01

    Proteinuria, inflammation, chronic hypoxia, and rarefaction of peritubular capillaries contribute to the progression of renal disease by affecting proximal tubular epithelial cells (PTECs). To study the transcriptional response that separates patients with a stable course from those with a progressive course of disease, we isolated PTECs by laser capture microdissection from cryocut tissue sections of patients with proteinuric glomerulopathies (stable n=20, progressive n=11) with a median clinical follow-up of 26 months. Gene-expression profiling and a systems biology analysis identified activation of intracellular vascular endothelial growth factor (VEGF) signaling and hypoxia response pathways in progressive patients, which was associated with upregulation of hypoxia-inducible-factor (HIF)-1alpha and several HIF target genes, such as transferrin, transferrin-receptor, p21, and VEGF-receptor 1, but downregulation of VEGF-A. The inverse expression levels of HIF-1alpha and VEGF-A were significantly superior in predicting clinical outcome as compared with proteinuria, renal function, and degree of tubular atrophy and interstitial fibrosis at the time of biopsy. Interactome analysis showed the association of attenuated VEGF-A expression with the downregulation of genes that usually stimulate VEGF-A expression, such as epidermal growth factor (EGF), insulin-like growth factor-1 (IGF-1), and HIF-2alpha. In vitro experiments confirmed the positive regulatory effect of EGF and IGF-1 on VEGF-A transcription in human proximal tubular cells. Thus, in progressive but not in stable proteinuric kidney disease, human PTECs show an attenuated VEGF-A expression despite an activation of intracellular hypoxia response and VEGF signaling pathways, which might be due to a reduced expression of positive coregulators, such as EGF and IGF-1.

  1. Scintillator-fiber charged-particle track-imaging detector

    NASA Technical Reports Server (NTRS)

    Binns, W. R.; Israel, M. H.; Klarmann, J.

    1983-01-01

    A scintillator-fiber charged-particle track-imaging detector has been developed using a bundle of square cross-section plastic scintillator fiber optics, proximity focused onto an image intensified Charge Injection Device (CID) camera. Detector to beams of 15 MeV protons and relativistic Neon, Manganese, and Gold nuclei have been exposed and images of their tracks are obtained. This paper presents details of the detector technique, properties of the tracks obtained, and range measurements of 15 MeV protons stopping in the fiber bundle.

  2. Proximal tubule-derived Colony Stimulating Factor-1 mediates polarization of renal macrophages and dendritic cells, and recovery in acute kidney injury

    PubMed Central

    Wang, Yinqiu; Chang, Jian; Yao, Bing; Niu, Aolei; Kelly, Emily; Breeggemann, Matthew C.; Abboud Werner, Sherry L.; Harris, Raymond C.; Zhang, Ming-Zhi

    2015-01-01

    Infiltrating cells play an important role in both the development of and recovery from acute kidney injury (AKI). Macrophages and renal dendritic cells are of particular interest because they can exhibit distinctly different functional phenotypes, broadly characterized as proinflammatory (M1) or tissue reparative (M2). Resident renal macrophages and dendritic cells participate in recovery from AKI in response to either ischemia/reperfusion or a model of selective proximal tubule injury induced by diphtheria toxin-induced apoptosis in transgenic mice expressing the human diphtheria toxin receptor on proximal tubule cells. Colony Stimulating Factor-1 (CSF-1) is an important factor mediating the recovery from AKI, and CSF-1 can stimulate macrophage and dendritic cell proliferation and polarization during the recovery phase of AKI. The kidney, and specifically the proximal tubule, is a major source of intrarenal CSF-1 production in response to AKI. We induced selective deletion of proximal tubule CSF-1 to determine its role in expansion and proliferation of renal macrophages and dendritic cells and in recovery from AKI. In both models of AKI, there was decreased M2 polarization, delayed functional and structural recovery and increased tubulointerstitial fibrosis. Thus, intrarenal CSF-1 is an important mediator of macrophage/dendritic cell polarization and recovery from AKI. PMID:26422503

  3. Effects of amyloid-β plaque proximity on the axon initial segment of pyramidal cells.

    PubMed

    León-Espinosa, Gonzalo; DeFelipe, Javier; Muñoz, Alberto

    2012-01-01

    The output of cortical pyramidal cells reflects the balance between excitatory inputs of cortical and subcortical origin, and inhibitory inputs from distinct populations of cortical GABAergic interneurons, each of which selectively innervate different domains of neuronal pyramidal cells (i.e., dendrites, soma and axon initial segment [AIS]). In Alzheimer's disease (AD), the presence of amyloid-β (Aβ) plaques alters the synaptic input to pyramidal cells in a number of ways. However, the effects of Aβ plaques on the AIS have still not been investigated to date. This neuronal domain is involved in input integration, as well as action potential initiation and propagation, and it exhibits Ca2+- and activity-dependent structural plasticity. The AIS is innervated by GABAergic axon terminals from chandelier cells, which are thought to exert a strong influence on pyramidal cell output. In the AβPP/PS1 transgenic mouse model of AD, we have investigated the effects of Aβ plaques on the morphological and neurochemical features of the AIS, including the cisternal organelle, using immunocytochemistry and confocal microscopy, as well as studying the innervation of the AIS by chandelier cell axon terminals. There is a strong reduction in GABAergic terminals that appose AIS membrane surfaces that are in contact with Aβ plaques, indicating altered inhibitory synapsis at the AIS. Thus, despite a lack of gross structural alterations in the AIS, this decrease in GABAergic innervation may deregulate AIS activity and contribute to the hyperactivity of neurons in contact with Aβ plaques.

  4. Variation in sorbitol accumulation and polyol-pathway activity in cultured human proximal tubule cells.

    PubMed

    Flath, M C; Bylander, J E; Sens, D A

    1992-09-01

    The polyol pathway is present in tissues of several organs where its activation may participate in the development of diabetic complications. We measured the accumulation of polyol-pathway intermediates in HPT cells isolated from 21 different human kidneys from nondiabetic individuals. When exposed to 27.5 mM glucose in the growth media, cells isolated from approximately 75% of individuals (accumulators) accumulated sorbitol within 1-4 days, whereas 25% (nonaccumulators) accumulated only negligible amounts, even when the period of exposure was extended to 2 wk. Surprisingly, measurement of the activities of the polyol-pathway enzymes showed no difference in the levels of either AR or SDH between accumulators and nonaccumulators, even when the conversion of galactose to galactitol was used to measure AR activity in intact cells independently of SDH. Measurement of sorbitol in the growth media indicated that nonaccumulators were not releasing sorbitol into the growth media. Fructose levels in the conditioned growth media were 4 times higher in the sorbitol-accumulating cells. Together, these results indicate that the tendency of cells from an individual to accumulate significant amounts of sorbitol may reflect the cells' ability to metabolize sorbitol in steps subsequent to the polyol pathway.

  5. Live-Cell Labeling of Specific Protein Glycoforms by Proximity-Enhanced Bioorthogonal Ligation

    PubMed Central

    Robinson, Peter V.; de Almeida-Escobedo, Gabriela; de Groot, Amber E.; McKechnie, Julia L.

    2016-01-01

    Reagents for detecting post-translational modifications in the context of their protein scaffold are powerful tools, but are challenging to develop for glycosylated epitopes. We describe a strategy for detecting protein-specific glycosylation through the use of cyclooctyne-aptamer conjugates. These molecules selectively ligate to azidosugar-labeled glycans exclusively on a target protein on live cells. We characterized aptamer conjugates against two different cell surface glycoproteins and show that these reagents are amenable to detecting protein sialoforms by mass spectrometry, Western blotting, and flow cytometry. Given the abundance of aptamers that bind cell surface targets, we expect this technology will be a useful platform for investigating the roles of protein-specific glycosylation in various cellular contexts. PMID:26280358

  6. Bilirubin activates transcription of HIF-1α in human proximal tubular cells cultured in the physiologic oxygen content.

    PubMed

    Kim, Sung Gyun; Ahn, Shin-Young; Lee, Eun Seong; Kim, Sejoong; Na, Ki Young; Chae, Dong-Wan; Chin, Ho Jun

    2014-09-01

    The expression of hypoxia-inducible factor (HIF) is influenced by reactive oxygen species (ROS). Effect of bilirubin on HIF-1 expression in proximal tubular cells was investigated under physiological oxygen concentration, which is relative hypoxic condition mimicking oxygen content in the medulla of renal tissue. The human kidney (HK2) cells were cultured in 5% oxygen with or without bilirubin. HIF-1α protein expression was increased by bilirubin treatment at 0.01-0.2 mg/dL concentration. The messenger RNA expression of HIF-1α was increased by 1.69±0.05 folds in the cells cultured with 0.1 mg/dL bilirubin, compared to the control cells. The inhibitors of PI3K/mTOR, PI3K/AKT, and ERK 1/2 pathways did not attenuate increased HIF-1α expression by bilirubin. HIF-1α expression decreased by 10 µM exogenous hydrogen peroxide (H2O2); scavenger of ROS with or without bilirubin in the HK2 cells increased HIF-1α concentration more than that in the cells without bilirubin. Exogenous H2O2 decreased the phosphorylation of P70S6 kinase, which was completely reversed by bilirubin treatment. Knockdown of NOX4 gene by small interfering RNA (siRNA) increased HIF-1α mRNA expression. In coonclusion, bilirubin enhances HIF-1α transcription as well as the up-regulation of HIF-1α protein translation through the attenuation of ROS and subunits of NADPH oxidase.

  7. Model of very fast (>75 Hz) network oscillations generated by electrical coupling between the proximal axons of cerebellar Purkinje cells

    PubMed Central

    Traub, Roger D; Middleton, Steven J; Knöpfel, Thomas; Whittington, Miles A

    2009-01-01

    Very fast oscillations (VFO, >75 Hz) occur transiently in vivo, in the cerebellum of mice genetically modified to model Angelman syndrome, and in a mouse model of fetal alcohol syndrome. We recently reported VFO in slices of mouse cerebellar cortex (Crus I and II of ansiform and paramedian lobules), either in association with gamma oscillations (~40 Hz, evoked by nicotine), or in isolation (evoked by nicotine in combination with GABAA receptor blockade). The experimental data suggest a role for electrical coupling between Purkinje cells (blockade of VFO by drugs reducing gap junction conductance, and spikelets in some Purkinje cells); and the data suggest the specific involvement of Purkinje cell axons (because of field oscillation maxima in the granular layer). We show here that a detailed network model (1,000 multicompartment Purkinje cells) replicates the experimental data, when gap junctions are located on the proximal axons of Purkinje cells, provided sufficient spontaneous firing is present. Unlike other VFO models, most somatic spikelets do not correspond to axonal spikes in the parent axon, but reflect spikes in electrically coupled axons. The model predicts gating of VFO frequency by gNa inactivation, and experiments prolonging this inactivation time constant, with β-pompilidotoxin, are consistent with this prediction. The model also predicts that cerebellar VFO can be explained as an electrically coupled system of axons which are not intrinsic oscillators: the electrically uncoupled cells do not individually oscillate (in the model), and axonal firing rates are much lower in the uncoupled state than in the coupled state. PMID:18973579

  8. Direct Contacts Between Extracellular Membrane-Proximal Domains are Required for VEGF Receptor Activation and Cell Signaling

    SciTech Connect

    Yang, Y.; Xie, P; Opatowsky, Y; Schlessinger, J

    2010-01-01

    Structural analyses of the extracellular region of stem cell factor (SCF) receptor (also designated KIT) in complex with SCF revealed a sequence motif in a loop in the fourth Ig-like domain (D4) that is responsible for forming homotypic receptor contacts and for ligand-induced KIT activation and cell signaling. An identical motif was identified in the most membrane-proximal seventh Ig-like domain (D7) of vascular endothelial growth factor receptor 1 (VEGFR1), VEGFR2, and VEGFR3. In this report we demonstrate that ligand-induced tyrosine autophosphorylation and cell signaling via VEGFR1 or VEGFR2 harboring mutations in critical residues (Arg726 or Asp731) in D7 are strongly impaired. We also describe the crystal structure of D7 of VEGFR2 to a resolution of 2.7 {angstrom}. The structure shows that homotypic D7 contacts are mediated by salt bridges and van der Waals contacts formed between Arg726 of one protomer and Asp731 of the other protomer. The structure of D7 dimer is very similar to the structure of D4 dimers seen in the crystal structure of KIT extracellular region in complex with SCF. The high similarity between VEGFR D7 and KIT D4 in both structure and function provides further evidence for common ancestral origins of type III and type V RTKs. It also reveals a conserved mechanism for RTK activation and a novel target for pharmacological intervention of pathologically activated RTKs.

  9. Capacitive proximity sensor

    DOEpatents

    Kronberg, J.W.

    1994-05-31

    A proximity sensor based on a closed field circuit is disclosed. The circuit comprises a ring oscillator using a symmetrical array of plates that creates an oscillating displacement current. The displacement current varies as a function of the proximity of objects to the plate array. Preferably the plates are in the form of a group of three pair of symmetric plates having a common center, arranged in a hexagonal pattern with opposing plates linked as a pair. The sensor produces logic level pulses suitable for interfacing with a computer or process controller. The proximity sensor can be incorporated into a load cell, a differential pressure gauge, or a device for measuring the consistency of a characteristic of a material where a variation in the consistency causes the dielectric constant of the material to change. 14 figs.

  10. Capacitive proximity sensor

    DOEpatents

    Kronberg, James W.

    1994-01-01

    A proximity sensor based on a closed field circuit. The circuit comprises a ring oscillator using a symmetrical array of plates that creates an oscillating displacement current. The displacement current varies as a function of the proximity of objects to the plate array. Preferably the plates are in the form of a group of three pair of symmetric plates having a common center, arranged in a hexagonal pattern with opposing plates linked as a pair. The sensor produces logic level pulses suitable for interfacing with a computer or process controller. The proximity sensor can be incorporated into a load cell, a differential pressure gauge, or a device for measuring the consistency of a characteristic of a material where a variation in the consistency causes the dielectric constant of the material to change.

  11. Scintillator Waveguide For Sensing Radiation

    DOEpatents

    Bliss, Mary; Craig, Richard A.; Reeder; Paul L.

    2003-04-22

    The present invention is an apparatus for detecting ionizing radiation, having: a waveguide having a first end and a second end, the waveguide formed of a scintillator material wherein the therapeutic ionizing radiation isotropically generates scintillation light signals within the waveguide. This apparatus provides a measure of radiation dose. The apparatus may be modified to permit making a measure of location of radiation dose. Specifically, the scintillation material is segmented into a plurality of segments; and a connecting cable for each of the plurality of segments is used for conducting scintillation signals to a scintillation detector.

  12. Scintillator requirements for medical imaging

    SciTech Connect

    Moses, William W.

    1999-09-01

    Scintillating materials are used in a variety of medical imaging devices. This paper presents a description of four medical imaging modalities that make extensive use of scintillators: planar x-ray imaging, x-ray computed tomography (x-ray CT), SPECT (single photon emission computed tomography) and PET (positron emission tomography). The discussion concentrates on a description of the underlying physical principles by which the four modalities operate. The scintillator requirements for these systems are enumerated and the compromises that are made in order to maximize imaging performance utilizing existing scintillating materials are discussed, as is the potential for improving imaging performance by improving scintillator properties.

  13. Carcinogens induce loss of the primary cilium in human renal proximal tubular epithelial cells independently of effects on the cell cycle

    PubMed Central

    Radford, Robert; Slattery, Craig; Jennings, Paul; Blacque, Oliver; Pfaller, Walter; Gmuender, Hans; Van Delft, Joost; Ryan, Michael P.

    2012-01-01

    The primary cilium is an immotile sensory and signaling organelle found on the majority of mammalian cell types. Of the multitude of roles that the primary cilium performs, perhaps some of the most important include maintenance of differentiation, quiescence, and cellular polarity. Given that the progression of cancer requires disruption of all of these processes, we have investigated the effects of several carcinogens on the primary cilium of the RPTEC/TERT1 human proximal tubular epithelial cell line. Using both scanning electron microscopy and immunofluorescent labeling of the ciliary markers acetylated tubulin and Arl13b, we confirmed that RPTEC/TERT1 cells express primary cilium upon reaching confluence. Treatment with the carcinogens ochratoxin A (OTA) and potassium bromate (KBrO3) caused a significant reduction in the number of ciliated cells, while exposure to nifedipine, a noncarcinogenic renal toxin, had no effect on primary cilium expression. Flow cytometric analysis of the effects of all three compounds on the cell cycle revealed that only KBrO3 resulted in an increase in the proportion of cells entering the cell cycle. Microarray analysis revealed dysregulation of multiple pathways affecting ciliogenesis and ciliary maintenance following OTA and KBrO3 exposure, which were unaffected by nifedipine exposure. The primary cilium represents a unique physical checkpoint with relevance to carcinogenesis. We have shown that the renal carcinogens OTA and KBrO3 cause significant deciliation in a model of the proximal tubule. With KBrO3, this was followed by reentry into the cell cycle; however, deciliation was not found to be associated with reentry into the cell cycle following OTA exposure. Transcriptomic analysis identified dysregulation of Wnt signaling and ciliary trafficking in response to OTA and KBrO3 exposure. PMID:22262483

  14. SCINTILLATION EXPOSURE RATE DETECTOR

    DOEpatents

    Spears, W.G.

    1960-11-01

    A radiation detector for gamma and x rays is described. The detector comprises a scintillation crystal disposed between a tantalum shield and the input of a photomultiplier tube, the crystal and the shield cooperating so that their combined response to a given quantity of radiation at various energy levels is substantially constant.

  15. Boron loaded scintillator

    SciTech Connect

    Bell, Zane William; Brown, Gilbert Morris; Maya, Leon; Sloop, Jr., Frederick Victor; Sloop, Jr., Frederick Victor

    2009-10-20

    A scintillating composition for detecting neutrons and other radiation comprises a phenyl containing silicone rubber with carborane units and at least one phosphor molecule. The carbonate units can either be a carborane molecule dispersed in the rubber with the aid of a compatibilization agent or can be covalently bound to the silicone.

  16. Polysiloxane scintillator composition

    DOEpatents

    Walker, J.K.

    1992-05-05

    A plastic scintillator useful for detecting ionizing radiation comprising a matrix which comprises an optically transparent polysiloxane having incorporated therein at least one ionizing radiation-hard fluor capable of converting electromagnetic energy produced in the polysiloxane upon absorption of ionizing radiation to detectable light.

  17. Polysiloxane scintillator composition

    DOEpatents

    Walker, James K.

    1992-01-01

    A plastic scintillator useful for detecting ionizing radiation comprising a matrix which comprises an optically transparent polysiloxane having incorporated therein at least one ionizing radiation-hard fluor capable of converting electromagnetic energy produced in the polysiloxane upon absorption of ionizing radiation to detectable light.

  18. Quenching equation for scintillation

    NASA Astrophysics Data System (ADS)

    Kato, Takahisa

    1980-06-01

    A mathematical expression is postulated showing the relationship between counting rate and quenching agent concentration in a liquid scintillation solution. The expression is more suited to a wider range of quenching agent concentrations than the Stern-Volmer equation. An estimation of the quenched correction is demonstrated using the expression.

  19. In vitro membrane reconstitution of the T cell receptor proximal signaling network

    PubMed Central

    Hui, Enfu; Vale, Ronald D.

    2014-01-01

    T-cell receptor (TCR) phosphorylation is controlled by a complex network that includes Lck, a Src family kinase (SFK), the tyrosine phosphatase CD45, and the Lck-inhibitory kinase Csk. How these competing phosphorylation and dephosphorylation reactions are modulated to produce T-cell triggering is not fully understood. Here we reconstituted this signaling network using purified enzymes on liposomes, recapitulating the membrane environment in which they normally interact. We demonstrate that Lck's enzymatic activity can be regulated over a ~10-fold range by controlling its phosphorylation state. By varying kinase and phosphatase concentrations, we constructed phase diagrams that reveal ultrasensitivity in the transition from the quiescent to the phosphorylated state and demonstrate that coclustering TCR-Lck or detaching Csk from the membrane can trigger TCR phosphorylation. Our results provide insight into the mechanism of TCR signaling as well as other signaling pathways involving SFKs. PMID:24463463

  20. Antenatal Betamethasone Attenuates the Angiotensin-(1-7)/Mas Receptor/Nitric Oxide Axis in Isolated Proximal Tubule Cells.

    PubMed

    Su, Yixin; Bi, Jianli; Pulgar, Victor M; Chappell, Mark C; Rose, James C

    2017-02-22

    We previously reported a sex-specific effect of antenatal treatment with betamethasone (Beta) on sodium (Na(+)) excretion in adult sheep whereby treated males but not females had an attenuated natriuretic response to Ang-(1-7). The present study determined the Na(+) uptake and nitric oxide (NO) response to low dose Ang-(1-7) (1 pM) in renal proximal tubule cells (RPTC) from adult male and female sheep antenatally exposed to Beta or vehicle. Data were expressed as % of basal uptake or area under the curve (AUC) for Na(+) or % of control for NO. Male Beta RPTC exhibited greater Na(+) uptake than male vehicle cells (433±28% vs. 330±26%; p<0.05); however, Beta exposure had no effect on Na(+) uptake in the female cells (255±16% vs. 255±14%; p>0.05). Ang-(1-7) significantly inhibited Na(+) uptake in RPTC from vehicle male (214±11%) and from both vehicle (190±14%) and Beta (209±11%) females, but failed to attenuate Na+ uptake in Beta male cells. Beta exposure also abolished stimulation of NO by Ang-(1-7) in male, but not female RPTC. Both the Na+ and NO responses to Ang-(1-7) were blocked by Mas receptor antagonist [D-Ala7]-Ang-(1-7). We conclude that the tubular Ang-(1-7)-Mas-NO pathway is attenuated in males and not females by antenatal Beta exposure. Moreover, since primary cultures of RPTC retain both the sex and Beta-induced phenotype of the adult kidney in vivo they appear to be an appropriate cell model to examine the effects of fetal programming on Na+ handling by the renal tubules.

  1. Investigating the Anisotropic Scintillation Response in Organic Crystal Scintillator Detectors

    NASA Astrophysics Data System (ADS)

    Schuster, Patricia Frances

    This dissertation presents several studies that experimentally characterize the scintillation anisotropy in organic crystal scintillators. These include measurements of neutron, gamma-ray and cosmic muon interactions in anthracene, a historical benchmark among organic scintillator materials, to confirm and extend measurements previously available in the literature. The gamma-ray and muon measurements provide new experimental confirmation that no scintillation anisotropy is present in their interactions. Observations from these measurements have updated the hypothesis for the physical mechanism that is responsible for the scintillation anisotropy concluding that a relatively high dE/dx is required in order to produce a scintillation anisotropy. The directional dependence of the scintillation output in liquid and plastic materials was measured to experimentally confirm that no scintillation anisotropy correlated to detector orientation exists in amorphous materials. These observations confirm that the scintillation anisotropy is not due to an external effect on the measurement system, and that a fixed, repeating structure is required for a scintillation anisotropy. The directional dependence of the scintillation output in response to neutron interactions was measured in four stilbene crystals of various sizes and growth-methods. The scintillation anisotropy in these materials was approximately uniform, indicating that the crystal size, geometry, and growth method do not significantly impact the effect. Measurements of three additional pure crystals and two mixed crystals were made. These measurements showed that 1) the magnitude of the effect varies with energy and material, 2) the relationship between the light output and pulse shape anisotropy varies across materials, and 3) the effect in mixed materials is very complex. These measurements have informed the hypothesis of the mechanism that produces the directional dependence. By comparing the various relationships

  2. A pharmacologically-based array to identify targets of cyclosporine A-induced toxicity in cultured renal proximal tubule cells

    SciTech Connect

    Sarró, Eduard; Jacobs-Cachá, Conxita; Itarte, Emilio; Meseguer, Anna

    2012-01-15

    Mechanisms of cyclosporine A (CsA)-induced nephrotoxicity were generally thought to be hemodynamic in origin; however, there is now accumulating evidence of a direct tubular effect. Although genomic and proteomic experiments by our group and others provided overall information on genes and proteins up- or down-regulated by CsA in proximal tubule cells (PTC), a comprehensive view of events occurring after CsA exposure remains to be described. For this purpose, we applied a pharmacologic approach based on the use of known activities of a large panel of potentially protective compounds and evaluated their efficacy in preventing CsA toxicity in cultured mouse PTC. Our results show that compounds that blocked protein synthesis and apoptosis, together with the CK2 inhibitor DMAT and the PI3K inhibitor apigenin, were the most efficient in preventing CsA toxicity. We also identified GSK3, MMPs and PKC pathways as potential targets to prevent CsA damage. Additionally, heparinase-I and MAPK inhibitors afforded partial but significant protection. Interestingly, antioxidants and calcium metabolism-related compounds were unable to ameliorate CsA-induced cytotoxicity. Subsequent experiments allowed us to clarify the hierarchical relationship of targeted pathways after CsA treatment, with ER stress identified as an early effector of CsA toxicity, which leads to ROS generation, phenotypical changes and cell death. In summary, this work presents a novel experimental approach to characterizing cellular responses to cytotoxics while pointing to new targets to prevent CsA-induced toxicity in proximal tubule cells. Highlights: ► We used a novel pharmacological approach to elucidate cyclosporine (CsA) toxicity. ► The ability of a broad range of compounds to prevent CsA toxicity was evaluated. ► CsA toxicity was monitored using LDH release assay and PARP cleavage. ► Protein synthesis, PI3K, GSK3, MMP, PKC and caspase inhibitors prevented CsA toxicity. ► We also identified ER

  3. A 350 bp region of the proximal promoter of Rds drives cell-type specific gene expression

    PubMed Central

    Cai, Xue; Conley, Shannon M.; Cheng, Tong; Al-Ubaidi, Muayyad R.; Naash, Muna I.

    2010-01-01

    RDS (retinal degeneration slow) is a photoreceptor-specific tetraspanin protein required for the biogenesis and maintenance of rod and cone outer segments. Mutations in the Rds gene are associated with multiple forms of rod- and cone-dominant retinal degeneration. To gain more insight into the mechanisms underlying the regulation of this gene the identification of regulatory sequences within the promoter of Rds was undertaken. A 3.5kb fragment of the 5′ flanking region of the mouse Rds gene was isolated and binding sites for Crx, Otx2, Nr2e3, RXR family members, Mef2C, Esrrb, NF1, AP1, and SP1 in addition to several E-boxes, GC-boxes and GAGA-boxes were identified. Crx binding sequences were conserved in all mammalian species examined. Truncation expression analysis of the Rds promoter region in Y-79 retinoblastoma cells showed maximal activity in the 350bp proximal promoter region. We also show that inclusion of more distal fragments reduced promoter activity to the basal level, and that the promoter activities are cell-type and direction specific. Co-transfection with Nrl increased promoter activity, suggesting that this gene positively regulates Rds expression. Based on these findings, a relatively small fragment of the Rds promoter may be useful in future gene transfer studies to drive gene expression in photoreceptors. PMID:20447394

  4. Interleukin-1β suppresses activity of an inwardly rectifying K+ channel in human renal proximal tubule cells.

    PubMed

    Nakamura, Kazuyoshi; Komagiri, You; Kubokawa, Manabu

    2013-09-01

    We investigated the effect of interleukin-1β (IL-1β) on activity of an inwardly rectifying K+ channel in cultured human proximal tubule cells (RPTECs), using the patch-clamp technique and Fura-2 Ca2+ imaging. IL-1β (15 pg/ml) acutely reduced K+ channel activity in cell-attached patches. This effect was blocked by the IL-1 receptor antagonist (20 ng/ml), an inhibitor of phospholipase C, neomycin (300 μM), and an inhibitor of protein kinase C (PKC), GF109203X (500 nM). The Fura-2 Ca2+ imaging revealed that IL-1β increased intracellular Ca2+ concentration even after removal of extracellular Ca2+, which was blocked by an inhibitor of inositol 1,4,5-trisphosphate receptors, 2-aminoethoxydiphenyl borate (2-APB, 1 μM). Moreover, IL-1β suppressed channel activity in the presence of 2-APB without extracellular Ca2+. These results suggest that IL-1β suppresses K+ channel activity in RPTECs through binding to its specific receptor and activation of the PKC pathway even though intracellular Ca2+ does not increase.

  5. Scintillator-fiber charged particle track-imaging detector

    NASA Technical Reports Server (NTRS)

    Binns, W. R.; Israel, M. H.; Klarmann, J.

    1983-01-01

    A scintillator-fiber charged-particle track-imaging detector was developed using a bundle of square cross section plastic scintillator fiber optics, proximity focused onto an image intensified charge injection device (CID) camera. The tracks of charged particle penetrating into the scintillator fiber bundle are projected onto the CID camera and the imaging information is read out in video format. The detector was exposed to beams of 15 MeV protons and relativistic Neon, Manganese, and Gold nuclei and images of their tracks were obtained. Details of the detector technique, properties of the tracks obtained, and preliminary range measurements of 15 MeV protons stopping in the fiber bundle are presented.

  6. Receptor-mediated endocytosis and endosomal acidification is impaired in proximal tubule epithelial cells of Dent disease patients.

    PubMed

    Gorvin, Caroline M; Wilmer, Martijn J; Piret, Sian E; Harding, Brian; van den Heuvel, Lambertus P; Wrong, Oliver; Jat, Parmjit S; Lippiat, Jonathan D; Levtchenko, Elena N; Thakker, Rajesh V

    2013-04-23

    Receptor-mediated endocytosis, involving megalin and cubilin, mediates renal proximal-tubular reabsorption and is decreased in Dent disease because of mutations of the chloride/proton antiporter, chloride channel-5 (CLC-5), resulting in low-molecular-weight proteinuria, hypercalciuria, nephrolithiasis, and renal failure. To facilitate studies of receptor-mediated endocytosis and the role of CLC-5, we established conditionally immortalized proximal-tubular epithelial cell lines (ciPTECs) from three patients with CLC-5 mutations (30:insH, R637X, and del132-241) and a normal male. Confocal microscopy using the tight junction marker zona occludens-1 (ZO-1) and end-binding protein-1 (EB-1), which is specific for the plus end of microtubules demonstrated that the ciPTECs polarized. Receptor-mediated endocytic uptake of fluorescent albumin and transferrin in 30:insH and R637X ciPTECs was significantly decreased, compared with normal ciPTECs, and could be further reduced by competition with 10-fold excess of unlabeled albumin and transferrin, whereas in the del132-241 ciPTEC, receptor-mediated endocytic uptake was abolished. Investigation of endosomal acidification by live-cell imaging of pHluorin-VAMP2 (vesicle-associated membrane protein-2), a pH-sensitive-GFP construct, revealed that the endosomal pH in normal and 30:insH ciPTECs was similar, whereas in del132-241 and R637X ciPTECs, it was significantly more alkaline, indicating defective acidification in these ciPTECs. The addition of bafilomycin-A1, a V-ATPase inhibitor, raised the pH significantly in all ciPTECs, demonstrating that the differences in acidification were not due to alterations in the V-ATPase, but instead to abnormalities of CLC-5. Thus, our studies, which have established human Dent disease ciPTECs that will facilitate studies of mechanisms in renal reabsorption, demonstrate that Dent disease-causing CLC-5 mutations have differing effects on endosomal acidification and receptor-mediated endocytosis

  7. A functional study of proximal goat β-casein promoter and intron 1 in immortalized goat mammary epithelial cells.

    PubMed

    Kung, M H; Lee, Y J; Hsu, J T; Huang, M C; Ju, Y T

    2015-06-01

    Goat β-casein (CSN2) promoter has been extensively used to derive expression of recombinant therapeutic protein in transgenic goats; however, little direct evidence exists for signaling molecules and the cis-elements of goat CSN2 promoter in response to lactogenic hormone stimulation in goat mammary epithelial cells. Here, we use an immortalized caprine mammary epithelial cell line (CMC) to search for evidence of the above. Serial 5'-flanking regions deleted of promoter and intron 1 in goat CSN2 (-4,047 to +2,054) driven by firefly luciferase reporter gene were constructed and applied to measure promoter activity in CMC. The intron 1 region (+393 to +501) significantly decreased basal activity of the promoter. This finding contradicts other studies of the role of intron 1. The signal transducer and activator of transcription (STAT)5a played a significant role in activating promoter activity by prolactin stimulation. Hydrocortisone enhanced and prolonged the activity of STAT5a and promoter in CMC, but was independent of the glucocorticoid receptor response element. The minimum length of the CSN2 promoter segment in response to lactogenic stimulation was confirmed by 5' serial deletions. A cis-element located from -300 to -90 in proximal goat CSN2 promoter that is absent in bovine and human CSN2 promoter was newly identified. We demonstrated the presence of a STAT5a binding site (-102 to -82) and preservation of the guanosine nucleotide at position -90 based on responses to the presence of lactogenic hormone using internal deletions and point mutations of the predicted STAT5a binding site, and chromatin immunoprecipitation assay. Together, these findings demonstrate that the proximal -300 bp of goat CSN2 promoter containing the STAT5a binding site (-102 to -82) is the response element for lactogenic hormone stimulation. Additionally, intron 1 may be required for tissue or developmental stage-specific expression in mammary gland. The role of the far-distal regions of

  8. Molecular mechanism of kNBC1-carbonic anhydrase II interaction in proximal tubule cells.

    PubMed

    Pushkin, Alexander; Abuladze, Natalia; Gross, Eitan; Newman, Debra; Tatishchev, Sergei; Lee, Ivan; Fedotoff, Olga; Bondar, Galyna; Azimov, Rustam; Ngyuen, Matt; Kurtz, Ira

    2004-08-15

    We have recently shown that carbonic anhydrase II (CAII) binds in vitro to the C-terminus of the electrogenic sodium bicarbonate cotransporter kNBC1 (kNBC1-ct). In the present study we determined the molecular mechanisms for the interaction between the two proteins and whether kNBC1 and CAII form a transport metabolon in vivo wherein bicarbonate is transferred from CAII directly to the cotransporter. Various residues in the C-terminus of kNBC1 were mutated and the effect of these mutations on both the magnitude of CAII binding and the function of kNBC1 expressed in mPCT cells was determined. Two clusters of acidic amino acids, L(958)DDV and D(986)NDD in the wild-type kNBC1-ct involved in CAII binding were identified. In both acidic clusters, the first aspartate residue played a more important role in CAII binding than others. A significant correlation between the magnitude of CAII binding and kNBC1-mediated flux was shown. The results indicated that CAII activity enhances flux through the cotransporter when the enzyme is bound to kNBC1. These data are the first direct evidence that a complex of an electrogenic sodium bicarbonate cotransporter with CAII functions as a transport metabolon.

  9. Proximity fuze

    DOEpatents

    Harrison, Thomas R.

    1989-08-22

    A proximity fuze system includes an optical ranging apparatus, a detonation circuit controlled by the optical ranging apparatus, and an explosive charge detonated by the detonation cirtcuit. The optical ranging apparatus includes a pulsed laser light source for generating target ranging light pulses and optical reference light pulses. A single lens directs ranging pulses to a target and collects reflected light from the target. An optical fiber bundle is used for delaying the optical reference pulses to correspond to a predetermined distance from the target. The optical ranging apparatus includes circuitry for providing a first signal depending upon the light pulses reflected from the target, a second signal depending upon the light pulses from the optical delay fiber bundle, and an output signal when the first and second signals coincide with each other. The output signal occurs when the distance from the target is equal to the predetermined distance form the target. Additional circuitry distinguishes pulses reflected from the target from background solar radiation.

  10. Proximity fuze

    DOEpatents

    Harrison, T.R.

    1987-07-10

    A proximity fuze system includes an optical ranging apparatus, a detonation circuit controlled by the optical ranging apparatus, and an explosive charge detonated by the detonation circuit. The optical ranging apparatus includes a pulsed laser light source for generating target ranging light pulses and optical reference light pulses. A single lens directs ranging pulses to a target and collects reflected light from the target. An optical fiber bundle is used for delaying the optical reference pulses to correspond to a predetermined distance from the target. The optical ranging apparatus includes circuitry for providing a first signal depending upon the light pulses reflected from the target, a second signal depending upon the light pulses from the optical delay fiber bundle, and an output signal when the first and second signals coincide with each other. The output signal occurs when the distance from the target is equal to the predetermined distance from the target. Additional circuitry distinguishes pulses reflected from the target from background solar radiation. 3 figs.

  11. Proximity fuze

    SciTech Connect

    Harrison, T.R.

    1989-08-22

    A proximity fuze system is described. It includes an optical ranging apparatus, a detonation circuit controlled by the optical ranging apparatus, and an explosive charge detonated by the detonation circuit. The optical ranging apparatus includes a pulsed laser light source for generating target ranging light pulses and optical reference light pulses. A single lens directs ranging pulses to a target and collects reflected light from the target. An optical fiber bundle is used for delaying the optical reference pulses to correspond to a predetermined distance from the target. The optical ranging apparatus includes circuitry for providing a first signal depending upon the light pulses reflected from the target, a second signal depending upon the light pulses from the optical delay fiber bundle, and an output signal when the first and second signals coincide with each other. The output signal occurs when the distance from the target is equal to the predetermined distance from the target. Additional circuitry distinguishes pulses reflected from the target from background solar radiation.

  12. Dominance of the proximal coordinate frame in determining the locations of hippocampal place cell activity during navigation

    PubMed Central

    Siegel, Jennifer J.; Neunuebel, Joshua P.; Knierim, James J.

    2007-01-01

    The place-specific activity of hippocampal cells provides downstream structures with information regarding an animal's position within an environment, and perhaps the location of goals within that environment. In rodents, recent research has suggested that distal cues primarily set the orientation of the spatial representation, whereas the boundaries of the behavioral apparatus determine the locations of place activity. The current study was designed to address possible biases in some previous research that may have minimized the likelihood of observing place activity bound to distal cues. Hippocampal single-unit activity was recorded from 6 freely moving rats as they were trained to perform a tone-initiated place preference task on an open field platform. To investigate whether place activity was bound to the room- or platform-based coordinate frame (or both), the platform was translated within the room at an “early” and at a “late” phase of task acquisition (Shift 1 and Shift 2). At both time points, CA1 and CA3 place cells demonstrated room-associated and/or platform-associated activity, or remapped in response to the platform-shift. Shift 1 revealed place activity that reflected an interaction between a dominant platform-based (proximal) coordinate frame and a weaker room-based (distal) frame, as many CA1 and CA3 place fields shifted to a location intermediate to the two reference frames. Shift 2 resulted in place activity that became more strongly bound to either the platform- or room-based coordinate frame, suggesting the emergence of two independent spatial frames of reference (with many more cells participating in platform-based than room-based representations). PMID:17959742

  13. Angiotensin type 1 receptor resistance to blockade in the opossum proximal tubule cell due to variations in the binding pocket.

    PubMed

    Nistala, Ravi; Andresen, Bradley T; Pulakat, Lakshmi; Meuth, Alex; Sinak, Catherine; Mandavia, Chirag; Thekkumkara, Thomas; Speth, Robert C; Whaley-Connell, Adam; Sowers, James R

    2013-04-15

    Blockade of the angiotensin (ANG) II receptor type 1 (AT(1)R) with angiotensin receptor blockers (ARBs) is widely used in the treatment of hypertension. However, ARBs are variably effective in reducing blood pressure, likely due, in part, to polymorphisms in the ARB binding pocket of the AT(1)R. Therefore, we need a better understanding of variations/polymorphisms that alter binding of ARBs in heterogeneous patient populations. The opossum proximal tubule cell (OKP) line is commonly used in research to evaluate renal sodium handling and therefore blood pressure. Investigating this issue, we found natural sequence variations in the opossum AT(1)R paralleling those observed in the human AT(1)R. Therefore, we posited that these sequence variations may explain ARB resistance. We demonstrate that OKP cells express AT(1)R mRNA, bind (125)I-labeled ANG II, and exhibit ANG II-induced phosphorylation of Jak2. However, Jak2 phosphorylation is not inhibited by five different ARBs commonly used to treat hypertension. Additionally, nonradioactive ANG II competes (125)I-ANG II efficiently, whereas a 10-fold molar excess of olmesartan and the ANG II receptor type 2 blocker PD-123319 is unable to block (125)I-ANG II binding. In contrast, ANG II binding to OKP cells stably expressing rat AT(1A)Rs, which have a conserved AT(1)R-binding pocket with human AT(1)R, is efficiently inhibited by olmesartan. A novel observation was that resistance to ARB binding to opossum AT(1)Rs correlates with variations from the human receptor at positions 108, 163, 192, and 198 within the ARB-binding pocket. These observations highlight the potential utility of evaluating AT(1)R polymorphisms within the ARB-binding pocket in various hypertensive populations.

  14. Composite scintillator screen

    DOEpatents

    Zeman, Herbert D.

    1994-01-01

    A scintillator screen for an X-ray system includes a substrate of low-Z material and bodies of a high-Z material embedded within the substrate. By preselecting the size of the bodies embedded within the substrate, the spacial separation of the bodies and the thickness of the screen, the sensitivity of the screen to X-rays within a predetermined energy range can be predicted.

  15. Evaluation of “Dream Herb,” Calea zacatechichi, for Nephrotoxicity Using Human Kidney Proximal Tubule Cells

    PubMed Central

    Flynn, Thomas J.; Vohra, Sanah; Wiesenfeld, Paddy; Sprando, Robert L.

    2016-01-01

    A recent surge in the use of dietary supplements, including herbal remedies, necessitates investigations into their safety profiles. “Dream herb,” Calea zacatechichi, has long been used in traditional folk medicine for a variety of purposes and is currently being marketed in the US for medicinal purposes, including diabetes treatment. Despite the inherent vulnerability of the renal system to xenobiotic toxicity, there is a lack of safety studies on the nephrotoxic potential of this herb. Additionally, the high frequency of diabetes-associated kidney disease makes safety screening of C. zacatechichi for safety especially important. We exposed human proximal tubule HK-2 cells to increasing doses of this herb alongside known toxicant and protectant control compounds to examine potential toxicity effects of C. zacatechichi relative to control compounds. We evaluated both cellular and mitochondrial functional changes related to toxicity of this dietary supplement and found that even at low doses evidence of cellular toxicity was significant. Moreover, these findings correlated with significantly elevated levels of nephrotoxicity biomarkers, lending further support for the need to further scrutinize the safety of this herbal dietary supplement. PMID:27703475

  16. Hypoxia reduces constitutive and TNF-{alpha}-induced expression of monocyte chemoattractant protein-1 in human proximal renal tubular cells

    SciTech Connect

    Li Xuan; Kimura, Hideki . E-mail: hkimura@fmsrsa.fukui-med.ac.jp; Hirota, Kiichi; Sugimoto, Hidehiro; Yoshida, Haruyoshi

    2005-10-07

    Chronic hypoxia has been reported to be associated with macrophage infiltration in progressive forms of kidney disease. Here, we investigated the regulatory effects of hypoxia on constitutive and TNF-{alpha}-stimulated expression of monocyte chemoattractant protein-1 (MCP-1) in cultured human proximal renal tubular cells (HPTECs). Hypoxia reduced constitutive MCP-1 expression at the mRNA and protein levels in a time-dependent fashion for up to 48 h. Hypoxia also inhibited MCP-1 up-regulation by TNF-{alpha}. Treatment with actinomycin D showed that hypoxic down-regulation of MCP-1 expression resulted mainly from a decrease in the transcription but not the mRNA stability. Immunoblot and immunofluorescence analyses revealed that treatment with hypoxia or an iron chelator, desferrioxamine, induced nuclear accumulation of hypoxia-inducible factor-1{alpha} (HIF-1{alpha}) in HPTECs. Desferrioxamine mimicked hypoxia in the reduction of MCP-1 expression. However, overexpression of a dominant negative form of HIF-1{alpha} did not abolish the hypoxia-induced reduction of MCP-1 expression in HPTECs. These results suggest that hypoxia is an important negative regulator of monocyte chemotaxis to the renal inflamed interstitium, by reducing MCP-1 expression partly via hypoxia-activated signals other than the HIF-1 pathway.

  17. Neutron crosstalk between liquid scintillators

    SciTech Connect

    Verbeke, J. M.; Prasad, M. K.; Snyderman, N. J.

    2015-05-01

    We propose a method to quantify the fractions of neutrons scattering between liquid scintillators. Using a spontaneous fission source, this method can be utilized to quickly characterize an array of liquid scintillators in terms of crosstalk. The point model theory due to Feynman is corrected to account for these multiple scatterings. Using spectral information measured by the liquid scintillators, fractions of multiple scattering can be estimated, and mass reconstruction of fissile materials under investigation can be improved. Monte Carlo simulations of mono-energetic neutron sources were performed to estimate neutron crosstalk. A californium source in an array of liquid scintillators was modeled to illustrate the improvement of the mass reconstruction.

  18. Neutron crosstalk between liquid scintillators

    NASA Astrophysics Data System (ADS)

    Verbeke, J. M.; Prasad, M. K.; Snyderman, N. J.

    2015-09-01

    A method is proposed to quantify the fractions of neutrons scattering between liquid scintillators. Using a spontaneous fission source, this method can be utilized to quickly characterize an array of liquid scintillators in terms of crosstalk. The point model theory due to Feynman is corrected to account for these multiple scatterings. Using spectral information measured by the liquid scintillators, fractions of multiple scattering can be estimated, and mass reconstruction of fissile materials under investigation can be improved. Monte Carlo simulations of mono-energetic neutron sources were performed to estimate neutron crosstalk. A californium source in an array of liquid scintillators was modeled to illustrate the improvement of the mass reconstruction.

  19. De novo expression of sodium-glucose cotransporter SGLT2 in Bowman's capsule coincides with replacement of parietal epithelial cell layer with proximal tubule-like epithelium.

    PubMed

    Tabatabai, Niloofar M; North, Paula E; Regner, Kevin R; Kumar, Suresh N; Duris, Christine B; Blodgett, Amy B

    2014-08-01

    In kidney nephron, parietal epithelial cells line the Bowman's capsule and function as a permeability barrier for the glomerular filtrate. Bowman's capsule cells with proximal tubule epithelial morphology have been found. However, the effects of tubular metaplasia in Bowman's capsule on kidney function remain poorly understood. Sodium-glucose cotransporter 2 (SGLT2) plays a major role in reabsorption of glucose in the kidney and is expressed on brush border membrane (BBM) of epithelial cells in the early segment of the proximal tubule. We hypothesized that SGLT2 is expressed in tubularized Bowman's capsule and used our novel antibody to test this hypothesis. Immunohistochemical analysis was performed with our SGLT2 antibody on C57BL/6 mouse kidney prone to have tubularized Bowman's capsules. Cell membrane was examined with periodic acid-Schiff (PAS) stain. The results showed that SGLT2 was localized on BBM of the proximal tubules in young and adult mice. Bowman's capsules were lined mostly with normal brush border-less parietal epithelial cells in young mice, while they were almost completely covered with proximal tubule-like cells in adult mice. Regardless of age, SGLT2 was expressed on BBM of the tubularized Bowman's capsule but did not co-localize with nephrin in the glomerulus. SGLT2-expressing tubular cells expanded from the urinary pole toward the vascular pole of the Bowman's capsule. This study identified the localization of SGLT2 in the Bowman's capsule. Bowman's capsules with tubular metaplasia may acquire roles in reabsorption of filtered glucose and sodium.

  20. The lipid composition of high-density lipoprotein affects its re-absorption in the kidney by proximal tubule epithelial cells.

    PubMed Central

    Breznan, Dalibor; Veereswaran, Vasanthi; Viau, France J; Neville, Tracey A-M; Sparks, Daniel L

    2004-01-01

    The kidney is believed to play a major role in the clearance of apoA-I (apolipoprotein A-I) and HDL (high-density lipoprotein) particles from the bloodstream. Proximal tubule epithelial cells of the kidney appear to prevent the loss of these proteins in the urine by re-absorbing them from the urinary filtrate. Experiments were undertaken to investigate the factors that regulate the renal re-absorption of apoA-I and small HDL in a transformed human proximal tubule epithelial (HKC-8) cell line. Fluorescent microscopic studies show that HKC-8 cells can readily bind and take up HDL particles. Intracellular localization of fluorescently labelled native HDL shows its accumulation in endocytotic vesicles, in a perinuclear region after 1 h. Binding studies reveal a saturable cell association of (125)I-HDL with the HKC-8 cell surface after 2 h. HKC-8 cells do not degrade apoA-I or other HDL-apoproteins. The specific cell association of lipid-free apoA-I is approx. 2-fold less than that observed for native HDL. Similarly, reconstituted HDL prepared from HDL-apoproteins and pure phospholipids also exhibits a low cell association with the HKC-8 cells. In contrast, reconstituted HDL prepared with the extracted lipids of HDL and pure apoA-I exhibits an even higher cell association than that observed with the native lipoprotein. A detailed characterization of the major lipid classes in reconstituted HDL shows that only cholesteryl ester increases the cell association of the recombinant particles. These results show that the cholesteryl ester content of HDL may play an important role in the re-absorptive salvage of HDL by the proximal tubule cells of the kidney. PMID:14711371

  1. Cobalt Chloride Induces Expression and Function of Breast Cancer Resistance Protein (BCRP/ABCG2) in Human Renal Proximal Tubular Epithelial Cell Line HK-2.

    PubMed

    Nishihashi, Katsuki; Kawashima, Kei; Nomura, Takami; Urakami-Takebayashi, Yumiko; Miyazaki, Makoto; Takano, Mikihisa; Nagai, Junya

    2017-01-01

    The human breast cancer resistance protein (BCRP/ABCG2), a member of the ATP-binding cassette transporter family, is a drug transporter restricting absorption and enhancing excretion of many compounds including anticancer drugs. The cis-regulatory elements in the BCRP promoter include a hypoxia response element, i.e., the DNA binding site for hypoxia-inducible factor-1 (HIF-1). In this study, we investigated the effect of cobalt chloride, a chemical inducer of HIF-1α, on the expression and function of BCRP in human renal proximal tubular cell line HK-2. Cobalt chloride treatment significantly increased the mRNA expression of not only glucose transporter 1 (GLUT1), a typical HIF-1 target gene mRNA, but also ABCG2 mRNA in HK-2 cells. The BCRP inhibitor Ko143-sensitive accumulation of BCRP substrates such as Hoechst33342 and mitoxantrone was significantly enhanced by cobalt chloride treatment. In addition, treatment with cobalt chloride significantly increased the Ko143-sensitive accumulation of fluorescein isothiocyanate-labeled methotrexate in HK-2 cells. Furthermore, cobalt chloride treatment attenuated the cytotoxicity induced by mitoxantrone and methotrexate, which might be, at least in part, due to the increase in BCRP-mediated transport activity via HIF-1 activation. These findings indicate that HIF-1 activation protects renal proximal tubular cells against BCRP substrate-induced cytotoxicity by enhancing the expression and function of BCRP in renal proximal tubular cells.

  2. YES, a Src Family Kinase, Is a Proximal Glucose-specific Activator of Cell Division Cycle Control Protein 42 (Cdc42) in Pancreatic Islet β Cells*

    PubMed Central

    Yoder, Stephanie M.; Dineen, Stacey L.; Wang, Zhanxiang; Thurmond, Debbie C.

    2014-01-01

    Second-phase insulin secretion sustains insulin release in the face of hyperglycemia associated with insulin resistance, requiring the continued mobilization of insulin secretory granules to the plasma membrane. Cdc42, the small Rho family GTPase recognized as the proximal glucose-specific trigger to elicit second-phase insulin secretion, signals downstream to activate the p21-activated kinase (PAK1), which then signals to Raf-1/MEK/ERK to induce filamentous actin (F-actin) remodeling, to ultimately mobilize insulin granules to the plasma membrane. However, the steps required to initiate Cdc42 activation in a glucose-specific manner in β cells have remained elusive. Toward this, we identified the involvement of the Src family kinases (SFKs), based upon the ability of SFK inhibitors to block glucose-stimulated Cdc42 and PAK1 activation events as well as the amplifying pathway of glucose-stimulated insulin release, in MIN6 β cells. Indeed, subsequent studies performed in human islets revealed that SFK phosphorylation was induced only by glucose and within 1 min of stimulation before the activation of Cdc42 at 3 min. Furthermore, pervanadate treatment validated the phosphorylation event to be tyrosine-specific. Although RT-PCR showed β cells to express five different SFK proteins, only two of these, YES and Fyn kinases, were found localized to the plasma membrane, and of these two, only YES kinase underwent glucose-stimulated tyrosine phosphorylation. Immunodetection and RNAi analyses further established YES kinase as a proximal glucose-specific signal in the Cdc42-signaling cascade. Identification of YES kinase provides new insight into the mechanisms underlying the sustainment of insulin secretion via granule mobilization/replenishment and F-actin remodeling. PMID:24610809

  3. YES, a Src family kinase, is a proximal glucose-specific activator of cell division cycle control protein 42 (Cdc42) in pancreatic islet β cells.

    PubMed

    Yoder, Stephanie M; Dineen, Stacey L; Wang, Zhanxiang; Thurmond, Debbie C

    2014-04-18

    Second-phase insulin secretion sustains insulin release in the face of hyperglycemia associated with insulin resistance, requiring the continued mobilization of insulin secretory granules to the plasma membrane. Cdc42, the small Rho family GTPase recognized as the proximal glucose-specific trigger to elicit second-phase insulin secretion, signals downstream to activate the p21-activated kinase (PAK1), which then signals to Raf-1/MEK/ERK to induce filamentous actin (F-actin) remodeling, to ultimately mobilize insulin granules to the plasma membrane. However, the steps required to initiate Cdc42 activation in a glucose-specific manner in β cells have remained elusive. Toward this, we identified the involvement of the Src family kinases (SFKs), based upon the ability of SFK inhibitors to block glucose-stimulated Cdc42 and PAK1 activation events as well as the amplifying pathway of glucose-stimulated insulin release, in MIN6 β cells. Indeed, subsequent studies performed in human islets revealed that SFK phosphorylation was induced only by glucose and within 1 min of stimulation before the activation of Cdc42 at 3 min. Furthermore, pervanadate treatment validated the phosphorylation event to be tyrosine-specific. Although RT-PCR showed β cells to express five different SFK proteins, only two of these, YES and Fyn kinases, were found localized to the plasma membrane, and of these two, only YES kinase underwent glucose-stimulated tyrosine phosphorylation. Immunodetection and RNAi analyses further established YES kinase as a proximal glucose-specific signal in the Cdc42-signaling cascade. Identification of YES kinase provides new insight into the mechanisms underlying the sustainment of insulin secretion via granule mobilization/replenishment and F-actin remodeling.

  4. AID-targeting and hypermutation of non-immunoglobulin genes does not correlate with proximity to immunoglobulin genes in germinal center B cells.

    PubMed

    Gramlich, Hillary Selle; Reisbig, Tara; Schatz, David G

    2012-01-01

    Upon activation, B cells divide, form a germinal center, and express the activation induced deaminase (AID), an enzyme that triggers somatic hypermutation of the variable regions of immunoglobulin (Ig) loci. Recent evidence indicates that at least 25% of expressed genes in germinal center B cells are mutated or deaminated by AID. One of the most deaminated genes, c-Myc, frequently appears as a translocation partner with the Ig heavy chain gene (Igh) in mouse plasmacytomas and human Burkitt's lymphomas. This indicates that the two genes or their double-strand break ends come into close proximity at a biologically relevant frequency. However, the proximity of c-Myc and Igh has never been measured in germinal center B cells, where many such translocations are thought to occur. We hypothesized that in germinal center B cells, not only is c-Myc near Igh, but other mutating non-Ig genes are deaminated by AID because they are near Ig genes, the primary targets of AID. We tested this "collateral damage" model using 3D-fluorescence in situ hybridization (3D-FISH) to measure the distance from non-Ig genes to Ig genes in germinal center B cells. We also made mice transgenic for human MYC and measured expression and mutation of the transgenes. We found that there is no correlation between proximity to Ig genes and levels of AID targeting or gene mutation, and that c-Myc was not closer to Igh than were other non-Ig genes. In addition, the human MYC transgenes did not accumulate mutations and were not deaminated by AID. We conclude that proximity to Ig loci is unlikely to be a major determinant of AID targeting or mutation of non-Ig genes, and that the MYC transgenes are either missing important regulatory elements that allow mutation or are unable to mutate because their new nuclear position is not conducive to AID deamination.

  5. GPS Scintillation Analysis.

    DTIC Science & Technology

    2007-11-02

    Rev. 2-89) Prescribed by ANSI Std. Z39-1 298-102 TABLE OF CONTENTS 1. INTRODUCTION 1 2. GPS COMPARISON WITH ALL-SKY IMAGES OVER AGUA VERDE...Depletions from 1 October 1994 2 3. GPS data from Agua Verde, Chile on the night of 1 October 1994 3 4. PL-SCINDA display of GPS ionospheric...comparison of GPS measurements with GOES8 L-band scintillation data, are discussed. 2. GPS COMPARISON WITH ALL-SKY IMAGES OVER AGUA VERDE, CHILE As

  6. Receptor-Targeted Nipah Virus Glycoproteins Improve Cell-Type Selective Gene Delivery and Reveal a Preference for Membrane-Proximal Cell Attachment

    PubMed Central

    Bender, Ruben R.; Muth, Anke; Schneider, Irene C.; Friedel, Thorsten; Hartmann, Jessica; Plückthun, Andreas; Maisner, Andrea; Buchholz, Christian J.

    2016-01-01

    Receptor-targeted lentiviral vectors (LVs) can be an effective tool for selective transfer of genes into distinct cell types of choice. Moreover, they can be used to determine the molecular properties that cell surface proteins must fulfill to act as receptors for viral glycoproteins. Here we show that LVs pseudotyped with receptor-targeted Nipah virus (NiV) glycoproteins effectively enter into cells when they use cell surface proteins as receptors that bring them closely enough to the cell membrane (less than 100 Å distance). Then, they were flexible in receptor usage as demonstrated by successful targeting of EpCAM, CD20, and CD8, and as selective as LVs pseudotyped with receptor-targeted measles virus (MV) glycoproteins, the current standard for cell-type specific gene delivery. Remarkably, NiV-LVs could be produced at up to two orders of magnitude higher titers compared to their MV-based counterparts and were at least 10,000-fold less effectively neutralized than MV glycoprotein pseudotyped LVs by pooled human intravenous immunoglobulin. An important finding for NiV-LVs targeted to Her2/neu was an about 100-fold higher gene transfer activity when particles were targeted to membrane-proximal regions as compared to particles binding to a more membrane-distal epitope. Likewise, the low gene transfer activity mediated by NiV-LV particles bound to the membrane distal domains of CD117 or the glutamate receptor subunit 4 (GluA4) was substantially enhanced by reducing receptor size to below 100 Å. Overall, the data suggest that the NiV glycoproteins are optimally suited for cell-type specific gene delivery with LVs and, in addition, for the first time define which parts of a cell surface protein should be targeted to achieve optimal gene transfer rates with receptor-targeted LVs. PMID:27281338

  7. Development of radiation hard scintillators

    SciTech Connect

    Markley, F.; Davidson, M.; Keller, J.; Foster, G.; Pla-Dalmau, A.; Harmon, J.; Biagtan, E.; Schueneman, G.; Senchishin, V.; Gustfason, H.; Rivard, M.

    1993-11-01

    The authors have demonstrated that the radiation stability of scintillators made from styrene polymer is very much improved by compounding with pentaphenyltrimethyltrisiloxane (DC 705 vacuum pump oil). The resulting scintillators are softer than desired, so they decided to make the scintillators directly from monomer where the base resin could be easily crosslinked to improve the mechanical properties. They can now demonstrate that scintillators made directly from the monomer, using both styrene and 4-methyl styrene, are also much more radiation resistant when modified with DC705 oil. In fact, they retain from 92% to 95% of their original light output after gamma irradiation to 10 Mrads in nitrogen with air annealing. When these scintillators made directly from monomer are compared with scintillators of the same composition made from polymer the latter have much higher light outputs. They commonly reach 83% while those made form monomer give only 50% to 60% relative to the reference, BC408. When oil modified scintillators using both p-terphenyl and tetraphenylbutadiene are compared with identical scintillators except that they use 3 hydroxy-flavone as the only luminophore the radiation stability is the same. However the 3HF system gives only 30% as much light as BC408 instead of 83% when both are measured with a green extended Phillips XP2081B phototube.

  8. Extruding plastic scintillator at Fermilab

    SciTech Connect

    Anna Pla-Dalmau; Alan D. Bross; Victor V. Rykalin

    2003-10-31

    An understanding of the costs involved in the production of plastic scintillators and the development of a less expensive material have become necessary with the prospects of building very large plastic scintillation detectors. Several factors contribute to the high cost of plastic scintillating sheets, but the principal reason is the labor-intensive nature of the manufacturing process. In order to significantly lower the costs, the current casting procedures had to be abandoned. Since polystyrene is widely used in the consumer industry, the logical path was to investigate the extrusion of commercial-grade polystyrene pellets with dopants to yield high quality plastic scintillator. This concept was tested and high quality extruded plastic scintillator was produced. The D0 and MINOS experiments are already using extruded scintillator strips in their detectors. An extrusion line has recently been installed at Fermilab in collaboration with NICADD (Northern Illinois Center for Accelerator and Detector Development). This new facility will serve to further develop and improve extruded plastic scintillator. This paper will discuss the characteristics of extruded plastic scintillator and its raw materials, the different manufacturing techniques and the current R&D program at Fermilab.

  9. Development of intrinsic IPT scintillator

    SciTech Connect

    Bross, A.D.

    1989-07-31

    We report on the development of a new polystyrene based plastic scintillator. Optical absorption, fluorescence and light output measurements are presented. Preliminary results of radiation damage effects are also given and compared to the effects on a commercial plastic scintillator, NE 110. 6 refs., 12 figs.

  10. TGF-β1 stimulates movement of renal proximal tubular epithelial cells in a three-dimensional cell culture via an autocrine TGF-β2 production.

    PubMed

    Luo, Deyi; Guan, Qiunong; Wang, Kunjie; Nguan, Christopher Y C; Du, Caigan

    2017-01-01

    TGF-βs are multifunctional cytokines, but their roles in human renal homeostasis are not fully understood. This study investigated the role of TGF-β1 in the movement of human renal proximal tubular epithelial cells (PTECs) in a three-dimensional (3D) model. HKC-8 cells, a human PTEC line, were grown in a 3D collagen culture system. Cell movement was observed under a microscope. The gene expression was examined using PCR Arrays or qRT-PCR, and protein levels by Western blot. Here, we showed that the tight junction structure formed between adjacent cells of a HKC-8 cell colony in 3D cultures, and TGF-β1 stimulated their movement, evidenced by the appearance of fingerlike pseudopodia in the leader cells at the edge of the colonies. The cell movement of these human PTECs was correlated with up-regulation of both MMP2 and MMP9 and down-regulation or inactivation of PLAUR and PTK2B. Analysis of TGF-β signaling targets confirmed autocrine production of TGF-β2 and its cleaving enzyme furin as well as SNAI1 by TGF-β1stimulation. Knockdown of TGF-β2 expression disrupted TGF-β1-stimulated PTEC invasiveness, which was correlated with the down-regulation of MMP2 and MMP9. In conclusion, the activation of TGF-β receptor autocrine signaling by up-regulated TGF-β2 may play a pivotal role in TGF-β1-induced human PTEC movement, which could be mediated at least by both MMP2 and MMP9.

  11. Discrete scintillator coupled mercuric iodide photodetector arrays for breast imaging

    SciTech Connect

    Tornai, M.P.; Levin, C.S.; Hoffman, E.J.

    1996-12-31

    Multi-element (4x4) imaging arrays with high resolution collimators, size matched to discrete CsI(Tl) scintillator arrays and mercuric iodide photodetector arrays (HgI{sub 2} PDA) are under development as prototypes for larger 16 x 16 element arrays. The compact nature of the arrays allows detector positioning in proximity to the breast to eliminate activity not in the line-of-sight of the collimator, thus reducing image background. Short collimators, size matched to {le}1.5 x 1.5 mm{sup 2} scintillators show a factor of 2 and 3.4 improvement in spatial resolution and efficiency, respectively, compared to high resolution collimated gamma cameras for the anticipated compressed breast geometries. Monte Carlo simulations, confirmed by measurements, demonstrated that scintillator length played a greater role in efficiency and photofraction for 140 keV gammas than cross sectional area, which affects intrinsic spatial resolution. Simulations also demonstrated that an increase in the ratio of scintillator area to length corresponds to an improvement in light collection. Electronic noise was below 40 e{sup -} RMS indicating that detector resolution was not noise limited. The high quantum efficiency and spectral match of prototype unity gain HgI{sub 2} PDAs coupled to 1 x 1 x 2.5 mm{sup 3} and 2 x 2 x 4 mm{sup 3} CsI(Tl) scintillators demonstrated energy resolutions of 9.4% and 8.8% FWHM at 140 keV, respectively, without the spectral tailing observed in standard high-Z, compound semi-conductor detectors. Line spread function measurements matched the scintillator size and pitch, and small, complex phantoms were easily imaged.

  12. Phosphorylation of rat kidney Na-K pump at Ser938 is required for rapid angiotensin II-dependent stimulation of activity and trafficking in proximal tubule cells.

    PubMed

    Massey, Katherine J; Li, Quanwen; Rossi, Noreen F; Keezer, Susan M; Mattingly, Raymond R; Yingst, Douglas R

    2016-02-01

    How angiotensin (ANG) II acutely stimulates the Na-K pump in proximal tubules is only partially understood, limiting insight into how ANG II increases blood pressure. First, we tested whether ANG II increases the number of pumps in plasma membranes of native rat proximal tubules under conditions of rapid activation. We found that exposure to 100 pM ANG II for 2 min, which was previously shown to increase affinity of the Na-K pump for Na and stimulate activity threefold, increased the amount of the Na-K pump in plasma membranes of native tubules by 33%. Second, we tested whether previously observed increases in phosphorylation of the Na-K pump at Ser(938) were part of the stimulatory mechanism. These experiments were carried out in opossum kidney cells, cultured proximal tubules stably coexpressing the ANG type 1 (AT1) receptor, and either wild-type or a S938A mutant of rat kidney Na-K pump under conditions found by others to stimulate activity. We found that 10 min of incubation in 10 pM ANG II stimulated activity of wild-type pumps from 2.3 to 3.5 nmol K · mg protein(-1) · min(-1) and increased the amount of the pump in the plasma membrane by 80% but had no effect on cells expressing the S938A mutant. We conclude that acute stimulation of Na-K pump activity in native rat proximal tubules includes increased trafficking to the plasma membrane and that phosphorylation at Ser(938) is part of the mechanism by which ANG II directly stimulates activity and trafficking of the rat kidney Na-K pump in opossum kidney cells.

  13. Global transcription regulation by DNA topoisomerase I in exponentially growing Saccharomyces cerevisiae cells: activation of telomere-proximal genes by TOP1 deletion.

    PubMed

    Lotito, Luca; Russo, Alessandra; Chillemi, Giovanni; Bueno, Susana; Cavalieri, Duccio; Capranico, Giovanni

    2008-03-21

    To establish the cellular functions of DNA topoisomerase I-B (Top1p) at a global level, we have determined the expression profiles and histone modification patterns affected by TOP1 gene deletion (DeltaTOP1) in Saccharomyces cerevisiae. In exponentially growing cells, DeltaTOP1 specifically increases transcription of telomere-proximal genes and decreases glucose utilization and energy production pathways. Immunoprecipitation data demonstrate that Top1p can bind to and is catalytically active at telomeric DNA repeats, and that both DeltaTOP1 and an inactive Y727F Top1p mutant increase H4 histone acetylation at telomere-proximal regions. Interestingly, while the Y727F mutation has no influence on enzyme recruitment to chromatin sites, it has a marked effect on H4 K16 acetylation at subtelomeric regions. The Top1p mutation also increases H3 histone K4 dimethylation, which has been associated with gene transcription, at 3' termini of subtelomeric genes. No major effect of DeltaTOP1 or mutation was detected on Sir3p recruitment; however, DeltaTOP1 has an effect on transcript levels of genes known to regulate telomeric silencing. Thus, the findings indicate that Top1p activity can favor both a repressed chromatin organization and a reduced gene expression level at telomere-proximal regions in yeast. As telomere-proximal regions are known to be enriched for stress-activated genes, our findings show that Top1p can optimize transcript levels for cell growth in exponentially growing cells under a synthetic medium with glucose.

  14. Proton recoil scintillator neutron rem meter

    DOEpatents

    Olsher, Richard H.; Seagraves, David T.

    2003-01-01

    A neutron rem meter utilizing proton recoil and thermal neutron scintillators to provide neutron detection and dose measurement. In using both fast scintillators and a thermal neutron scintillator the meter provides a wide range of sensitivity, uniform directional response, and uniform dose response. The scintillators output light to a photomultiplier tube that produces an electrical signal to an external neutron counter.

  15. Proximal Tibial Bone Graft

    MedlinePlus

    ... All Site Content AOFAS / FootCareMD / Treatments Proximal Tibial Bone Graft Page Content What is a bone graft? Bone grafts may be needed for various ... the proximal tibia. What is a proximal tibial bone graft? Proximal tibial bone graft (PTBG) is a ...

  16. Liquid scintillator tiles for calorimetry

    SciTech Connect

    Amouzegar, M.; Belloni, A.; Bilki, B.; Calderon, J.; Barbaro, P. De; Eno, S. C.; Hatakeyama, K.; Hirschauer, J.; Jeng, G. Y.; Pastika, N. J.; Pedro, K.; Rumerio, Paolo; Samuel, J.; Sharp, E.; Shin, Y. H.; Tiras, E.; Vishnevskiy, D.; Wetzel, J.; Yang, Z.; Yao, Y.; Youn, S. W.

    2016-11-28

    Future experiments in high energy and nuclear physics may require large, inexpensive calorimeters that can continue to operate after receiving doses of 50 Mrad or more. Also, the light output of liquid scintillators suffers little degradation under irradiation. However, many challenges exist before liquids can be used in sampling calorimetry, especially regarding developing a packaging that has sufficient efficiency and uniformity of light collection, as well as suitable mechanical properties. We present the results of a study of a scintillator tile based on the EJ-309 liquid scintillator using cosmic rays and test beam on the light collection efficiency and uniformity, and some preliminary results on radiation hardness.

  17. Liquid scintillator tiles for calorimetry

    NASA Astrophysics Data System (ADS)

    Amouzegar, M.; Belloni, A.; Bilki, B.; Calderon, J.; De Barbaro, P.; Eno, S. C.; Hatakeyama, K.; Hirschauer, J.; Jeng, G. Y.; Pastika, N. J.; Pedro, K.; Rumerio, Paolo; Samuel, J.; Sharp, E.; Shin, Y. H.; Tiras, E.; Vishnevskiy, D.; Wetzel, J.; Yang, Z.; Yao, Y.; Youn, S. W.

    2016-11-01

    Future experiments in high energy and nuclear physics may require large, inexpensive calorimeters that can continue to operate after receiving doses of 50 Mrad or more. The light output of liquid scintillators suffers little degradation under irradiation. However, many challenges exist before liquids can be used in sampling calorimetry, especially regarding developing a packaging that has sufficient efficiency and uniformity of light collection, as well as suitable mechanical properties. We present the results of a study of a scintillator tile based on the EJ-309 liquid scintillator using cosmic rays and test beam on the light collection efficiency and uniformity, and some preliminary results on radiation hardness.

  18. Scintillator materials containing lanthanum fluorides

    DOEpatents

    Moses, William W.

    1991-01-01

    An improved radiation detector containing a crystalline mixture of LaF.sub.3 and CeF.sub.3 as the scintillator element is disclosed. Scintillators made with from 25% to 99.5% LaF.sub.3 and the remainder CeF.sub.3 have been found to provide a balance of good stopping power, high light yield and short decay constant that is equal to or superior to other known scintillator materials, and which may be processed from natural starting materials containing both rare earth elements. The radiation detectors disclosed are favorably suited for use in general purpose detection and in positron emission tomography.

  19. High energy resolution plastic scintillator

    NASA Astrophysics Data System (ADS)

    van Loef, Edgar V.; Feng, Patrick; Markosyan, Gary; Shirwadkar, Urmila; Doty, Patrick; Shah, Kanai S.

    2016-09-01

    In this paper we present results on a novel tin-loaded plastic scintillator. We will show that this particular plastic scintillator has a light output similar to that of BGO, a fast scintillation decay (< 10 ns), exhibits good neutron/gamma PSD with a Figure-of-Merit of 1.3 at 2.5 MeVee cut-off energy, and excellent energy resolution of about 12% (FWHM) at 662 keV. Under X-ray excitation, the radioluminescence spectrum exhibits a broad band between 350 and 500 nm peaking at 420 nm which is well-matched to bialkali photomultiplier tubes and UV-enhanced photodiodes.

  20. Ionospheric scintillation studies

    NASA Technical Reports Server (NTRS)

    Rino, C. L.; Freemouw, E. J.

    1973-01-01

    The diffracted field of a monochromatic plane wave was characterized by two complex correlation functions. For a Gaussian complex field, these quantities suffice to completely define the statistics of the field. Thus, one can in principle calculate the statistics of any measurable quantity in terms of the model parameters. The best data fits were achieved for intensity statistics derived under the Gaussian statistics hypothesis. The signal structure that achieved the best fit was nearly invariant with scintillation level and irregularity source (ionosphere or solar wind). It was characterized by the fact that more than 80% of the scattered signal power is in phase quadrature with the undeviated or coherent signal component. Thus, the Gaussian-statistics hypothesis is both convenient and accurate for channel modeling work.

  1. Evaluation of biomarkers for in vitro prediction of drug-induced nephrotoxicity: comparison of HK-2, immortalized human proximal tubule epithelial, and primary cultures of human proximal tubular cells

    PubMed Central

    Huang, Johnny X; Kaeslin, Geraldine; Ranall, Max V; Blaskovich, Mark A; Becker, Bernd; Butler, Mark S; Little, Melissa H; Lash, Lawrence H; Cooper, Matthew A

    2015-01-01

    There has been intensive effort to identify in vivo biomarkers that can be used to monitor drug-induced kidney damage and identify injury before significant impairment occurs. Kidney injury molecule-1 (KIM-1), neutrophil gelatinase-associated lipocalin (NGAL), and human macrophage colony stimulating factor (M-CSF) have been validated as urinary and plasma clinical biomarkers predictive of acute and chronic kidney injury and disease. Similar validation of a high throughput in vitro assay predictive of nephrotoxicity could potentially be implemented early in drug discovery lead optimization to reduce attrition at later stages of drug development. To assess these known in vivo biomarkers for their potential for in vitro screening of drug-induced nephrotoxicity, we selected a panel of nephrotoxic agents and examined their effects on the overexpression of nephrotoxicity biomarkers in immortalized (HK-2) and primary (commercially available and freshly in-house produced) human renal proximal tubule epithelial cells. Traditional cytotoxicity was contrasted with expression levels of KIM-1, NGAL, and M-CSF assessed using ELISA and real-time quantitative reverse transcription PCR. Traditional cytotoxicity assays and biomarker assays using HK-2 cells were both unsuitable for prediction of nephrotoxicity. However, increases in protein levels of KIM-1 and NGAL in primary cells were well correlated with dose levels of known nephrotoxic compounds, with limited correlation seen in M-CSF protein and mRNA levels. These results suggest that profiling compounds against primary cells with monitoring of biomarker protein levels may have potential as in vitro predictive assays of drug-induced nephrotoxicity. PMID:26171227

  2. Properties of scintillator solutes

    SciTech Connect

    Fluornoy, J.M.

    1998-06-01

    This special report summarizes measurements of the spectroscopic and other properties of the solutes that were used in the preparation of several new liquid scintillators developed at EG and G/Energy Measurements/Santa Barbara Operations (the precursor to Bechtel Nevada/Special Technologies Laboratory) on the radiation-to-light converter program. The data on the individual compounds are presented in a form similar to that used by Prof. Isadore Berlman in his classic handbook of fluorescence spectra. The temporal properties and relative efficiencies of the new scintillators are presented in Table 1, and the efficiencies as a function of wavelength are presented graphically in Figure 1. In addition, there is a descriptive glossary of the abbreviations used herein. Figure 2 illustrates the basic structures of some of the compounds and of the four solvents reported in this summary. The emission spectra generally exhibit more structure than the absorption spectra, with the result that the peak emission wavelength for a given compound may lie several nm away from the wavelength, {lambda}{sub avg}, at the geometric center of the emission spectrum. Therefore, the author has chosen to list absorption peaks, {lambda}{sub max}, and emission {lambda}{sub avg} values in Figures 3--30, as being most illustrative of the differences between the compounds. The compounds, BHTP, BTPB, ADBT, and DPTPB were all developed on this program. P-terphenyl, PBD, and TPB are commercially available blue emitters. C-480 and the other longer-wavelength emitters are laser dyes available commercially from Exciton Corporation. 1 ref., 30 figs.

  3. A retinoid responsive cytokine gene, MK, is preferentially expressed in the proximal tubules of the kidney and human tumor cell lines.

    PubMed Central

    Kitamura, M.; Shirasawa, T.; Mitarai, T.; Muramatsu, T.; Maruyama, N.

    1993-01-01

    The aim of this study was to survey the expression of an embryonic cytokine gene, MK, in the normal organs and neoplastic tissues of adults. Northern analysis showed that MK mRNA was exclusively expressed in the kidney among murine organs including thymus, lung, heart, spleen, liver, and kidney. In situ hybridization analysis revealed that MK expression was localized in the proximal tubules and metaplastic Bowman's epithelium, but not in other nephron segments such as glomeruli, loop of Henle, distal tubules, and collecting ducts. To investigate whether MK expression is a marker of tubular cell lineage, several cell lines originating from renal tubules were tested. No expression of MK was detected in PtK1 and LLC-PK1 cells derived from marsupial and porcine proximal tubules or in MDBK and MDCK cells from bovine and canine distal/collecting tubules. Unexpectedly, the MK gene was expressed in a human renal cell carcinoma line, VMRC-RCW, and the expression was up-regulated in the presence of retinoic acid. To elucidate the involvement of MK in the development of tumors, we further examined its expression in a variety of human neoplastic cell lines: YMB-1-C (breast cancer), EBC-1 (lung squamous cell carcinoma), RERF-LC-OK (lung adenocarcinoma), SBC-3 (lung small cell carcinoma), HSC-2 (mouth squamous cell carcinoma), NUGC-2 (gastric cancer), COLO201 (colon cancer), HepG2 (hepatoma), MIA PaCa-2 (pancreatic cancer), MCAS (ovarian cancer), HeLa (cervical cancer), BeWo (chorionic carcinoma), ITO-II (testicular tumor), T24 (urinary bladder tumor), and G-401 (Wilms' tumor). Strong signals were detected in COLO201, HepG2, ITO-II, T24, G-401, and weaker but distinct signals were detected in YMB-1-C, HSC-2, and MCAS cells. The MK gene was, therefore, widely expressed in neoplastic cells originating from genital organs, intestinal tract, liver, mammary gland, and urinary tract, and the expression was not restricted to adenocarcinomas, but was also observed in other types of

  4. Scintillator fiber optic long counter

    DOEpatents

    McCollum, T.; Spector, G.B.

    1994-03-29

    A flat response position sensitive neutron detector capable of providing neutron spectroscopic data utilizing scintillator fiber optic filaments embedded in a neutron moderating housing having an open end through which neutrons enter to be detected is described. 11 figures.

  5. Neutron crosstalk between liquid scintillators

    DOE PAGES

    Verbeke, J. M.; Prasad, M. K.; Snyderman, N. J.

    2015-05-01

    We propose a method to quantify the fractions of neutrons scattering between liquid scintillators. Using a spontaneous fission source, this method can be utilized to quickly characterize an array of liquid scintillators in terms of crosstalk. The point model theory due to Feynman is corrected to account for these multiple scatterings. Using spectral information measured by the liquid scintillators, fractions of multiple scattering can be estimated, and mass reconstruction of fissile materials under investigation can be improved. Monte Carlo simulations of mono-energetic neutron sources were performed to estimate neutron crosstalk. A californium source in an array of liquid scintillators wasmore » modeled to illustrate the improvement of the mass reconstruction.« less

  6. Scintillator fiber optic long counter

    DOEpatents

    McCollum, Tom; Spector, Garry B.

    1994-01-01

    A flat response position sensitive neutron detector capable of providing neutron spectroscopic data utilizing scintillator fiber optic filaments embedded in a neutron moderating housing having an open end through which neutrons enter to be detected.

  7. Protective effect of 1α,25-dihydroxyvitamin D3 on effector CD4+ T cell induced injury in human renal proximal tubular epithelial cells

    PubMed Central

    Chung, Byung Ha; Kim, Bo-Mi; Doh, Kyoung Chan; Cho, Mi-La

    2017-01-01

    Background The aim of this study was to investigate the protective effect of 1α,25-dihydroxyvitamin D3 [1,25(OH)2D3] on effector CD4+ T cells or on inflammatory cytokine-induced injury in human renal proximal tubular epithelial cells (HRPTEpiC). Methods First, we investigated the effect of 1,25(OH)2D3 on CD4+ T cell proliferation. Second, we examined the effect of 1,25(OH)2D3 on inflammatory cytokine secretion or fibrosis in HRPTEpiC induced by inflammatory cytokines or activated CD4+ T cells using ELISA and real-time PCR. Lastly, we compared urine inflammatory-cytokine (IL-6, IL-8) or KIM-1 levels in kidney transplant recipients low serum 25-hydroxyvitamin D (25(OH)D) group (< 20 ng/mL) (n = 40) and normal 25(OH)D group (n = 50). Results Pre-incubation with 1,25(OH)2D3 significantly reduced the percentages of Th1 and Th17 cells compared to that of Th0 condition (P < 0.05 for each). In contrast, 1,25(OH)2D3 increased the proportion of Th2 and Treg cells in a dose-dependent manner (P < 0.05 for each). Treatment of HRPTEpiC with inflammatory cytokines (TNF-α, IL-17, and TGF-β) or effector CD4+ T cells resulted in increased production of IL-6, IL-8, or KIM-1 from HRPTEpiC in a dose-dependent manner. However, treatment with 1,25(OH)2D3 significantly reduced the level of these cytokines (P < 0.05 for all). Western blot analysis demonstrated that the mTOR/STAT3/ERK pathway was downregulated by 1,25(OH)2D3 in HRPTEpiC. Furthermore, the concentrations of urine IL-6/creatinine (P < 0.05) and Kim-1/creatinine (P < 0.05) were higher in the low 25(OH)D group than in the normal 25(OH)D group in kidney transplant recipients. Conclusion The results of this study suggests that vitamin D may have a significant role in the regulation of inflammation in allograft tissue in kidney transplant recipients. Trial registration All participants provided written informed consent in accordance with the Declaration of Helsinki. This study was approved by the Institutional Review Board of

  8. About NICADD extruded scintillating strips

    SciTech Connect

    Dyshkant, A.; Beznosko, D.; Blazey, G.; Chakraborty, D.; Francis, K.; Kubik, D.; Lima, J.G.; Rykalin, V.; Zutshi, v.; Baldina, E.; Bross, A.; Deering, P.; Nebel, T.; Pla-Dalmau, A.; Schellpfeffer, J.; Serritella, C.; Zimmerman, J.; /Fermilab

    2005-04-01

    The results of control measurements of extruded scintillating strip responses to a radioactive source Sr-90 are provided, and details of strip choice, preparation, and method of measurement are included. About four hundred one meter long extruded scintillating strips were measured at four different points. These results were essential for prototyping a tail catcher and muon tracker for a future international electron positron linear collider detector.

  9. Serum level of proximal renal tubular epithelial cell-binding immunoglobulin G in patients with lupus nephritis.

    PubMed

    Yap, D Y H; Yung, S; Zhang, Q; Tang, C; Chan, T M

    2016-01-01

    In vitro data showed that immunoglobulin G (IgG) from lupus nephritis (LN) patients could bind to proximal renal tubular epithelial cells (PTEC), but the clinical relevance of such binding remained unclear. Binding of IgG and subclasses to PTEC was measured by cellular ELISA (expressed as OD index) in 189 serial serum samples from 23 Class III/IV ± V LN patients who had repeated renal flares (48 during renal flares, 141 during low level disease activity (LLDA)), and compared with 64 patients with non-lupus glomerular diseases (NLGD) and 23 healthy individuals. Total IgG PTEC-binding index was 0.34 ± 0.16, 0.29 ± 0.16, 0.62 ± 0.27 and 0.83 ± 0.38 in healthy controls, NLGD, LN patients during LLDA, and LN patients during nephritic flare, respectively (p < 0.001, LLDA vs. renal flare; p < 0.001, healthy controls or NLGD vs. LN during LLDA or renal flare). PTEC-binding index for IgG1 was 0.09 ± 0.05, 0.16 ± 0.12, 0.44 ± 0.34 and 0.71 ± 0.46 for the corresponding groups (p < 0.001, LLDA vs. renal flare; p < 0.001, healthy controls or NLGD vs. LN during LLDA or renal flare). Sixteen of 48 episodes (33.3%) of nephritic flare showed persistent PTEC-binding IgG seropositivity for more than 9.4 ± 3.1 months, despite clinical response to immunosuppressive treatment. Total IgG and IgG1 PTEC-binding correlated with anti-dsDNA level (r = 0.34 and 0.52, respectively, p < 0.001 for both), and inversely with C3 level (r = -0.26 and -0.50, respectively, p = 0.002 and<0.001). Sensitivity/specificity of PTEC-binding index in detecting renal flares was 45.8%/80.1% for total IgG (ROC AUC 0.630, p = 0.007) and 87.5%/35.5% for IgG1 (ROC AUC 0.615, p = 0.018). IgG1 PTEC-binding index correlated with tubulo-interstitial inflammation score in renal biopsy from corresponding patients. Our data suggested that total IgG and IgG1 PTEC-binding index in serum of LN patients correlate with serological activity, and in combination could predict renal flares. The correlation between IgG1

  10. Tofogliflozin, A Highly Selective Inhibitor of SGLT2 Blocks Proinflammatory and Proapoptotic Effects of Glucose Overload on Proximal Tubular Cells Partly by Suppressing Oxidative Stress Generation.

    PubMed

    Ishibashi, Y; Matsui, T; Yamagishi, S

    2016-03-01

    Ninety percent of glucose filtered by the glomerulus is reabsorbed by a sodium-glucose cotransporter 2 (SGLT2), which is mainly expressed on S1 and S2 segment of renal proximal tubules. Since SGLT-2-mediated glucose reabsorption is increased under diabetic conditions, selective inhibition of SGLT2 is a potential therapeutic target for the treatment of diabetes. We have recently shown that an inhibitor of SGLT2 has anti-inflammatory and antifibrotic effects on experimental diabetic nephropathy partly by suppressing advanced glycation end products formation and oxidative stress generation in the kidney. However, the direct effects of SGLT2 inhibitor on tubular cell damage remain unclear. In this study, we investigated the effects of tofogliflozin, a highly selective inhibitor of SGLT2 on oxidative stress generation, inflammatory and proapoptotic reactions in cultured human proximal tubular cells exposed to high glucose. Tofogliflozin dose-dependently suppressed glucose entry into tubular cells. High glucose exposure (30 mM) for 4 and 24 h significantly increased oxidative stress generation in tubular cells, which were suppressed by the treatment of tofogliflozin or an antioxidant N-acetylcysteine (NAC). Monocyte chemoattractant protein-1 (MCP-1) gene expression and apoptotic cell death were induced by 4 h- and 8 day-exposure to high glucose, respectively, both of which were also blocked by tofogliflozin or NAC. The present study suggests that SGLT2-mediated glucose entry into tubular cells could stimulate oxidative stress and evoke inflammatory and proapoptotic reactions in this cell type. Blockade of glucose reabsorption in tubular cells by SGLT2 inhibitor might exert beneficial effects on tubulointerstitial damage in diabetic nephropathy.

  11. CBFB and MYH11 in inv(16)(p13q22) of Acute Myeloid Leukemia Display Close Spatial Proximity in Interphase Nuclei of Human Hematopoietic Stem Cells

    PubMed Central

    Weckerle, Allison B.; Santra, Madhumita; Ng, Maggie C.Y.; Koty, Patrick P.; Wang, Yuh-Hwa

    2013-01-01

    To gain a better understanding of the mechanism of chromosomal translocations in cancer, we investigated the spatial proximity between CBFB and MYH11 genes involved in inv(16)(p13q22) found in acute myeloid leukemia patients. Previous studies have demonstrated a role for spatial genome organization in the formation of tumorigenic abnormalities. The non-random localization of chromosomes and, more specifically, of genes appears to play a role in the mechanism of chromosomal translocations. Here, two-color fluorescence in situ hybridization and confocal microscopy were used to measure the interphase distance between CBFB and MYH11 in hematopoietic stem cells, where inv(16)(p13q22) is believed to occur, leading to leukemia development. The measured distances in hematopoietic stem cells were compared to mesenchymal stem cells, peripheral blood lymphocytes and fibroblasts, as spatial genome organization is determined to be cell-type specific. Results indicate that CBFB and MYH11 are significantly closer in hematopoietic stem cells compared to all other cell types examined. Furthermore, the CBFB-MYH11 distance is significantly reduced compared to CBFB and a control locus in hematopoietic stem cells, although separation between CBFB and the control is ~70% of that between CBFB and MYH11 on metaphase chromosomes. Hematopoietic stem cells were also treated with fragile site-inducing chemicals since both genes contain translocation breakpoints within these regions. However, treatment with fragile site-inducing chemicals did not significantly affect the interphase distance. Consistent with previous studies, our results suggest that gene proximity may play a role in the formation of cancer-causing rearrangements, providing insight into the mechanism of chromosomal abnormalities in human tumors. PMID:21638519

  12. Extruded plastic scintillator including inorganic powders

    DOEpatents

    Bross, Alan D.; Mellott, Kerry L.; Pla-Dalmau, Anna

    2006-06-27

    A method for producing a plastic scintillator is disclosed. A plurality of nano-sized particles and one or more dopants can be combined with a plastic material for the formation of a plastic scintillator thereof. The nano-sized particles, the dopant and the plastic material can be combined within the dry inert atmosphere of an extruder to produce a reaction that results in the formation of a plastic scintillator thereof and the deposition of energy within the plastic scintillator, such that the plastic scintillator produces light signifying the detection of a radiative element. The nano-sized particles can be treated with an inert gas prior to processing the nano-sized particles, the dopant and the plastic material utilizing the extruder. The plastic scintillator can be a neutron-sensitive scintillator, x-ray sensitive scintillator and/or a scintillator for the detection of minimum ionizing particles.

  13. Renal cysteine conjugate C-S lyase mediated toxicity of halogenated alkenes in primary cultures of human and rat proximal tubular cells.

    PubMed

    McGoldrick, Trevor A; Lock, Edward A; Rodilla, Vicente; Hawksworth, Gabrielle M

    2003-07-01

    Proximal tubular cells from human (HPT) and rat (RPT) kidneys were isolated, grown to confluence and incubated with S-(1,2-dichlorovinyl)- l-cysteine (DCVC), S-(1,2,2-trichlorovinyl)- l-cysteine (TCVC), S-(1,1,2,2-tetrafluoroethyl)- l-cysteine (TFEC) and S-(2-chloro-1,1-difluorethyl)- l-cysteine (CDFEC), the cysteine conjugates of nephrotoxicants. The cultures were exposed to the conjugates for 12, 24 and 48 h and the toxicity determined using the MTT assay. All four conjugates caused dose-dependent toxicity to RPT cells over the range 50-1,000 microM, the order of toxicity being DCVC>TCVC>TFEC=CDFEC. The inclusion of aminooxyacetic acid (AOAA; 250 microM), an inhibitor of pyridoxal phosphate-dependent enzymes such as C-S lyase, afforded protection, indicating that C-S lyase has a role in the bioactivation of these conjugates. In HPT cultures only DCVC caused significant time- and dose-dependent toxicity. Exposure to DCVC (500 microM) for 48 h decreased cell viability to 7% of control cell values, whereas co-incubation of DCVC (500 microM) with AOAA (250 microM) resulted in cell viability of 71%. Human cultures were also exposed to S-(1,2-dichlorovinyl)-glutathione (DCVG). DCVG was toxic to HPT cells, but the onset of toxicity was delayed compared with the corresponding cysteine conjugate. AOAA afforded almost complete protection from DCVG toxicity. Acivicin (250 microM), an inhibitor of gamma-glutamyl transferase (gamma-GT), partially protected against DCVG (500 microM)-induced toxicity at 48 h (5% viability and 53% viability in the absence and presence of acivicin, respectively). These results suggest that DCVG requires processing by gamma-GT prior to bioactivation by C-S lyase in HPT cells. The activity of C-S lyase, using TFEC as a substrate, and glutamine transaminase K (GTK) was measured in rat and human cells with time in culture. C-S lyase activity in RPT and HPT cells decreased to approximately 30% of fresh cell values by the time the cells reached

  14. RNA Whole-Mount In Situ Hybridization Proximity Ligation Assay (rISH-PLA), an Assay for Detecting RNA-Protein Complexes in Intact Cells.

    PubMed

    Roussis, Ioannis M; Myers, Fiona A; Scarlett, Garry P

    2017-03-03

    Techniques for studying RNA-protein interactions have lagged behind those for DNA-protein interactions as a consequence of the complexities associated with working with RNA. This unit describes a method for the adaptation of the In Situ Hybridization-Proximity Ligation Assay (ISH-PLA) to the study of RNA regulation (rISH-PLA). The rISH-PLA assay allows the identification of a given RNA-protein complex at subcellular and single-cell resolution, thus avoiding the lack of spatial resolution and sensitivity associated with assaying heterogeneous cell populations from which conventional RNA-protein interaction detection techniques suffer. This technique will be particularly usefully for studying the activity of RNA binding proteins (RBPs) in complex mixtures of cells, for example tissue sections or whole embryos. © 2017 by John Wiley & Sons, Inc.

  15. Proximal Tubule Epithelial Cell Specific Ablation of the Spermidine/Spermine N1-Acetyltransferase Gene Reduces the Severity of Renal Ischemia/Reperfusion Injury

    PubMed Central

    Zahedi, Kamyar; Barone, Sharon; Wang, Yang; Murray-Stewart, Tracy; Roy-Chaudhury, Prabir; Smith, Roger D.; Casero, Robert A.; Soleimani, Manoocher

    2014-01-01

    Background Expression and activity of spermidine/spermine N1-acetyltransferase (SSAT) increases in kidneys subjected to ischemia/reperfusion (I/R) injury, while its ablation reduces the severity of such injuries. These results suggest that increased SSAT levels contribute to organ injury; however, the role of SSAT specifically expressed in proximal tubule epithelial cells, which are the primary targets of I/R injury, in the mediation of renal damage remains unresolved. Methods Severity of I/R injury in wt and renal proximal tubule specific SSAT-ko mice (PT-SSAT-Cko) subjected to bilateral renal I/R injury was assessed using cellular and molecular biological approaches. Results Severity of the loss of kidney function and tubular damage are reduced in PT-SSAT-Cko- compared to wt-mice after I/R injury. In addition, animals treated with MDL72527, an inhibitor of polyamine oxidases, had less severe renal damage than their vehicle treated counter-parts. The renal expression of HMGB 1 and Toll like receptors (TLR) 2 and 4 were also reduced in PT-SSAT-Cko- compared to wt mice after I/R injury. Furthermore, infiltration of neutrophils, as well as expression of tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1) and interleukin-6 (IL-6) transcripts were lower in the kidneys of PT-SSAT-Cko compared to wt mice after I/R injury. Finally, the activation of caspase3 was more pronounced in the wt compared to PT-SSAT-Cko animals. Conclusions Enhanced SSAT expression by proximal tubule epithelial cells leads to tubular damage, and its deficiency reduces the severity of renal I/R injury through reduction of cellular damage and modulation of the innate immune response. PMID:25390069

  16. Molecular origins of scintillation in organic scintillators (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Feng, Patrick; Mengesha, Wondwosen; Myllenbeck, Nicholas

    2016-09-01

    Organic-based scintillators are indispensable materials for radiation detection owing to their high sensitivity to fast neutrons, low cost, and tailorable properties. There has been a recent resurgence of interest in organic scintillators due to exciting discoveries related to neutron discrimination and gamma-ray spectroscopy, which represent capabilities previously thought not possible in these materials. I will discuss our development of crystalline and polymer-based scintillators for these applications. Structure-property relationships related to intermolecular interactions and host-guest electronic exchange will be discussed in the context of energy-transfer pathways relevant to scintillation. An emphasis will be placed on the rational design of these materials, as guided by first principles and DFT calculations. Two related topics will be discussed: 1) Incorporation of organometallic triplet-harvesting additives to plastic scintillator matrices to confer a 'two-state' (singlet and triplet) luminescence signature to different types of ionizing radiation. This approach relies upon energetic and spatial overlap between the donor and acceptor excited states for efficient electronic exchange. Key considerations also include synthetic modification of the luminescence spectra and kinetics, as well as the addition of secondary additives to increase the recombination efficiency. 2) Design of organotin-containing plastic scintillators as a route towards gamma-ray spectroscopy. Organometallic compounds were selected on the basis of distance-dependent quenching relationships, phase compatibility with the polymer matrix, and the gamma-ray cross sections. This approach is guided by molecular modeling and radiation transport modeling to achieve the highest possible detection sensitivity luminescence intensity.

  17. FNAL-NICADD extruded scintillator

    SciTech Connect

    Beznosko, D.; Bross, A.; Dyshkant, A.; Pla-Dalmau, A.; Rykalin, V.; /Northern Illinois U.

    2005-09-01

    The possibility to produce a scintillator that satisfies the demands of physicists from different science areas has emerged with the installation of an extrusion line at Fermi National Accelerator Laboratory (FNAL). The extruder is the product of the fruitful collaboration between FNAL and Northern Illinois Center for Accelerator and Detector Development (NICADD) at Northern Illinois University (NIU). The results from the light output, light attenuation length and mechanical tolerance indicate that FNAL-NICADD scintillator is of high quality. Improvements in the extrusion die will yield better scintillator profiles and decrease the time needed for initial tuning. This paper will present the characteristics of the FNAL-NICADD scintillator based on the measurements performed. They include the response to MIPs from cosmic rays for individual extruded strips and irradiation studies where extruded samples were irradiated up to 1 Mrad. We will also discuss the results achieved with a new die design. The attractive perspective of using the extruded scintillator with MRS (Metal Resistive Semiconductor) photodetector readout will also be shown.

  18. Sp7/Osterix induces the mouse pro-α2(I) collagen gene (Col1a2) expression via the proximal promoter in osteoblastic cells.

    PubMed

    Yano, Hiroyuki; Hamanaka, Ryoji; Nakamura-Ota, Miki; Adachi, Sawako; Zhang, Juan Juan; Matsuo, Noritaka; Yoshioka, Hidekatsu

    2014-09-26

    Bone is essentially composed of two components, hydroxyapatite and extracellular matrix proteins. The extracellular matrix of bone is primary composed of collagen, mostly type I collagen, with lesser amounts of other types of collagen such as type V collagen. Osteoblast differentiation is a multi-step process in which many classes of factors function in a coordinated manner. Sp7/Osterix, which binds to G/C-rich sequences, is a transcription factor that contributes to osteoblast differentiation. The present study aimed to clarify the involvement of Sp7/Osterix with the proximal promoter region of the mouse Col1a2 gene containing multiple G/C-rich sequences exist. Consequently, a functional analysis of the proximal mouse Col1a2 promoter showed that a substitution mutation of the second G/C-rich sequence from the transcription site specifically decreased the activity of osteoblastic cells. In addition, the experiments of overexpression of Sp7/Osterix and treatment with its specific siRNA showed that this G/C-rich sequence is responsible for the specific expression in osteoblastic cells. Consistent with these data, Sp7/Osterix bound to the region and increased the expression of the Col1a2 gene in association with osteoblast differentiation in the culture system.

  19. Heat shock protein 27 expression in human proximal tubule cells exposed to lethal and sublethal concentrations of CdCl2.

    PubMed Central

    Somji, S; Sens, D A; Garrett, S H; Sens, M A; Todd, J H

    1999-01-01

    The expression of hsp 27 mRNA and protein was determined in cultured human proximal tubule (HPT) cells exposed to lethal and sublethal concentrations of Cd2+ under both acute and extended conditions. Initial procedures demonstrated that HPT cells display the classic stress response following physical and chemical stress. Heat stress (42.5 degrees C for 1 hr) caused an increase in both hsp 27 mRNA and protein as well as a shift in the protein to a more phosphorylated state. Results were similar when the cells were subjected to chemical stress (exposure to 100 microM sodium arsenite for 4 hr). Acute exposure to 53 microM CdCl2 for 4 hr also resulted in an increase in hsp 27 mRNA and protein and a shift to the more phosphorylated protein isoform. Extended Cd2+ exposure involved continuous treatment with Cd2+ at both lethal and sublethal levels over a 16-day time course. The results of this treatment showed that chronic exposure to Cd2+ failed to increase either hsp 27 mRNA or protein expression in HPT cells, even at lethal Cd2+ concentrations. In fact, hsp 27 protein levels decreased as compared to controls at both lethal and sub-lethal exposure to Cd2+. These findings imply that hsp 27 expression in human proximal tubule cells may have two distinct modes depending on the nature (acute vs. chronic) of the stress. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 PMID:10379001

  20. Transforming growth factor-β1 reduces megalin- and cubilin-mediated endocytosis of albumin in proximal-tubule-derived opossum kidney cells

    PubMed Central

    Gekle, Michael; Knaus, Petra; Nielsen, Rikke; Mildenberger, Sigrid; Freudinger, Ruth; Wohlfarth, Verena; Sauvant, Christoph; Christensen, Erik I

    2003-01-01

    Transforming growth factor (TGF)-β1 is a member of a superfamily of multifunctional cytokines involved in several pathological processes of the kidney, including fibrogenesis, apoptosis and epithelial-mesenchymal transition. These events lead to tubulointerstitial fibrosis and glomerulosclerosis. Less is known about TGF-β1-induced alterations of cell function. An important function of proximal tubular cells is reabsorption of filtered proteins, including albumin, via megalin-cubilin-dependent receptor-mediated endocytosis. In this study we used a well established cell culture model (proximal-tubule-derived opossum kidney (OK) cells) in order to test the hypothesis that TGF-β1 reduces megalin-cubilin-mediated endocytosis. Previously we have shown that albumin endocytosis in OK cells is mediated by megalin/cubulin. TGF-β1 led to a time- and dose-dependent downregulation of megalin-cubilin-mediated endocytosis without affecting two other transport systems tested. Binding, internalization and intracellular trafficking of the ligand albumin were affected. Decreased binding resulted from reduced cubilin and megalin expression in the 200 000 g membrane fraction. The underlying mechanism of TGF-β1 action does not involve mitogen-activated protein kinases, protein kinase C or A, or reactive oxygen species. In contrast, TGF-β1-induced downregulation of megalin-cubilin-mediated endocytosis was sensitive to inhibition of translation and transcription and was preceded by Smad2 and 3 phosphorylation. Dominant negative Smad2/3 constructs prevented the effect of TGF-β1. In conclusion our data indicate that enhanced levels of TGF-β1 occurring in various nephropathies can lead to downregulation of megalin-cubilin-dependent endocytosis. Probably, TGF-β1 leads to Smad2- and Smad3-dependent expression of negative regulators of receptor-mediated endocytosis. PMID:14561830

  1. Transforming growth factor-beta1 reduces megalin- and cubilin-mediated endocytosis of albumin in proximal-tubule-derived opossum kidney cells.

    PubMed

    Gekle, Michael; Knaus, Petra; Nielsen, Rikke; Mildenberger, Sigrid; Freudinger, Ruth; Wohlfarth, Verena; Sauvant, Christoph; Christensen, Erik I

    2003-10-15

    Transforming growth factor (TGF)-beta1 is a member of a superfamily of multifunctional cytokines involved in several pathological processes of the kidney, including fibrogenesis, apoptosis and epithelial-mesenchymal transition. These events lead to tubulointerstitial fibrosis and glomerulosclerosis. Less is known about TGF-beta1-induced alterations of cell function. An important function of proximal tubular cells is reabsorption of filtered proteins, including albumin, via megalin-cubilin-dependent receptor-mediated endocytosis. In this study we used a well established cell culture model (proximal-tubule-derived opossum kidney (OK) cells) in order to test the hypothesis that TGF-beta1 reduces megalin-cubilin-mediated endocytosis. Previously we have shown that albumin endocytosis in OK cells is mediated by megalin/cubulin. TGF-beta1 led to a time- and dose-dependent downregulation of megalin-cubilin-mediated endocytosis without affecting two other transport systems tested. Binding, internalization and intracellular trafficking of the ligand albumin were affected. Decreased binding resulted from reduced cubilin and megalin expression in the 200 000 g membrane fraction. The underlying mechanism of TGF-beta1 action does not involve mitogen-activated protein kinases, protein kinase C or A, or reactive oxygen species. In contrast, TGF-beta1-induced downregulation of megalin-cubilin-mediated endocytosis was sensitive to inhibition of translation and transcription and was preceded by Smad2 and 3 phosphorylation. Dominant negative Smad2/3 constructs prevented the effect of TGF-beta1. In conclusion our data indicate that enhanced levels of TGF-beta1 occurring in various nephropathies can lead to downregulation of megalin-cubilin-dependent endocytosis. Probably, TGF-beta1 leads to Smad2- and Smad3-dependent expression of negative regulators of receptor-mediated endocytosis.

  2. Radiopure metal-loaded liquid scintillator

    SciTech Connect

    Rosero, Richard; Yeh, Minfang

    2015-08-17

    Metal-loaded liquid scintillator plays a key role in particle and nuclear physics experiments. The applications of metal ions in various neutrino experiments and the purification methods for different scintillator components are discussed in this paper.

  3. Radiopure Metal-Loaded Liquid Scintillator

    SciTech Connect

    Rosero, Richard; Yeh, Minfang

    2015-03-18

    Metal-loaded liquid scintillator plays a key role in particle and nuclear physics experiments. The applications of metal ions in various neutrino experiments and the purification methods for different scintillator components are discussed in this paper.

  4. Advances in scintillators for medical imaging applications

    NASA Astrophysics Data System (ADS)

    van Loef, Edgar V.; Shah, Kanai S.

    2014-09-01

    A review is presented of some recent work in the field of inorganic scintillator research for medical imaging applications, in particular scintillation detectors for Single-Photon Emission Computed Tomography (SPECT) and Positron Emission Tomography (PET).

  5. A SILAC-Based Approach Elicits the Proteomic Responses to Vancomycin-Associated Nephrotoxicity in Human Proximal Tubule Epithelial HK-2 Cells.

    PubMed

    Li, Zhi-Ling; Zhou, Shu-Feng

    2016-01-29

    Vancomycin, a widely used antibiotic, often induces nephrotoxicity, however, the molecular targets and underlying mechanisms of this side effect remain unclear. The present study aimed to examine molecular interactome and analyze the signaling pathways related to the vancomycin-induced nephrotoxicity in human proximal tubule epithelial HK-2 cells using the stable isotope labeling by amino acids in cell culture (SILAC) approach. The quantitative proteomic study revealed that there were at least 492 proteins interacting with vancomycin and there were 290 signaling pathways and cellular functions potentially regulated by vancomycin in HK-2 cells. These proteins and pathways played a critical role in the regulation of cell cycle, apoptosis, autophagy, EMT, and ROS generation. These findings suggest that vancomycin-induced proteomic responses in HK-2 cells involvefunctional proteins and pathways that regulate cell cycle, apoptosis, autophagy, and redox homeostasis. This is the first systemic study revealed the networks of signaling pathways and proteomic responses to vancomycin treatment in HK-2 cells, and the data may be used to discriminate the molecular and clinical subtypes and to identify new targets and biomarkers for vancomycin-induced nephrotoxic effect. Further studies are warranted to explore the potential of quantitative proteomic analysis in the identification of new targets and biomarkers for drug-induced renal toxicity.

  6. Scintillating glass fiber neutron senors

    SciTech Connect

    Abel, K.H.; Arthur, R.J.; Bliss, M.

    1994-04-01

    Cerium-doped lithium-silicate glass fibers have been developed at Pacific Northwest Laboratory (PNL) for use as thermal neutron detectors. By using highly-enriched {sup 6} Li , these fibers efficiently capture thermal neutrons and produce scintillation light that can be detected at the ends of the fibers. Advantages of scintillating fibers over {sup 3}He or BF{sub 3} proportional tubes include flexibility in geometric configuration, ruggedness in high-vibration environments, and less detector weight for the same neutron sensitivity. This paper describes the performance of these scintillating fibers with regard to count rates, pulse height spectra, absolute efficiencies, and neutron/gamma discrimination. Fibers with light transmission lengths (1/e) of greater than 2 m have been produced at PNL. Neutron sensors in fiber form allow development of a variety of neutron detectors packaged in previously unavailable configurations. Brief descriptions of some of the devices already produced are included to illustrate these possibilities.

  7. Scintillation-Hardened GPS Receiver

    NASA Technical Reports Server (NTRS)

    Stephens, Donald R.

    2015-01-01

    CommLargo, Inc., has developed a scintillation-hardened Global Positioning System (GPS) receiver that improves reliability for low-orbit missions and complies with NASA's Space Telecommunications Radio System (STRS) architecture standards. A software-defined radio (SDR) implementation allows a single hardware element to function as either a conventional radio or as a GPS receiver, providing backup and redundancy for platforms such as the International Space Station (ISS) and high-value remote sensing platforms. The innovation's flexible SDR implementation reduces cost, weight, and power requirements. Scintillation hardening improves mission reliability and variability. In Phase I, CommLargo refactored an open-source GPS software package with Kalman filter-based tracking loops to improve performance during scintillation and also demonstrated improved navigation during a geomagnetic storm. In Phase II, the company generated a new field-programmable gate array (FPGA)-based GPS waveform to demonstrate on NASA's Space Communication and Navigation (SCaN) test bed.

  8. Liquid scintillator tiles for calorimetry

    DOE PAGES

    Amouzegar, M.; Belloni, A.; Bilki, B.; ...

    2016-11-28

    Future experiments in high energy and nuclear physics may require large, inexpensive calorimeters that can continue to operate after receiving doses of 50 Mrad or more. Also, the light output of liquid scintillators suffers little degradation under irradiation. However, many challenges exist before liquids can be used in sampling calorimetry, especially regarding developing a packaging that has sufficient efficiency and uniformity of light collection, as well as suitable mechanical properties. We present the results of a study of a scintillator tile based on the EJ-309 liquid scintillator using cosmic rays and test beam on the light collection efficiency and uniformity,more » and some preliminary results on radiation hardness.« less

  9. Unitary scintillation detector and system

    DOEpatents

    McElhaney, Stephanie A.; Chiles, Marion M.

    1994-01-01

    The invention is a unitary alpha, beta, and gamma scintillation detector and system for sensing the presence of alpha, beta, and gamma radiations selectively or simultaneously. The scintillators are mounted in a light-tight housing provided with an entrance window for admitting alpha, beta, and gamma radiation and excluding ambient light from the housing. Light pulses from each scintillator have different decay constants that are converted by a photosensitive device into corresponding differently shaped electrical pulses. A pulse discrimination system identifies the electrical pulses by their respective pulse shapes which are determined by decay time. The identified electrical pulses are counted in separate channel analyzers to indicate the respective levels of sensed alpha, beta, and gamma radiations.

  10. Unitary scintillation detector and system

    DOEpatents

    McElhaney, S.A.; Chiles, M.M.

    1994-05-31

    The invention is a unitary alpha, beta, and gamma scintillation detector and system for sensing the presence of alpha, beta, and gamma radiations selectively or simultaneously. The scintillators are mounted in a light-tight housing provided with an entrance window for admitting alpha, beta, and gamma radiation and excluding ambient light from the housing. Light pulses from each scintillator have different decay constants that are converted by a photosensitive device into corresponding differently shaped electrical pulses. A pulse discrimination system identifies the electrical pulses by their respective pulse shapes which are determined by decay time. The identified electrical pulses are counted in separate channel analyzers to indicate the respective levels of sensed alpha, beta, and gamma radiations. 10 figs.

  11. Fracture-resistant lanthanide scintillators

    DOEpatents

    Doty, F. Patrick

    2011-01-04

    Lanthanide halide alloys have recently enabled scintillating gamma ray spectrometers comparable to room temperature semiconductors (<3% FWHM energy resolutions at 662 keV). However brittle fracture of these materials upon cooling hinders the growth of large volume crystals. Efforts to improve the strength through non-lanthanide alloy substitution, while preserving scintillation, have been demonstrated. Isovalent alloys having nominal compositions of comprising Al, Ga, Sc, Y, and In dopants as well as aliovalent alloys comprising Ca, Sr, Zr, Hf, Zn, and Pb dopants were prepared. All of these alloys exhibit bright fluorescence under UV excitation, with varying shifts in the spectral peaks and intensities relative to pure CeBr.sub.3. Further, these alloys scintillate when coupled to a photomultiplier tube (PMT) and exposed to .sup.137Cs gamma rays.

  12. Development of radiation hard scintillators

    SciTech Connect

    Markley, F.; Woods, D.; Pla-Dalmau, A.; Foster, G. ); Blackburn, R. )

    1992-05-01

    Substantial improvements have been made in the radiation hardness of plastic scintillators. Cylinders of scintillating materials 2.2 cm in diameter and 1 cm thick have been exposed to 10 Mrads of gamma rays at a dose rate of 1 Mrad/h in a nitrogen atmosphere. One of the formulations tested showed an immediate decrease in pulse height of only 4% and has remained stable for 12 days while annealing in air. By comparison a commercial PVT scintillator showed an immediate decrease of 58% and after 43 days of annealing in air it improved to a 14% loss. The formulated sample consisted of 70 parts by weight of Dow polystyrene, 30 pbw of pentaphenyltrimethyltrisiloxane (Dow Corning DC 705 oil), 2 pbw of p-terphenyl, 0.2 pbw of tetraphenylbutadiene, and 0.5 pbw of UVASIL299LM from Ferro.

  13. Nanophosphor composite scintillator with a liquid matrix

    DOEpatents

    McKigney, Edward Allen; Burrell, Anthony Keiran; Bennett, Bryan L.; Cooke, David Wayne; Ott, Kevin Curtis; Bacrania, Minesh Kantilal; Del Sesto, Rico Emilio; Gilbertson, Robert David; Muenchausen, Ross Edward; McCleskey, Thomas Mark

    2010-03-16

    An improved nanophosphor scintillator liquid comprises nanophosphor particles in a liquid matrix. The nanophosphor particles are optionally surface modified with an organic ligand. The surface modified nanophosphor particle is essentially surface charge neutral, thereby preventing agglomeration of the nanophosphor particles during dispersion in a liquid scintillator matrix. The improved nanophosphor scintillator liquid may be used in any conventional liquid scintillator application, including in a radiation detector.

  14. Coenzyme Q10 protects renal proximal tubule cells against nicotine-induced apoptosis through induction of p66(shc)-dependent antioxidant responses.

    PubMed

    Arany, Istvan; Carter, Anthony; Hall, Samuel; Fulop, Tibor; Dixit, Mehul

    2017-02-01

    Chronic nicotine exposure (via smoking, E-cigarettes) increases oxidative stress in the kidney that sensitizes it to additional injury in experimental models and in the renal patient. The pro-apoptotic p66(shc) protein-via serine36 phosphorylation that facilitates its mitochondrial translocation and therein cytochrome c binding-generates oxidative stress that leads to injury of renal proximal tubule cells during chronic nicotine exposure. Coenzyme Q10-a clinically safe antioxidant-has been used against nicotine/smoke extract-associated oxidative stress in various non-renal cells. This study explored the anti-oxidant/anti-apoptotic effect of Coenzyme Q10 on nicotine-induced oxidative stress and its impact on p66shc in cultured rat renal proximal tubule cells (NRK52E). We studied the anti-oxidant effect of 10 µM Coenzyme Q10 using various mutants of the p66shc gene and also determined the induction of selected anti-oxidant entities (antioxidant response element, promoter of the manganese superoxide dismutase gene) in reporter luciferase assay during oxidative stress induced by 200 µM nicotine. Our studies revealed that Coenzyme Q10 strongly inhibits nicotine-mediated production of reactive oxygen species and consequent apoptosis that requires serine36 phosphorylation but not mitochondrial translocation/cytochrome c binding of p66(shc). While both nicotine and Coenzyme Q10 stimulates the p66shc promoter, only nicotine exposure results in mitochondrial translocation of p66(shc). In contrast, the Coenzyme Q10-stimulated and non-mitochondrial p66(shc) activates the anti-oxidant manganese superoxide dismutase promoter via the antioxidant response elements and hence, rescues cells from nicotine-induced oxidative stress and consequent apoptosis.

  15. Genetic and epigenetic regulation of AHR gene expression in MCF-7 breast cancer cells: role of the proximal promoter GC-rich region

    PubMed Central

    Englert, Neal A.; Turesky, Robert J.; Han, Weiguo; Bessette, Erin E.; Spivack, Simon D.; Caggana, Michele; Spink, David C.; Spink, Barbara C.

    2014-01-01

    The aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor, contributes to carcinogenesis through its role in the regulation of cytochrome P450 1 (CYP1)-catalyzed metabolism of carcinogens. Here, we investigated genetic and epigenetic mechanisms that affect AhR expression. Analyses of the human AHR proximal promoter in MCF-7 human breast cancer cells using luciferase assays and electrophoretic mobility shift assays revealed multiple specificity protein (Sp) 1 binding sequences that are transcriptional activators in vitro. The regulation of AhR expression was evaluated in long-term estrogen exposed (LTEE) MCF-7 cells, which showed increased AhR expression, enhanced CYP1 inducibility, and increased capacity to form DNA adducts when exposed to the dietary carcinogen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine. The increased AhR expression in LTEE cells was found not to result from increased mRNA stability, differential RNA processing, or decreased DNA methylation. Analysis of the AHR proximal promoter region using chromatin immunoprecipitation confirmed that enhanced expression of AhR in LTEE cells involves changes in histone modifications, notably decreased trimethylation of histone 3, lysine 27. Upon further examination of the GC-rich Sp1-binding region, we confirmed that it contains a polymorphic (GGGGC)n repeat. In a population of newborns from New York State, the allele frequency of (GGGGC)n was n = 4>5≫6, 2. Circular dichroism spectroscopy revealed the ability of sequences of this GC-rich region to form guanine-quadruplex structures in vitro. These studies revealed multiple levels at which AhR expression may be controlled, and offer additional insights into mechanisms regulating AhR expression that can ultimately impact carcinogenesis. PMID:22728919

  16. Genetic and epigenetic regulation of AHR gene expression in MCF-7 breast cancer cells: role of the proximal promoter GC-rich region.

    PubMed

    Englert, Neal A; Turesky, Robert J; Han, Weiguo; Bessette, Erin E; Spivack, Simon D; Caggana, Michele; Spink, David C; Spink, Barbara C

    2012-09-01

    The aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor, contributes to carcinogenesis through its role in the regulation of cytochrome P450 1 (CYP1)-catalyzed metabolism of carcinogens. Here, we investigated genetic and epigenetic mechanisms that affect AhR expression. Analyses of the human AHR proximal promoter in MCF-7 human breast cancer cells using luciferase assays and electrophoretic mobility shift assays revealed multiple specificity protein (Sp) 1 binding sequences that are transcriptional activators in vitro. The regulation of AhR expression was evaluated in long-term estrogen exposed (LTEE) MCF-7 cells, which showed increased AhR expression, enhanced CYP1 inducibility, and increased capacity to form DNA adducts when exposed to the dietary carcinogen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine. The increased AhR expression in LTEE cells was found not to result from increased mRNA stability, differential RNA processing, or decreased DNA methylation. Analysis of the AHR proximal promoter region using chromatin immunoprecipitation confirmed that enhanced expression of AhR in LTEE cells involves changes in histone modifications, notably decreased trimethylation of histone 3, lysine 27. Upon further examination of the GC-rich Sp1-binding region, we confirmed that it contains a polymorphic (GGGGC)(n) repeat. In a population of newborns from New York State, the allele frequency of (GGGGC)(n) was n = 4 > 5 ≫ 6, 2. Circular dichroism spectroscopy revealed the ability of sequences of this GC-rich region to form guanine-quadruplex structures in vitro. These studies revealed multiple levels at which AhR expression may be controlled, and offer additional insights into mechanisms regulating AhR expression that can ultimately impact carcinogenesis.

  17. CBFB and MYH11 in inv(16)(p13q22) of acute myeloid leukemia displaying close spatial proximity in interphase nuclei of human hematopoietic stem cells.

    PubMed

    Weckerle, Allison B; Santra, Madhumita; Ng, Maggie C Y; Koty, Patrick P; Wang, Yuh-Hwa

    2011-09-01

    To gain a better understanding of the mechanism of chromosomal translocations in cancer, we investigated the spatial proximity between CBFB and MYH11 genes involved in inv(16)(p13q22) found in patients with acute myeloid leukemia. Previous studies have demonstrated a role for spatial genome organization in the formation of tumorigenic abnormalities. The nonrandom localization of chromosomes and, more specifically, of genes appears to play a role in the mechanism of chromosomal translocations. Here, two-color fluorescence in situ hybridization and confocal microscopy were used to measure the interphase distance between CBFB and MYH11 in hematopoietic stem cells (HSCs), where inv(16)(p13q22) is believed to occur, leading to leukemia development. The measured distances in HSCs were compared with mesenchymal stem cells, peripheral blood lymphocytes, and fibroblasts, as spatial genome organization is determined to be cell-type specific. Results indicate that CBFB and MYH11 are significantly closer in HSCs compared with all other cell types examined. Furthermore, the CBFB-MYH11 distance is significantly reduced compared with CBFB and a control locus in HSCs, although separation between CBFB and the control is ∼70% of that between CBFB and MYH11 on metaphase chromosomes. HSCs were also treated with fragile site-inducing chemicals because both the genes contain translocation breakpoints within these regions. However, treatment with fragile site-inducing chemicals did not significantly affect the interphase distance. Consistent with previous studies, our results suggest that gene proximity may play a role in the formation of cancer-causing rearrangements, providing insight into the mechanism of chromosomal abnormalities in human tumors.

  18. Hygroscopicity Evaluation of Halide Scintillators

    SciTech Connect

    Zhuravleva, M; Stand, L; Wei, H; Hobbs, C. L.; Boatner, Lynn A; Ramey, Joanne Oxendine; Burger, Arnold; Rowe, E; Bhattacharya, P.; Tupitsyn, E; Melcher, Charles L

    2014-01-01

    A collaborative study of relative hygroscopicity of anhydrous halide scintillators grown at various laboratories is presented. We have developed a technique to evaluate moisture sensitivity of both raw materials and grown crystals, in which the moisture absorption rate is measured using a gravimetric analysis. Degradation of the scintillation performance was investigated by recording gamma-ray spectra and monitoring the photopeak position, count rate and energy resolution. The accompanying physical degradation of the samples exposed to ambient atmosphere was photographically recorded as well. The results were compared with ben

  19. Characteristics of High Latitude Ionosphere Scintillations

    NASA Astrophysics Data System (ADS)

    Morton, Y.

    2012-12-01

    As we enter a new solar maximum period, global navigation satellite systems (GNSS) receivers, especially the ones operating in high latitude and equatorial regions, are facing an increasing threat from ionosphere scintillations. The increased solar activities, however, also offer a great opportunity to collect scintillation data to characterize scintillation signal parameters and ionosphere irregularities. While there are numerous GPS receivers deployed around the globe to monitor ionosphere scintillations, most of them are commercial receivers whose signal processing mechanisms are not designed to operate under ionosphere scintillation. As a result, they may distort scintillation signal parameters or lose lock of satellite signals under strong scintillations. Since 2008, we have established and continuously improved a unique GNSS receiver array at HAARP, Alaska. The array contains high ends commercial receivers and custom RF front ends which can be automatically triggered to collect high quality GPS and GLONASS satellite signals during controlled heating experiments and natural scintillation events. Custom designed receiver signal tracking algorithms aim to preserve true scintillation signatures are used to process the raw RF samples. Signal strength, carrier phase, and relative TEC measurements generated by the receiver array since its inception have been analyzed to characterize high latitude scintillation phenomena. Daily, seasonal, and solar events dependency of scintillation occurrence, spectral contents of scintillation activities, and plasma drifts derived from these measurements will be presented. These interesting results demonstrate the feasibility and effectiveness of our experimental data collection system in providing insightful details of ionosphere responses to active perturbations and natural disturbances.

  20. Composite scintillators for detection of ionizing radiation

    DOEpatents

    Dai, Sheng [Knoxville, TN; Stephan, Andrew Curtis [Knoxville, TN; Brown, Suree S [Knoxville, TN; Wallace, Steven A [Knoxville, TN; Rondinone, Adam J [Knoxville, TN

    2010-12-28

    Applicant's present invention is a composite scintillator having enhanced transparency for detecting ionizing radiation comprising a material having optical transparency wherein said material comprises nano-sized objects having a size in at least one dimension that is less than the wavelength of light emitted by the composite scintillator wherein the composite scintillator is designed to have selected properties suitable for a particular application.

  1. Photonic crystal scintillators and methods of manufacture

    SciTech Connect

    Torres, Ricardo D.; Sexton, Lindsay T.; Fuentes, Roderick E.; Cortes-Concepcion, Jose

    2015-08-11

    Photonic crystal scintillators and their methods of manufacture are provided. Exemplary methods of manufacture include using a highly-ordered porous anodic alumina membrane as a pattern transfer mask for either the etching of underlying material or for the deposition of additional material onto the surface of a scintillator. Exemplary detectors utilizing such photonic crystal scintillators are also provided.

  2. Blockade of excitatory synaptogenesis with proximal dendrites of dentate granule cells following rapamycin treatment in a mouse model of temporal lobe epilepsy

    PubMed Central

    Yamawaki, Ruth; Thind, Khushdev; Buckmaster, Paul S.

    2014-01-01

    Inhibiting the mTOR signaling pathway with rapamycin blocks granule cell axon (mossy fiber) sprouting after epileptogenic injuries, including pilocarpine-induced status epilepticus. However, it remains unclear whether axons from other types of neurons sprout into the inner molecular layer and synapse with granule cell dendrites despite rapamycin treatment. If so, other aberrant positive-feedback networks might develop. To test this possibility stereological electron microscopy was used to estimate numbers of excitatory synapses in the inner molecular layer per hippocampus in pilocarpine-treated control mice, in mice 5 d after pilocarpine-induced status epilepticus, and after status epilepticus and daily treatment beginning 24 h later with rapamycin or vehicle for 2 months. The optical fractionator method was used to estimate numbers of granule cells in Nissl-stained sections so that numbers of excitatory synapses in the inner molecular layer per granule cell could be calculated. Control mice had an average of 2280 asymmetric synapses in the inner molecular layer per granule cell, which was reduced to 63% of controls 5 d after status epilepticus, recovered to 93% of controls in vehicle-treated mice 2 months after status epilepticus, but remained at only 63% of controls in rapamycin-treated mice. These findings reveal that rapamycin prevented excitatory axons from synapsing with proximal dendrites of granule cells and raise questions about the recurrent excitation hypothesis of temporal lobe epilepsy. PMID:25234294

  3. Synthesis of plastic scintillation microspheres: Evaluation of scintillators

    NASA Astrophysics Data System (ADS)

    Santiago, L. M.; Bagán, H.; Tarancón, A.; Garcia, J. F.

    2013-01-01

    The use of plastic scintillation microspheres (PSm) appear to be an alternative to liquid scintillation for the quantification of alpha and beta emitters because it does not generate mixed wastes after the measurement (organic and radioactive). In addition to routine radionuclide determinations, PSm can be used for further applications, e.g. for usage in a continuous monitoring equipment, for measurements of samples with a high salt concentration and for an extractive scintillation support which permits the separation, pre-concentration and measurement of the radionuclides without additional steps of elution and sample preparation. However, only a few manufacturers provide PSm, and the low number of regular suppliers reduces its availability and restricts the compositions and sizes available. In this article, a synthesis method based on the extraction/evaporation methodology has been developed and successfully used for the synthesis of plastic scintillation microspheres. Seven different compositions of plastic scintillation microspheres have been synthesised; PSm1 with polystyrene, PSm2 with 2,5-Diphenyloxazol(PPO), PSm3 with p-terphenyl (pT), PSm4 with PPO and 1,4-bis(5-phenyloxazol-2-yl) (POPOP), PSm5 pT and (1,4-bis [2-methylstyryl] benzene) (Bis-MSB), PSm6 with PPO, POPOP and naphthalene and PSm7 with pT, Bis-MSB and naphthalene. The synthesised plastic scintillation microspheres have been characterised in terms of their morphology, detection capabilities and alpha/beta separation capacity. The microspheres had a median diameter of approximately 130 μm. Maximum detection efficiency values were obtained for the PSm4 composition as follows 1.18% for 3H, 51.2% for 14C, 180.6% for 90Sr/90Y and 76.7% for 241Am. Values of the SQP(E) parameter were approximately 790 for PSm4 and PSm5. These values show that the synthesised PSm exhibit good scintillation properties and that the spectra are at channel numbers higher than in commercial PSm. Finally, the addition of

  4. Differential Response of the Human Renal Proximal Tubular Epithelial Cell Line HK-2 to Shiga Toxin Types 1 and 2 ▿

    PubMed Central

    Lentz, Erin K.; Leyva-Illades, Dinorah; Lee, Moo-Seung; Cherla, Rama P.; Tesh, Vernon L.

    2011-01-01

    Shiga toxins (Stxs) are expressed by the enteric pathogens Shigella dysenteriae serotype 1 and certain serotypes of Escherichia coli. Stx-producing bacteria cause bloody diarrhea with the potential to progress to acute renal failure. Stxs are potent protein synthesis inhibitors and are the primary virulence factors responsible for renal damage that may follow diarrheal disease. We explored the use of the immortalized human proximal tubule epithelial cell line HK-2 as an in vitro model of Stx-induced renal damage. We showed that these cells express abundant membrane Gb3 and are differentially susceptible to the cytotoxic action of Stxs, being more sensitive to Shiga toxin type 1 (Stx1) than to Stx2. At early time points (24 h), HK-2 cells were significantly more sensitive to Stxs than Vero cells; however, by 72 h, Vero cell monolayers were completely destroyed while some HK-2 cells survived toxin challenge, suggesting that a subpopulation of HK-2 cells are relatively toxin resistant. Fluorescently labeled Stx1 B subunits localized to both lysosomal and endoplasmic reticulum (ER) compartments in HK-2 cells, suggesting that differences in intracellular trafficking may play a role in susceptibility to Stx-mediated cytotoxicity. Although proinflammatory cytokines were not upregulated by toxin challenge, Stx2 selectively induced the expression of two chemokines, macrophage inflammatory protein-1α (MIP-1α) and MIP-1β. Stx1 and Stx2 differentially activated components of the ER stress response in HK-2 cells. Finally, we demonstrated significant poly(ADP-ribose) polymerase (PARP) cleavage after exposure to Stx1 or Stx2. However, procaspase 3 cleavage was undetectable, suggesting that HK-2 cells may undergo apoptosis in response to Stxs in a caspase 3-independent manner. PMID:21708996

  5. Nifedipine inhibits advanced glycation end products (AGEs) and their receptor (RAGE) interaction-mediated proximal tubular cell injury via peroxisome proliferator-activated receptor-gamma activation

    SciTech Connect

    Matsui, Takanori; Yamagishi, Sho-ichi; Takeuchi, Masayoshi; Ueda, Seiji; Fukami, Kei; Okuda, Seiya

    2010-07-23

    Research highlights: {yields} Nifedipine inhibited the AGE-induced up-regulation of RAGE mRNA levels in tubular cells, which was prevented by GW9662, an inhibitor of peroxisome proliferator-activated receptor-{gamma}. {yields} GW9662 treatment alone increased RAGE mRNA levels in tubular cells. {yields} Nifedipine inhibited the AGE-induced reactive oxygen species generation, NF-{kappa}B activation and increases in intercellular adhesion molecule-1 and transforming growth factor-{beta} gene expression in tubular cells, all of which were blocked by GW9662. -- Abstract: There is a growing body of evidence that advanced glycation end products (AGEs) and their receptor (RAGE) interaction evokes oxidative stress generation and subsequently elicits inflammatory and fibrogenic reactions, thereby contributing to the development and progression of diabetic nephropathy. We have previously found that nifedipine, a calcium-channel blocker (CCB), inhibits the AGE-induced mesangial cell damage in vitro. However, effects of nifedipine on proximal tubular cell injury remain unknown. We examined here whether and how nifedipine blocked the AGE-induced tubular cell damage. Nifedipine, but not amlodipine, a control CCB, inhibited the AGE-induced up-regulation of RAGE mRNA levels in tubular cells, which was prevented by the simultaneous treatment of GW9662, an inhibitor of peroxisome proliferator-activated receptor-{gamma} (PPAR{gamma}). GW9662 treatment alone was found to increase RAGE mRNA levels in tubular cells. Further, nifedipine inhibited the AGE-induced reactive oxygen species generation, NF-{kappa}B activation and increases in intercellular adhesion molecule-1 and transforming growth factor-beta gene expression in tubular cells, all of which were blocked by GW9662. Our present study provides a unique beneficial aspect of nifedipine on diabetic nephropathy; it could work as an anti-oxidative and anti-inflammatory agent against AGEs in tubular cells by suppressing RAGE expression

  6. Parathyroid hormone induces epithelial-to-mesenchymal transition via the Wnt/β-catenin signaling pathway in human renal proximal tubular cells.

    PubMed

    Guo, Yunshan; Li, Zhen; Ding, Raohai; Li, Hongdong; Zhang, Lei; Yuan, Weijie; Wang, Yanxia

    2014-01-01

    Epithelial-to-mesenchymal transition (EMT) has been shown to play an important role in renal fibrogenesis. Recent studies suggested parathyroid hormone (PTH) could accelerate EMT and subsequent organ fibrosis. However, the precise molecular mechanisms underlying PTH-induced EMT remain unknown. The present study was to investigate whether Wnt/β-catenin signaling pathway is involved in PTH-induced EMT in human renal proximal tubular cells (HK-2 cells) and to determine the profile of gene expression associated with PTH-induced EMT. PTH could induce morphological changes and gene expression characteristic of EMT in cultured HK-2 cells. Suppressing β-catenin expression or DKK1 limited gene expression characteristic of PTH-induced EMT. Based on the PCR array analysis, PTH treatment resulted in the up-regulation of 18 genes and down-regulation of 9 genes compared with the control. The results were further supported by a western blot analysis, which showed the increased Wnt4 protein expression. Wnt4 overexpression also promotes PTH-induced EMT in HK-2 cells. The findings demonstrated that PTH-induced EMT in HK-2 cells is mediated by Wnt/β-catenin signal pathway, and Wnt4 might be a key gene during PTH-induced EMT.

  7. Calcium oxalate crystals and oxalate induce an epithelial-to-mesenchymal transition in the proximal tubular epithelial cells: Contribution to oxalate kidney injury

    PubMed Central

    Convento, Marcia Bastos; Pessoa, Edson Andrade; Cruz, Edgar; da Glória, Maria Aparecida; Schor, Nestor; Borges, Fernanda Teixeira

    2017-01-01

    TGF-β1 is the main mediator of epithelial-to-mesenchymal transition (EMT). Hyperoxaluria induces crystalluria, interstitial fibrosis, and progressive renal failure. This study analyzed whether hyperoxaluria is associated with TGF-β1 production and kidney fibrosis in mice and if oxalate or calcium oxalate (CaOx) could induce EMT in proximal tubule cells (HK2) and therefore contribute to the fibrotic process. Hyperoxaluria was induced by adding hydroxyproline and ethylene glycol to the mice’s drinking water for up to 60 days. Renal function and oxalate and urinary crystals were evaluated. Kidney collagen production and TGF-β1 expression were assessed. EMT was analyzed in vitro according to TGF-β1 production, phenotypic characterization, invasion, cell migration, gene and protein expression of epithelial and mesenchymal markers. Hyperoxaluric mice showed a decrease in renal function and an increase in CaOx crystals and Ox urinary excretion. The deposition of collagen in the renal interstitium was observed. HK2 cells stimulated with Ox and CaOx exhibited a decreased expression of epithelial as well as increased expression mesenchymal markers; these cells presented mesenchymal phenotypic changes, migration, invasiveness capability and TGF-β1 production, characterizing EMT. Treatment with BMP-7 or its overexpression in HK2 cells was effective at preventing it. This mechanism may contribute to the fibrosis observed in hyperoxaluria. PMID:28387228

  8. Extruded plastic scintillator for MINERvA

    SciTech Connect

    Pla-Dalmau, Anna; Bross, Alan D.; Rykalin, Victor V.; Wood, Brian M.; /NICADD, DeKalb

    2005-11-01

    An extrusion line has recently been installed at Fermilab in collaboration with NICADD (Northern Illinois Center for Accelerator and Detector Development). This new facility will serve to further develop and improve extruded plastic scintillator. Since polystyrene is widely used in the consumer industry, the logical path was to investigate the extrusion of commercial-grade polystyrene pellets with dopants to yield high quality plastic scintillator. The D0 and MINOS experiments are already using extruded scintillator strips in their detectors. A new experiment at Fermilab is pursuing the use of extruded plastic scintillator. A new plastic scintillator strip is being tested and its properties characterized. The initial results are presented here.

  9. SNO+ Scintillator Purification and Assay

    SciTech Connect

    Ford, R.; Vazquez-Jauregui, E.; Chen, M.; Chkvorets, O.; Hallman, D.

    2011-04-27

    We describe the R and D on the scintillator purification and assay methods and technology for the SNO+ neutrino and double-beta decay experiment. The SNO+ experiment is a replacement of the SNO heavy water with liquid scintillator comprised of 2 g/L PPO in linear alkylbenzene (LAB). During filling the LAB will be transported underground by rail car and purified by multi-stage distillation and steam stripping at a flow rate of 19 LPM. While the detector is operational the scintillator can be recirculated at 150 LPM (full detector volume in 4 days) to provide repurification as necessary by either water extraction (for Ra, K, Bi) or by functional metal scavenger columns (for Pb, Ra, Bi, Ac, Th) followed by steam stripping to remove noble gases and oxygen (Rn, O{sub 2}, Kr, Ar). The metal scavenger columns also provide a method for scintillator assay for ex-situ measurement of the U and Th chain radioactivity. We have developed ''natural'' radioactive spikes of Pb and Ra in LAB and use these for purification testing. Lastly, we present the planned operating modes and purification strategies and the plant specifications and design.

  10. SNO+ Scintillator Purification and Assay

    NASA Astrophysics Data System (ADS)

    Ford, R.; Chen, M.; Chkvorets, O.; Hallman, D.; Vázquez-Jáuregui, E.

    2011-04-01

    We describe the R&D on the scintillator purification and assay methods and technology for the SNO+ neutrino and double-beta decay experiment. The SNO+ experiment is a replacement of the SNO heavy water with liquid scintillator comprised of 2 g/L PPO in linear alkylbenzene (LAB). During filling the LAB will be transported underground by rail car and purified by multi-stage distillation and steam stripping at a flow rate of 19 LPM. While the detector is operational the scintillator can be recirculated at 150 LPM (full detector volume in 4 days) to provide repurification as necessary by either water extraction (for Ra, K, Bi) or by functional metal scavenger columns (for Pb, Ra, Bi, Ac, Th) followed by steam stripping to remove noble gases and oxygen (Rn, O2, Kr, Ar). The metal scavenger columns also provide a method for scintillator assay for ex-situ measurement of the U and Th chain radioactivity. We have developed "natural" radioactive spikes of Pb and Ra in LAB and use these for purification testing. Lastly, we present the planned operating modes and purification strategies and the plant specifications and design.

  11. Application of scintillating properties of liquid xenon and silicon photomultiplier technology to medical imaging

    NASA Astrophysics Data System (ADS)

    Gomez-Cadenas, J. J.; Benlloch-Rodriguez, J. M.; Ferrario, Paola

    2016-04-01

    We describe a new positron emission time-of-flight apparatus using liquid xenon. The detector is based in a liquid xenon scintillating cell. The cell shape and dimensions can be optimized depending on the intended application. In its simplest form, the liquid xenon scintillating cell is a box in which two faces are covered by silicon photomultipliers and the others by a reflecting material such as Teflon. It is a compact, homogenous and highly efficient detector which shares many of the desirable properties of monolithic crystals, with the added advantage of high yield and fast scintillation offered by liquid xenon. Our initial studies suggest that good energy and spatial resolution comparable with that achieved by lutetium oxyorthosilicate crystals can be obtained with a detector based in liquid xenon scintillating cells. In addition, the system can potentially achieve an excellent coincidence resolving time of better than 100 ps.

  12. Indoxyl sulfate-induced activation of (pro)renin receptor is involved in expression of TGF-β1 and α-smooth muscle actin in proximal tubular cells.

    PubMed

    Saito, Shinichi; Shimizu, Hidehisa; Yisireyili, Maimaiti; Nishijima, Fuyuhiko; Enomoto, Atsushi; Niwa, Toshimitsu

    2014-05-01

    Activation of (pro)renin receptor (PRR) is involved in the progression of chronic kidney disease. However, the role of indoxyl sulfate, a uremic toxin, in the activation of PRR is not clear. The present study aimed to clarify the role of indoxyl sulfate in activation of PRR, in relation to renal expression of fibrotic genes. Renal expression of PRR and renin/prorenin was up-regulated in chronic kidney disease rats compared with normal rats, whereas AST-120 suppressed these expression by reducing serum levels of indoxyl sulfate. Furthermore, administration of indoxyl sulfate to normotensive and hypertensive rats increased renal expression of PRR and renin/prorenin. Indoxyl sulfate induced expression of PRR and prorenin in cultured human proximal tubular cells (HK-2 cells). Indoxyl sulfate-induced PRR expression was inhibited by small interfering RNAs of signal transducer and activator of transcription 3 (Stat3) and nuclear factor-κB p65 in proximal tubular cells. N-acetylcysteine, an antioxidant, and diphenyleneiodonium, an inhibitor of nicotinamide adenine dinucleotide phosphate oxidase, suppressed indoxyl sulfate-induced PRR expression in proximal tubular cells. N-acetylcysteine prevented indoxyl sulfate-induced phosphorylation of Stat3 in proximal tubular cells. PRR small interfering RNA inhibited indoxyl sulfate-induced expression of TGF-β1 and α-smooth muscle actin in proximal tubular cells. Taken together, indoxyl sulfate-induced up-regulation of prorenin expression and activation of PRR through production of reactive oxygen species and activation of Stat3 and nuclear factor-κB play an important role in the expression of TGF-β1 and α-smooth muscle actin in proximal tubular cells. Thus, indoxyl sulfate-induced activation of prorenin/PRR might be involved in renal fibrosis.

  13. Restoration of autophagy by puerarin in lead-exposed primary rat proximal tubular cells via regulating AMPK-mTOR signaling.

    PubMed

    Song, Xiangbin; Li, Zifa; Liu, Fei; Wang, Zhenyong; Wang, Lin

    2017-03-01

    Previous study has demonstrated that puerarin (PU) exerts nephroprotective effect against Pb-induced cytotoxicity in primary rat proximal tubular (rPT) cells. Autophagy can protect cells from various cytotoxic stimuli, but its role in the process of PU against Pb-induced nephrotoxicity is still unknown. This study aims to investigate whether PU can alleviate Pb-induced renal damage by recovering autophagy. Data showed that Pb inhibited the autophagic flux, as evidenced by the accumulation of LC3-II and p62 as well as the confocal microscopy analysis of GFP-LC3 puncta and punctate spots of monodansylcadaverine staining, whereas coadministration of PU could restore Pb-induced autophagy inhibition. Moreover, PU dramatically enhanced the phosphorylation of 5'AMP-activated protein kinase (AMPK) and inhibited the phosphorylation of mammalian target of rapamycin (mTOR) and its target proteins p70S6 kinase (p70S6K) and 4E-binding protein 1 (4E-BP1) in Pb-exposed rPT cells. Collectively, these evidence suggested that PU restored the impaired autophagic flux in Pb-treated rPT cells partly by activating autophagy via AMPK/mTOR-mediated signaling pathway.

  14. Fenofibrate, a PPARα agonist, protect proximal tubular cells from albumin-bound fatty acids induced apoptosis via the activation of NF-kB.

    PubMed

    Zuo, Nan; Zheng, Xiaoyu; Liu, Hanzhe; Ma, Xiaoli

    2015-01-01

    Albumin-bound fatty acids is the main cause of renal damage, PPARα is responsible in the metabolism of fatty acids. Previous study found that PPARα played a protective role in fatty acids overload associated tubular injury. The aim of the present study is to investigate whether fenofibrate, a PPARα ligands, could contribute to the renoprotective action in fatty acids overload proximal tubule epithelial cells. We observed in HK-2 cells that fenofibrate significantly inhibited fatty acids bound albumin (FA-BSA) induced up-regulation of MCP-1 and IL-8. Treatment with fenofibrate attenuated renal oxidative stress induced by FA-BSA as evidenced by decreased MDA level, increased SOD activity and catalase, GPx-1 expression. FA-BSA induced apoptosis of HK-2 cells were also obviously prevented by fenofibrate. Furthermore, fenofibrate significantly increased the expression of PPARα mRNA and protein in FA-BSA treated cells. Finally, the activation of NF-kB induced by FA-BSA was markedly suppressed by fenofibrate. Taken together, our study describes a renoprotective role of fenofibrate in fatty acids associated tubular toxicity, and the transcriptional activation of PPARα and suppression of NF-kB were at least partially involved.

  15. Cobalt chloride attenuates oxidative stress and inflammation through NF-κB inhibition in human renal proximal tubular epithelial cells.

    PubMed

    Oh, Se Won; Lee, Yun-Mi; Kim, Sejoong; Chin, Ho Jun; Chae, Dong-Wan; Na, Ki Young

    2014-09-01

    We evaluated the effect of cobalt chloride (CoCl2) on TNF-α and IFN-γ-induced-inflammation and reactive oxygen species (ROS) in renal tubular epithelial cells (HK-2 cells). We treated HK-2 cells with CoCl2 before the administration of TNF-α/IFN-γ. To regulate hemeoxygenase-1 (HO-1) expression, the cells were treated CoCl2 or HO-1 siRNA. CoCl2 reduced the generation of ROS induced by TNF-α/IFN-γ. TNF-α/IFN-γ-treated-cells showed an increase in the nuclear translocation of phosphorylated NF-κBp65 protein, the DNA-binding activity of NF-κBp50 and NF-κB transcriptional activity and a decrease in IκBα protein expression. These changes were restored by CoCl2. We noted an intense increase in monocyte chemoattractant protein-1 (MCP-1) and regulated on activation normal T cell expressed and secreted (RANTES) production in TNF-α/IFN-γ-treated cells. We demonstrated that this effect was mediated through NF-κB signaling because an NF-κB inhibitor significantly reduced MCP-1 and RANTES production. CoCl2 effectively reduced MCP-1 and RANTES production. The expression of HO-1 was increased by CoCl2 and decreased by HO-1 siRNA. However, knockdown of HO-1 by RNA interference did not affect MCP-1 or RANTES production. We suggest that CoCl2 has a protective effect on TNF-α/IFN-γ-induced inflammation through the inhibition of NF-κB and ROS in HK-2 cells. However, CoCl2 appears to act in an HO-1-independent manner.

  16. Nickel (II)-induced cytotoxicity and apoptosis in human proximal tubule cells through a ROS- and mitochondria-mediated pathway

    SciTech Connect

    Wang, Yi-Fen; Shyu, Huey-Wen; Chang, Yi-Chuang; Tseng, Wei-Chang; Huang, Yeou-Lih; Lin, Kuan-Hua; Chou, Miao-Chen; Liu, Heng-Ling; Chen, Chang-Yu

    2012-03-01

    Nickel compounds are known to be toxic and carcinogenic in kidney and lung. In this present study, we investigated the roles of reactive oxygen species (ROS) and mitochondria in nickel (II) acetate-induced cytotoxicity and apoptosis in the HK-2 human renal cell line. The results showed that the cytotoxic effects of nickel (II) involved significant cell death and DNA damage. Nickel (II) increased the generation of ROS and induced a noticeable reduction of mitochondrial membrane potential (MMP). Analysis of the sub-G1 phase showed a significant increase in apoptosis in HK-2 cells after nickel (II) treatment. Pretreatment with N-acetylcysteine (NAC) not only inhibited nickel (II)-induced cell death and DNA damage, but also significantly prevented nickel (II)-induced loss of MMP and apoptosis. Cell apoptosis triggered by nickel (II) was characterized by the reduced protein expression of Bcl-2 and Bcl-xL and the induced the protein expression of Bad, Bcl-Xs, Bax, cytochrome c and caspases 9, 3 and 6. The regulation of the expression of Bcl-2-family proteins, the release of cytochrome c and the activation of caspases 9, 3 and 6 were inhibited in the presence of NAC. These results suggest that nickel (II) induces cytotoxicity and apoptosis in HK-2 cells via ROS generation and that the mitochondria-mediated apoptotic signaling pathway may be involved in the positive regulation of nickel (II)-induced renal cytotoxicity.

  17. A Human Renal Proximal Tubule Cell Line with Stable Organic Anion Transporter 1 and 3 Expression Predictive for Antiviral-Induced Toxicity.

    PubMed

    Nieskens, Tom T G; Peters, Janny G P; Schreurs, Marieke J; Smits, Niels; Woestenenk, Rob; Jansen, Katja; van der Made, Thom K; Röring, Melanie; Hilgendorf, Constanze; Wilmer, Martijn J; Masereeuw, Rosalinde

    2016-03-01

    Drug-induced nephrotoxicity still hampers drug development, because current translation from in vitro or animal studies to human lacks high predictivity. Often, renal adverse effects are recognized only during clinical stages of drug development. The current study aimed to establish a robust and a more complete human cell model suitable for screening of drug-related interactions and nephrotoxicity. In addition to endogenously expressed renal organic cation transporters and efflux transporters, conditionally immortalized proximal tubule epithelial cells (ciPTEC) were completed by transduction of cells with the organic anion transporter (OAT) 1 or OAT3. Fluorescence-activated cell sorting upon exposure to the OAT substrate fluorescein successfully enriched transduced cells. A panel of organic anions was screened for drug-interactions in ciPTEC-OAT1 and ciPTEC-OAT3. The cytotoxic response to the drug-interactions with antivirals was further examined by cell viability assays. Upon subcloning, concentration-dependent fluorescein uptake was found with a higher affinity for ciPTEC-OAT1 (Km = 0.8 ± 0.1 μM) than ciPTEC-OAT3 (Km = 3.7 ± 0.5 μM). Co-exposure to known OAT1 and/or OAT3 substrates (viz. para-aminohippurate, estrone sulfate, probenecid, furosemide, diclofenac, and cimetidine) in cultures spanning 29 passage numbers revealed relevant inhibitory potencies, confirming the robustness of our model for drug-drug interactions studies. Functional OAT1 was directly responsible for cytotoxicity of adefovir, cidofovir, and tenofovir, while a drug interaction with zidovudine was not associated with decreased cell viability. Our data demonstrate that human-derived ciPTEC-OAT1 and ciPTEC-OAT3 are promising platforms for highly predictive drug screening during early phases of drug development.

  18. The role of HIF-1 in up-regulating MICA expression on human renal proximal tubular epithelial cells during hypoxia/reoxygenation

    PubMed Central

    2010-01-01

    Background Human major histocompatibility complex class I-related chain A (MICA) plays a dual role in adaptive and innate immune responses. Increasing evidence demonstrates that MICA is closely correlated with acute and chronic kidney allograft rejection. Therefore, understanding the activation mechanisms of MICA is important in kidney transplantation. We previously demonstrated that ischemia/reperfusion injury (IRI) could up-regulate MICA expression on mouse kidney allografts. Since hypoxia-inducible factor-1 (HIF-1) is the master regulator of cellular adaptive responses to hypoxia during IRI, here we investigate whether HIF-1 could up-regulate MICA expression and its influence on NK cell cytotoxicity. Results We find that HIF-1alpha plays an important role in up-regulating MICA expression, inducing IFNgamma secretion and NK cell cytotoxicity during hypoxia/reoxygenation. First, we generated a HIF-1alphaDELTAODD-expressing adenovirus to stably and functionally express HIF-1alpha in human renal proximal tubular epithelial (HK-2) cells under normoxia conditions. HIF-1alpha over-expression in HK-2 cells induces MICA expression and enhances NK cell cytotoxic activity towards cells that express HIF-1alpha. Second, we used a hypoxia/reoxygenation cell model to simulate IRI in vitro and found that the suppression of HIF-1alpha by RNAi induces down-regulation of MICA expression and inhibits NK cytotoxicity. In antibody blocking experiments, an anti-MICA mAb was able to down-regulate NK cell cytotoxic activity towards HK-2 cells that over-expressed HIF-1alpha. Moreover, when NK cells were co-cultured with the HK-2 cells expressing MICA, which was up-regulated by over-expression of HIF-1alpha, there was a significant increase in the secretion of IFNgamma. In the presence of the blocking MICA mAb, IFNgamma secretion was significantly decreased. Conclusions These results demonstrate that hypoxia/reoxygenation-promoted MICA expression on HK-2 cells is through a HIF-1 pathway

  19. Expression of the constitutive and inducible forms of heat shock protein 70 in human proximal tubule cells exposed to heat, sodium arsenite, and CdCl(2).

    PubMed Central

    Somji, S; Todd, J H; Sens, M A; Garrett, S H; Sens, D A

    1999-01-01

    We determined the expression of the constitutive (hsc 70) and inducible (hsp 70) forms of heat shock protein 70 mRNA and protein in human proximal tubule (HPT) cells exposed to lethal and sublethal concentrations of Cd(+2) under both acute and extended conditions of exposure. The HPT cells exhibited the classic heat shock response when subjected to a physical (heat) or chemical stress (sodium arsenite); hsc 70 mRNA and protein levels were constant or slightly increased, whereas hsp 70 mRNA and protein were greatly elevated. Acute exposure to 53.4 microM CdCl(2) for 4 hr failed to increase either hsc 70 mRNA or protein, a finding similar to that observed under classic conditions of stress. However, under identical conditions of acute exposure to Cd(2+), the expected increase in hsp 70 protein level was suppressed as compared to that found under classic conditions of physical or chemical stress. The decrease in hsp 70 protein level correlated to the reduced expression of mRNA from the hsp 70B gene. The expression of mRNA from the hsp 70A and hsp 70C genes was similar to that found when the cells were treated with heat shock or sodium arsenite. We modeled an extended exposure to Cd(2+) by treating the cells continuously with Cd(2+) at both lethal and sublethal levels over a 16-day time course. Chronic exposure to Cd(2+) failed to increase either hsc 70 mRNA or protein levels in the HPT cells at a nonlethal dosage level and decreased hsc 70 mRNA and protein levels late in the time course of lethal exposure. Under identical conditions, the expression of hsp 70 protein remained at basal levels that were only marginally detectable throughout the time course. Hsp 70A and hsp 70C mRNA levels were unaltered by extended exposure to Cd(2+), and hsp 70B mRNA was not detected during the 16-day time course. Cd(2+) is a poor inducer of hsc 70 and hsp 70 in the proximal tubule under both acute and long-term exposure. These results reinforce the fact that the expression of hsp 70

  20. Role of metabolites in MDMA (ecstasy)-induced nephrotoxicity: an in vitro study using rat and human renal proximal tubular cells.

    PubMed

    Carvalho, Márcia; Hawksworth, Gabrielle; Milhazes, Nuno; Borges, Fernanda; Monks, Terrence J; Fernandes, Eduarda; Carvalho, Félix; Bastos, Maria Lourdes

    2002-10-01

    The metabolism of 3,4-methylenedioxymethamphetamine (MDMA, ecstasy) has recently been implicated in the mechanisms underlying ecstasy-induced neurotoxicity and hepatotoxicity. However, its potential role in ecstasy-induced kidney toxicity has yet to be investigated. Thus, primary cultures of rat and human renal proximal tubular cells (PTCs) were used to investigate the cytotoxicity induced by MDMA and its metabolites methylenedioxyamphetamine (MDA), alpha-methyldopamine (alpha-MeDA), and the glutathione (GSH) conjugates 5-(glutathion- S-yl)-alpha-MeDA and 2,5- bis(glutathion- S-yl)-alpha-MeDA. Cell viability was evaluated using the mitochondrial MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. MDMA and MDA were not found to be toxic to either rat or human PTCs at any concentration tested (100-800 micro M). In contrast, 800 micro M alpha-MeDA caused 60% and 40% cell death in rat and human PTCs, respectively. Conjugation of alpha-MeDA with GSH resulted in the formation of even more potent nephrotoxicants. Thus, exposure of rat and human PTC monolayers to 400 micro M 5-(glutathion- S-yl)-alpha-MeDA caused approximately 80% and 70% cell death, respectively. 5-(Glutathion- S-yl)-alpha-MeDA (400 micro M) was more toxic than 2,5- bis(glutathion- S-yl)-alpha-MeDA to rat renal PTCs but equally potent in human renal PTCs. Pre-incubation of rat PTCs with either acivicin, an inhibitor of gamma-glutamyl transpeptidase (gamma-GT), or bestatin, an inhibitor of aminopeptidase M, resulted in increased toxicity of 5-(glutathion- S-yl)-alpha-MeDA but had no effect on 2,5- bis(glutathion- S-yl)-alpha-MeDA-mediated cytotoxicity. The present data provide evidence that metabolism is required for the expression of MDMA-induced renal toxicity in vitro. In addition, metabolism of 5-(glutathion- S-yl)-alpha-MeDA by gamma-GT and aminopeptidase M to the corresponding cystein- S-yl-glycine and/or cystein- S-yl conjugates is likely to be associated with detoxication of

  1. Characterization of the scintillation anisotropy in crystalline stilbene scintillator detectors

    SciTech Connect

    Schuster, P.; Brubaker, E.

    2016-11-23

    This study reports a series of measurements that characterize the directional dependence of the scintillation response of crystalline melt-grown and solution-grown trans-stilbene to incident DT and DD neutrons. These measurements give the amplitude and pulse shape dependence on the proton recoil direction over one hemisphere of the crystal, confirming and extending previous results in the literature for melt-grown stilbene and providing the first measurements for solution-grown stilbene. In similar measurements of liquid and plastic detectors, no directional dependence was observed, confirming the hypothesis that the anisotropy in stilbene and other organic crystal scintillators is a result of internal effects due to the molecular or crystal structure and not an external effect on the measurement system.

  2. Characterization of the scintillation anisotropy in crystalline stilbene scintillator detectors

    DOE PAGES

    Schuster, P.; Brubaker, E.

    2016-11-23

    This study reports a series of measurements that characterize the directional dependence of the scintillation response of crystalline melt-grown and solution-grown trans-stilbene to incident DT and DD neutrons. These measurements give the amplitude and pulse shape dependence on the proton recoil direction over one hemisphere of the crystal, confirming and extending previous results in the literature for melt-grown stilbene and providing the first measurements for solution-grown stilbene. In similar measurements of liquid and plastic detectors, no directional dependence was observed, confirming the hypothesis that the anisotropy in stilbene and other organic crystal scintillators is a result of internal effects duemore » to the molecular or crystal structure and not an external effect on the measurement system.« less

  3. Pravastatin inhibits advanced glycation end products (AGEs)-induced proximal tubular cell apoptosis and injury by reducing receptor for AGEs (RAGE) level.

    PubMed

    Ishibashi, Yuji; Yamagishi, Sho-ichi; Matsui, Takanori; Ohta, Keisuke; Tanoue, Ryuichiro; Takeuchi, Masayoshi; Ueda, Seiji; Nakamura, Kei-ichiro; Okuda, Seiya

    2012-08-01

    Advanced glycation end products (AGEs) and their receptor (RAGE) axis play a role in diabetic nephropathy. Statins have been shown to ameliorate renal function and reduce proteinuria in patients with chronic kidney disease. However, the effects of statin on AGEs-induced tubular cell damage remain unknown. We examined here whether and how pravastatin could block the AGEs-RAGE-elicited tubular cell injury in vitro. Gene expression level was evaluated by real-time reverse-transcription polymerase chain reactions. Reactive oxygen species (ROS) generation was measured with dihydroethidium staining. Apoptosis was analyzed in an enzyme-linked immunosorbent assay. Asymmetric dimethylarginine (ADMA) expression was evaluated by immunostaining. Pravastatin dose-dependently inhibited the AGEs-induced up-regulation of RAGE mRNA level, ROS generation and apoptosis in human renal proximal tubular cells. Further, AGEs decreased mRNA level of dimethylarginine dimethylaminohydrolase-2, an enzyme that mainly degrades asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase and subsequently increased ADMA generation in tubular cells, both of which were also prevented by pravastatin. Geranylgeranyl pyrophosphate (GGPP) treatment blocked all of the effects of pravastatin on tubular cells. We found that rosuvastatin also significantly blocked the AGEs-induced increase in RAGE mRNA level and ROS generation, both of which were prevented by GGPP. Our present study suggests that pravastatin could inhibit the AGEs-induced apoptosis and ADMA generation in tubular cells by suppressing RAGE expression probably via inhibition of GGPP synthesis. Pravastatin may exert beneficial effects on tubular damage in diabetic nephropathy by blocking the AGEs-RAGE axis.

  4. Analysis of Altered MicroRNA Expression Profiles in Proximal Renal Tubular Cells in Response to Calcium Oxalate Monohydrate Crystal Adhesion: Implications for Kidney Stone Disease

    PubMed Central

    Wang, Bohan; Wu, Bolin; Liu, Jun; Yao, Weimin; Xia, Ding; Li, Lu; Chen, Zhiqiang; Ye, Zhangqun; Yu, Xiao

    2014-01-01

    Background Calcium oxalate monohydrate (COM) is the major crystalline component in kidney stones and its adhesion to renal tubular cells leads to tubular injury. However, COM-induced toxic effects in renal tubular cells remain ambiguous. MicroRNAs (miRNAs) play an important role in gene regulation at the posttranscriptional levels. Objective The present study aimed to assess the potential changes in microRNAs of proximal renal tubular cells in response to the adhesion of calcium oxalate monohydrate (COM) crystals. Methodology Lactate dehydrogenase (LDH) activity and DAPI staining were used to measure the toxic effects of HK-2 cells exposed to COM crystals. MicroRNA microarray and mRNA microarray were applied to evaluate the expression of HK-2 cells exposed to COM crystals. Quantitative real-time PCR (qRT-PCR) technology was used to validate the microarray results. Target prediction, Gene Ontology (GO) analysis and pathway analysis were applied to predict the potential roles of microRNAs in biological processes. Principal Findings Our study showed that COM crystals significantly altered the global expression profile of miRNAs in vitro. After 24 h treatment with a dose (1 mmol/L), 25 miRNAs were differentially expressed with a more than 1.5-fold change, of these miRNAs, 16 were up-regulated and 9 were down-regulated. A majority of these differentially expressed miRNAs were associated with cell death, mitochondrion and metabolic process. Target prediction and GO analysis suggested that these differentially expressed miRNAs potentially targeted many genes which were related to apoptosis, regulation of metabolic process, intracellular signaling cascade, insulin signaling pathway and type 2 diabetes. Conclusion Our study provides new insights into the role of miRNAs in the pathogenesis associated with nephrolithiasis. PMID:24983625

  5. Boiogito Increases the Metabolism of Fatty Acids in Proximal Tubular Cells through Peroxisome Proliferators-Activated Receptor (PPAR) α Agonistic Activity.

    PubMed

    Kobayashi, Kyoko; Matsuyama, Wakana; Arai, Yuhei; Koizumi, Saho; Shimizu, Tatsuya; Tomioka, Rie; Sasaki, Kenroh

    2016-01-01

    The promotion of fatty acid metabolism, to which peroxisome proliferators-activated receptor (PPAR) α contributes, has been suggested to participate in maintaining the function of renal proximal tubular epithelial cells (PTECs). The loading of fatty acids to PTECs could result in cell inflammation and cell death. A "Kampo" medicine, Boiogito (BO), is used to treat overweight women exhibiting chronic fatigue and edema in the lower extremities or knees. BO improves renal function by reducing the portion of fatty acids, thereby preventing damage to PTECs. In this study, BO and Astragalus Root (AsR), a constituent crude drug of BO, were administered orally to intravenously bovine serum albumin (BSA)-administered mice to evaluate the PPARα-cAMP responsive element binding protein (CREB) binding protein (CBP) complex binding activity and/or mRNA expression of PPARα, as quantified by enzyme-linked immunosorbent assay (ELISA) and/or polymerase chain reaction (PCR). Increases in PPARα-CBP complex binding activity and the expression of PPARα mRNA were observed not only in BO-administered mice but also in AsR-administered mice, accompanied by a decrease in the amount of renal fatty acid.

  6. Methods of Fabricating Scintillators with Radioisotopes for Beta Battery Applications

    NASA Technical Reports Server (NTRS)

    Rensing, Noa M.; Squillante, Michael R.; Tieman, Timothy C.; Higgins, William; Shiriwadkar, Urmila

    2013-01-01

    Technology has been developed for a class of self-contained, long-duration power sources called beta batteries, which harvest the energy contained in the radioactive emissions from beta decay isotopes. The new battery is a significant improvement over the conventional phosphor/solar cell concept for converting this energy in three ways. First, the thin phosphor is replaced with a thick scintillator that is transparent to its own emissions. By using a scintillator sufficiently thick to completely stop all the beta particles, efficiency is greatly improved. Second, since the energy of the beta particles is absorbed in the scintillator, the semiconductor photodetector is shielded from radiation damage that presently limits the performance and lifetime of traditional phosphor converters. Finally, instead of a thin film of beta-emitting material, the isotopes are incorporated into the entire volume of the thick scintillator crystal allowing more activity to be included in the converter without self-absorption. There is no chemical difference between radioactive and stable strontium beta emitters such as Sr-90, so the beta emitter can be uniformly distributed throughout a strontium based scintillator crystal. When beta emitter material is applied as a foil or thin film to the surface of a solar cell or even to the surface of a scintillator, much of the radiation escapes due to the geometry, and some is absorbed within the layer itself, leading to inefficient harvesting of the energy. In contrast, if the emitting atoms are incorporated within the scintillator, the geometry allows for the capture and efficient conversion of the energy of particles emitted in any direction. Any gamma rays associated with secondary decays or Bremsstrahlung photons may also be absorbed within the scintillator, and converted to lower energy photons, which will in turn be captured by the photocell or photodiode. Some energy will be lost in this two-stage conversion process (high-energy particle

  7. Role of ARF6 in internalization of metal-binding proteins, metallothionein and transferrin, and cadmium-metallothionein toxicity in kidney proximal tubule cells

    SciTech Connect

    Wolff, Natascha A.; Lee, Wing-Kee; Abouhamed, Marouan

    2008-07-01

    Filtered metal-protein complexes, such as cadmium-metallothionein-1 (CdMT-1) or transferrin (Tf) are apically endocytosed partly via megalin/cubilin by kidney proximal tubule (PT) cells where CdMT-1 internalization causes apoptosis. Small GTPase ARF (ADP-ribosylation factor) proteins regulate endocytosis and vesicular trafficking. We investigated roles of ARF6, which has been shown to be involved in internalization of ligands and endocytic trafficking in PT cells, following MT-1/CdMT-1 and Tf uptake by PT cells. WKPT-0293 Cl.2 cells derived from rat PT S1 segment were transfected with hemagglutinin-tagged wild-type (ARF6-WT) or dominant negative (ARF6-T27N) forms of ARF6. Using immunofluorescence, endogenous ARF6 was associated with the plasma membrane (PM) as well as juxtanuclear and co-localized with Rab5a and Rab11 involved in early and recycling endosomal trafficking. Immunofluorescence staining of megalin showed reduced surface labelling in ARF6 dominant negative (ARF6-DN) cells. Intracellular Alexa Fluor 546-conjugated MT-1 uptake was reduced in ARF6-DN cells and CdMT-1 (14.8 {mu}M for 24 h) toxicity was significantly attenuated from 27.3 {+-} 3.9% in ARF6-WT to 11.1 {+-} 4.0% in ARF6-DN cells (n = 6, P < 0.02). Moreover, reduced Alexa Fluor 546-conjugated Tf uptake was observed in ARF-DN cells (75.0 {+-} 4.6% versus 3.9 {+-} 3.9% of ARF6-WT cells, n = 3, P < 0.01) and/or remained near the PM (89.3 {+-} 5. 6% versus 45.2 {+-} 14.3% of ARF6-WT cells, n = 3, P < 0.05). In conclusion, the data support roles for ARF6 in receptor-mediated endocytosis and trafficking of MT-1/Tf to endosomes/lysosomes and CdMT-1 toxicity of PT cells.

  8. ZAP-70, CTLA-4 and proximal T cell receptor signaling in cows infected with Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Paratuberculosis is a chronic intestinal disease of ruminant animals caused by Mycobacterium avium subsp. paratuberculosis (MAP). A hallmark of paratuberculosis is a transition from a cell-mediated Th1 type response to a humoral Th2 response with the progression of disease from a subclinical to clin...

  9. ZAP-70, CTLA-4, and proximal T cell receptor signaling in cows infected with Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Paratuberculosis is a chronic intestinal disease of ruminant animals caused by Mycobacterium avium subsp. paratuberculosis (MAP). A hallmark of paratuberculosis is a transition from a cell-mediated Th1 type response to a humoral Th2 response with the progression of disease from a subclinical to clin...

  10. LHCb Upgrade: Scintillating Fibre Tracker

    NASA Astrophysics Data System (ADS)

    Tobin, Mark

    2016-07-01

    The LHCb detector will be upgraded during the Long Shutdown 2 (LS2) of the LHC in order to cope with higher instantaneous luminosities and to read out the data at 40 MHz using a trigger-less read-out system. All front-end electronics will be replaced and several sub-detectors must be redesigned to cope with higher occupancy. The current tracking detectors downstream of the LHCb dipole magnet will be replaced by the Scintillating Fibre (SciFi) Tracker. The SciFi Tracker will use scintillating fibres read out by Silicon Photomultipliers (SiPMs). State-of-the-art multi-channel SiPM arrays are being developed to read out the fibres and a custom ASIC will be used to digitise the signals from the SiPMs. The evolution of the design since the Technical Design Report in 2014 and the latest R & D results are presented.

  11. Scintillation Monitoring Using Asymmetry Index

    NASA Astrophysics Data System (ADS)

    Shaikh, Muhammad Mubasshir; Mahrous, Ayman; Abdallah, Amr; Notarpietro, Riccardo

    Variation in electron density can have significant effect on GNSS signals in terms of propagation delay. Ionospheric scintillation can be caused by rapid change of such delay, specifically, when they last for a longer period of time. Ionospheric irregularities that account for scintillation may vary significantly in spatial range and drift with the background plasma at speeds of 45 to 130 m/sec. These patchy irregularities may occur several times during night, e.g. in equatorial region, with the patches move through the ray paths of the GNSS satellite signals. These irregularities are often characterized as either ‘large scale’ (which can be as large as several hundred km in East-West direction and many times that in the North-South direction) or ‘small scale’ (which can be as small as 1m). These small scale irregularities are regarded as the main cause of scintillation [1,2]. In normal solar activity conditions, the mid-latitude ionosphere is not much disturbed. However, during severe magnetic storms, the aurora oval extends towards the equator and the equator anomaly region may stretched towards poles extending the scintillation phenomena more typically associated with those regions into mid-latitudes. In such stormy conditions, the predicted TEC may deviate largely from the true value of the TEC both at low and mid-latitudes due to which GNSS applications may be strongly degraded. This work is an attempt to analyze ionospheric scintillation (S4 index) using ionospheric asymmetry index [3]. The asymmetry index is based on trans-ionospheric propagation between GPS and LEO satellites in a radio occultation (RO) scenario, using background ionospheric data provided by MIDAS [4]. We attempted to simulate one of the recent geomagnetic storms (NOAA scale G4) occurred over low/mid-latitudes. The storm started on 26 September 2011 at UT 18:00 and lasted until early hours of 27 September 2011. The scintillation data for the storm was taken from an ionospheric

  12. Proximal humerus fractures.

    PubMed

    Price, Matthew C; Horn, Pamela L; Latshaw, James C

    2013-01-01

    Proximal humerus fractures are among the most common fractures associated with osteoporosis. With an aging population, incidence of these fractures will only increase. The proximal humerus not only forms the lateral portion of the shoulder articulation but also has significant associations with musculoskeletal and neurovascular structures. As a result, fractures of the proximal humerus can significantly impact not only the function of the shoulder joint, but the health and function of the entire upper extremity as well. Understanding of these fractures, the management options, and associated nursing care, can help reduce morbidity rate and improve functional outcomes.

  13. Detecting scintillations in liquid helium

    NASA Astrophysics Data System (ADS)

    Huffman, P. R.; McKinsey, D. N.

    2013-09-01

    We review our work in developing a tetraphenyl butadiene (TPB)-based detection system for a measurement of the neutron lifetime using magnetically confined ultracold neutrons (UCN). As part of the development of the detection system for this experiment, we studied the scintillation properties of liquid helium itself, characterized the fluorescent efficiencies of different fluors, and built and tested three detector geometries. We provide an overview of the results from these studies as well as references for additional information.

  14. Characterization of the Interaction of Staphylococcal Entertoxin B with CD1d Expressed in Human Renal Proximal Tubule Epithelial Cells

    DTIC Science & Technology

    2015-02-04

    typical antigen recognition process that recruits the major histocompatibility complex class II (MHC II) molecules. The immunological importance of CD1d is...underscored by its influences on the performances of natural killer T-cells and thereby mediates the innate and adaptive immune systems. Results...release of many cytokines, and thereby systematically in- fluences the hosts’ adaptive and innate immune systems [35,36]. The involvement of CD1d in

  15. Beneficial effects of metformin and irbesartan on advanced glycation end products (AGEs)-RAGE-induced proximal tubular cell injury.

    PubMed

    Ishibashi, Yuji; Matsui, Takanori; Takeuchi, Masayoshi; Yamagishi, Sho-ichi

    2012-03-01

    Advanced glycation end products (AGEs) and their receptor (RAGE) axis contributes to diabetic nephropathy. An oral hypoglycemic agent, metformin may have a potential effect on the inhibition of glycation reactions. Further, since a pathophysiological crosstalk between renin-angiotensin system (RAS) and AGEs-RAGE axis is involved in diabetic nephropathy, it is conceivable that metformin and irbesartan additively could protect against the AGEs-RAGE-induced tubular cell injury. In this study, we addressed the issues. Metformin dose-dependently inhibited the formation of AGEs modification of bovine serum albumin (BSA). Compared with AGEs-modified BSA prepared without metformin (AGEs-MF0), those prepared in the presence of 30 mM or 100 mM metformin (AGEs-MF30 or AGEs-MF100) significantly reduced RAGE mRNA level, reactive oxygen species (ROS) generation, apoptosis, monocyte chemoattractant protein-1 and transforming growth factor-β mRNA level in tubular cells. Irbesartan further inhibited the harmful effects of AGEs-MF0 or AGEs-MF30 on tubular cells. Our present study suggests that combination therapy with metformin and irbesartan may have therapeutic potential in diabetic nephropathy; it could play a protective role against tubular injury in diabetes not only by inhibiting AGEs formation, but also by attenuating the deleterious effects of AGEs via down-regulating RAGE expression and subsequently suppressing ROS generation.

  16. Simultaneous targeted activation of Notch1 and Vhl-disruption in the kidney proximal epithelial tubular cells in mice

    PubMed Central

    Johansson, Elinn; Rönö, Birgitte; Johansson, Martin; Lindgren, David; Möller, Christina; Axelson, Håkan; Smith, Emma M. K.

    2016-01-01

    Clear cell renal cell carcinoma (ccRCC) is the most common subtype of kidney cancer, representing approximately 75% of all renal neoplasms. ccRCC is known to be strongly associated with silencing of the von Hippel Lindau (VHL) tumor suppressor gene, yet VHL deficiency alone does not seem to be sufficient to drive the oncogenic transformation of normal renal epithelium and induce renal tumorigenesis. We, and others, have previously suggested that constitutive activation of the Notch signaling pathway, alongside with VHL loss, contribute to the oncogenic features of ccRCC. Here we report a prevailing hyperactivation of the Notch1 receptor in human ccRCC relative to the healthy counterpart. To explore the consequences of the elevated Notch1 signaling observed in ccRCC patient material, we made use of a conditional mouse model based on concurrent ectopic expression of constitutively active Notch1 (NICD1) and deletion of the Vhl gene. Histological examination of the kidneys of the conditional mice demonstrate the existence of nests of dysplastic cells with a clear cytoplasm as a consequence of lipid accumulation, thus displaying a one important hallmark of human ccRCC. PMID:27491826

  17. Simultaneous targeted activation of Notch1 and Vhl-disruption in the kidney proximal epithelial tubular cells in mice.

    PubMed

    Johansson, Elinn; Rönö, Birgitte; Johansson, Martin; Lindgren, David; Möller, Christina; Axelson, Håkan; Smith, Emma M K

    2016-08-05

    Clear cell renal cell carcinoma (ccRCC) is the most common subtype of kidney cancer, representing approximately 75% of all renal neoplasms. ccRCC is known to be strongly associated with silencing of the von Hippel Lindau (VHL) tumor suppressor gene, yet VHL deficiency alone does not seem to be sufficient to drive the oncogenic transformation of normal renal epithelium and induce renal tumorigenesis. We, and others, have previously suggested that constitutive activation of the Notch signaling pathway, alongside with VHL loss, contribute to the oncogenic features of ccRCC. Here we report a prevailing hyperactivation of the Notch1 receptor in human ccRCC relative to the healthy counterpart. To explore the consequences of the elevated Notch1 signaling observed in ccRCC patient material, we made use of a conditional mouse model based on concurrent ectopic expression of constitutively active Notch1 (NICD1) and deletion of the Vhl gene. Histological examination of the kidneys of the conditional mice demonstrate the existence of nests of dysplastic cells with a clear cytoplasm as a consequence of lipid accumulation, thus displaying a one important hallmark of human ccRCC.

  18. Extracellular pH alkalinization by Cl-/HCO3- exchanger is crucial for TASK2 activation by hypotonic shock in proximal cell lines from mouse kidney.

    PubMed

    L'Hoste, S; Barriere, H; Belfodil, R; Rubera, I; Duranton, C; Tauc, M; Poujeol, C; Barhanin, J; Poujeol, P

    2007-02-01

    We have previously shown that K(+)-selective TASK2 channels and swelling-activated Cl(-) currents are involved in a regulatory volume decrease (RVD; Barriere H, Belfodil R, Rubera I, Tauc M, Lesage F, Poujeol C, Guy N, Barhanin J, Poujeol P. J Gen Physiol 122: 177-190, 2003; Belfodil R, Barriere H, Rubera I, Tauc M, Poujeol C, Bidet M, Poujeol P. Am J Physiol Renal Physiol 284: F812-F828, 2003). The aim of this study was to determine the mechanism responsible for the activation of TASK2 channels during RVD in proximal cell lines from mouse kidney. For this purpose, the patch-clamp whole-cell technique was used to test the effect of pH and the buffering capacity of external bath on Cl(-) and K(+) currents during hypotonic shock. In the presence of a high buffer concentration (30 mM HEPES), the cells did not undergo RVD and did not develop outward K(+) currents (TASK2). Interestingly, the hypotonic shock reduced the cytosolic pH (pH(i)) and increased the external pH (pH(e)) in wild-type but not in cftr (-/-) cells. The inhibitory effect of DIDS suggests that the acidification of pH(i) and the alkalinization of pH(e) induced by hypotonicity in wild-type cells could be due to an exit of HCO(3)(-). In conclusion, these results indicate that Cl(-) influx will be the driving force for HCO(3)(-) exit through the activation of the Cl(-)/HCO(3)(-) exchanger. This efflux of HCO(3)(-) then alkalinizes pH(e), which in turn activates TASK2 channels.

  19. Polyamine-modulated c-Myc expression in normal intestinal epithelial cells regulates p21Cip1 transcription through a proximal promoter region

    PubMed Central

    Liu, Lan; Guo, Xin; Rao, Jaladanki N.; Zou, Tongtong; Marasa, Bernard S.; Chen, Jie; Greenspon, Jose; Casero, Robert A.; Wang, Jian-Ying

    2006-01-01

    Maintenance of intestinal mucosal epithelial integrity requires cellular polyamines that regulate expression of various genes involved in cell proliferation, growth arrest and apoptosis. Our previous studies have shown that polyamines are essential for expression of the c-myc gene and that polyamine-induced c-Myc plays a critical role in stimulation of normal IEC (intestinal epithelial cell) proliferation, but the exact downstream targets of induced c-Myc are still unclear. The p21Cip1 protein is a major player in cell cycle control, which is primarily regulated at the transcriptional level. The current study was designed to determine whether induced c-Myc stimulates normal IEC proliferation by repressing p21Cip1 transcription following up-regulation of polyamines. Overexpression of the ODC (ornithine decarboxylase) gene increased levels of cellular polyamines, induced c-Myc expression and inhibited p21Cip1 transcription, as indicated by repression of p21Cip1 promoter activity and a decrease in p21Cip1 protein levels. In contrast, depletion of cellular polyamines by inhibiting ODC enzyme activity with α-difluoromethylornithine decreased c-Myc, but increased p21Cip1 transcription. Ectopic expression of wild-type c-myc not only inhibited basal levels of p21Cip1 transcription in control cells, but also prevented increased p21Cip1 in polyamine-deficient cells. Experiments using different p21Cip1 promoter mutants showed that transcriptional repression of p21Cip1 by c-Myc was mediated through Miz-1- and Sp1-binding sites within the proximal region of the p21Cip1 promoter in normal IECs. These findings confirm that p21Cip1 is one of the direct mediators of induced c-Myc following increased polyamines and that p21Cip1 repression by c-Myc is implicated in stimulation of normal IEC proliferation. PMID:16706751

  20. Scintillation Forecasting Using NPOESS Data

    NASA Astrophysics Data System (ADS)

    Basu, B.; Retterer, J.; Demajistre, R.; de La Beaujardiere, O.; Scro, K.

    2005-12-01

    We have conducted a theoretical study of the use of NPOESS data for the forecasting of equatorial radio scintillation using knowledge of the equatorial Appleton anomaly, e.g., the peak-to-valley ratio of TEC (Total Electron Content) between the anomaly crests and the magnetic equator. The peak-to-valley ratio can be obtained from the UV (ultraviolet) imagery of the anomaly region that will be provided by the NPOESS sensors. The post-sunset enhancement of the upward drift velocity of the equatorial plasma has been shown, both theoretically and observationally, to be an important determinant of both the onset of scintillation and the strength of the anomaly. The technical approach is to run PBMOD, the AFRL low-latitude ionosphere model, with a range of post-sunset vertical drift velocities to determine the quantitative relationship between the peak-to-valley ratio and the maximum value of the pot-sunset upward drift velocity of equatorial plasma. Once the relationship is validated, it will be used to estimate the maximum value of the drift velocity from the peak-to-valley ratio, which is derived from the UV imagery data provided by NPOESS-like sensor, such as GUVI on TIMED satellite. The drift velocity will then be used in PBMOD to simulate the formation and evolution of equatorial plasma `bubbles' and calculate the distribution of the amplitude scintillation index S4. Results of the study will be discussed.

  1. A Review of Ionospheric Scintillation Models.

    PubMed

    Priyadarshi, S

    This is a general review of the existing climatological models of ionospheric radio scintillation for high and equatorial latitudes. Trans-ionospheric communication of radio waves from transmitter to user is affected by the ionosphere which is highly variable and dynamic in both time and space. Scintillation is the term given to irregular amplitude and phase fluctuations of the received signals and related to the electron density irregularities in the ionosphere. Key sources of ionospheric irregularities are plasma instabilities; every irregularities model is based on the theory of radio wave propagation in random media. It is important to understand scintillation phenomena and the approach of different theories. Therefore, we have briefly discussed the theories that are used to interpret ionospheric scintillation data. The global morphology of ionospheric scintillation is also discussed briefly. The most important (in our opinion) analytical and physical models of scintillation are reviewed here.

  2. Caspase-Dependent and Caspase-Independent Pathways Are Involved in Cadmium-Induced Apoptosis in Primary Rat Proximal Tubular Cell Culture

    PubMed Central

    Long, Mengfei; Bian, Jianchun; Liu, Xuezhong; Gu, Jianhong; Yuan, Yan; Song, Ruilong; Wang, Yi; Zhu, Jiaqiao; Liu, Zongping

    2016-01-01

    We designed this study to investigate whether cadmium induces caspase-independent apoptosis and to investigate the relationship between the caspase-dependent and caspase-independent apoptotic pathways. Cadmium (1.25–2.5 μM) induced oxidative stress in rat proximal tubular (rPT) cells, as seen in the reactive oxygen species levels; N-acetylcysteine prevented this. Cyclosporin A (CsA) prevented mitochondrial permeability transition pore opening and apoptosis; there was mitochondrial ultrastructural disruption, mitochondrial cytochrome c (cyt c) translocation to the cytoplasm, and subsequent caspase-9 and caspase-3 activation. Z-VAD-FMK prevented caspase-3 activation and apoptosis and decreased BNIP-3 (Bcl-2/adenovirus E1B 19-kDa interacting protein 3) expression levels and apoptosis-inducing factor/endonuclease G (AIF/Endo G) translocation. Simultaneously, cadmium induced prominent BNIP-3 expression in the mitochondria and cytoplasmic AIF/Endo G translocation to the nucleus. BNIP-3 silencing significantly prevented AIF and Endo G translocation and decreased the apoptosis rate, cyt c release, and caspase-9 and caspase-3 activation. These results suggest that BNIP-3 is involved in the caspase-independent apoptotic pathway and is located upstream of AIF/Endo G; both the caspase-dependent and caspase-independent pathways are involved in cadmium-induced rPT cell apoptosis and act synergistically. PMID:27861627

  3. Recording of relativistic particles in thin scintillators

    SciTech Connect

    Tolstukhin, I A.; Somov, Alexander S.; Somov, S. V.; Bolozdynya, A. I.

    2014-11-01

    Results of investigating an assembly of thin scintillators and silicon photomultipliers for registering relativistic particles with the minimum ionization are presented. A high efficiency of registering relativistic particles using an Ej-212 plastic scintillator, BSF-91A wavelength-shifting fiber (Saint-Gobain), and a silicon photomultiplier (Hamamtsu) is shown. The measurement results are used for creating a scintillation hodoscope of the magnetic spectrometer for registering γ quanta in the GlueX experiment.

  4. Scintillator tiles read out with silicon photomultipliers

    NASA Astrophysics Data System (ADS)

    Pooth, O.; Radermacher, T.; Weingarten, S.; Weinstock, L.

    2015-10-01

    A detector prototype based on a fast plastic scintillator read out with silicon photomultipliers is presented. All studies have been done with cosmic muons and focus on parameter optimization such as coupling the SiPM to the scintillator or wrapping the scintillator with reflective material. The prototype shows excellent results regarding the light-yield and offers a detection efficiency of 99.5% with a signal purity of 99.9% for cosmic muons.

  5. Spacecraft Radio Scintillation and Solar System Exploration

    NASA Technical Reports Server (NTRS)

    Woo, Richard

    1993-01-01

    When a wave propagates through a turbulent medium, scattering by the random refractive index inhomogeneities can lead to a wide variety of phenomena that have been the subject of extensive study. The observed scattering effects include amplitude or intensity scintillation, phase scintillation, angular broadening, and spectral broadening, among others. In this paper, I will refer to these scattering effects collectively as scintillation. Although the most familiar example is probably the twinkling of stars (light wave intensity scintillation by turbulence in the Earth's atmosphere), scintillation has been encountered and investigated in such diverse fields as ionospheric physics, oceanography, radio astronomy, and radio and optical communications. Ever since planetary spacecraft began exploring the solar system, scintillation has appeared during the propagation of spacecraft radio signals through planetary atmospheres, planetary ionospheres, and the solar wind. Early studies of these phenomena were motivated by the potential adverse effects on communications and navigation, and on experiments that use the radio link to conduct scientific investigations. Examples of the latter are radio occultation measurements (described below) of planetary atmospheres to deduce temperature profiles, and the search for gravitational waves. However,these concerns soon gave way to the emergence of spacecraft radio scintillation as a new scientific tool for exploring small-scale dynamics in planetary atmospheres and structure in the solar wind, complementing in situ and other remote sensing spacecraft measurements, as well as scintillation measurements using natural (celestial) radio sources. The purpose of this paper is to briefly describe and review the solar system spacecraft radio scintillation observations, to summarize the salient features of wave propagation analyses employed in interpreting them, to underscore the unique remote sensing capabilities and scientific relevance of

  6. The Effect of Neutron and Gamma Ray Cross Talk Between Plastic Scintillating Detectors

    SciTech Connect

    Pozzi, S.A.

    2000-11-06

    In this paper a method is developed, using higher order statistics, to identify the type and degree of neutron and gamma ray cross talk between detectors that are placed in proximity to one another. A set of measurements was performed using the Nuclear Materials Identification System (NMIS) to acquire the time-dependent bicovariance of the pulses in fast plastic scintillating detectors. These signatures were analyzed to infer the degree and type of false coincidences (cross talk) in relation to true coincidences.

  7. Structural design of a high energy particle detector using liquid scintillator

    SciTech Connect

    Berg, Timothy John; /Minnesota U.

    1997-02-01

    This thesis presents a design for a 10,000 ton liquid scintillator neutrino detector being considered for the MINOS project at Fermilab. Details of designing, manufacturing, and assembling the active detector components are presented. The detector consists of 1080 magnetized steel absorber planes alternating with 1080 active detector planes. Each active plane is made up of plastic extrusions divided into nearly 400 cells for positional resolution. Life tests on the plastic extrusions determine their feasibility for containing the scintillator. The extrusions are sealed at the bottom, filled with liquid scintillator, and have an optical fiber running the entire length of each cell. The fibers terminate at the top of each extrusion in a manifold. An optical-fiber-light-guide connects the fibers in each manifold to a photo-detector. The photo-detector converts the light signals from the scintillator and optical fibers into electrical impulses for computer analysis.

  8. Alisol A 24-Acetate and Alisol B 23-Acetate Induced Autophagy Mediates Apoptosis and Nephrotoxicity in Human Renal Proximal Tubular Cells

    PubMed Central

    Wang, Chunfei; Feng, Liang; Ma, Liang; Chen, Haifeng; Tan, Xiaobin; Hou, Xuefeng; Song, Jie; Cui, Li; Liu, Dan; Chen, Juan; Yang, Nan; Wang, Jing; Liu, Ying; Zhao, Bingjie; Wang, Gang; Zhou, Yuanli; Jia, Xiaobin

    2017-01-01

    Two natural compounds alisol A 24-acetate (24A) and alisol B 23-acetate (23B) are abundant in Rhizoma alismatis. In the present study, we evaluated the induction of 24A and 23B on apoptosis and possible nephrotoxicity of human renal proximal tubular (HK-2) cells by activating autophagy and also explored its regulation on PI3K/Akt/mTOR signaling pathway. Presently, Clusterin, Kim-1, and TFF-3 were considered to be new bioindicators of nephrotoxicity. Interestingly, the protein expression and mRNA levels of Clusterin, Kim-1 and TFF-3 could be significantly increased by 23B and 24A in vivo and in vitro. Furthermore, cell apoptosis could be triggered by 23B and 24A via significantly decreasing the protein expression and mRNA levels of Bcl-2 and Bcl-xl. Autophagy of HK-2 cells could be induced by both 23B and 24A via significantly enhancing the ratio of LC3II/LC3I, the protein expression of Beclin-1 as well as the mRNA levels of LC3 and Beclin-1. Meanwhile, PI3K/Akt/mTOR signaling pathway could be inhibited by these two compounds. An autophagy inhibitor, 3-methyladenine, could partially reverse cell viability and conversely change the ratio of LC3II/LC3I and the protein expression of Bcl-2 and Kim-1. Thus this study helped to understand that 23B and 24A induced autophagy resulted in apoptosis and nephrotoxicity through inhibiting PI3K/Akt/mTOR signaling pathway, facilitating further studies for nephrotoxicity induced by these two compounds and could be beneficial for safe use of Rhizoma alismatis in clinic.

  9. New Scintillators for Photosensitive Gaseous Detectors

    NASA Astrophysics Data System (ADS)

    Charpak, G.; Peskov, V.; Scigocki, D.; Valbis, J.

    A new family of scintillators are presented. Their properties are similar to those of barium fluoride, and the spectrum of the scintillation emission is between 140 and 300 nm. Our latest efficiency measurements of ethyl ferrocene and triethylamine liquid or caesium iodide solid photocathodes, in parallel-plate avalanche chambers (PPACs) at high electric field, are also presented. We discuss the revolutionary consequences of the combination of the new scintillators with PPACs with semitransparent photocathodes deposited on the crystals, such as high speed, high resistance to radiation damage, compacity, high gamma efficiency, and applications to tracking devices with scintillation optical fibres.

  10. Divalent fluoride doped cerium fluoride scintillator

    DOEpatents

    Anderson, David F.; Sparrow, Robert W.

    1991-01-01

    The use of divalent fluoride dopants in scintillator materials comprising cerium fluoride is disclosed. The preferred divalent fluoride dopants are calcium fluoride, strontium fluoride, and barium fluoride. The preferred amount of divalent fluoride dopant is less than about two percent by weight of the total scintillator. Cerium fluoride scintillator crystals grown with the addition of a divalent fluoride have exhibited better transmissions and higher light outputs than crystals grown without the addition of such dopants. These scintillators are useful in radiation detection and monitoring applications, and are particularly well suited for high-rate applications such as positron emission tomography (PET).

  11. Waveshifters and Scintillators for Ionizing Radiation Detection

    SciTech Connect

    B.Baumgaugh; J.Bishop; D.Karmgard; J.Marchant; M.McKenna; R.Ruchti; M.Vigneault; L.Hernandez; C.Hurlbut

    2007-12-11

    Scintillation and waveshifter materials have been developed for the detection of ionizing radiation in an STTR program between Ludlum Measurements, Inc. and the University of Notre Dame. Several new waveshifter materials have been developed which are comparable in efficiency and faster in fluorescence decay than the standard material Y11 (K27) used in particle physics for several decades. Additionally, new scintillation materials useful for fiber tracking have been developed which have been compared to 3HF. Lastly, work was done on developing liquid scintillators and paint-on scintillators and waveshifters for high radiation environments.

  12. Radio wave scintillations at equatorial regions

    NASA Technical Reports Server (NTRS)

    Poularikas, A. D.

    1972-01-01

    Radio waves, passing through the atmosphere, experience amplitude and phase fluctuations know as scintillations. A characterization of equatorial scintillation, which has resulted from studies of data recorded primarily in South America and equatorial Africa, is presented. Equatorial scintillation phenomena are complex because they appear to vary with time of day (pre-and postmidnight), season (equinoxes), and magnetic activity. A wider and more systematic geographical coverage is needed for both scientific and engineering purposes; therefore, it is recommended that more observations should be made at earth stations (at low-geomagnetic latitudes) to record equatorial scintillation phenomena.

  13. Generation of Long-Lived Bone Marrow Plasma Cells Secreting Antibodies Specific for the HIV-1 gp41 Membrane-Proximal External Region in the Absence of Polyreactivity

    PubMed Central

    Donius, Luke R.; Cheng, Yuxing; Choi, Jaewon; Sun, Zhen-Yu J.; Hanson, Melissa; Zhang, Michael; Gierahn, Todd M.; Marquez, Susanna; Uduman, Mohammed; Kleinstein, Steven H.; Irvine, Darrell; Love, J. Christopher; Reinherz, Ellis L.

    2016-01-01

    ABSTRACT An effective preventive vaccine is highly sought after in order to stem the current HIV-1 pandemic. Both conservation of contiguous gp41 membrane-proximal external region (MPER) amino acid sequences across HIV-1 clades and the ability of anti-MPER broadly neutralizing antibodies (BNAbs) to block viral hemifusion/fusion establish the MPER as a prime vaccination target. In earlier studies, we described the development of an MPER vaccine formulation that takes advantage of liposomes to array the MPER on a lipid bilayer surface, paralleling its native configuration on the virus membrane while also incorporating molecular adjuvant and CD4 T cell epitope cargo. Here we demonstrate that several immunizations with MPER/liposomes induce high levels of bone marrow long-lived plasma cell (LLPC) antibody production. Single-cell immunoglobulin gene retrieval analysis shows that these plasma cells are derived from a germ line repertoire of B cells with a diverse representation of immunoglobulin genes, exhibiting antigen-driven positive selection. Characterization of LLPC recombinant monoclonal antibodies (rMAbs) indicates that antigen recognition is achieved through convergence on a common epitopic focus by utilizing various complementarity-determining region H3 (CDRH3) lengths. Importantly, the vast majority of rMAbs produced from these cells lack polyreactivity yet manifest antigen specificity in the context of lipids, shaping MPER-specific paratopes through selective pressure. Taken together, these findings demonstrate that the MPER is a vaccine target with minimal risk of generating off-target autoimmunity. IMPORTANCE A useful vaccine must generate desired long-term, antigen-specific antibody responses devoid of polyreactivity or autoreactivity. The common polyreactive features of some HIV-1 BNAbs have raised concern about elicitation of anti-MPER antibodies. Utilizing single-LLPC repertoire analysis and biophysical characterization of anti-MPER rMAbs, we show that

  14. Characterization of the Mouse and Human Monoacylglycerol O-Acyltransferase 1 (Mogat1) Promoter in Human Kidney Proximal Tubule and Rat Liver Cells

    PubMed Central

    Sankella, Shireesha; Garg, Abhimanyu; Agarwal, Anil K.

    2016-01-01

    Monoacylglycerol acyltransferase 1 (Mogat1) catalyzes the conversion of monoacylglycerols (MAG) to diacylglycerols (DAG), the precursor of several physiologically important lipids such as phosphatidylcholine, phosphatidylethanolamine and triacylglycerol (TAG). Expression of Mogat1 is tissue restricted and it is highly expressed in the kidney, stomach and adipose tissue but minimally in the normal adult liver. To understand the transcriptional regulation of Mogat1, we characterized the mouse and human Mogat1 promoters in human kidney proximal tubule-2 (HK-2) cells. In-silico analysis revealed several peroxisome proliferator response element (PPRE) binding sites in the promoters of both human and mouse Mogat1. These sites responded to all three peroxisome proliferator activated receptor (PPAR) isoforms such that their respective agonist or antagonist activated or inhibited the expression of Mogat1. PPRE site mutagenesis revealed that sites located at -592 and -2518 are very effective in decreasing luciferase reporter gene activity. Chromatin immunoprecipitation (ChIP) assay using PPARα antibody further confirmed the occupancy of these sites by PPARα. While these assays revealed the core promoter elements necessary for Mogat1 expression, there are additional elements required to regulate its tissue specific expression. Chromosome conformation capture (3C) assay revealed additional cis-elements located ~10–15 kb upstream which interact with the core promoter. These chromosomal regions are responsive to both PPARα agonist and antagonist. PMID:27611931

  15. Gastrin decreases Na+,K+-ATPase activity via a PI 3-kinase- and PKC-dependent pathway in human renal proximal tubule cells.

    PubMed

    Liu, Tianbing; Konkalmatt, Prasad R; Yang, Yu; Jose, Pedro A

    2016-04-01

    The natriuretic effect of gastrin suggests a role in the coordinated regulation of sodium balance by the gastrointestinal tract and the kidney. The renal molecular targets and signal transduction pathways for such an effect of gastrin are largely unknown. Recently, we reported that gastrin induces NHE3 phosphorylation and internalization via phosphatidylinositol (PI) 3-kinase and PKCα. In this study, we show that gastrin induced the phosphorylation of human Na(+),K(+)-ATPase at serine 16, resulting in its endocytosis via Rab5 and Rab7 endosomes. The gastrin-stimulated phosphorylation of Na(+),K(+)-ATPase was dependent on PI 3-kinase because the phosphorylation was blocked by the PI 3-kinase inhibitor wortmannin. The phosphorylation of Na(+),K(+)-ATPase was also blocked by chelerythrine, a pan-PKC inhibitor, Gö-6976, a conventional PKC (cPKC) inhibitor, and BAPTA-AM, an intracellular calcium chelator, suggesting the importance of cPKC and intracellular calcium in the gastrin signaling pathway. The gastrin-mediated phosphorylation of Na(+),K(+)-ATPase was also inhibited by U-73122, a phospholipase C (PLC) inhibitor. These results suggest that gastrin regulates sodium hydrogen exchanger and pump in renal proximal tubule cells at the apical and basolateral membranes.

  16. Iodinated contrast media inhibit oxygen consumption in freshly isolated proximal tubular cells from elderly humans and diabetic rats: Influence of nitric oxide.

    PubMed

    Liss, Per; Hansell, Peter; Fasching, Angelica; Palm, Fredrik

    2016-01-01

    Objectives Mechanisms underlying contrast medium (CM)-induced nephropathy remain elusive, but recent attention has been directed to oxygen availability. The purpose of this study was to evaluate the effect of the low-osmolar CM iopromide and the iso-osmolar CM iodixanol on oxygen consumption (QO2) in freshly isolated proximal tubular cells (PTC) from kidneys ablated from elderly humans undergoing nephrectomy for renal carcinomas and from normoglycemic or streptozotocin-diabetic rats. Materials PTC were isolated from human kidneys, or kidneys of normoglycemic or streptozotocin-diabetic rats. QO2 was measured with Clark-type microelectrodes in a gas-tight chamber with and without each CM (10 mg I/mL medium). L-NAME was used to inhibit nitric oxide (NO) production caused by nitric oxide synthase. Results Both CM reduced QO2 in human PTC (about -35%) which was prevented by L-NAME. PTC from normoglycemic rats were unaffected by iopromide, whereas iodixanol decreased QO2 (-34%). Both CM decreased QO2 in PTC from diabetic rats (-38% and -36%, respectively). L-NAME only prevented the effect of iopromide in the diabetic rat PTC. Conclusions These observations demonstrate that CM can induce NO release from isolated PTC in vitro, which affects QO2. Our results suggest that the induction of NO release and subsequent effect on the cellular oxygen metabolism are dependent on several factors, including CM type and pre-existing risk factors for the development of CM-induced nephropathy.

  17. Accelerated discovery of elpasolite scintillators

    SciTech Connect

    Doty, F. Patrick; Yang, Pin; Zhou, Xiaowang

    2014-12-01

    Elpasolite scintillators are a large family of halides which includes compounds reported to meet the NA22 program goals of <3% energy resolution at 662 keV1. This work investigated the potential to produce quality elpasolite compounds and alloys of useful sizes at reasonable cost, through systematic experimental and computational investigation of crystal structure and properties across the composition space. Discovery was accelerated by computational methods and models developed previously to efficiently identify cubic members of the elpasolite halides, and to evaluate stability of anion and cation exchange alloys.

  18. Photodetectors for Scintillator Proportionality Measurement

    SciTech Connect

    Moses, William W.; Choong, Woon-Seng; Hull, Giulia; Payne, Steve; Cherepy, Nerine; Valentine, J.D.

    2010-10-18

    We evaluate photodetectors for use in a Compton Coincidence apparatus designed for measuring scintillator proportionality. There are many requirements placed on the photodetector in these systems, including active area, linearity, and the ability to accurately measure low light levels (which implies high quantum efficiency and high signal-to-noise ratio). Through a combination of measurement and Monte Carlo simulation, we evaluate a number of potential photodetectors, especially photomultiplier tubes and hybrid photodetectors. Of these, we find that the most promising devices available are photomultiplier tubes with high ({approx}50%) quantum efficiency, although hybrid photodetectors with high quantum efficiency would be preferable.

  19. Current status on plastic scintillators modifications

    SciTech Connect

    Hamel, Matthieu; Bertrand, Guillaume H.V.; Carrel, Frederick; Coulon, Romain; Dumazert, Jonathan; Montbarbon, Eva; Sguerra, Fabien

    2015-07-01

    Recent developments of plastic scintillators are reviewed, from 2000 to March 2015. All examples are distributed into the main purpose, i.e. the nature of the radionuclide provided with the scope of detection of various radiation particles. The main characteristics of these newly created scintillators and their detection properties are given. (authors)

  20. Binderless composite scintillator for neutron detection

    DOEpatents

    Hodges, Jason P [Knoxville, TN; Crow, Jr; Lowell, M [Oak Ridge, TN; Cooper, Ronald G [Oak Ridge, TN

    2009-03-10

    Composite scintillator material consisting of a binderless sintered mixture of a Lithium (Li) compound containing .sup.6Li as the neutron converter and Y.sub.2SiO.sub.5:Ce as the scintillation phosphor, and the use of this material as a method for neutron detection. Other embodiments of the invention include various other Li compounds.

  1. Epoxy resins produce improved plastic scintillators

    NASA Technical Reports Server (NTRS)

    Markley, F. W.

    1967-01-01

    Plastic scintillator produced by the substitution of epoxy resins for the commonly used polystyrene is easy to cast, stable at room temperature, and has the desirable properties of a thermoset or cross-linked system. Such scintillators can be immersed directly in strong solvents, an advantage in many chemical and biological experiments.

  2. Scintillator handbook with emphasis on cesium iodide

    NASA Technical Reports Server (NTRS)

    Tidd, J. L.; Dabbs, J. R.; Levine, N.

    1973-01-01

    This report provides a background of reasonable depth and reference material on scintillators in general. Particular attention is paid to the cesium iodide scintillators as used in the High Energy Astronomy Observatory (HEAO) experiments. It is intended especially for use by persons such as laboratory test personnel who need to obtain a working knowledge of these materials and their characteristics in a short time.

  3. Proximal Hamstring Repair Strength

    PubMed Central

    Harvey, Margaret Ann; Singh, Hardeep; Obopilwe, Elifho; Charette, Ryan; Miller, Suzanne

    2015-01-01

    Background: Proximal hamstring repair for complete ruptures has become a common treatment. There is no consensus in the literature about postoperative rehabilitation protocols following proximal hamstring repair. Some protocols describe bracing to prevent hip flexion or knee extension while others describe no immobilization. There are currently no biomechanical studies evaluating proximal hamstring repairs; nor are there any studies evaluating the effect of different hip flexion angles on these repairs. Hypothesis: As hip flexion increases from 0° to 90°, there will be a greater gap with cyclical loading. Study Design: Controlled laboratory study. Methods: Proximal hamstring insertions were detached from the ischial tuberosity in 24 cadavers and were repaired with 3 single-loaded suture anchors in the hamstring footprint with a Krakow suture technique. Cyclic loading from 10 to 125 N at 1 Hz was then performed for 0°, 45°, and 90° of hip flexion for 1500 cycles. Gap formation, stiffness, yield load, ultimate load, and energy to ultimate load were compared between groups using paired t tests. Results: Cyclic loading demonstrated the least amount of gap formation (P < .05) at 0° of hip flexion (2.39 mm) and most at 90° of hip flexion (4.19 mm). There was no significant difference in ultimate load between hip flexion angles (326, 309, and 338 N at 0°, 45°, and 90°, respectively). The most common mode of failure occurred with knot/suture failure (n = 17). Conclusion: Increasing hip flexion from 0° to 90° increases the displacement across proximal hamstring repairs. Postoperative bracing that limits hip flexion should be considered. Clinical Relevance: Repetitive motion involving hip flexion after a proximal hamstring repair may cause compromise of the repair. PMID:26665049

  4. Role of proximal tubules in the pathogenesis of kidney disease.

    PubMed

    Nakhoul, Nazih; Batuman, Vecihi

    2011-01-01

    The proximal tubules make up a significant portion of the kidneys; proximal tubule epithelial cells are the most populous cell type in the kidney, and carry out diverse regulatory and endocrine functions where numerous transporters are located. Under normal circumstances, more than two thirds of filtered salt and water, and all filtered bicarbonate is reabsorbed in the proximal tubule. A number of inherited and acquired acid-base and tubule disorders are linked to impaired transporters in the proximal tubule cells. Equally important is the intrinsic immune characteristics of proximal tubule cells that give them the ability to also function as immune responders to a wide range of immunologic, ischemic or toxic injury. It is therefore not surprising that proximal tubule-related phenomena are closely related to the pathogenesis of a vast array of kidney diseases. Many kidney diseases, acute and chronic, first manifest with proximal tubule disorders. Recent insight into molecular characteristics of transport functions in the proximal tubules, and the recognition that proximal tubule cells possess intrinsic immune responses have contributed to an improved understanding of important areas in nephrology, such as Fanconi's syndrome, renal tubular acidosis, phosphate wasting syndromes, Dent's disease, cystinuria and other amino acid transport disorders, acute kidney injury, and the role of proximal tubules in progressive kidney disease. Megalin/ cubilin-mediated endocytosis by proximal tubule cells of increased quantities of filtered proteins (protein overloading) in glomerular diseases appears to evoke cell stress responses resulting in increased inflammatory cytokines leading to tubulointerstitial inflammation and fibrosis. Finally, the proximal tubule may be the site of both active vitamin D synthesis through the action of 1-α-hydroxylase, and the site where erythropoietin synthesis takes place. Thus, proximal tubule injury also contributes to two distressing

  5. Spleen tyrosine kinase mediates high glucose-induced transforming growth factor-{beta}1 up-regulation in proximal tubular epithelial cells

    SciTech Connect

    Yang, Won Seok; Chang, Jai Won; Han, Nam Jeong; Lee, Sang Koo; Park, Su-Kil

    2012-09-10

    The role of spleen tyrosine kinase (Syk) in high glucose-induced intracellular signal transduction has yet to be elucidated. We investigated whether Syk is implicated in high glucose-induced transforming growth factor-{beta}1 (TGF-{beta}1) up-regulation in cultured human proximal tubular epithelial cells (HK-2 cell). High glucose increased TGF-{beta}1 gene expression through Syk, extracellular signal-regulated kinase (ERK), AP-1 and NF-{kappa}B. High glucose-induced AP-1 DNA binding activity was decreased by Syk inhibitors and U0126 (an ERK inhibitor). Syk inhibitors suppressed high glucose-induced ERK activation, whereas U0126 had no effect on Syk activation. High glucose-induced NF-{kappa}B DNA binding activity was also decreased by Syk inhibitors. High glucose increased nuclear translocation of p65 without serine phosphorylation of I{kappa}B{alpha} and without degradation of I{kappa}B{alpha}, but with an increase in tyrosine phosphorylation of I{kappa}B{alpha} that may account for the activation of NF-{kappa}B. Both Syk inhibitors and Syk-siRNA attenuated high glucose-induced I{kappa}B{alpha} tyrosine phosphorylation and p65 nuclear translocation. Depletion of p21-activated kinase 2 (Pak2) by transfection of Pak2-siRNA abolished high glucose-induced Syk activation. In summary, high glucose-induced TGF-{beta}1 gene transcription occurred through Pak2, Syk and subsequent ERK/AP-1 and NF-{kappa}B pathways. This suggests that Syk might be implicated in the diabetic kidney disease.

  6. Cyclophilin B Interacts with Sodium-Potassium ATPase and Is Required for Pump Activity in Proximal Tubule Cells of the Kidney

    PubMed Central

    Suñé, Guillermo; Sarró, Eduard; Puigmulé, Marta; López-Hellín, Joan; Zufferey, Madeleine; Pertel, Thomas; Luban, Jeremy; Meseguer, Anna

    2010-01-01

    Cyclophilins (Cyps), the intracellular receptors for Cyclosporine A (CsA), are responsible for peptidyl-prolyl cis-trans isomerisation and for chaperoning several membrane proteins. Those functions are inhibited upon CsA binding. Albeit its great benefits as immunosuppressant, the use of CsA has been limited by undesirable nephrotoxic effects, including sodium retention, hypertension, hyperkalemia, interstial fibrosis and progressive renal failure in transplant recipients. In this report, we focused on the identification of novel CypB-interacting proteins to understand the role of CypB in kidney function and, in turn, to gain further insight into the molecular mechanisms of CsA-induced toxicity. By means of yeast two-hybrid screens with human kidney cDNA, we discovered a novel interaction between CypB and the membrane Na/K-ATPase β1 subunit protein (Na/K-β1) that was confirmed by pull-down, co-immunoprecipitation and confocal microscopy, in proximal tubule-derived HK-2 cells. The Na/K-ATPase pump, a key plasma membrane transporter, is responsible for maintenance of electrical Na+ and K+ gradients across the membrane. We showed that CypB silencing produced similar effects on Na/K-ATPase activity than CsA treatment in HK-2 cells. It was also observed an enrichment of both alpha and beta subunits in the ER, what suggested a possible failure on the maturation and routing of the pump from this compartment towards the plasma membrane. These data indicate that CypB through its interaction with Na/K-β1 might regulate maturation and trafficking of the pump through the secretory pathway, offering new insights into the relationship between cyclophilins and the nephrotoxic effects of CsA. PMID:21085665

  7. Extruded scintillator for the calorimetry applications

    SciTech Connect

    Dyshkant, A.; Rykalin, V.; Pla-Dalmau, A.; Beznosko, D.; /SUNY, Stony Brook

    2006-08-01

    An extrusion line has been installed and successfully operated at FNAL (Fermi National Accelerator Laboratory) in collaboration with NICADD (Northern Illinois Center for Accelerator and Detector Development). This new Facility will serve to further develop and improve extruded plastic scintillator. Recently progress has been made in producing co-extruded plastic scintillator, thus increasing the potential HEP applications of this Facility. The current R&D work with extruded and co-extruded plastic scintillator for a potential ALICE upgrade, the ILC calorimetry program and the MINERvA experiment show the attractiveness of the chosen strategy for future experiments and calorimetry. We extensively discuss extruded and co-extruded plastic scintillator in calorimetry in synergy with new Solid State Photomultipliers. The characteristics of extruded and co-extruded plastic scintillator will be presented here as well as results with non-traditional photo read-out.

  8. Extruded scintillator for the Calorimetry applications

    NASA Astrophysics Data System (ADS)

    Dyshkant, A.; Rykalin, V.; Pla-Dalmau, A.; Beznosko, D.

    2006-10-01

    An extrusion line has been installed and successfully operated at FNAL (Fermi National Accelerator Laboratory) in collaboration with NICADD (Northern Illinois Center for Accelerator and Detector Development). This new Facility will serve to further develop and improve extruded plastic scintillator. Recently progress has been made in producing co-extruded plastic scintillator, thus increasing the potential HEP applications of this Facility. The current R&D work with extruded and co-extruded plastic scintillator for a potential ALICE upgrade, the ILC calorimetry program and the MINERvA experiment show the attractiveness of the chosen strategy for future experiments and calorimetry. We extensively discuss extruded and co-extruded plastic scintillator in calorimetry in synergy with new Solid State Photomultipliers. The characteristics of extruded and co-extruded plastic scintillator will be presented here as well as results with non-traditional photo read-out.

  9. Scintillation Effects on Space Shuttle GPS Data

    NASA Technical Reports Server (NTRS)

    Goodman, John L.; Kramer, Leonard

    2001-01-01

    Irregularities in ionospheric electron density result in variation in amplitude and phase of Global Positioning System (GPS) signals, or scintillation. GPS receivers tracking scintillated signals may lose carrier phase or frequency lock in the case of phase sc intillation. Amplitude scintillation can cause "enhancement" or "fading" of GPS signals and result in loss of lock. Scintillation can occur over the equatorial and polar regions and is a function of location, time of day, season, and solar and geomagnetic activity. Mid latitude regions are affected only very rarely, resulting from highly disturbed auroral events. In the spring of 1998, due to increasing concern about scintillation of GPS signals during the upcoming solar maximum, the Space Shuttle Program began to assess the impact of scintillation on Collins Miniaturized Airborne GPS Receiver (MAGR) units that are to replace Tactical Air Control and Navigation (TACAN) units on the Space Shuttle orbiters. The Shuttle Program must determine if scintillation effects pose a threat to safety of flight and mission success or require procedural and flight rule changes. Flight controllers in Mission Control must understand scintillation effects on GPS to properly diagnose "off nominal" GPS receiver performance. GPS data from recent Space Shuttle missions indicate that the signals tracked by the Shuttle MAGR manifest scintillation. Scintillation is observed as anomalous noise in velocity measurements lasting for up to 20 minutes on Shuttle orbit passes and are not accounted for in the error budget of the MAGR accuracy parameters. These events are typically coincident with latitude and local time occurrence of previously identified equatorial spread F within about 20 degrees of the magnetic equator. The geographic and seasonal history of these events from ground-based observations and a simple theoretical model, which have potential for predicting events for operational purposes, are reviewed.

  10. Simulation of optical interstellar scintillation

    NASA Astrophysics Data System (ADS)

    Habibi, F.; Moniez, M.; Ansari, R.; Rahvar, S.

    2013-04-01

    Aims: Stars twinkle because their light propagates through the atmosphere. The same phenomenon is expected on a longer time scale when the light of remote stars crosses an interstellar turbulent molecular cloud, but it has never been observed at optical wavelengths. The aim of the study described in this paper is to fully simulate the scintillation process, starting from the molecular cloud description as a fractal object, ending with the simulations of fluctuating stellar light curves. Methods: Fast Fourier transforms are first used to simulate fractal clouds. Then, the illumination pattern resulting from the crossing of background star light through these refractive clouds is calculated from a Fresnel integral that also uses fast Fourier transform techniques. Regularisation procedure and computing limitations are discussed, along with the effect of spatial and temporal coherency (source size and wavelength passband). Results: We quantify the expected modulation index of stellar light curves as a function of the turbulence strength - characterised by the diffraction radius Rdiff - and the projected source size, introduce the timing aspects, and establish connections between the light curve observables and the refractive cloud. We extend our discussion to clouds with different structure functions from Kolmogorov-type turbulence. Conclusions: Our study confirms that current telescopes of ~4 m with fast-readout, wide-field detectors have the capability of discovering the first interstellar optical scintillation effects. We also show that this effect should be unambiguously distinguished from any other type of variability through the observation of desynchronised light curves, simultaneously measured by two distant telescopes.

  11. Brief report: Using global positioning system (GPS) enabled cell phones to examine adolescent travel patterns and time in proximity to alcohol outlets.

    PubMed

    Byrnes, Hilary F; Miller, Brenda A; Morrison, Christopher N; Wiebe, Douglas J; Remer, Lillian G; Wiehe, Sarah E

    2016-07-01

    As adolescents gain freedom to explore new environments unsupervised, more time in proximity to alcohol outlets may increase risks for alcohol and marijuana use. This pilot study: 1) Describes variations in adolescents' proximity to outlets by time of day and day of the week, 2) Examines variations in outlet proximity by drinking and marijuana use status, and 3) Tests feasibility of obtaining real-time data to study adolescent proximity to outlets. U.S. adolescents (N = 18) aged 16-17 (50% female) carried GPS-enabled smartphones for one week with their locations tracked. The geographic areas where adolescents spend time, activity spaces, were created by connecting GPS points sequentially and adding spatial buffers around routes. Proximity to outlets was greater during after school and evening hours. Drinkers and marijuana users were in proximity to outlets 1½ to 2 times more than non-users. Findings provide information about where adolescents spend time and times of greatest risk, informing prevention efforts.

  12. Traumatic proximal tibiofibular dislocation.

    PubMed

    Burgos, J; Alvarez-Montero, R; Gonzalez-Herranz, P; Rapariz, J M

    1997-01-01

    Proximal tibiofibular dislocation is an exceptional lesion. Rarer still is its presentation in childhood. We describe the clinical case of a 6-year-old boy, the victim of a road accident. He had a tibiofibular dislocation associated with a metaphyseal fracture of the tibia.

  13. Steerable Capacitive Proximity Sensor

    NASA Technical Reports Server (NTRS)

    Jenstrom, Del T.; Mcconnell, Robert L.

    1994-01-01

    Steerable capacitive proximity sensor of "capaciflector" type based partly on sensing units described in GSC-13377 and GSC-13475. Position of maximum sensitivity adjusted without moving sensor. Voltage of each driven shield adjusted separately to concentrate sensing electric field more toward one side or other.

  14. Proximate Analysis of Coal

    ERIC Educational Resources Information Center

    Donahue, Craig J.; Rais, Elizabeth A.

    2009-01-01

    This lab experiment illustrates the use of thermogravimetric analysis (TGA) to perform proximate analysis on a series of coal samples of different rank. Peat and coke are also examined. A total of four exercises are described. These are dry exercises as students interpret previously recorded scans. The weight percent moisture, volatile matter,…

  15. Subacute diabetic proximal neuropathy

    NASA Technical Reports Server (NTRS)

    Pascoe, M. K.; Low, P. A.; Windebank, A. J.; Litchy, W. J.

    1997-01-01

    OBJECTIVE: To evaluate the clinical, electrophysiologic, autonomic, and neuropathologic characteristics and the natural history of subacute diabetic proximal neuropathy and its response to immunotherapy. MATERIAL AND METHODS: For the 12-year period from 1983 to 1995, we conducted a retrospective review of medical records of Mayo Clinic patients with diabetes who had subacute onset and progression of proximal weakness. The responses of treated versus untreated patients were compared statistically. RESULTS: During the designated study period, 44 patients with subacute diabetic proximal neuropathy were encountered. Most patients were middle-aged or elderly, and no sex preponderance was noted. The proximal muscle weakness often was associated with reduced or absent lower extremity reflexes. Associated weight loss was a common finding. Frequently, patients had some evidence of demyelination on nerve conduction studies, but it invariably was accompanied by concomitant axonal degeneration. The cerebrospinal fluid protein concentration was usually increased. Diffuse and substantial autonomic failure was generally present. In most cases, a sural nerve biopsy specimen suggested demyelination, although evidence of an inflammatory infiltrate was less common. Of 12 patients who received treatment (with prednisone, intravenous immune globulin, or plasma exchange), 9 had improvement of their conditions, but 17 of 29 untreated patients (59%) with follow-up also eventually had improvement, albeit at a much slower rate. Improvement was usually incomplete. CONCLUSION: We suggest that the entity of subacute diabetic proximal neuropathy is an extensive and severe variant of bilateral lumbosacral radiculoplexopathy, with some features suggestive of an immune-mediated cause. It differs from chronic inflammatory demyelinating polyradiculoneuropathy in that most cases have a more restricted distribution and seem to be monophasic and self-limiting. The efficacy of immunotherapy is unproved

  16. Measurement of radiation damage of water-based liquid scintillator and liquid scintillator

    DOE PAGES

    Bignell, L. J.; Diwan, M. V.; Hans, S.; ...

    2015-10-19

    Liquid scintillating phantoms have been proposed as a means to perform real-time 3D dosimetry for proton therapy treatment plan verification. We have studied what effect radiation damage to the scintillator will have upon this application. We have performed measurements of the degradation of the light yield and optical attenuation length of liquid scintillator and water-based liquid scintillator after irradiation by 201 MeV proton beams that deposited doses of approximately 52 Gy, 300 Gy, and 800 Gy in the scintillator. Liquid scintillator and water-based liquid scintillator (composed of 5% scintillating phase) exhibit light yield reductions of 1.74 ± 0.55 % andmore » 1.31 ± 0.59 % after ≈ 800 Gy of proton dose, respectively. Some increased optical attenuation was observed in the irradiated samples, the measured reduction to the light yield is also due to damage to the scintillation light production. Based on our results and conservative estimates of the expected dose in a clinical context, a scintillating phantom used for proton therapy treatment plan verification would exhibit a systematic light yield reduction of approximately 0.1% after a year of operation.« less

  17. Measurement of radiation damage of water-based liquid scintillator and liquid scintillator

    SciTech Connect

    Bignell, L. J.; Diwan, M. V.; Hans, S.; Jaffe, D. E.; Rosero, R.; Vigdor, S.; Viren, B.; Worcester, E.; Yeh, M.; Zhang, C.

    2015-10-19

    Liquid scintillating phantoms have been proposed as a means to perform real-time 3D dosimetry for proton therapy treatment plan verification. We have studied what effect radiation damage to the scintillator will have upon this application. We have performed measurements of the degradation of the light yield and optical attenuation length of liquid scintillator and water-based liquid scintillator after irradiation by 201 MeV proton beams that deposited doses of approximately 52 Gy, 300 Gy, and 800 Gy in the scintillator. Liquid scintillator and water-based liquid scintillator (composed of 5% scintillating phase) exhibit light yield reductions of 1.74 ± 0.55 % and 1.31 ± 0.59 % after ≈ 800 Gy of proton dose, respectively. Some increased optical attenuation was observed in the irradiated samples, the measured reduction to the light yield is also due to damage to the scintillation light production. Based on our results and conservative estimates of the expected dose in a clinical context, a scintillating phantom used for proton therapy treatment plan verification would exhibit a systematic light yield reduction of approximately 0.1% after a year of operation.

  18. An angiotensin-(1-7) peptidase in the kidney cortex, proximal tubules, and human HK-2 epithelial cells that is distinct from insulin-degrading enzyme.

    PubMed

    Wilson, Bryan A; Cruz-Diaz, Nildris; Marshall, Allyson C; Pirro, Nancy T; Su, Yixin; Gwathmey, TanYa M; Rose, James C; Chappell, Mark C

    2015-03-15

    Angiotensin 1-7 [ANG-(1-7)] is expressed within the kidney and exhibits renoprotective actions that antagonize the inflammatory, fibrotic, and pro-oxidant effects of ANG II. We previously identified an peptidase that preferentially metabolized ANG-(1-7) to ANG-(1-4) in the brain medulla and cerebrospinal fluid (CSF) of sheep (Marshall AC, Pirro NT, Rose JC, Diz DI, Chappell MC. J Neurochem 130: 313-323, 2014); thus the present study established the expression of the peptidase in the kidney. Utilizing a sensitive HPLC-based approach, we demonstrate a peptidase activity that hydrolyzed ANG-(1-7) to ANG-(1-4) in the sheep cortex, isolated tubules, and human HK-2 renal epithelial cells. The peptidase was markedly sensitive to the metallopeptidase inhibitor JMV-390; human HK-2 cells expressed subnanomolar sensitivity (IC50 = 0.5 nM) and the highest specific activity (123 ± 5 fmol·min(-1)·mg(-1)) compared with the tubules (96 ± 12 fmol·min(-1)·mg(-1)) and cortex (107 ± 9 fmol·min(-1)·mg(-1)). The peptidase was purified 41-fold from HK-2 cells; the activity was sensitive to JMV-390, the chelator o-phenanthroline, and the mercury-containing compound p-chloromercuribenzoic acid (PCMB), but not to selective inhibitors against neprilysin, neurolysin and thimet oligopeptidase. Both ANG-(1-7) and its endogenous analog [Ala(1)]-ANG-(1-7) (alamandine) were preferentially hydrolyzed by the peptidase compared with ANG II, [Asp(1)]-ANG II, ANG I, and ANG-(1-12). Although the ANG-(1-7) peptidase and insulin-degrading enzyme (IDE) share similar inhibitor characteristics of a metallothiolendopeptidase, we demonstrate marked differences in substrate specificity, which suggest these peptidases are distinct. We conclude that an ANG-(1-7) peptidase is expressed within the renal proximal tubule and may play a potential role in the renal renin-angiotensin system to regulate ANG-(1-7) tone.

  19. An angiotensin-(1–7) peptidase in the kidney cortex, proximal tubules, and human HK-2 epithelial cells that is distinct from insulin-degrading enzyme

    PubMed Central

    Wilson, Bryan A.; Cruz-Diaz, Nildris; Marshall, Allyson C.; Pirro, Nancy T.; Su, Yixin; Gwathmey, TanYa M.; Rose, James C.

    2015-01-01

    Angiotensin 1–7 [ANG-(1–7)] is expressed within the kidney and exhibits renoprotective actions that antagonize the inflammatory, fibrotic, and pro-oxidant effects of ANG II. We previously identified an peptidase that preferentially metabolized ANG-(1–7) to ANG-(1–4) in the brain medulla and cerebrospinal fluid (CSF) of sheep (Marshall AC, Pirro NT, Rose JC, Diz DI, Chappell MC. J Neurochem 130: 313–323, 2014); thus the present study established the expression of the peptidase in the kidney. Utilizing a sensitive HPLC-based approach, we demonstrate a peptidase activity that hydrolyzed ANG-(1–7) to ANG-(1–4) in the sheep cortex, isolated tubules, and human HK-2 renal epithelial cells. The peptidase was markedly sensitive to the metallopeptidase inhibitor JMV-390; human HK-2 cells expressed subnanomolar sensitivity (IC50 = 0.5 nM) and the highest specific activity (123 ± 5 fmol·min−1·mg−1) compared with the tubules (96 ± 12 fmol·min−1·mg−1) and cortex (107 ± 9 fmol·min−1·mg−1). The peptidase was purified 41-fold from HK-2 cells; the activity was sensitive to JMV-390, the chelator o-phenanthroline, and the mercury-containing compound p-chloromercuribenzoic acid (PCMB), but not to selective inhibitors against neprilysin, neurolysin and thimet oligopeptidase. Both ANG-(1–7) and its endogenous analog [Ala1]-ANG-(1–7) (alamandine) were preferentially hydrolyzed by the peptidase compared with ANG II, [Asp1]-ANG II, ANG I, and ANG-(1–12). Although the ANG-(1–7) peptidase and insulin-degrading enzyme (IDE) share similar inhibitor characteristics of a metallothiolendopeptidase, we demonstrate marked differences in substrate specificity, which suggest these peptidases are distinct. We conclude that an ANG-(1–7) peptidase is expressed within the renal proximal tubule and may play a potential role in the renal renin-angiotensin system to regulate ANG-(1–7) tone. PMID:25568136

  20. Protein kinase C-α interaction with iHSP70 in mitochondria promotes recovery of mitochondrial function after injury in renal proximal tubular cells.

    PubMed

    Nowak, Grazyna; Soundararajan, Sridharan; Mestril, Ruben

    2013-09-01

    This study determined the role of PKC-α and associated inducible heat shock protein 70 (iHSP70) in the repair of mitochondrial function in renal proximal tubular cells (RPTCs) after oxidant injury. Wild-type PKC-α (wtPKC-α) and an inactive PKC-α [dominant negative dn; PKC-α] mutant were overexpressed in primary cultures of RPTCs, and iHSP70 levels and RPTC regeneration were assessed after treatment with the oxidant tert-butylhydroperoxide (TBHP). TBHP exposure increased ROS production and induced RPTC death, which was prevented by ferrostatin and necrostatin-1 but not by cyclosporin A. Overexpression of wtPKC-α maintained mitochondrial levels of active PKC-α, reduced cell death, and accelerated proliferation without altering ROS production in TBHP-injured RPTCs. In contrast, dnPKC-α blocked proliferation and monolayer regeneration. Coimmunoprecipitation and proteomic analysis demonstrated an association between inactive, but not active, PKC-α and iHSP70 in mitochondria. Mitochondrial iHSP70 levels increased as levels of active PKC-α decreased after injury. Overexpression of dnPKC-α augmented, whereas overexpression of wtPKC-α abrogated, oxidant-induced increases in mitochondrial iHSP70 levels. iHSP70 overexpression (1) maintained mitochondrial levels of phosphorylated PKC-α, (2) improved the recovery of state 3 respiration and ATP content, (3) decreased RPTC death (an effect abrogated by cyclosporine A), and (4) accelerated proliferation after oxidant injury. In contrast, iHSP70 inhibition blocked the recovery of ATP content and exacerbated RPTC death. Inhibition of PKC-α in RPTC overexpressing iHSP70 blocked the protective effects of iHSP70. We conclude that active PKC-α maintains mitochondrial function and decreases cell death after oxidant injury. iHSP70 is recruited to mitochondria in response to PKC-α dephosphorylation and associates with and reactivates inactive PKC-α, which promotes the recovery of mitochondrial function, decreases RPTC

  1. Regulation of SGLT expression and localization through Epac/PKA-dependent caveolin-1 and F-actin activation in renal proximal tubule cells.

    PubMed

    Lee, Yu Jin; Kim, Mi Ok; Ryu, Jung Min; Han, Ho Jae

    2012-04-01

    This study demonstrated that exchange proteins directly activated by cAMP (Epac) and protein kinase A (PKA) by 8-bromo (8-Br)-adenosine 3',5'-cyclic monophosphate (cAMP) stimulated [(14)C]-α-methyl-D-glucopyranoside (α-MG) uptake through increased sodium-glucose cotransporters (SGLTs) expression and translocation to lipid rafts in renal proximal tubule cells (PTCs). In PTCs, SGLTs were colocalized with lipid raft caveolin-1 (cav-1), disrupted by methyl-β-cyclodextrin (MβCD). Selective activators of Epac or PKA, 8-Br-cAMP, and forskolin stimulated expressions of SGLTs and α-MG uptake in PTCs. In addition, 8-Br-cAMP-induced PKA and Epac activation increased phosphorylation of extracellular signal-regulated kinase (ERK), p38 mitogen-activated protein kinase (MAPK), and nuclear factor kappa B (NF-κB), which were involved in expressions of SGLTs. Furthermore, 8-Br-cAMP stimulated SGLTs translocation to lipid rafts via filamentous actin (F-actin) organization, which was blocked by cytochalasin D. In addition, cav-1 and SGLTs stimulated by 8-Br-cAMP were detected in lipid rafts, which were blocked by cytochalasin D. Furthermore, 8-Br-cAMP-induced SGLTs translocation and α-MG uptake were attenuated by inhibition of cav-1 activation with cav-1 small interfering RNA (siRNA) and inhibition of F-actin organization with TRIO and F-actin binding protein (TRIOBP). In conclusion, 8-Br-cAMP stimulated α-MG uptake via Epac and PKA-dependent SGLTs expression and trafficking through cav-1 and F-actin in PTCs.

  2. A novel role for c-Myc in G protein-coupled receptor kinase 4 (GRK4) transcriptional regulation in human kidney proximal tubule cells.

    PubMed

    Gildea, John J; Tran, Hanh T; Van Sciver, Robert E; Bigler Wang, Dora; Carlson, Julia M; Felder, Robin A

    2013-05-01

    The G protein-coupled receptor kinase 4 (GRK4) negatively regulates the dopaminergic system by desensitizing the dopamine-1-receptor. The expressional control of GRK4 has not been reported, but here we show that the transcription factor c-Myc binds to the promoter of GRK4 and positively regulates GRK4 protein expression in human renal proximal tubule cells (RPTCs). Addition of phorbol esters to RPTCs not only increased c-Myc binding to the GRK4 promoter but also increased both phospho-c-Myc and GRK4 expression. The phorbol ester-mediated increase in GRK4 expression was completely blocked by the c-Myc inhibitor, 10074-G5, indicating that GRK4 is downstream of phospho-c-Myc. The autocrine production of angiotensin II (Ang II) in RPTCs increased the phosphorylation and activation of c-Myc and subsequently GRK4 expression. 3-Amino-4-thio-butyl sulfonate, an inhibitor of aminopeptidase A, increased RPTC secretion of Ang II. 3-Amino-4-thio-butyl sulfonate or Ang II increased the expression of both phospho-c-Myc and GRK4, which was blocked by 10074-G5. Blockade of the Ang II type 1 receptor with losartan decreased phospho-c-Myc and GRK4 expression. Both inhibition of c-Myc activity and blockade of Ang II type 1 receptor restored the coupling of dopamine-1-receptor to adenylyl cyclase stimulation in uncoupled RPTCs, whereas phorbol esters or Ang II caused the uncoupling of normally coupled RPTCs. We suggest that the Ang II type 1 receptor impairs dopamine-1-receptor function via c-Myc activation of GRK4. This novel pathway may be involved in the increase in blood pressure in hypertension that is mediated by increased activity of the renin-angiotensin system and decreased activity of the renal dopaminergic system.

  3. The differential role of Smad2 and Smad3 in the regulation of pro-fibrotic TGFβ1 responses in human proximal-tubule epithelial cells

    PubMed Central

    Phanish, Mysore K.; Wahab, Nadia A.; Colville-Nash, Paul; Hendry, Bruce M.; Dockrell, Mark E. C.

    2005-01-01

    In chronic renal diseases, progressive loss of renal function correlates with advancing tubulo-interstitial fibrosis. TGFβ1-Smad (transforming growth factor-β1–Sma and Mad protein) signalling plays an important role in the development of renal tubulo-interstitial fibrosis. Secretion of CTGF (connective-tissue growth factor; CCN2) by PTECs (proximal-tubule epithelial cells) and EMT (epithelial–mesenchymal transdifferentiation) of PTECs to myofibroblasts in response to TGFβ are critical Smad-dependent events in the development of tubulo-interstitial fibrosis. In the present study we have investigated the distinct contributions of Smad2 and Smad3 to expression of CTGF, E-cadherin, α-SMA (α-smooth-muscle actin) and MMP-2 (matrix-metalloproteinase-2) in response to TGFβ1 treatment in an in vitro culture model of HKC-8 (transformed human PTECs). RNA interference was used to achieve selective and specific knockdown of Smad2 and Smad3. Cellular E-cadherin, α-SMA as well as secreted CTGF and MMP-2 were assessed by Western immunoblotting. TGFβ1 treatment induced a fibrotic phenotype with increased expression of CTGF, MMP-2 and α-SMA, and decreased expression of E-cadherin. TGFβ1-induced increases in CTGF and decreases in E-cadherin expression were Smad3-dependent, whereas increases in MMP-2 expression were Smad2-dependent. Increases in α-SMA expression were dependent on both Smad2 and Smad3 and were abolished by combined knockdown of both Smad2 and Smad3. In conclusion, we have demonstrated distinct roles for Smad2 and Smad3 in TGFβ1-induced CTGF expression and markers of EMT in human PTECs. This can be of therapeutic value in designing targeted anti-fibrotic therapies for tubulo-interstitial fibrosis. PMID:16253118

  4. Scintillation Breakdowns in Chip Tantalum Capacitors

    NASA Technical Reports Server (NTRS)

    Teverovsky, Alexander

    2008-01-01

    Scintillations in solid tantalum capacitors are momentarily local breakdowns terminated by a self-healing or conversion to a high-resistive state of the manganese oxide cathode. This conversion effectively caps the defective area of the tantalum pentoxide dielectric and prevents short-circuit failures. Typically, this type of breakdown has no immediate catastrophic consequences and is often considered as nuisance rather than a failure. Scintillation breakdowns likely do not affect failures of parts under surge current conditions, and so-called "proofing" of tantalum chip capacitors, which is a controllable exposure of the part after soldering to voltages slightly higher than the operating voltage to verify that possible scintillations are self-healed, has been shown to improve the quality of the parts. However, no in-depth studies of the effect of scintillations on reliability of tantalum capacitors have been performed so far. KEMET is using scintillation breakdown testing as a tool for assessing process improvements and to compare quality of different manufacturing lots. Nevertheless, the relationship between failures and scintillation breakdowns is not clear, and this test is not considered as suitable for lot acceptance testing. In this work, scintillation breakdowns in different military-graded and commercial tantalum capacitors were characterized and related to the rated voltages and to life test failures. A model for assessment of times to failure, based on distributions of breakdown voltages, and accelerating factors of life testing are discussed.

  5. Estimation of Fano factor in inorganic scintillators

    PubMed Central

    Bora, Vaibhav; Barrett, Harrison H.; Fastje, David; Clarkson, Eric; Furenlid, Lars; Bousselham, Abdelkader; Shah, Kanai S.; Glodo, Jarek

    2015-01-01

    The Fano factor of an integer-valued random variable is defined as the ratio of its variance to its mean. Correlation between the outputs of two photomultiplier tubes on opposite faces of a scintillation crystal was used to estimate the Fano factor of photoelectrons and scintillation photons. Correlations between the integrals of the detector outputs were used to estimate the photoelectron and photon Fano factor for YAP:Ce, SrI2:Eu and CsI:Na scintillator crystals. At 662 keV, SrI2:Eu was found to be sub-Poisson, while CsI:Na and YAP:Ce were found to be super-Poisson. An experiment setup inspired from the Hanbury Brown and Twiss experiment was used to measure the correlations as a function of time between the outputs of two photomultiplier tubes looking at the same scintillation event. A model of the scintillation and the detection processes was used to generate simulated detector outputs as a function of time for different values of Fano factor. The simulated outputs from the model for different Fano factors was compared to the experimentally measured detector outputs to estimate the Fano factor of the scintillation photons for YAP:Ce, LaBr3:Ce scintillator crystals. At 662 keV, LaBr3:Ce was found to be sub-Poisson, while YAP:Ce was found to be close to Poisson. PMID:26644631

  6. Equatorial scintillations: advances since ISEA-6

    SciTech Connect

    Not Available

    1985-01-01

    Our understanding of the morphology of equatorial scintillations has advanced due to more intensive observations at the equatorial anomaly locations in the different longitude zones. The unmistakable effect of the sunspot cycle in controlling irregularity belt width and electron concentration responsible for strong scintillation in the controlling the magnitude of scintillations has been recognized by interpreting scintillation observations inthe light of realistic models of total electron content at various longitudes. A hypothesis based on the alignment of the solar terminator with the geomagnetic flux tubes as an indicator of enhanced scintillation occurrence and another based on the influence of a transequatorial thermospheric neutral wind have been postulated to describe the observed longitudinal variation. A distinct class of equatorial irregularities known as the bottomside sinusoidal (BSS) type was identified. These irregularities occur in very large patches, sometimes in excess of several thousand kilometers in the E-W direction and are associated with frequency spread on ionograms. Scintillations caused by such irregularities exist only in the VHF band, exhibit Fresnel oscillations in intensity spectra and are found to give rise to extremely long durations (approx. several hours) of uninterrrupted scintillations.

  7. Protein Neighbors and Proximity Proteomics.

    PubMed

    Rees, Johanna S; Li, Xue-Wen; Perrett, Sarah; Lilley, Kathryn S; Jackson, Antony P

    2015-11-01

    Within cells, proteins can co-assemble into functionally integrated and spatially restricted multicomponent complexes. Often, the affinities between individual proteins are relatively weak, and proteins within such clusters may interact only indirectly with many of their other protein neighbors. This makes proteomic characterization difficult using methods such as immunoprecipitation or cross-linking. Recently, several groups have described the use of enzyme-catalyzed proximity labeling reagents that covalently tag the neighbors of a targeted protein with a small molecule such as fluorescein or biotin. The modified proteins can then be isolated by standard pulldown methods and identified by mass spectrometry. Here we will describe the techniques as well as their similarities and differences. We discuss their applications both to study protein assemblies and to provide a new way for characterizing organelle proteomes. We stress the importance of proteomic quantitation and independent target validation in such experiments. Furthermore, we suggest that there are biophysical and cell-biological principles that dictate the appropriateness of enzyme-catalyzed proximity labeling methods to address particular biological questions of interest.

  8. Effect of exogenous TGF-β1 on the cadmium-induced nephrotoxicity by inhibiting apoptosis of proximal tubular cells through PI3K-AKT-mTOR signaling pathway.

    PubMed

    Huang, Minyi; Su, Li; Yang, Limin; Zhu, Liangliang; Liu, Zhaowen; Duan, Renyan

    2017-03-22

    Heavy metal polluted soils have been a serious problem for the global ecological balance and people's health. Cadmium (Cd), one of the heavy metals, could induce apoptosis of proximal tubular cells in many experimental models and lead to damage the human kidney. Here, we reported a potent chemokine TGF-β1 which could ameliorate cadmium-induced nephrotoxicity. Interestingly, western blotting and TUNEL staining assays indicated that PI3K-AKT-mTOR signaling pathway was involved in the protective mechanism of TGF-β1 in vitro and in vivo. Moreover, TGF-β1 could alleviate Cd-induced nephrotoxicity by inhibiting apoptosis of proximal tubular cells through detecting the level of caspase 3, 8 and 9. Therefore, up-regulation of exogenous TGF-β1 may be a potential strategy to reverse cadumium-induced nephrotoxicity.

  9. Scintillation counter with WLS fiber readout

    NASA Astrophysics Data System (ADS)

    Bukin, D. A.; Druzhinin, V. P.; Golubev, V. B.; Serednyakov, S. I.

    1997-02-01

    The parameters of a cylindrical scintillation counter of 126 mm in diameter and 370 mm in length with wavelength shifter (WLS) fiber readout are presented. The fibers are glued into machined grooves along the scintillator. Light from both ends of the WLS fibers is transmitted to separate photomultipliers by 1 m long clear optical fibers. The average total signal, collected from both sides of the counter is equivalent to 8 photoelectrons per minimum ionizing particle. The described cylindrical scintillation counter is a part of inner system of collider detector SND.

  10. Scintillating Track Image Camera-SCITIC

    NASA Astrophysics Data System (ADS)

    Sato, Akira; Asai, Jyunkichi; Ieiri, Masaharu; Iwata, Soma; Kadowaki, Tetsuhito; Kurosawa, Maki; Nagae, Tomohumi; Nakai, Kozi

    2004-04-01

    A new type of track detector, scintillating track image camera (SCITIC) has been developed. Scintillating track images of particles in a scintillator are focused by an optical lens system on a photocathode on image intesifier tube (IIT). The image signals are amplified by an IIT-cascade and stored by a CCD camera. The performance of the detector has been tested with cosmic-ray muons and with pion- and proton-beams from the KEK 12-GeV proton synchrotron. Data of the test experiments have shown promising features of SCITIC as a triggerable track detector with a variety of possibilities.

  11. Measurement of light emission in scintillation vials

    SciTech Connect

    Duran Ramiro, M. Teresa; Garcia-Torano, Eduardo

    2005-09-15

    The efficiency and energy resolution of liquid scintillation counting (LSC) systems are strongly dependent on the optical characteristics of scintillators, vials, and reflectors. This article presents the results of measurements of the light-emission profile of scintillation vials. Two measurement techniques, autoradiographs and direct measurements with a photomultiplier tube, have been used to obtain light-emission distribution for standard vials of glass, etched glass and polyethylene. Results obtained with both techniques are in good agreement. For the first time, the effect of the meniscus in terms of light contribution has been numerically estimated. These results can help design LSC systems that are more efficient in terms of light collection.

  12. Advanced plastic scintillators for fast neutron discrimination

    SciTech Connect

    Feng, Patrick L; Anstey, Mitchell; Doty, F. Patrick; Mengesha, Wondwosen

    2014-09-01

    The present work addresses the need for solid-state, fast neutron discriminating scintillators that possess higher light yields and faster decay kinetics than existing organic scintillators. These respective attributes are of critical importance for improving the gamma-rejection capabilities and increasing the neutron discrimination performance under high-rate conditions. Two key applications that will benefit from these improvements include large-volume passive detection scenarios as well as active interrogation search for special nuclear materials. Molecular design principles were employed throughout this work, resulting in synthetically tailored materials that possess the targeted scintillation properties.

  13. Large volume flow-through scintillating detector

    DOEpatents

    Gritzo, Russ E.; Fowler, Malcolm M.

    1995-01-01

    A large volume flow through radiation detector for use in large air flow situations such as incinerator stacks or building air systems comprises a plurality of flat plates made of a scintillating material arranged parallel to the air flow. Each scintillating plate has a light guide attached which transfers light generated inside the scintillating plate to an associated photomultiplier tube. The output of the photomultiplier tubes are connected to electronics which can record any radiation and provide an alarm if appropriate for the application.

  14. Proximity Networks and Epidemics

    NASA Astrophysics Data System (ADS)

    Guclu, Hasan; Toroczkai, Zoltán

    2007-03-01

    We presented the basis of a framework to account for the dynamics of contacts in epidemic processes, through the notion of dynamic proximity graphs. By varying the integration time-parameter T, which is the period of infectivity one can give a simple account for some of the differences in the observed contact networks for different diseases, such as smallpox, or AIDS. Our simplistic model also seems to shed some light on the shape of the degree distribution of the measured people-people contact network from the EPISIM data. We certainly do not claim that the simplistic graph integration model above is a good model for dynamic contact graphs. It only contains the essential ingredients for such processes to produce a qualitative agreement with some observations. We expect that further refinements and extensions to this picture, in particular deriving the link-probabilities in the dynamic proximity graph from more realistic contact dynamics should improve the agreement between models and data.

  15. A Distal to Proximal Gradient of Human Choroid Plexus Development, with Antagonistic Expression of Glut1 and AQP1 in Mature Cells vs. Calbindin and PCNA in Proliferative Cells

    PubMed Central

    Castañeyra-Ruiz, Leandro; González-Marrero, Ibrahim; Hernández-Abad, Luis G.; Carmona-Calero, Emilia M.; Meyer, Gundela; Castañeyra-Perdomo, Agustín

    2016-01-01

    The choroid plexuses (ChP) are highly vascularized tissues suspended from each of the cerebral ventricles. Their main function is to secret cerebrospinal fluid (CSF) that fills the ventricles and the subarachnoid spaces, forming a crucial system for the development and maintenance of the CNS. However, despite the essential role of the ChP–CSF system to regulate the CNS in a global manner, it still remains one of the most understudied areas in neurobiology. Here we define by immunohistochemistry the expression of different proteins involved in the maturation and functionality of the ChP from the late embryological period to maturity. We found an opposite gradient of expression between aquaporin 1 (AQP1) and glucose transporter 1 (Glut 1) that define functional maturation in the ChP periphery, and proliferating cell nuclear antigen (PCNA) and calbindin (CB), present in the ChP root zone with proliferative activity. We conclude that the maturation of the ChP matures from distal to proximal, starting in the areas nearest to the cortex, expressing in the distal, mature areas AQP1 and Glut1 (related to ChP functionality to support cortex development), and in the proximal immature areas (ChP root) CB and PCNA related to progenitor activity and proliferation. PMID:27721744

  16. Glucosamine-induced Sp1 O-GlcNAcylation ameliorates hypoxia-induced SGLT dysfunction in primary cultured renal proximal tubule cells.

    PubMed

    Suh, Han Na; Lee, Yu Jin; Kim, Mi Ok; Ryu, Jung Min; Han, Ho Jae

    2014-10-01

    The aim of this study is to determine whether GlcN could recover the endoplasmic reticulum (ER) stress-induced dysfunction of Na(+) /glucose cotransporter (SGLT) in renal proximal tubule cells (PTCs) under hypoxia. With the rabbit model, the renal ischemia induced tubulointerstitial abnormalities and decreased SGLTs expression in tubular brush-border, which were recovered by GlcN. Thus, the protective mechanism of GlcN against renal ischemia was being examined by using PTCs. Hypoxia decreased the level of protein O-GlcNAc and the expression of O-GlcNAc transferase (OGT) while increased O-GlcNAcase (OGA) and these were reversed by GlcN. Hypoxia also decreased the expression of SGLTs (SGLT1 and 2) and [(14) C]-α-methyl-D-glucopyranoside (α-MG) uptake which were recovered by GlcN and PUGNAc (OGA inhibitor). Hypoxia enhanced reactive oxygen species (ROS) and then ER stress proteins, glucose-regulated protein 78 (GRP78), and C/EBP-homologous protein (CHOP). However, the expression of GRP78 increased till 6 h and then decreased whereas CHOP increased gradually. Moreover, decreased GRP78 and increased CHOP were reversed by NAC (antioxidant) and GlcN. GlcN ameliorated hypoxia-induced decrease of O-GlcNAc modification of Sp1 but OGT or Sp1 siRNAs blocked the recovery effect of GlcN on SGLT expression and α-MG uptake. In addition, hypoxia-decreased GRP78 and HIF-1α expression was reversed by GlcN but OGT siRNA or Sp1 siRNA ameliorated the effect of GlcN. When PTCs were transfected with GRP78 siRNA or HIF-1α siRNA, SGLT expression and α-MG uptake was decreased. Taken together, these data suggest that GlcN-induced O-GlcNAc modified Sp1 with stimulating GRP78 and HIF-1α activity ameliorate hypoxia-induced SGLT dysfunction in renal PTCs. J. Cell. Physiol. 229: 1557-1568, 2014. © 2014 Wiley Periodicals, Inc.

  17. Lack of formic acid production in rat hepatocytes and human renal proximal tubule cells exposed to chloral hydrate or trichloroacetic acid.

    PubMed

    Lock, Edward A; Reed, Celia J; McMillan, Joellyn M; Oatis, John E; Schnellmann, Rick G

    2007-02-12

    The industrial solvent trichloroethylene (TCE) and its major metabolites have been shown to cause formic aciduria in male rats. We have examined whether chloral hydrate (CH) and trichloroacetic acid (TCA), known metabolites of TCE, produce an increase in formic acid in vitro in cultures of rat hepatocytes or human renal proximal tubule cells (HRPTC). The metabolism and cytotoxicity of CH was also examined to establish that the cells were metabolically active and not compromised by toxicity. Rat hepatocytes and HRPTC were cultured in serum-free medium and then treated with 0.3-3mM CH for 3 days or 0.03-3mM CH for 10 days, respectively and formic acid production, metabolism to trichloroethanol (TCE-OH) and TCA and cytotoxicity determined. No increase in formic acid production in rat hepatocytes or HRPTC exposed to CH was observed over and above that due to chemical degradation, neither was formic acid production observed in rat hepatocytes exposed to TCA. HRPTC metabolized CH to TCE-OH and TCA with a 12-fold greater capacity to form TCE-OH versus TCA. Rat hepatocytes exhibited a 1.6-fold and three-fold greater capacity than HRPTC to form TCE-OH and TCA, respectively. CH and TCA were not cytotoxic to rat hepatocytes at concentrations up to 3mM/day for 3 days. With HRPTC, one sample showed no cytotoxicity to CH at concentrations up to 3mM/day for 10 days, while in another cytotoxicity was seen at 1mM/day for 3 days. In summary, increased formic acid production was not observed in rat hepatocytes or HRPTC exposed to TCE metabolites, suggesting that the in vivo response cannot be modelled in vitro. CH was toxic to HRPTC at millimolar concentrations/day over 10 days, while glutathione derived metabolites of TCE were toxic at micromolar concentrations/day over 10 days [Lock, E.A., Reed, C.J., 2006. Trichloroethylene: mechanisms of renal toxicity and renal cancer and relevance to risk assessment. Toxicol. Sci. 19, 313-331] supporting the view that glutathione derived

  18. Mouse amnionless, which is required for primitive streak assembly, mediates cell-surface localization and endocytic function of cubilin on visceral endoderm and kidney proximal tubules.

    PubMed

    Strope, Sharon; Rivi, Roberta; Metzger, Thomas; Manova, Katia; Lacy, Elizabeth

    2004-10-01

    Impaired primitive streak assembly in the mouse amnionless (amn) mutant results in the absence of non-axial trunk mesoderm, a derivative of the middle region of the primitive streak. In addition, the epiblast of amn mutants fails to increase significantly in size after E7.0, indicating that middle primitive streak assembly is mechanistically tied to the growth of the embryo during gastrulation. Amn, a novel transmembrane protein, is expressed exclusively in an extra-embryonic tissue, visceral endoderm (VE), during the early post-implantation stages. We show that Amn is also expressed in kidney proximal tubules (KPT) and intestinal epithelium, which, like the VE, are polarized epithelia specialized for resorption and secretion. To explore whether Amn participates in the development or function of KPT and intestinal epithelia and to gain insight into the function of Amn during gastrulation, we constructed Amn(-/-) ES cell<-->+/+ blastocyst chimeras. While chimeras form anatomically normal kidneys and intestine, they exhibit variable, selective proteinuria, a sign of KPT malfunction. In humans, AMN has been genetically connected to Cubilin (CUBN), a multi-ligand scavenger receptor expressed by KPT, intestine and yolk sac. Loss of CUBN, the intestinal intrinsic factor (IF)-vitamin B12 receptor, results in hereditary megaloblastic anemia (MGA1), owing to vitamin B12 malabsorption. The recent report of MGA1 families with mutations in AMN suggests that AMN functions in the same pathway as CUBN. We demonstrate that Cubn is not properly localized to the cell surface in Amn(-/-) tissues in the embryo and adult mouse, and that adult chimeras exhibit selective proteinuria of Cubn ligands. This study demonstrates that Amn is an essential component of the Cubn receptor complex in vivo and suggests that Amn/Cubn is required for endocytosis/transcytosis of one or more ligands in the VE during gastrulation to coordinate growth and patterning of the embryo. Furthermore, as AMN is

  19. Tumor Cell-Derived Microvesicles Induced Not Epithelial-Mesenchymal Transition but Apoptosis in Human Proximal Tubular (HK-2) Cells: Implications for Renal Impairment in Multiple Myeloma

    PubMed Central

    Zhao, Aiqi; Kong, Fancong; Liu, Chun-Jie; Yan, Guoxin; Gao, Fei; Guo, Hao; Guo, An-Yuan; Chen, Zhichao; Li, Qiubai

    2017-01-01

    Renal impairment (RI) is one of the hallmarks of multiple myeloma (MM) and carries a poor prognosis. Microvesicles (MVs) are membrane vesicles and play an important role in disease progression. Here, we investigated the role of MVs derived from MM cells (MM-MVs) in RI of MM. We found that MM-MVs significantly inhibited viability and induced apoptosis, but not epithelial-mesenchymal transition in human kidney-2 (HK-2), a human renal tubular epithelial cell line. The protein levels of cleaved caspase-3, 8, and 9, and E-cadherin, were increased, but vementin levels were decreased in the HK-2 cells treated with MM-MVs. Through a comparative sequencing and analysis of RNA content between the MVs from RPMI8226 MM cells (RPMI8226-MVs) and K562 leukemia cells, RPMI8226-MVs were enriched with more renal-pathogenic miRNAs, in which the selective miRNAs may participate in the up-regulation of the levels of cleaved caspase-3. Furthermore, the levels of CD138+ circulating MVs (cirMVs) in the peripheral blood were positively correlated with the severity of RI in newly-diagnosed MM. Our study supports MM-MVs representing a previously undescribed factor and playing a potential role in the development of RI of MM patients, and sheds light on the potential application of CD138+ cirMV counts in precise diagnosis of RI in MM and exploring MM-MVs as a therapeutic target. PMID:28264449

  20. Research and Development of Scintillation fiber Trackers

    SciTech Connect

    Kobayashi, A.; ITO, H.; Kawai, H.; Kodama, S.; Kaneko, N.; Han, S.

    2015-07-01

    We are developing the scintillation fiber trackers. This detector is consist of 0.5 mm diameter scintillation fibers and PPDs. This detector has the doughnut shape with outer diameter of 50 cm and inner diameter of 10 cm and thickness of 2 mm. The position resolution is 70 μm. There are no ineffective area. And the cost is several million yen. (authors)

  1. Liquid scintillators for optical fiber applications

    DOEpatents

    Franks, Larry A.; Lutz, Stephen S.

    1982-01-01

    A multicomponent liquid scintillator solution for use as a radiation-to-light converter in conjunction with a fiber optic transmission system. The scintillator includes a quantity of 1, 2, 4, 5, 3H, 6H, 1 OH, tetrahydro-8-trifluoromethyl (1) benzopyrano (9, 9a, 1-gh) quinolizin-10-one (Coumarin) as a solute in a fluor solvent such as benzyl alcohol or pseudo-cumene. The use of BIBUQ as an additional or primary solute is also disclosed.

  2. Liquid scintillators for optical fiber applications

    SciTech Connect

    Franks, L.A.; Lutz, S.S.

    1982-11-16

    A multicomponent liquid scintillator solution for use as a radiation-to-light converter in conjunction with a fiber optic transmission system. The scintillator includes a quantity of 1, 2 , 4, 5, 3H, 6H, 1 OH, tetrahydro-8-trifluoromethyl (1) benzopyrano (9, 9a, 1-gh) quinolizin-10-one (Coumarin) as a solute in a fluor solvent such as benzyl alcohol or pseudocumene. The use of bibuq as an additional or primary solute is also disclosed.

  3. Ternary liquid scintillator for optical fiber applications

    DOEpatents

    Franks, Larry A.; Lutz, Stephen S.

    1982-01-01

    A multicomponent liquid scintillator solution for use as a radiation-to-light converter in conjunction with a fiber optic transmission system. The scintillator includes a quantity of 5-amino-9-diethylaminobenz (a) phenoxazonium nitrate (Nile Blue Nitrate) as a solute in a fluor solvent such as benzyl alcohol. The use of PPD as an additional solute is also disclosed. The system is controllable by addition of a suitable quenching agent, such as phenol.

  4. Current status on plastic scintillators modifications.

    PubMed

    Bertrand, Guillaume H V; Hamel, Matthieu; Sguerra, Fabien

    2014-11-24

    Recent developments of plastic scintillators are reviewed, from 2000 to March 2014, distributed in two different chapters. First chapter deals with the chemical modifications of the polymer backbone, whereas modifications of the fluorescent probe are presented in the second chapter. All examples are provided with the scope of detection of various radiation particles. The main characteristics of these newly created scintillators and their detection properties are given.

  5. Real-time volumetric scintillation dosimetry

    NASA Astrophysics Data System (ADS)

    Beddar, S.

    2015-01-01

    The goal of this brief review is to review the current status of real-time 3D scintillation dosimetry and what has been done so far in this area. The basic concept is to use a large volume of a scintillator material (liquid or solid) to measure or image the dose distributions from external radiation therapy (RT) beams in three dimensions. In this configuration, the scintillator material fulfills the dual role of being the detector and the phantom material in which the measurements are being performed. In this case, dose perturbations caused by the introduction of a detector within a phantom will not be at issue. All the detector configurations that have been conceived to date used a Charge-Coupled Device (CCD) camera to measure the light produced within the scintillator. In order to accurately measure the scintillation light, one must correct for various optical artefacts that arise as the light propagates from the scintillating centers through the optical chain to the CCD chip. Quenching, defined in its simplest form as a nonlinear response to high-linear energy transfer (LET) charged particles, is one of the disadvantages when such systems are used to measure the absorbed dose from high-LET particles such protons. However, correction methods that restore the linear dose response through the whole proton range have been proven to be effective for both liquid and plastic scintillators. Volumetric scintillation dosimetry has the potential to provide fast, high-resolution and accurate 3D imaging of RT dose distributions. Further research is warranted to optimize the necessary image reconstruction methods and optical corrections needed to achieve its full potential.

  6. Multi-GNSS for Ionospheric Scintillation Studies

    NASA Astrophysics Data System (ADS)

    Morton, Y.

    2015-12-01

    GNSS have been widely used for ionospheric monitoring. We anticipate over 160 GNSS satellites broadcasting 400 signals by 2023, nearly double the number today. With their well-defined signal structures, high spatial density and spectral diversity, GNSS offers low cost and distributed passive sensing of ionosphere effects. There are, however, many challenges to utilize GNSS resources to characterize and forecast ionospheric scintillation. Originally intended for navigation purposes, GNSS receivers are designed to filter out nuisance effects due to ionosphere effects. GNSS measurements are plagued with errors from multipath, oscillator jitters, processing artifacts, and neutral atmosphere effects. Strong scintillation events are often characterized by turbulent structures in ionosphere, causing simultaneous deep amplitude fading and abrupt carrier phase changes. The combined weak signal and high carrier dynamics imposes conflicting requirements for GNSS receiver design. Therefore, GNSS receivers often experience cycle slips and loss of lock of signals during strong scintillation events. High quality, raw GNSS signals bearing space weather signatures and robust receiver algorithms designed to capture these signatures are needed in order for GNSS to be a reliable and useful agent for scintillation monitoring and forecasting. Our event-driven, reconfigurable data collection system is designed to achieve this purpose. To date, our global network has collected ~150TB of raw GNSS data during space weather events. A suite of novel receiver processing algorithms has been developed by exploitating GNSS spatial, frequency, temporal, and constellation diversity to process signals experiencing challenging scintillation impact. The algorithms and data have advanced our understanding of scintillation impact on GNSS, lead to more robust receiver technologies, and enabled high spatial and temporal resolution depiction of ionosphere responses to solar and geomagnetic conditions. This

  7. Metabolic alkalosis transition in renal proximal tubule cells facilitates an increase in CYP27B1, while blunting responsiveness to PTH

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Parathyroid hormone (PTH) is the central activator of renal proximal 1-alpha-hydroxylase (CYP27B1), the enzyme responsible for synthesis of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3). Past studies have documented a disruption of CYP27B1 activity in chronic metabolic acidosis in vivo, while simulated ac...

  8. Gallic acid ameliorates renal functions by inhibiting the activation of p38 MAPK in experimentally induced type 2 diabetic rats and cultured rat proximal tubular epithelial cells.

    PubMed

    Ahad, Amjid; Ahsan, Haseeb; Mujeeb, Mohd; Siddiqui, Waseem Ahmad

    2015-10-05

    Diabetic nephropathy (DN) is one of the leading causes of morbidity and mortality in diabetic patients that accounts for about 40% of deaths in type 2 diabetes. p38 mitogen activated protein kinase (p38 MAPK), a serine-threonine kinase, plays an important role in tissue inflammation and is known to be activated under conditions of oxidative stress and hyperglycemia. The role of p38 MAPK has been demonstrated in DN, and its inhibition has been suggested as an alternative approach in the treatment of DN. In the present study, we investigated the nephroprotective effects of an anti-inflammatory phenolic compound, gallic acid (GA, 3,4,5-trihydroxybenzoic acid), in high fat diet/streptozotocin (HFD/STZ) induce type 2 diabetic wistar albino rats. GA (25 mg/kgbw and 50 mg/kgbw, p.o.) treatment for 16 weeks post induction of diabetes led to a significant reduction in the levels of blood glucose, HbA1c, serum creatinine, blood urea nitrogen and proteinuria as well as a significant reduction in the levels of creatinine clearance. GA significantly inhibited the renal p38 MAPK and nuclear factor kappa B (N-κB) activation as well as significantly reduced the levels of renal transforming growth factor beta (TGF-β) and fibronectin. Treatment with GA resulted in a significant reduction in the serum levels of proinflammatory cytokines viz. interleukin 1 beta (IL-1β), IL-6 and tumor necrosis factor alpha (TNF-α). Moreover, GA significantly lowered renal pathology and attenuated renal oxidative stress. In cultured rat NRK 52E proximal tubular epithelial cells, GA treatment inhibited high glucose induced activation of p38 MAPK and NF-κB as well as suppressed proinflammatory cytokine synthesis. The results of the present study provide in vivo and in vitro evidences that the p38 MAPK pathway plays an important role in the pathogenesis of DN, and GA attenuates the p38 MAPK-mediated renal dysfunction in HFD/STZ induced type 2 diabetic rats.

  9. Development of Novel Polycrystalline Ceramic Scintillators

    SciTech Connect

    Wisniewska, Monika; Boatner, Lynn A; Neal, John S; Jellison Jr, Gerald Earle; Ramey, Joanne Oxendine; North, Andrea L; Wisniewski, Monica; Payzant, E Andrew; Howe, Jane Y; Lempicki, Aleksander; Brecher, Charlie; Glodo, J.

    2008-01-01

    For several decades most of the efforts to develop new scintillator materials have concentrated on high-light-yield inorganic single-crystals while polycrystalline ceramic scintillators, since their inception in the early 1980 s, have received relatively little attention. Nevertheless, transparent ceramics offer a promising approach to the fabrication of relatively inexpensive scintillators via a simple mechanical compaction and annealing process that eliminates single-crystal growth. Until recently, commonly accepted concepts restricted the polycrystalline ceramic approach to materials exhibiting a cubic crystal structure. Here, we report our results on the development of two novel ceramic scintillators based on the non-cubic crystalline materials: Lu SiO:Ce (LSO:Ce) and LaBr:Ce. While no evidence for texturing has been found in their ceramic microstructures, our LSO:Ce ceramics exhibit a surprisingly high level of transparency/ translucency and very good scintillation characteristics. The LSO:Ce ceramic scintillation reaches a light yield level of about 86% of that of a good LSO:Ce single crystal, and its decay time is even faster than in single crystals. Research on LaBr:Ce shows that translucent ceramics of the high-light-yield rare-earth halides can also be synthesized. Our LaBr:Ce ceramics have light yields above 42 000 photons/MeV (i.e., 70%of the single-crystal light yield).

  10. Crystal growth and scintillation properties of strontium iodide scintillators

    SciTech Connect

    van Loef, Edgar; Wilson, Cody; Cherepy, Nerine; Payne, Steven; Choong, Woon-Seng; Moses, William W.; Shah, Kanai

    2009-06-01

    Single crystals of SrI{sub 2}:Eu and SrI{sub 2}:Ce/Na were grown from anhydrous iodides by the vertical Bridgman technique in evacuated silica ampoules. Growth rates were of the order of 5-30 mm/day. Radioluminescence spectra of SrI{sub 2}:Eu and SrI{sub 2}:Ce/Na exhibit a broad band due to Eu{sup 2+} and Ce{sup 3+} emission, respectively. The maximum in the luminescence spectrum of SrI{sub 2}:Eu is found at 435 nm. The spectrum of SrI{sub 2}:Ce/Na exhibits a doublet peaking at 404 and 435 nm attributed to Ce{sup 3+} emission, while additional impurity - or defected - related emission is present at approximately 525 nm. The strontium iodide scintillators show very high light yields of up to 120,000 photons/MeV, have energy resolutions down to 3% at 662 keV (Full Width Half Maximum) and exhibit excellent light yield proportionality with a standard deviation of less than 5% between 6 and 460 keV.

  11. Validating the use of scintillation proxies to study ionospheric scintillation over the Ugandan region

    NASA Astrophysics Data System (ADS)

    Amabayo, Emirant B.; Jurua, Edward; Cilliers, Pierre J.

    2015-06-01

    In this study, we compare the standard scintillation indices (S4 and σΦ) from a SCINDA receiver with scintillation proxies (S4p and | sDPR |) derived from two IGS GPS receivers. Amplitude (S4) and phase (σΦ) scintillation data were obtained from the SCINDA installed at Makerere University (0.34°N, 32.57°E). The corresponding amplitude (S4p) and phase (| sDPR |) scintillation proxies were derived from data archived by IGS GPS receivers installed at Entebbe (0.04°N, 32.44°E) and Mbarara (0.60°S, 30.74°E). The results show that for most of the cases analysed in this study, σΦ and | sDPR | are in agreement. Amplitude scintillation occurrence estimated using the S4p are fairly consistent with the standard S4, mainly between 17:00 UT and 21:00 UT, despite a few cases of over and under estimation of scintillation levels by S4p. Correlation coefficients between σΦ and the | sDPR | proxy revealed positive correlation. Generally, S4p and S4 exhibits both moderate and strong positive correlation. TEC depletions associated with equatorial plasma bubbles are proposed as the cause of the observed scintillation over the region. These equatorial plasma bubbles were evident along the ray paths to satellites with PRN 2, 15, 27 and 11 as observed from MBAR and EBBE. In addition to equatorial plasma bubbles, atmospheric gravity waves with periods similar to those of large scale traveling ionospheric disturbances were also observed as one of the mechanisms for scintillation occurrence. The outcome of this study implies that GPS derived scintillation proxies can be used to quantify scintillation levels in the absence of standard scintillation data in the equatorial regions.

  12. CFTR is restricted to a small population of high expresser cells that provide a forskolin-sensitive transepithelial Cl- conductance in the proximal colon of the possum, Trichosurus vulpecula.

    PubMed

    Fan, Shujun; Harfoot, Natalie; Bartolo, Ray C; Butt, A Grant

    2012-04-01

    The cystic fibrosis transmembrane conductance regulator (CFTR) is central to anion secretion in both the possum and eutherian small intestine. Here, we investigated its role in the possum proximal colon, which has novel transport properties compared with the eutherian proximal colon. Despite considerable CFTR expression, high doses of the CFTR activator forskolin (EC(50)≈10 μmol l(-1)) were required for a modest, CFTR-dependent increase in short-circuit current (I(sc)) in the proximal colon. Presumably, this is because CFTR is restricted to the apical membrane of a small population of CFTR high expresser (CHE) cells in the surface and upper crypt epithelium. Furthermore, although the forskolin-stimulated I(sc) was dependent on serosal Na(+), Cl(-) and HCO(3)(-), consistent with anion secretion, inhibition of the basolateral Na-K-2Cl(-) (NKCC1) or Na-HCO(3) (pNBCe1) cotransporters did not prevent it. Therefore, although NKCC1 and pNBCe1 are expressed in the colonic epithelium they do not appear to be expressed in CHE cells. At low doses (IC(50)≈1 μmol l(-1)), forskolin also decreased the transepithelial conductance (G(T)) of the colon through inhibition of a 4,4'-diisothiocyano-2,2'-stilbenedisulphonic acid-sensitive anion conductance in the basolateral membrane of the CHE cells. This conductance is arranged in series with CFTR in the CHE cells and, therefore, the CHE cells provide a transepithelial Cl(-) conductance for passive Cl(-) absorption across the epithelium. Inhibition of the basolateral Cl(-) conductance of the CHE cells by forskolin will inhibit Na(+) absorption by restricting the movement of its counter-ion Cl(-), assisting in the conversion of the tissue from an absorptive to a secretory state.

  13. Echosonography with proximity sensors

    NASA Astrophysics Data System (ADS)

    Thaisiam, W.; Laithong, T.; Meekhun, S.; Chaiwathyothin, N.; Thanlarp, P.; Danworaphong, S.

    2013-03-01

    We propose the use of a commercial ultrasonic proximity sensor kit for profiling an altitude-varying surface by employing echosonography. The proximity sensor kit, two identical transducers together with its dedicated operating circuit, is used as a profiler for the construction of an image. Ultrasonic pulses are emitted from one of the transducers and received by the other. The time duration between the pulses allows us to determine the traveling distance of each pulse. In the experiment, the circuit is used with the addition of two copper wires for directing the outgoing and incoming signals to an oscilloscope. The time of flight of ultrasonic pulses can thus be determined. Square grids of 5 × 5 cm2 are made from fishing lines, forming pixels in the image. The grids are designed to hold the detection unit in place, about 30 cm above a flat surface. The surface to be imaged is constructed to be height varying and placed on the flat surface underneath the grids. Our result shows that an image of the profiled surface can be created by varying the location of the detection unit along the grid. We also investigate the deviation in relation to the time of flight of the ultrasonic pulse. Such an experiment should be valuable for conveying the concept of ultrasonic imaging to physical and medical science undergraduate students. Due to its simplicity, the setup could be made in any undergraduate laboratory relatively inexpensively and it requires no complex parts. The results illustrate the concept of echosonography.

  14. A capture-gated neutron calorimeter using plastic scintillators and 3He drift tubes

    SciTech Connect

    Wang, Zhehui; Morris, Christopher L; Spaulding, Randy J; Bacon, Jeffrey D; Borozdin, Konstantin N; Chung, Kiwhan; Clark, Deborah J; Green, Jesse A; Greene, Steven J; Hogan, Gary E; Jason, Andrew; Lisowski, Paul W; Makela, Mark F; Mariam, Fessaha G; Miyadera, Haruo; Murray, Matthew M; Saunders, Alexander; Wysocki, Frederick J; Gray, Frederick E

    2010-01-01

    A segmented neutron calorimeter using nine 4-inch x 4-inch x 48-inch plastic scintillators and sixteen 2-inch-diameter 48-inch-long 200-mbar-{sup 3}He drift tubes is described. The correlated scintillator and neutron-capture events provide a means for n/{gamma} discrimination, critical to the neutron calorimetry when the {gamma} background is substantial and the {gamma} signals are comparable in amplitude to the neutron signals. A single-cell prototype was constructed and tested. It can distinguish between a {sup 17}N source and a {sup 252}Cf source when the {gamma} and the thermal neutron background are sufficiently small. The design and construction of the nine-cell segmented detector assembly follow the same principle. By recording the signals from individual scintillators, additional {gamma}-subtraction schemes, such as through the time-of-flight between two scintillators, may also be used. The variations of the light outputs from different parts of a scintillator bar are less than 10%.

  15. Noble gas excimer scintillation following neutron capture in boron thin films

    SciTech Connect

    McComb, Jacob C.; Al-Sheikhly, Mohamad; Coplan, Michael A.; Thompson, Alan K.; Vest, Robert E.; Clark, Charles W.

    2014-04-14

    Far-ultraviolet scintillation signals have been measured in heavy noble gases (argon, krypton, xenon) following boron-neutron capture ({sup 10}B(n,α){sup 7}Li) in {sup 10}B thin films. The observed scintillation yields are comparable to the yields from some liquid and solid neutron scintillators. At noble gas pressures of 107 kPa, the number of photons produced per neutron absorbed following irradiation of a 1200 nm thick {sup 10}B film was 14 000 for xenon, 11 000 for krypton, and 6000 for argon. The absolute scintillation yields from the experimental configuration were calculated using data from (1) experimental irradiations, (2) thin-film characterizations, (3) photomultiplier tube calibrations, and (4) photon collection modeling. Both the boron films and the photomultiplier tube were characterized at the National Institute of Standards and Technology. Monte Carlo modeling of the reaction cell provided estimates of the photon collection efficiency and the transport behavior of {sup 10}B(n,α){sup 7}Li reaction products escaping the thin films. Scintillation yields increased with gas pressure due to increased ionization and excitation densities of the gases from the {sup 10}B(n,α){sup 7}Li reaction products, increased frequency of three-body, excimer-forming collisions, and reduced photon emission volumes (i.e., larger solid angle) at higher pressures. Yields decreased for thicker {sup 10}B thin films due to higher average energy loss of the {sup 10}B(n,α){sup 7}Li reaction products escaping the films. The relative standard uncertainties in the measurements were determined to lie between 14% and 16%. The observed scintillation signal demonstrates that noble gas excimer scintillation is promising for use in practical neutron detectors.

  16. Mercuric iodide photodetectors for scintillation spectroscopy

    SciTech Connect

    Markakis, J.; Dabrowski, A.; Iwanczyk, J.; Ortale, C.; Schnepple, W.

    1985-02-01

    We have measured the responses to /sup 137/Cs (662 keV) of both a 1-inch-diam by 2-inch-thick NaI(Tl) scintillator optically coupled to a 1-inch-diam by 800-..mu..mthick mercuric iodide (HgI/sub 2/) photodetector, and a 1-cmdiam by 1-cm-thick CaWO/sub 4/ scintillator coupled to a 1.3-cm-diam by 600-..mu..m-thick HgI/sub 2/ photodetector. Best spectral resolution to /sup 137/Cs was 7.8% FWHM for the NaI(Tl)-HgI/sub 2/ and 12.5% FWHM for the CaWO/sub 4/-HgI/sub 2/ detectors; peak-to-valley ratios were 26:1 and 16:1, respectively. HgI/sub 2/ detectors operate at room temperature and their use in scintillation spectroscopy presents the ultimate miniaturization of scintillation detectors, limited mainly by the size of the scintillation crystal.

  17. Mercuric iodide photodetectors for scintillation spectroscopy

    SciTech Connect

    Markakis, J.; Ortale, C.; Schnepple, W.; Iwanczyk, J.; Dabrowski, A.

    1984-01-01

    We have measured the responses to /sup 137/Cs (662 keV) of both a 1-inch-diam by 2-inch-thick NaI(Tl) scintillator optically coupled to a 1-inch-diam by 800-..mu..m-thick mercuric iodide (HgI/sub 2/) photodetector, and a 1-cm-diam by 1-cm-thick CaWO/sub 4/ scintillator coupled to a 1.3-cm-diam by 600-..mu..m-thick HgI/sub 2/ photodetector. Best spectral resolution to /sup 137/Cs was 7.8% FWHM for the NaI(Tl)-HgI/sub 2/ and 12.5% FWHM for the CaWO/sub 4/-HgI/sub 2/ detectors; peak-to-valley ratios were 26:1 and 16:1, respectively. HgI/sub 2/ detectors operate at room temperature and their use in scintillation spectroscopy presents the ultimate miniaturization of scintillation detectors, limited mainly by the size of the scintillation crystal.

  18. Some Properties of Fuzzy Soft Proximity Spaces

    PubMed Central

    Demir, İzzettin; Özbakır, Oya Bedre

    2015-01-01

    We study the fuzzy soft proximity spaces in Katsaras's sense. First, we show how a fuzzy soft topology is derived from a fuzzy soft proximity. Also, we define the notion of fuzzy soft δ-neighborhood in the fuzzy soft proximity space which offers an alternative approach to the study of fuzzy soft proximity spaces. Later, we obtain the initial fuzzy soft proximity determined by a family of fuzzy soft proximities. Finally, we investigate relationship between fuzzy soft proximities and proximities. PMID:25793224

  19. Some properties of fuzzy soft proximity spaces.

    PubMed

    Demir, İzzettin; Özbakır, Oya Bedre

    2015-01-01

    We study the fuzzy soft proximity spaces in Katsaras's sense. First, we show how a fuzzy soft topology is derived from a fuzzy soft proximity. Also, we define the notion of fuzzy soft δ-neighborhood in the fuzzy soft proximity space which offers an alternative approach to the study of fuzzy soft proximity spaces. Later, we obtain the initial fuzzy soft proximity determined by a family of fuzzy soft proximities. Finally, we investigate relationship between fuzzy soft proximities and proximities.

  20. Isotopic response with small scintillator based gamma-ray spectrometers

    DOEpatents

    Madden, Norman W [Sparks, NV; Goulding, Frederick S [Lafayette, CA; Asztalos, Stephen J [Oakland, CA

    2012-01-24

    The intrinsic background of a gamma ray spectrometer is significantly reduced by surrounding the scintillator with a second scintillator. This second (external) scintillator surrounds the first scintillator and has an opening of approximately the same diameter as the smaller central scintillator in the forward direction. The second scintillator is selected to have a higher atomic number, and thus has a larger probability for a Compton scattering interaction than within the inner region. Scattering events that are essentially simultaneous in coincidence to the first and second scintillators, from an electronics perspective, are precluded electronically from the data stream. Thus, only gamma-rays that are wholly contained in the smaller central scintillator are used for analytic purposes.

  1. Scintillating-glass-fiber neutron sensors

    NASA Astrophysics Data System (ADS)

    Abel, K. H.; Arthur, R. J.; Bliss, M.; Brite, D. W.; Brodzinski, R. L.; Craig, R. A.; Geelhood, B. D.; Goldman, D. S.; Griffin, J. W.; Perkins, R. W.; Reeder, P. L.; Richey, W. R.; Stahl, K. A.; Sunberg, D. S.; Warner, R. A.; Wogman, N. A.; Weber, M. J.

    1994-12-01

    Cerium-doped lithium-silicate glass fibers have been developed at Pacific Northwest Laboratory (PNL) for use as thermal neutron detectors. By using highly-enriched 6Li, these fibers efficiently capture thermal neutrons and produce scintillation light that can be detected at the ends of the fibers. Advantages of scintillating fibers over 3He or BF 3 proportional tubes include flexibility in geometric configuration, ruggedness in high-vibration environments, and less detector weight for the same neutron sensitivity. This paper describes the performance of these scintillating fibers with regard to count rates, pulse height spectra, absolute efficiencies, and neutron/gamma discrimination. Fibers with light transmission lengths ( {1}/{e}) of greater than 2 m have been produced at PNL. Neutron sensors in fiber form allow development of a variety of neutron detectors packaged in previously unavailable configurations. Brief descriptions of some of the devices already produced are included to illustrate these possibilities.

  2. Purification of large liquid scintillators for Borexino

    SciTech Connect

    Benziger, J.B.; Calaprice, F.P.; Vogelaar, R.B.

    1993-10-01

    Distillation extraction and crystallization have been used on scintillator mixtures for solar neutrino physics to remove cosmo- genically produced impurities ({sup 7}Be) and naturally occurring impurities ({sup 238}U, {sup 232}Th, and {sup 40}K), and to improve the optical transmission. Distillation was effective at removing {sup 7}Be and other impurities from aromatic solvents (p-xylene and pseudocumene) used as scintillator solvents. Distillation also provided the greatest improvement in the optical clarity of the solvents. Commercially available fluors (PPO and PMP) have high levels of potassium, far in excess of those tolerable for Borexino. Extraction techniques have been found to be effective at removing radioactive impurities, particularly potassium, from the fluors. An overall strategy for on-line purification of the scintillator for Borexino will be presented.

  3. Current trends in scintillator detectors and materials

    SciTech Connect

    Moses, William W.

    2001-10-23

    The last decade has seen a renaissance in inorganic scintillator development for gamma ray detection. Lead tungstate (PbWO4) has been developed for high energy physics experiments, and possesses exceptionally high density and radiation hardness, albeit with low luminous efficiency. Lutetium orthosilicate or LSO (Lu2SiO5:Ce) possesses a unique combination of high luminous efficiency, high density, and reasonably short decay time, and is now incorporated in commercial positron emission tomography (PET) cameras. There have been advances in understanding the fundamental mechanisms that limit energy resolution, and several recently discovered materials (such as LaBr3:Ce) possess energy resolution that approaches that of direct solid state detectors. Finally, there are indications that a neglected class of scintillator materials that exhibit near band-edge fluorescence could provide scintillators with sub-nanosecond decay times and high luminescent efficiency.

  4. On the scintillation efficiency of carborane-loaded liquid scintillators for thermal neutron detection

    NASA Astrophysics Data System (ADS)

    Chang, Zheng; Okoye, Nkemakonam C.; Urffer, Matthew J.; Green, Alexander D.; Childs, Kyle E.; Miller, Laurence F.

    2015-01-01

    The scintillation efficiency in response to thermal neutrons was studied by loading different concentrations of carborane (0-8.5 wt%) and naphthalene (0 and 100 g/L) in five liquid organic scintillators. The sample was characterized in Pb and Cd shields under the irradiation of the thermal neutrons from a 252Cf source. A method was developed to extract the net neutron response from the pulse-height spectra. It was found that the order of scintillation efficiencies for both γ-rays and thermal neutrons is as follows: diisopropylnaphthalene>toluene (concentrated solutes)>toluene~pseudocumene~m-xylene. The quench constants, obtained by fitting the Stern-Volmer model to the plots of light output versus carborane concentration, are in the range of 0.35-1.4 M-1 for all the scintillators. The Birks factors, estimated using the specific energy loss profiles of the incident particles, are in the range of 9.3-14 mg cm-2 MeV-1 for all the samples. The light outputs are in the range of 63-86 keV electron equivalents (keVee) in response to thermal neutrons. Loading naphthalene generally promotes the scintillation efficiency of the scintillator with a benzene derivative solvent. Among all the scintillators tested, the diisopropylnaphthalene-based scintillator shows the highest scintillation efficiency, lowest Birks factor, and smallest quench constants. These properties are primarily attributed to the double fused benzene-ring structure of the solvent, which is more efficient to populate to the excited singlet state under ionizing radiation and to transfer the excitation energy to the fluorescent solutes.

  5. The design of the TASD (totally active scintillator detector) prototype

    SciTech Connect

    Mefodiev, A. V. Kudenko, Yu. G.

    2015-12-15

    Totally active and magnetic segmented scintillation neutrino detectors are developed for the nextgeneration accelerator neutrino experiments. Such detectors will incorporate scintillation modules with scintillation counters that form X and Y planes. A single counter is a 7 × 10 × 90 mm{sup 3} scintillation bar with gluedin wavelength-shifting fibers and micropixel avalanche photodiodes. The results of measurements of the parameters of these detectors are presented.

  6. Statistics of time averaged atmospheric scintillation

    SciTech Connect

    Stroud, P.

    1994-02-01

    A formulation has been constructed to recover the statistics of the moving average of the scintillation Strehl from a discrete set of measurements. A program of airborne atmospheric propagation measurements was analyzed to find the correlation function of the relative intensity over displaced propagation paths. The variance in continuous moving averages of the relative intensity was then found in terms of the correlation functions. An empirical formulation of the variance of the continuous moving average of the scintillation Strehl has been constructed. The resulting characterization of the variance of the finite time averaged Strehl ratios is being used to assess the performance of an airborne laser system.

  7. Plastic scintillator centrality detector for BRAHMS

    NASA Astrophysics Data System (ADS)

    Lee, Y. K.; Debbe, R.; Lee, J. H.; Ito, Hironori; Sanders, S. J.

    2004-01-01

    An array of 40 tiles of thin plastic scintillators is used to construct the outer layer of the charged particle multiplicity detector for the BRAHMS experiment at the Relativistic Heavy Ion Collider (RHIC). Each tile is a square with 12 cm long sides and 5 mm thickness. The light from each of the scintillators is collected by wavelength shifting fibers embedded on the periphery. The light collection is uniform within 5% over the tile with the edge effect limited to 4 mm along the edge. The response is found to be linear in the high-multiplicity environment at RHIC with Au+Au beams at s NN of 200 GeV.

  8. Quality study of the purified liquid scintillator

    NASA Astrophysics Data System (ADS)

    Konno, Y.; Nakajima, K.; Kibe, Y.

    2008-07-01

    We have been distilling the KamLAND liquid scintillator (LS) for the low energy solar neutrino observation. The distillation removes radioactive impurities from LS efficiently. We developed two types of high sensitivity radon detectors to monitor 222Rn contamination which causes a primary background source 210Pb. Their required sensitivity is several mBq/m3. The features and the measurement results of these detectors are presented. We also report the study of liquid scintillator properties after the distillation: attenuation length, light output and PPO density.

  9. Effects of radiation on scintillating fiber performance

    SciTech Connect

    Bauer, M.L.; Cohn, H.; Efremenko, Yu.; Gordeev, A.; Kamyshkov, Yu.; Onopienko, D.; Savin, S.; Shmakov, K.; Tarkovsky, E.; Young, K.G.; Carey, R.; Rothman, M.; Sulak, L.; Worstell, W.; Parr, H.

    1992-12-31

    Continued rapid improvements in formulations for scintillating fibers require the ability to parameterize and predict effects of radiation on detector performance. Experimental techniques necessary to obtain needed information and calculational procedures used in performing predications for hadron scintillating fiber calorimetry in the Superconducting Supercollider environment are described. The experimental techniques involve control of the testing environment, consideration of dose rate effects, and other factors. These calculations involve the behavior of particle showers in the detector, expected levels of radiation, and parameterization of the radiation effects. A summary of significant work is also presented.

  10. Effects of radiation on scintillating fiber performance

    SciTech Connect

    Young, K.G.; Bauer, M.L.; Cohn, H.; Efremenko, Yu.; Gordeev, A.; Kamyshkov, Yu.; Onopienko, D.; Savin, S.; Shmakov, K.; Tarkovsky, E. ); Carey, R.; Rothman, M.; Sulak, L.; Worstell, W. ); Paar, H. )

    1993-08-01

    Continued rapid improvements in formulations for scintillating fibers require the ability to parameterize and predict effects of radiation on detector performance. Experimental techniques necessary to obtain desired information and calculational procedures used in performing predictions for hadron scintillating fiber calorimetry in the Superconducting Supercollider environment are described. The experimental techniques involve control of the testing environment, consideration of dose rate effects, and other factors. The calculations involve the behavior of particle showers in the detector, expected levels of radiation, and parameterization of the radiation effects. A summary of significant work is also presented.

  11. Near-infrared scintillation of liquid argon

    SciTech Connect

    Alexander, T.; Escobar, C. O.; Lippincott, W. H.; Rubinov, P.

    2016-03-03

    Since the 1970s it has been known that noble gases scintillate in the near infrared (NIR) region of the spectrum (0.7 $\\mu$m < $\\lambda$; < 1.5$\\mu$m). More controversial has been the question of the NIR light yield for condensed noble gases. We first present the motivation for using the NIR scintillation in liquid argon detectors, then briefly review early as well as more recent efforts and finally show encouraging preliminary results of a test performed at Fermilab.

  12. The homestake surface-underground scintillations: Description

    NASA Technical Reports Server (NTRS)

    Cherry, M. L.; Corbato, S.; Daily, T.; Fenyves, E. J.; Kieda, D.; Lande, K.; Lee, C. K.

    1985-01-01

    Two new detectors are currently under construction at the Homestake Gold Mine a 140-ton Large Area Scintillation Detector (LASD) with an upper surface area of 130 square meters, a geometry factor (for an isotropic flux) of 1200 square meters, sr, and a depth of 4200 m.w.e.; and a surface air shower array consisting of 100 scintillator elements, each 3 square meters, spanning an area of approximately square kilometers. Underground, half of the LASD is currently running and collecting muon data; on the surface, the first section of the air shower array will begin operation in the spring of 1985. The detectors and their capabilities are described.

  13. Plastic scintillators modifications for a selective radiation detection

    SciTech Connect

    Hamel, Matthieu; Bertrand, Guillaume H.V.; Carrel, Frederick; Coulon, Romain; Dumazert, Jonathan; Montbarbon, Eva; Sguerra, Fabien

    2015-07-01

    Recent developments of plastic scintillators are reviewed, from January 2000 to June 2015. All examples are distributed into the main application, i.e. how the plastic scintillator was modified to enhance the detection towards a given radiation particle. The main characteristics of these newly created scintillators and their detection properties are given. (authors)

  14. New liquid scintillators for fiber-optic applications

    SciTech Connect

    Lutz, S.S.; Franks, L.A.; Flournoy, J.M.; Lyons, P.B.

    1981-01-01

    New long-wavelength-emitting, high-speed, liquid scintillators have been developed and tailored specifically for plasma diagnostic experiments employing fiber optics. These scintillators offer significant advantages over commercially available plastic scintillators in terms of sensitivity and bandwidth. FWHM response times as fast as 350 ps have been measured. Emission spectra, time response data, and relative sensitivity information are presented.

  15. Upconverting nanoparticles for optimizing scintillator based detection systems

    DOEpatents

    Kross, Brian; McKisson, John E; McKisson, John; Weisenberger, Andrew; Xi, Wenze; Zom, Carl

    2013-09-17

    An upconverting device for a scintillation detection system is provided. The detection system comprises a scintillator material, a sensor, a light transmission path between the scintillator material and the sensor, and a plurality of upconverting nanoparticles particles positioned in the light transmission path.

  16. Assessment of scintillation proxy maps for a scintillation study during geomagnetically quiet and disturbed conditions over Uganda

    NASA Astrophysics Data System (ADS)

    Amabayo, Emirant B.; Jurua, Edward; Cilliers, Pierre J.

    2017-02-01

    The objective of this paper is demonstrate the validity and usefulness of scintillation proxies derived from IGS data, through its comparison with data from dedicated scintillation monitors and its application to GNSS scintillation patterns. The paper presents scintillation patterns developed by using data from the dedicated scintillation monitors of the scintillation network decision aid (SCINDA) network, and proxy maps derived from IGS GPS data for 2011 and 2012 over low latitude stations in Uganda. The amplitude and phase scintillation indicies (S4 and σΦ) were obtained from the Novatel GSV4004B ionospheric scintillation and total electron content (TEC) monitor managed by SCINDA at Makerere (0.340N, 32.570E). The corresponding IGS GPS proxy data were obtained from the receivers at Entebbe (0.040N, 32.440E) and Mbarara (0.600S, 30.740E). The derived amplitude (S4p) and phase (sDPR) scintillation proxy maps were compared with maps of S4 and σΦ during geomagnetic storms (moderate and strong) and geomagnetically quiet conditions. The scintillation patterns using S4 and σΦ and their respective proxies revealed similar diurnal and seasonal patterns of strong scintillation occurrence. The peaks of scintillation occurrence with mean values in the range 0.3 < (S4p , sDPR) ≤ 0.6 were observed during nighttime (17:00-22:00 UT) and in the months of March-April and September-October. The results also indicate that high level scintillations occur during geomagnetically disturbed (moderate and strong) and quiet conditions over the Ugandan region. The results show that SCINDA and IGS based scintillation patterns reveal the same nighttime and seasonal occurrence of irregularities over Uganda irrespective of the geomagnetic conditions. Therefore, the amplitude and phase scintillation proxies presented here can be used to fill gaps in low-latitude data where there are no data available from dedicated scintillation receivers, irrespective of the geomagnetic conditions.

  17. Development of new Polysiloxane Based Liquid Scintillators

    SciTech Connect

    Dalla Palma, M.; Quaranta, A.; Gramegna, F.; Marchi, T.; Cinausero, M.; Carturan, S.; Collazuol, G.; Checchia, C.; Degerlier, M.

    2015-07-01

    In the last decade, attention toward neutron detection has been growing in the scientific community, driven by new requirements in different fields of application ranging from homeland security to medical and material analysis, from research physics, to nuclear energy production. So far neutron detection, with particular attention to fast neutrons, has been mainly based on organic liquid scintillators, owing to their good efficiency and pulse shape discrimination (PSD) capability. Most of these liquids have however some main drawbacks given by toxicity, flammability, volatility and sensitivity to dissolved oxygen that limits the duration and the quality of their performances with worse handiness and increased costs. Phenyl-substituted polysiloxanes could address most of these issues, being characterized by low toxicity, low volatility and low flammability. Their optical properties can be tailored by changing the phenyl distribution and concentration thus allowing to increase the solubility of organic dyes, to modify the fluorescence spectra and to vary the refractive index of the medium. Furthermore, polysiloxanes have been recently exploited for the production of plastic scintillators with very good chemical and thermal stability and very good radiation hardness and the development of polysiloxane liquid scintillators could allow to combine these interesting properties with the supremacy of liquid scintillators as regarding PSD over plastics. For these reasons, the properties of several phenyl-substituted polysiloxane with different phenyl amounts and different viscosities have been investigated, with particular attention to the scintillation response and the pulse shape discrimination capability, and the results of the investigation are reported in this work. More in details, the scintillation light yield towards gamma rays ({sup 60}Co and {sup 137}Cs) of several polysiloxane liquids has been analyzed and compared with the light yield of a commercial non

  18. Robust GPS carrier tracking under ionospheric scintillation

    NASA Astrophysics Data System (ADS)

    Susi, M.; Andreotti, M.; Aquino, M. H.; Dodson, A.

    2013-12-01

    Small scale irregularities present in the ionosphere can induce fast and unpredictable fluctuations of Radio Frequency (RF) signal phase and amplitude. This phenomenon, known as scintillation, can degrade the performance of a GPS receiver leading to cycle slips, increasing the tracking error and also producing a complete loss of lock. In the most severe scenarios, if the tracking of multiple satellites links is prevented, outages in the GPS service can also occur. In order to render a GPS receiver more robust under scintillation, particular attention should be dedicated to the design of the carrier tracking stage, that is the receiver's part most sensitive to these types of phenomenon. This paper exploits the reconfigurability and flexibility of a GPS software receiver to develop a tracking algorithm that is more robust under ionospheric scintillation. For this purpose, first of all, the scintillation level is monitored in real time. Indeed the carrier phase and the post correlation terms obtained by the PLL (Phase Locked Loop) are used to estimate phi60 and S4 [1], the scintillation indices traditionally used to quantify the level of phase and amplitude scintillations, as well as p and T, the spectral parameters of the fluctuations PSD. The effectiveness of the scintillation parameter computation is confirmed by comparing the values obtained by the software receiver and the ones provided by a commercial scintillation monitoring, i.e. the Septentrio PolarxS receiver [2]. Then the above scintillation parameters and the signal carrier to noise density are exploited to tune the carrier tracking algorithm. In case of very weak signals the FLL (Frequency Locked Loop) scheme is selected in order to maintain the signal lock. Otherwise an adaptive bandwidth Phase Locked Loop (PLL) scheme is adopted. The optimum bandwidth for the specific scintillation scenario is evaluated in real time by exploiting the Conker formula [1] for the tracking jitter estimation. The performance

  19. Scintillating fiber detector performance, detector geometries, trigger, and electronics issues for scintillating fiber tracking

    SciTech Connect

    Baumbaugh, A.E.

    1994-06-01

    Scintillating Fiber tracking technology has made great advances and has demonstrated great potential for high speed charged particle tracking and triggering. The small detector sizes and fast scintillation fluors available make them very promising for use at high luminosity experiments at today`s and tomorrow`s colliding and fixed target experiments where high rate capability is essential. This talk will discuss the current state of Scintillating fiber performance and current Visual Light Photon Counter (VLPC) characteristics. The primary topic will be some of the system design and integration issues which should be considered by anyone attempting to design a scintillating fiber tracking system which includes a high speed tracking trigger. Design. constraints placed upon the detector system by the electronics and mechanical sub-systems will be discussed. Seemingly simple and unrelated decisions can have far reaching effects on overall system performance. SDC and DO example system designs will be discussed.

  20. The SNO+ Scintillator Purification Plant and Projected Sensitivity to Solar Neutrinos in the Pure Scintillator Phase

    NASA Astrophysics Data System (ADS)

    Pershing, Teal; SNO+ Collaboration

    2016-03-01

    The SNO+ detector is a neutrino and neutrinoless double-beta decay experiment utilizing the renovated SNO detector. In the second phase of operation, the SNO+ detector will contain 780 tons of organic liquid scintillator composed of 2 g/L 2,5-diphenyloxazole (PPO) in linear alkylbenzene (LAB). In this phase, SNO+ will strive to detect solar neutrinos in the sub-MeV range, including CNO production neutrinos and pp production neutrinos. To achieve the necessary detector sensitivity, a four-part scintillator purification plant has been constructed in SNOLAB for the removal of ionic and radioactive impurities. We present an overview of the SNO+ scintillator purification plant stages, including distillation, water extraction, gas stripping, and metal scavenger columns. We also give the projected SNO+ sensitivities to various solar-produced neutrinos based on the scintillator plant's projected purification efficiency.

  1. Characterizing Scintillation and Cherenkov Light in Water-Based Liquid Scintillators

    NASA Astrophysics Data System (ADS)

    Land, Benjamin; Caravaca, Javier; Descamps, Freija; Orebi Gann, Gabriel

    2016-09-01

    The recent development of Water-based Liquid Scintillator (WbLS) has made it possible to produce scintillating materials with highly tunable light yields and excellent optical clarity. This allows for a straightforward combination of the directional properties of Cherenkov light with the greater energy resolution afforded by the typically brighter scintillation light which lends itself well to a broad program of neutrino physics. Here we explore the light yields and time profiles of WbLS materials in development for Theia (formerly ASDC) as measured in CheSS: our bench-top Cherenkov and scintillation separation R&D project at Berkeley Lab. This work was supported by the Laboratory Directed Research and Development Program of Lawrence Berkeley National Laboratory under U.S. Department of Energy Contract No. DE-AC02-05CH11231.

  2. Development of scintillator plates with high energy resolution for alpha particles made of GPS scintillator grains

    NASA Astrophysics Data System (ADS)

    Shimaoka, Takehiro; Kaneko, Junichi H.; Izaki, Kenji; Tsubota, Youichi; Higuchi, Mikio; Nishiyama, Shusuke

    2014-01-01

    A scintillator plate with high energy resolution was developed to produce an alpha particle monitor used in nuclear fuel reprocessing plants and mixed plutonium-uranium oxide (MOX) fuel plants. Grains of a Gd2Si2O7 (GPS) scintillator of several 10 to 550 μm were fixed on a glass substrate and were then mechanically polished. By increasing the size of scintillator grains and removing fine powders, the collected light yield and energy resolution for alpha particles were drastically improved. Energy resolution of 9.3% was achieved using average grain size of 91 μm. Furthermore, the ratios between counts in a peak and total counts were improved by more than 60% by the further increase of grain size and adoption of mechanically polished surfaces on both sides. Beta and gamma ray influences were suppressed sufficiently by the thin 100 μm scintillator plates.

  3. Telmisartan, a possible PPAR-δ agonist, reduces TNF-α-stimulated VEGF-C production by inhibiting the p38MAPK/HSP27 pathway in human proximal renal tubular cells.

    PubMed

    Kimura, Hideki; Mikami, Daisuke; Kamiyama, Kazuko; Sugimoto, Hidehiro; Kasuno, Kenji; Takahashi, Naoki; Yoshida, Haruyoshi; Iwano, Masayuki

    2014-11-14

    Vascular endothelial growth factor-C (VEGF-C) is a main inducer of inflammation-associated lymphangiogenesis in various inflammatory disorders including chronic progressive kidney diseases, for which angiotensin II receptor type 1 blockers (ARBs) are widely used as the main treatment. Although proximal renal tubular cells may affect the formation of lymphatic vessels in the interstitial area by producing VEGF-C, the molecular mechanisms of VEGF-C production and its manipulation by ARB have not yet been examined in human proximal renal tubular epithelial cells (HPTECs). In the present study, TNF-α dose-dependently induced the production of VEGF-C in HPTECs. The TNF-α-induced production of VEGF-C was mediated by the phosphorylation of p38MAPK and HSP27, but not by that of ERK or NFkB. Telmisartan, an ARB that can activate the peroxisome proliferator-activated receptor (PPAR), served as a PPAR-δ activator and reduced the TNF-α-stimulated production of VEGF-C. This reduction was partially attributed to a PPAR-δ-dependent decrease in p38MAPK phosphorylation. Our results indicate that TNF-α induced the production of VEGF-C in HPTECs by activating p38MAPK/HSP27, and this was partially inhibited by telmisartan in a PPAR-δ dependent manner. These results provide a novel insight into inflammation-associated lymphangiogenesis.

  4. Liquid Scintillator Production for the NOvA Experiment

    DOE PAGES

    Mufson, S.; Baugh, B.; Bower, C.; ...

    2015-04-15

    The NOvA collaboration blended and delivered 8.8 kt (2.72M gal) of liquid scintillator as the active detector medium to its near and far detectors. The composition of this scintillator was specifically developed to satisfy NOvA's performance requirements. A rigorous set of quality control procedures was put in place to verify that the incoming components and the blended scintillator met these requirements. The scintillator was blended commercially in Hammond, IN. The scintillator was shipped to the NOvA detectors using dedicated stainless steel tanker trailers cleaned to food grade.

  5. Liquid scintillator production for the NOvA experiment

    NASA Astrophysics Data System (ADS)

    Mufson, S.; Baugh, B.; Bower, C.; Coan, T. E.; Cooper, J.; Corwin, L.; Karty, J. A.; Mason, P.; Messier, M. D.; Pla-Dalmau, A.; Proudfoot, M.

    2015-11-01

    The NOvA collaboration blended and delivered 8.8 kt (2.72M gal) of liquid scintillator as the active detector medium to its near and far detectors. The composition of this scintillator was specifically developed to satisfy NOvA's performance requirements. A rigorous set of quality control procedures was put in place to verify that the incoming components and the blended scintillator met these requirements. The scintillator was blended commercially in Hammond, IN. The scintillator was shipped to the NOvA detectors using dedicated stainless steel tanker trailers cleaned to food grade.

  6. Light propagation and fluorescence quantum yields in liquid scintillators

    NASA Astrophysics Data System (ADS)

    Buck, C.; Gramlich, B.; Wagner, S.

    2015-09-01

    For the simulation of the scintillation and Cherenkov light propagation in large liquid scintillator detectors a detailed knowledge about the absorption and emission spectra of the scintillator molecules is mandatory. Furthermore reemission probabilities and quantum yields of the scintillator components influence the light propagation inside the liquid. Absorption and emission properties are presented for liquid scintillators using 2,5-Diphenyloxazole (PPO) and 4-bis-(2-Methylstyryl)benzene (bis-MSB) as primary and secondary wavelength shifter. New measurements of the quantum yields for various aromatic molecules are shown.

  7. Furin gene (fur) regulation in differentiating human megakaryoblastic Dami cells: involvement of the proximal GATA recognition motif in the P1 promoter and impact on the maturation of furin substrates.

    PubMed

    Laprise, Marie-Hélène; Grondin, Francine; Cayer, Pauline; McDonald, Patrick P; Dubois, Claire M

    2002-11-15

    The convertase furin is involved in the maturation of key growth/aggregation mediators synthesized by the platelet producers, megakaryocytes, but the regulation of furin in these cells remains unknown. Computer-assisted search of the furin promoter sequence revealed multiple potential binding motifs for GATA-1, suggesting that furin is expressed and regulated in these cells. Using megakaryoblastic Dami cells, we observed that fur mRNA expression increased gradually on phorbol 12-myristate 13-acetate-induced differentiation, reaching maximum levels (8.3-fold increase) at 10 days. Transient transfections with P1, P1A, or P1B fur-LUC-promoter constructs revealed that in Dami cells, the P1 promoter is the strongest and the most sensitive to forced expression of GATA-1. Coexpression of GATA-1 and its comodulator, Friend of GATA-1 (FOG-1), resulted in a cooperative increase in P1 activity. Deletion analysis indicated that important GATA-1-regulated sequences are located in the most proximal region of the P1 promoter. Further analysis revealed 2 potential GATA-binding motifs at positions -66 and +62. Point mutation of each of the 2 motifs indicated that the intactness of the first GATA site is required for full basal and GATA-1-stimulated promoter activity. Finally, the inhibition of furin activity through gene transfer of the inhibitor alpha1-AT-PDX led to a block in maturation of the furin substrates transforming growth factor-beta1 and platelet-derived growth factor. Taken together, these results indicate that the most proximal GATA element in the P1 promoter is needed for fur gene expression in megakaryoblastic cells. They also suggest that proper regulation of the fur gene in megakaryocytes has an impact on the activation of furin substrates involved in megakaryocyte maturation and platelet functions.

  8. High resolution scintillation detector with semiconductor readout

    DOEpatents

    Levin, Craig S.; Hoffman, Edward J.

    2000-01-01

    A novel high resolution scintillation detector array for use in radiation imaging such as high resolution Positron Emission Tomography (PET) which comprises one or more parallelepiped crystals with at least one long surface of each crystal being in intimate contact with a semiconductor photodetector such that photons generated within each crystal by gamma radiation passing therethrough is detected by the photodetector paired therewith.

  9. Fluorescent compounds for plastic scintillation applications

    SciTech Connect

    Pla-Dalmau, A.; Bross, A.D.

    1994-04-01

    Several 2-(2{prime}-hydroxyphenyl)benzothiazole, -benzoxazole, and -benzimidazole derivatives have been prepared. Transmittance, fluorescence, light yield, and decay time characteristics of these compounds have been studied in a polystyrene matrix and evaluated for use in plastic scintillation detectors. Radiation damage studies utilizing a {sup 60}C source have also been performed.

  10. Scintillator Development for the PROSPECT Experiment

    NASA Astrophysics Data System (ADS)

    Yeh, Minfang

    2014-03-01

    Doped scintillator is the target material of choice for antineutrino detection as it utilizes the time-delayed coincidence signature of the positron annihilation and neutron capture resulting from the Inverse Beta Decay (IBD) interaction. Additionally, the multiple gamma rays or heavy ions emitted after neutron capture on either Gd or 6Li respectively provide a distinct signal for the identification of antineutrino events and therefore significantly enhance accidental background reduction. The choice of scintillator and dopant depends on the detector requirements and scintillator performance criteria. Both Gd and 6Li doped scintillators have been used in past reactor antineutrino experiments such as Double Chooz, Daya Bay, RENO, and Bugey3 and are currently under investigation by the PROSPECT collaboration. Their properties in terms of light yield, optical transparency, chemical stability and background rejection efficiency using Pulse Shape Discrimination (PSD) will be reported. Research sponsored by the U.S. Department of Energy, Office of Nuclear Physics and Office of High Energy Physics, under contract with Brookhaven National Laboratory-Brookhaven Science Associates.

  11. Progress in studying scintillator proportionality: Phenomenological model

    SciTech Connect

    Bizarri, Gregory; Cherepy, Nerine; Choong, Woon-Seng; Hull, Giulia; Moses, William; Payne, Sephen; Singh, Jai; Valentine, John; Vasilev, Andrey; Williams, Richard

    2009-04-30

    We present a model to describe the origin of non-proportional dependence of scintillator light yield on the energy of an ionizing particle. The non-proportionality is discussed in terms of energy relaxation channels and their linear and non-linear dependences on the deposited energy. In this approach, the scintillation response is described as a function of the deposited energy deposition and the kinetic rates of each relaxation channel. This mathematical framework allows both a qualitative interpretation and a quantitative fitting representation of scintillation non-proportionality response as function of kinetic rates. This method was successfully applied to thallium doped sodium iodide measured with SLYNCI, a new facility using the Compton coincidence technique. Finally, attention is given to the physical meaning of the dominant relaxation channels, and to the potential causes responsible for the scintillation non-proportionality. We find that thallium doped sodium iodide behaves as if non-proportionality is due to competition between radiative recombinations and non-radiative Auger processes.

  12. Optimization of Shielded Scintillator for Neutron Detection

    NASA Astrophysics Data System (ADS)

    Belancourt, Patrick; Morrison, John; Akli, Kramer; Freeman, Richard; High Energy Density Physics Team

    2011-10-01

    The High Energy Density Physics group is interested in the basic science of creating a neutron and gamma ray source. The neutrons and gamma rays are produced by accelerating ions via a laser into a target and creating fusion neutrons and gamma rays. A scintillator and photomultiplier tube will be used to detect these neutrons. Neutrons and photons produce ionizing radiation in the scintillator which then activates metastable states. These metastable states have both short and long decay rates. The initial photon count is orders of magnitude higher than the neutron count and poses problems for accurately detecting the neutrons due to the long decay state that is activated by the photons. The effects of adding lead shielding on the temporal response and signal level of the neutron detector will be studied in an effort to minimize the photon count without significant reduction to the temporal resolution of the detector. MCNP5 will be used to find the temporal response and energy deposition into the scintillator by adding lead shielding. Results from the simulations will be shown. Optimization of our scintillator neutron detection system is needed to resolve the neutron energies and neutron count of a novel neutron and gamma ray source.

  13. Equitorial Scintillations: Advances Since ISEA-6.

    DTIC Science & Technology

    1985-01-01

    thermospheric neutral wind have been postulated to describe the observe l longitudinal variation._--.-, A distinct class of equatorial irregularities...Unclassified SECURITY CLASSIFICATION OF THIS PAGE associated with frequency spread on ionograms . Scintillations caused by such irregularities exist only...another based on the influence of a transequatorial thermospheric neutral wind have been postu- lated to describe the observed longitudinal variation. A

  14. Proximal monomelic amyotrophy of the upper limb.

    PubMed

    Amir, D; Magora, A; Vatine, J J

    1987-07-01

    A 30-year-old patient of Central European origin, suffering from monomelic amyotrophy, is presented. The disease was characterized by proximal weakness of one upper limb, mainly of the shoulder girdle, accompanied by atrophy. The electrodiagnostic examination revealed signs of partial denervation in the presence of normal motor and sensory conduction. The disease, which is probably of the anterior horn cells, had a benign course and good prognosis, as evident from repeated examinations during a follow-up of eight years.

  15. Outward atmospheric scintillation effects and inward atmospheric scintillation effects comparisons for direct detection ladar applications

    NASA Astrophysics Data System (ADS)

    Youmans, Douglas G.

    2014-06-01

    Atmospheric turbulence produces intensity modulation or "scintillation" effects on both on the outward laser-mode path and on the return backscattered radiation path. These both degrade laser radar (ladar) target acquisition, ranging, imaging, and feature estimation. However, the finite sized objects create scintillation averaging on the outgoing path and the finite sized telescope apertures produce scintillation averaging on the return path. We expand on previous papers going to moderate to strong turbulence cases by starting from a 20kft altitude platform and propagating at 0° elevation (with respect to the local vertical) for 100km range to a 1 m diameter diffuse sphere. The outward scintillation and inward scintillation effects, as measured at the focal plane detector array of the receiving aperture, will be compared. To eliminate hard-body surface speckle effects in order to study scintillation, Goodman's M-parameter is set to 106 in the analytical equations and the non-coherent imaging algorithm is employed in Monte Carlo realizations. The analytical equations of the signal-to-noise ratio (SNRp), or mean squared signal over a variance, for a given focal plane array pixel window of interest will be summarized and compared to Monte Carlo realizations of a 1m diffuse sphere.

  16. Systematic study of particle quenching in organic scintillators

    NASA Astrophysics Data System (ADS)

    Santiago, L. M.; Bagán, H.; Tarancón, A.; Rauret, G.; Garcia, J. F.

    2013-01-01

    Among the different factors that affect measurements by organic scintillators, the majority of attention has been focused on those related to the scintillator (i.e., ionization, chemical, color and optical quenching), and less attention has been paid to the loss of energy before the particle (i.e., alpha or beta) arrives at the scintillator (i.e., particle quenching). This study evaluates the effect of particle quenching in different scintillation methods (i.e., using two plastic scintillation microspheres (PSm1 and PSm2), liquid scintillator and gel scintillator) by measuring solutions that contain increasing concentrations of NaCl, BaCl2 and glycerin. The results show the importance of particle quenching in PSm measurements because detection efficiency decreases with increasing concentrations of the quenching component, although the spectrum position and external standard parameter remain constant. The results have shown evidence of particle quenching, although at a lower magnitude, in the liquid scintillation or gel scintillation measurements. Moreover, the use of two PSm with different diameters and salty compound that alters the equilibrium of the liquid and gel emulsions also exemplified the importance of the transmission of optical photons through different scintillation media (i.e., optical quenching). Improvement and deterioration of the optical conditions on the scintillation media is manifested as a movement of the spectrum to higher and lower energies, respectively. The results obtained with PSm were confirmed by Monte Carlo simulation.

  17. Non-Carbon Dyes For Platic Scintillators- Report

    SciTech Connect

    Teprovich, J.; Colon-Mercado, H.; Gaillard, J.; Sexton, L.; Washington, A.; Ward, P.; Velten, J.

    2015-10-19

    Scintillation based detectors are desirable for many radiation detection applications (portal and border monitoring, safeguards verification, contamination detection and monitoring). The development of next generation scintillators will require improved detection sensitivity for weak gamma ray sources, and fast and thermal neutron quantification. Radiation detection of gamma and neutron sources can be accomplished with organic scintillators, however, the single crystals are difficult to grow for large area detectors and subject to cracking. Alternatives to single crystal organic scintillators are plastic scintillators (PS) which offer the ability to be shaped and scaled up to produce large sized detectors. PS is also more robust than the typical organic scintillator and are ideally suited for deployment in harsh real-world environments. PS contain a mixture of dyes to down-convert incident radiation into visible light that can be detected by a PMT. This project will evaluate the potential use of nano-carbon dyes in plastic scintillators.

  18. The statistical distribution of the number of counted scintillation photons in digital silicon photomultipliers: model and validation.

    PubMed

    van Dam, Herman T; Seifert, Stefan; Schaart, Dennis R

    2012-08-07

    In the design and application of scintillation detectors based on silicon photomultipliers (SiPMs), e.g. in positron emission tomography imaging, it is important to understand and quantify the non-proportionality of the SiPM response due to saturation, crosstalk and dark counts. A new type of SiPM, the so-called digital silicon photomultiplier (dSiPM), has recently been introduced. Here, we develop a model of the probability distribution of the number of fired microcells, i.e. the number of counted scintillation photons, in response to a given amount of energy deposited in a scintillator optically coupled to a dSiPM. Based on physical and functional principles, the model elucidates the statistical behavior of dSiPMs. The model takes into account the photon detection efficiency of the detector; the light yield, excess variance and time profile of the scintillator; and the crosstalk probability, dark count rate, integration time and the number of microcells of the dSiPM. Furthermore, relations for the expectation value and the variance of the number of fired cells are deduced. These relations are applied in the experimental validation of the model using a dSiPM coupled to a LSO:Ce,Ca scintillator. Finally, we propose an accurate method for the correction of energy spectra measured with dSiPM-based scintillation detectors.

  19. Design and fabrication of a microplatform for the proximity effect study of localized ELF-EMF on the growth of in vitro HeLa and PC-12 cells

    NASA Astrophysics Data System (ADS)

    Chen, Y. C.; Chen, C. C.; Tu, W.; Cheng, Y. T.; Tseng, F. G.

    2010-12-01

    This paper presents a platform technology with experimental results that show the scientists and biologists a way to rapidly investigate and analyze the biological effects of localized extremely low frequency (ELF) electromagnetic field (EMF) on living cells. The proximity effect of the localized ELF-EMF on living cells is revealed using the bio-compatible microplatform on which an on-glass inductive coil array, the source of the localized ELF-EMF in micro scale, is designed, fabricated and operated with a field strength of 1.2 ± 0.1 mT at 60 Hz for cell culturing study. After a 72 h ELF-EMF exposure, HeLa (human cervical cancer) and PC-12 (rat pheochromocytoma) cells exhibit about 18.4% and 12.9% cell proliferation rate reduction, respectively. Furthermore, according to the presented dynamic model, the reduction of the proliferation can be attributed to the interference of signal transduction processes due to the tangential currents induced around the cells.

  20. The role of orientation processing in the scintillating grid illusion.

    PubMed

    Qian, Kun; Kawabe, Takahiro; Yamada, Yuki; Miura, Kayo

    2012-07-01

    In the scintillating grid illusion, illusory dark spots are perceived on white patches at the intersections of gray bars. Previous studies have suggested that processing related to the orientation of the bars plays a role in this illusion, but the specific underlying mechanisms are unclear. In the present study, we investigated the role of orientation processing across the intersection in generating the scintillating grid illusion. The results revealed that the illusion was attenuated when the patch was located at the intersection of short bars (Experiment 1), irrespective of the spatial distance between patches (Experiment 2). The local cruciform patterns determined the strength of the illusion, even when lateral offset of the patches was employed (Experiment 3). The illusion was observed even when a small spatial gap was introduced around the patches. A larger gap produced a weaker illusion (Experiment 4). Spatial offsets of the bars across the gapped intersection greatly reduced the illusion (Experiment 5). We discuss these findings with regard to the activity of S1-type simple cells that respond to the luminance along an oriented edge across the intersection.

  1. Electrokinetic removal of radionuclides contained in scintillation liquids absorbed in soil type Phaeozem.

    PubMed

    Valdovinos, V; Monroy-Guzmán, F; Bustos, E

    2016-10-01

    Control samples of scintillation liquids - Phaeozem soil mixtures were prepared with different scintillation liquids as the support electrolyte, Install Gel(®) XF, (Ultima Gold AB™ and Ultima Gold XR™), to construct the polarization curves, and to select the cell potential with the highest mass transfer to remove (24)Na (15 h) and (99m)Tc (6 h) as radiotracers from polluted Phaeozem soil. During the electrokinetic treatment (EKT), the removal of radionuclides contained in scintillation liquids absorbed in Phaeozem soil, liquid phase was characterized by Gas Chromatography coupled with a Flame Ionization Detector (GC-FID) and Fourier Transform Infrared Spectrometry (FTIR), solids by FTIR, before and after the application of cell potential. In this sense, the support electrolyte was selected based on the highest current generated (1 mA), as in the case of scintillation liquid 50% Ultima Gold XR™ + 50% Water (1:1), which was used for 6 h in the presence of a mesh and a titanium rod, as anode and cathode, respectively. Finally, the removal percentage accumulated in the liquid phase after the EKT of Phaeozem soil polluted by (99m)Tc was 61% close to the anode after 4 h. It was also 61% for (24)Na close to cathode after 2 h, and after 4 h it was 71.8%.

  2. Bio-maleimide-stained plasma microparticles can be purified in a native state and target human proximal tubular HK2 cells

    PubMed Central

    Alkhatatbeh, Mohammad J.; Lincz, Lisa F.; Thorne, Rick F.

    2017-01-01

    Plasma microparticles (MPs) are heterogeneously sized submicron extracellular vesicles that originate from the cell membrane as a result of cell activation or apoptosis. Circulating MPs express cell-specific molecules that reflect their cell of origin and they are increasingly investigated for their potential role in intercellular communication. The aim of the current study was to determine if size exclusion chromatography could be used to purify fluorescent-labeled MPs in sufficient concentrations to be used experimentally in cell binding assays. Bio-maleimide was used to stain plasma MPs in platelet free plasma before applying to size exclusion chromatography. Collected fractions were analyzed for protein content and MPs were enumerated by flow cytometry. Fractions were ultracentrifuged and MPs further confirmed by western blotting for the putative diabetic marker, cluster of differentiation (CD)36 and platelet-specific CD41 proteins. Fractions that contained MPs were incubated with HK2 cells to determine MP-cell binding. Bio-maleimide-stained MPs were detected across various fractions of size exclusion, and pellets of these fractions confirmed positivity for the MP markers, CD41 and CD36. The addition of the isolated MPs to HK2 renal tubular cells and analysis by epi-fluorescent imaging demonstrated that, in principle, the labeled MPs are able to bind to cells in vitro. Notably, only the first eluted MP fraction bound HK2 cells indicating a possible association between MP size and cell-targeting properties. PMID:28123709

  3. Real-time Scintillation Monitoring in Alaska from a Longitudinal Chain of ASTRA's SM-211 GPS TEC and Scintillation Receivers

    NASA Astrophysics Data System (ADS)

    Crowley, G.; Azeem, S. I.; Reynolds, A.; Santana, J.; Hampton, D. L.

    2013-12-01

    Amplitude and phase scintillation can cause serious difficulties for GPS receivers. Intense scintillation can cause loss of lock. High latitude studies generally show that phase scintillation can be severe, but the amplitude scintillation tends to be small. The reason for this is not yet understood. Furthermore, the actual causes of the ionospheric irregularities that produce high latitude scintillation are not well understood. While the gradient drift instability is thought to be important in the F-region, there may be other structures present in either the E- or F-regions. The role of particle precipitation is also not well understood. Four of ASTRA's CASES GPS receivers were deployed in Alaska to demonstrate our ability to map scintillation in realtime, to provide space weather services to GPS users, and to initiate a detailed investigation of these effects. These dual-frequency GPS receivers measure total electron content (TEC) and scintillation. The scintillation monitors were deployed in a longitudinal chain at sites in Kaktovic, Fort Yukon, Poker Flat, and Gakona. Scintillation statistics show phase scintillations to be largest at Kaktovic and smallest at Gakona. We present GPS phase scintillation and auroral emission results from the Alaska chain to characterize the correspondence between scintillation and auroral features, and to investigate the role of high latitude auroral features in driving the phase scintillations. We will also present data showing how phase scintillation can cause other GPS receivers to lose lock. The data and results are particularly valuable because they illustrate some of the challenges of using GPS systems for positioning and navigation in an auroral region like Alaska. These challenges for snowplough drivers were recently highlighted, along with the CASES SM-211 space weather monitor, in a special video in which ASTRA and three other small businesses were presented with an entrepreneurial award from William Shatner (http://youtu.be/bIVKEQH_YPk).

  4. Hirayama Disease with Proximal Involvement

    PubMed Central

    2016-01-01

    Hirayama disease is a slowly progressing benign motor neuron disease that affects the distal upper limb. A 29-year-old man visited the hospital with a 1-year history of weakened left proximal upper limb. He was diagnosed with Hirayama disease 9 years ago, while there was no further progression of the muscle weakness afterward. Atrophy and weakness was detected in proximal upper limb muscles. Magnetic resonance imaging and somatosensory evoked potentials were normal. Needle electromyography showed abnormal findings in proximal upper limb muscles. Our patient had Hirayama disease involving the proximal portion through secondary progression. Clinical manifestation and accurate electromyography may be useful for diagnosis. Rare cases with progression patterns as described here are helpful and have clinical meaning for clinicians. PMID:27550499

  5. Tools for proximal soil sensing

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Proximal soil sensing (i.e. near-surface geophysical methods) are used to study soil phenomena across spatial scales. Geophysical methods exploit contrasts in physical properties (dielectric permittivity, apparent electrical conductivity or resistivity, magnetic susceptibility) to indirectly measur...

  6. Nonproportionality of Scintillator Detectors: Theory and Experiment

    SciTech Connect

    Payne, Stephen; Cherepy, Nerine; Hull, Giulia; Valentine, John; Moses, William; Choong, Woon-Seng

    2009-08-17

    On the basis of nonproportionality data obtained for several scintillators, we have developed a theory to describe the carrier dynamics to fit the light yield versus electron energy. The theory of Onsager was adapted to explain how the carriers form excitons or sequentially arrive at the activators to promote the ion to an excited state, and the theory of Birks was employed to allow for exciton-exciton annihilation. We then developed a second theory to deduce the degradation in resolution that results from nonproportionality by evoking Landau fluctuations, which are essentially variations in the deposited energy density that occur as the high energy electron travels along its trajectory. In general there is good agreement with the data, in terms of fitting the nonproportionality curves and reproducing the literature values of nonproportionality's contribution to the scintillator resolution. With the resurgence of interest in developing scintillator detectors that have good energy resolution, an improved understanding of nonproportionality has become a crucial matter since it presents the fundamental limit to the achievable resolution. In order to hasten an improved understanding of scintillator nonproportionality, we have constructed an instrument referred to as SLYNCI (Scintillator Light Yield Nonproportionality Compton Instrument). This is a second-generation instrument to the original device developed by Valentine and coworkers, wherein several new principles of operation have served to increase the data rate by an order of magnitude as discussed in detail in References. In the present article, the focus is on a theory to describe the measured electron response, which is the light yield as a function of the electron energy. To do this, we account for transport of carriers and excitons, in terms of how they transfer their energy to the activators with competition from nonradiative decay pathways. This work builds on the original work of Murray and coworkers, and

  7. Neutron spectroscopy with scintillation detectors using wavelets

    NASA Astrophysics Data System (ADS)

    Hartman, Jessica

    The purpose of this research was to study neutron spectroscopy using the EJ-299-33A plastic scintillator. This scintillator material provided a novel means of detection for fast neutrons, without the disadvantages of traditional liquid scintillation materials. EJ-299-33A provided a more durable option to these materials, making it less likely to be damaged during handling. Unlike liquid scintillators, this plastic scintillator was manufactured from a non-toxic material, making it safer to use, as well as easier to design detectors. The material was also manufactured with inherent pulse shape discrimination abilities, making it suitable for use in neutron detection. The neutron spectral unfolding technique was developed in two stages. Initial detector response function modeling was carried out through the use of the MCNPX Monte Carlo code. The response functions were developed for a monoenergetic neutron flux. Wavelets were then applied to smooth the response function. The spectral unfolding technique was applied through polynomial fitting and optimization techniques in MATLAB. Verification of the unfolding technique was carried out through the use of experimentally determined response functions. These were measured on the neutron source based on the Van de Graff accelerator at the University of Kentucky. This machine provided a range of monoenergetic neutron beams between 0.1 MeV and 24 MeV, making it possible to measure the set of response functions of the EJ-299-33A plastic scintillator detector to neutrons of specific energies. The response of a plutonium-beryllium (PuBe) source was measured using the source available at the University of Nevada, Las Vegas. The neutron spectrum reconstruction was carried out using the experimentally measured response functions. Experimental data was collected in the list mode of the waveform digitizer. Post processing of this data focused on the pulse shape discrimination analysis of the recorded response functions to remove the

  8. Dietary fat composition influences glomerular and proximal convoluted tubule cell structure and autophagic processes in kidneys from calorie-restricted mice.

    PubMed

    Calvo-Rubio, Miguel; Burón, M Isabel; López-Lluch, Guillermo; Navas, Plácido; de Cabo, Rafael; Ramsey, Jon J; Villalba, José M; González-Reyes, José A

    2016-06-01

    Calorie restriction (CR) has been repeatedly shown to prevent cancer, diabetes, hypertension, and other age-related diseases in a wide range of animals, including non-human primates and humans. In rodents, CR also increases lifespan and is a powerful tool for studying the aging process. Recently, it has been reported in mice that dietary fat plays an important role in determining lifespan extension with 40% CR. In these conditions, animals fed lard as dietary fat showed an increased longevity compared with mice fed soybean or fish oils. In this paper, we study the effect of these dietary fats on structural and physiological parameters of kidney from mice maintained on 40% CR for 6 and 18 months. Analyses were performed using quantitative electron microcopy techniques and protein expression in Western blots. CR mitigated most of the analyzed age-related parameters in kidney, such as glomerular basement membrane thickness, mitochondrial mass in convoluted proximal tubules and autophagic markers in renal homogenates. The lard group showed improved preservation of several renal structures with aging when compared to the other CR diet groups. These results indicate that dietary fat modulates renal structure and function in CR mice and plays an essential role in the determination of health span in rodents.

  9. Clinical Impact of Proximal Autosomal Imbalances

    PubMed Central

    Hamid, AB; Weise, A; Voigt, M; Bucksch, M; Kosyakova, N; Liehr, T; Klein, E

    2012-01-01

    Centromere-near gain of copy number can be induced by intra- or inter-chromosomal rearrangements or by the presence of a small supernumerary marker chromosome (sSMC). Interestingly, partial trisomy to hexasomy of euchromatic material may be present in clinically healthy or affected individuals, depending on origin and size of chromosomal material involved. Here we report the known minimal sizes of all centromere-near, i.e., proximal auto-somal regions in humans, which are tolerated; over 100 Mb of coding DNA are comprised in these regions. Additionally, we have summarized the typical symptoms for nine proximal autosomal regions including genes obviously sensitive to copy numbers. Overall, studying the carriers of specific chromosomal imbalances using genomics-based medicine, combined with single cell analysis can provide the genotype-phenotype correlations and can also give hints where copy-number-sensitive genes are located in the human genome. PMID:24052727

  10. Structural Basis of the CD8[alpha beta]/MHC Class I Interaction: Focused Recognition Orients CD8[beta] to a T Cell Proximal Position[superscript 1,2

    SciTech Connect

    Wang, Rui; Natarajan, Kannan; Margulies, David H.

    2009-09-18

    In the immune system, B cells, dendritic cells, NK cells, and T lymphocytes all respond to signals received via ligand binding to receptors and coreceptors. Although the specificity of T cell recognition is determined by the interaction of T cell receptors with MHC/peptide complexes, the development of T cells in the thymus and their sensitivity to Ag are also dependent on coreceptor molecules CD8 (for MHC class I (MHCI)) and CD4 (for MHCII). The CD8{alpha}{beta} heterodimer is a potent coreceptor for T cell activation, but efforts to understand its function fully have been hampered by ignorance of the structural details of its interactions with MHCI. In this study we describe the structure of CD8{alpha}{beta} in complex with the murine MHCI molecule H-2D{sup d} at 2.6 {angstrom} resolution. The focus of the CD8{alpha}{beta} interaction is the acidic loop (residues 222-228) of the {alpha}3 domain of H-2D{sup d}. The {beta} subunit occupies a T cell membrane proximal position, defining the relative positions of the CD8{alpha} and CD8{beta} subunits. Unlike the CD8{alpha}{alpha} homodimer, CD8{alpha}{beta} does not contact the MHCI {alpha}{sub 2}- or {beta}{sub 2}-microglobulin domains. Movements of the CD8{alpha} CDR2 and CD8{beta} CDR1 and CDR2 loops as well as the flexibility of the H-2D{sup d} CD loop facilitate the monovalent interaction. The structure resolves inconclusive data on the topology of the CD8{alpha}{beta}/MHCI interaction, indicates that CD8{beta} is crucial in orienting the CD8{alpha}{beta} heterodimer, provides a framework for understanding the mechanistic role of CD8{alpha}{beta} in lymphoid cell signaling, and offers a tangible context for design of structurally altered coreceptors for tumor and viral immunotherapy.

  11. Monitoring and Forecasting Ionospheric Scintillation at High Latitudes (Invited)

    NASA Astrophysics Data System (ADS)

    Prikryl, P.; Jayachandran, P. T.; Chadwick, R.; Kelly, T.

    2013-12-01

    Ionospheric scintillation (rapid signal amplitude fading and phase fluctuation) poses a threat to reliable and safe operation of modern technology that relies on Global Navigation Satellite Systems (GNSS). Ionospheric scintillation of GNSS signal severely degrades positional accuracy, causes cycle slips leading to loss of lock that affects performance of radio communication and navigation systems. At high latitudes, the scintillation is caused by ionospheric irregularities produced through coupling between solar wind plasma and the magnetosphere. Climatology of GPS scintillation at high latitudes in both hemispheres shows that phase scintillation occurs predominantly on the dayside in the cusp and in the nightside auroral oval. Solar wind disturbances, in particular the co-rotating interaction regions (CIR) on the leading edge of high-speed streams (HSS) and interplanetary coronal mass ejections (ICME), have been closely correlated with the occurrence of scintillation at high latitudes. These results demonstrated a technique of probabilistic forecast of high-latitude phase scintillation occurrence relative to arrival times of HSS and ICME. The Canadian High Arctic Ionospheric Network (CHAIN) has been monitoring GPS ionospheric scintillation and total electron content (TEC) since November 2007. One-minute amplitude and phase scintillation indices from L1 GPS signals and TEC from L1 and L2 GPS signals are computed from amplitude and phase data sampled at 50 Hz. Since 2012, significant expansion of CHAIN has begun with installation of new receivers, each capable of tracking up to 30 satellites including GLONASS and Galileo. The receivers log the raw phase and amplitude of the signal up to a 100-Hz rate for scintillation measurements. We briefly review observations of ionospheric scintillation and highlight new results from CHAIN, including the climatology of scintillation occurrence, collocation with aurora and HF radar backscatter, correlation with CIRs and ICMEs

  12. Divalent europium doped and un-doped calcium iodide scintillators: Scintillator characterization and single crystal growth

    DOE PAGES

    Boatner, L. A.; Ramey, J. O.; Kolopus, J. A.; ...

    2015-02-21

    Initially, the alkaline-earth scintillator, CaI2:Eu2+, was discovered around 1964 by Hofstadter, Odell, and Schmidt. Serious practical problems quickly arose, however, that were associated with the growth of large monolithic single crystals of this material due to its lamellar, mica-like structure. As a result of its theoretically higher light yield, CaI2:Eu2+ has the potential to exceed the excellent scintillation performance of SrI2:Eu2+. In fact, theoretical predictions for the light yield of CaI2:Eu2+ scintillators suggested that an energy resolution approaching 2% at 662 keV could be achievable. Like the early SrI2:Eu2+ scintillator, the performance of CaI2:Eu2+ scintillators has traditionally suffered due, atmore » least in part, to outdated materials synthesis, component stoichiometry/purity, and single-crystal-growth techniques. Based on our recent work on SrI2:Eu2+ scintillators in single-crystal form, we have developed new techniques that are applied here to CaI2:Eu2+ and pure CaI2 with the goal of growing large un-cracked crystals and, potentially, realizing the theoretically predicted performance of the CaI2:Eu2+ form of this material. Calcium iodide does not adhere to modern glassy carbon Bridgman crucibles - so there should be no differential thermal-contraction-induced crystal/crucible stresses on cooling that would result in crystal cracking of the lamellar structure of CaI2. Here we apply glassy carbon crucible Bridgman growth, high-purity growth-charge compounds, our molten salt processing/filtration technique, and extended vacuum-melt-pumping methods to the growth of both CaI2:Eu2+ and un-doped CaI2. Moreover, large scintillating single crystals were obtained, and detailed characterization studies of the scintillation properties of CaI2:Eu2+ and pure CaI2 single crystals are presented that include studies of the effects of plastic deformation of the crystals on the scintillator performance.« less

  13. Divalent europium doped and un-doped calcium iodide scintillators: Scintillator characterization and single crystal growth

    SciTech Connect

    Boatner, L. A.; Ramey, J. O.; Kolopus, J. A.; Neal, John S.

    2015-02-21

    Initially, the alkaline-earth scintillator, CaI2:Eu2+, was discovered around 1964 by Hofstadter, Odell, and Schmidt. Serious practical problems quickly arose, however, that were associated with the growth of large monolithic single crystals of this material due to its lamellar, mica-like structure. As a result of its theoretically higher light yield, CaI2:Eu2+ has the potential to exceed the excellent scintillation performance of SrI2:Eu2+. In fact, theoretical predictions for the light yield of CaI2:Eu2+ scintillators suggested that an energy resolution approaching 2% at 662 keV could be achievable. Like the early SrI2:Eu2+ scintillator, the performance of CaI2:Eu2+ scintillators has traditionally suffered due, at least in part, to outdated materials synthesis, component stoichiometry/purity, and single-crystal-growth techniques. Based on our recent work on SrI2:Eu2+ scintillators in single-crystal form, we have developed new techniques that are applied here to CaI2:Eu2+ and pure CaI2 with the goal of growing large un-cracked crystals and, potentially, realizing the theoretically predicted performance of the CaI2:Eu2+ form of this material. Calcium iodide does not adhere to modern glassy carbon Bridgman crucibles - so there should be no differential thermal-contraction-induced crystal/crucible stresses on cooling that would result in crystal cracking of the lamellar structure of CaI2. Here we apply glassy carbon crucible Bridgman growth, high-purity growth-charge compounds, our molten salt processing/filtration technique, and extended vacuum-melt-pumping methods to the growth of both CaI2:Eu2+ and un-doped CaI2. Moreover, large scintillating single crystals were obtained, and detailed characterization studies of the

  14. Simulation of scintillation light output in LYSO scintillators through a full factorial design

    NASA Astrophysics Data System (ADS)

    Loignon-Houle, Francis; Bergeron, Mélanie; Pepin, Catherine M.; Charlebois, Serge A.; Lecomte, Roger

    2017-01-01

    Individually coupled scintillation detectors used in positron emission tomography (PET) imaging suffer from important signal losses due to the suboptimal light collection from crystals. As only a fraction of the light is generally extracted from long and thin scintillators, it is important to identify and understand the predominant causes of signal loss in order to eventually recover it. This simulation study investigates the multiple factors affecting the light transport in high-aspect ratio LYSO scintillators wrapped in specular reflectors through a full factorial design. By exploring various combinations of crystal geometry, readout conditions and wrapping conditions, it was found that an optimum light output can only be achieved through a careful selection of highly reflective material along with high-transmittance optical adhesive used to bond the reflector. Decreasing the adhesive thickness was also found to have a positive outcome in most explored configurations, however to a much lesser extent. Suboptimal reflectivity and adhesive transmittance also lead to an asymmetric light output distribution dependent on the depth of interaction of the radiation, potentially degrading energy resolution. By identifying the factors causing the most significant scintillation light losses through a factorial design, the most promising detector configurations have been identified in the quest for optimal light collection from scintillators.

  15. Codoped direct-gap semiconductor scintillators

    DOEpatents

    Derenzo, Stephen E.; Bourret-Courchesne, Edith; Weber, Marvin J.; Klintenberg, Mattias K.

    2006-05-23

    Fast, bright inorganic scintillators at room temperature are based on radiative electron-hole recombination in direct-gap semiconductors, e.g. CdS and ZnO. The direct-gap semiconductor is codoped with two different impurity atoms to convert the semiconductor to a fast, high luminosity scintillator. The codopant scheme is based on dopant band to dopant trap recombination. One dopant provides a significant concentration of carriers of one type (electrons or holes) and the other dopant traps carriers of the other type. Examples include CdS:In,Te; CdS:In,Ag; CdS:In,Na; ZnO:Ga,P; ZnO:Ga,N; ZnO:Ga,S; and GaN:Ge,Mg.

  16. Plastic scintillator detector for pulsed flux measurements

    NASA Astrophysics Data System (ADS)

    Kadilin, V. V.; Kaplun, A. A.; Taraskin, A. A.

    2017-01-01

    A neutron detector, providing charged particle detection capability, has been designed. The main purpose of the detector is to measure pulsed fluxes of both charged particles and neutrons during scientific experiments. The detector consists of commonly used neutron-sensitive ZnS(Ag) / 6LiF scintillator screens wrapping a layer of polystyrene based scintillator (BC-454, EJ-254 or equivalent boron loaded plastic). This type of detector design is able to log a spatial distribution of events and may be scaled to any size. Different variations of the design were considered and modelled in specialized toolkits. The article presents a review of the detector design features as well as simulation results.

  17. Transparent Ceramic Scintillator Fabrication, Properties and Applications

    SciTech Connect

    Cherepy, N J; Kuntz, J D; Roberts, J J; Hurst, T A; Drury, O B; Sanner, R D; Tillotson, T M; Payne, S A

    2008-08-24

    Transparent ceramics offer an alternative to single crystals for scintillator applications such as gamma ray spectroscopy and radiography. We have developed a versatile, scaleable fabrication method, using Flame Spray Pyrolysis (FSP) to produce feedstock which is readily converted into phase-pure transparent ceramics. We measure integral light yields in excess of 80,000 Ph/MeV with Cerium-doped Garnets, and excellent optical quality. Avalanche photodiode readout of Garnets provides resolution near 6%. For radiography applications, Lutetium Oxide offers a high performance metric and is formable by ceramics processing. Scatter in transparent ceramics due to secondary phases is the principal limitation to optical quality, and afterglow issues that affect the scintillation performance are presently being addressed.

  18. Scintillating 99Tc Selective Ion Exchange Resins

    SciTech Connect

    Mitchell Greenhalgh; Richard D. Tillotson

    2012-07-01

    Scintillating technetium (99Tc) selective ion exchange resins have been developed and evaluated for equilibrium capacities and detection efficiencies. These resins can be utilized for the in-situ concentration and detection of low levels of pertechnetate anions (99TcO4-) in natural waters. Three different polystyrene type resin support materials were impregnated with varying amounts of tricaprylmethylammonium chloride (Aliquat 336) extractant, several different scintillating fluors and wavelength shifters. The prepared resins were contacted batch-wise to equilibrium over a wide range of 99TcO4- concentrations in natural water. The measured capacities were used to develop Langmuir adsorption isotherms for each resin. 99Tc detection efficiencies were determined and up to 71.4 ± 2.6% was achieved with some resins. The results demonstrate that a low level detection limit for 99TcO4- in natural waters can be realized.

  19. Reflectance of polytetrafluoroethylene for xenon scintillation light

    SciTech Connect

    Silva, C.; Pinto da Cunha, J.; Pereira, A.; Chepel, V.; Lopes, M. I.; Solovov, V.; Neves, F.

    2010-03-15

    Gaseous and liquid xenon particle detectors are being used in a number of applications including dark matter search and neutrino-less double beta decay experiments. Polytetrafluoroethylene (PTFE) is often used in these detectors both as electrical insulator and as a light reflector to improve the efficiency of detection of scintillation photons. However, xenon emits in the vacuum ultraviolet (VUV) wavelength region ({lambda}{approx_equal}175 nm) where the reflecting properties of PTFE are not sufficiently known. In this work, we report on measurements of PTFE reflectance, including its angular distribution, for the xenon scintillation light. Various samples of PTFE, manufactured by different processes (extruded, expanded, skived, and pressed) have been studied. The data were interpreted with a physical model comprising both specular and diffuse reflections. The reflectance obtained for these samples ranges from about 47% to 66% for VUV light. Other fluoropolymers, namely, ethylene tetrafluoroethylene (ETFE), fluorinated ethylene propylene (FEP), and perfluoro-alkoxyalkane (PFA) were also measured.

  20. Plastic fiber scintillator response to fast neutrons

    SciTech Connect

    Danly, C. R.; Sjue, S.; Wilde, C. H.; Merrill, F. E.; Haight, R. C.

    2014-11-15

    The Neutron Imaging System at NIF uses an array of plastic scintillator fibers in conjunction with a time-gated imaging system to form an image of the neutron emission from the imploded capsule. By gating on neutrons that have scattered from the 14.1 MeV DT energy to lower energy ranges, an image of the dense, cold fuel around the hotspot is also obtained. An unmoderated spallation neutron beamline at the Weapons Neutron Research facility at Los Alamos was used in conjunction with a time-gated imaging system to measure the yield of a scintillating fiber array over several energy bands ranging from 1 to 15 MeV. The results and comparison to simulation are presented.

  1. Neutron detection with single crystal organic scintillators

    SciTech Connect

    Zaitseva, N; Newby, J; Hamel, S; Carman, L; Faust, M; Lordi, V; Cherepy, N; Stoeffl, W; Payne, S

    2009-07-15

    Detection of high-energy neutrons in the presence of gamma radiation background utilizes pulse-shape discrimination (PSD) phenomena in organics studied previously only with limited number of materials, mostly liquid scintillators and single crystal stilbene. The current paper presents the results obtained with broader varieties of luminescent organic single crystals. The studies involve experimental tools of crystal growth and material characterization in combination with the advanced computer modeling, with the final goal of better understanding the relevance between the nature of the organic materials and their PSD properties. Special consideration is given to the factors that may diminish or even completely obscure the PSD properties in scintillating crystals. Among such factors are molecular and crystallographic structures that determine exchange coupling and exciton mobility in organic materials and the impurity effect discussed on the examples of trans-stilbene, bibenzyl, 9,10-diphenylanthracene and diphenylacetylene.

  2. Codoped direct-gap semiconductor scintillators

    DOEpatents

    Derenzo, Stephen Edward; Bourret-Courchesne, Edith; Weber, Marvin J.; Klintenberg, Mattias K.

    2008-07-29

    Fast, bright inorganic scintillators at room temperature are based on radiative electron-hole recombination in direct-gap semiconductors, e.g. CdS and ZnO. The direct-gap semiconductor is codoped with two different impurity atoms to convert the semiconductor to a fast, high luminosity scintillator. The codopant scheme is based on dopant band to dopant trap recombination. One dopant provides a significant concentration of carriers of one type (electrons or holes) and the other dopant traps carriers of the other type. Examples include CdS:In,Te; CdS:In,Ag; CdS:In,Na; ZnO:Ga,P; ZnO:Ga,N; ZnO:Ga,S; and GaN:Ge,Mg.

  3. Development of High-Resolution Scintillator Systems

    SciTech Connect

    Larry A. Franks; Warnick J. Kernan

    2007-09-01

    Mercuric iodide (HgI2) is a well known material for the direct detection of gamma-rays; however, the largest volume achievable is limited by the thickness of the detector which needs to be a small fraction of the average trapping length for electrons. We report results of using HgI2 crystals to fabricate photocells used in the readout of scintillators. The optical spectral response and efficiency of these photocells were measured and will be reported. Nuclear response from an HgI2 photocell that was optically matched to a cerium-activated scintillator is presented and discussed. Further improvements can be expected by optimizing the transparent contact technology.

  4. Cell confluency-induced Stat3 activation regulates NHE3 expression by recruiting Sp1 and Sp3 to the proximal NHE3 promoter region during epithelial dome formation.

    PubMed

    Su, Hsiao-Wen; Wang, Shainn-Wei; Ghishan, Fayez K; Kiela, Pawel R; Tang, Ming-Jer

    2009-01-01

    Activation of signal transducer and activator of transcription-3 (Stat3) during cell confluency is related to its regulatory roles in cell growth arrest- or survival-related physiological or developmental processes. We previously demonstrated that this signaling event triggers epithelial dome formation by transcriptional augmentation of sodium hydrogen exchanger-3 (NHE3) expression. However, the detailed molecular mechanism remained unclear. By using serial deletions, site-directed mutagenesis, and EMSA analysis, we now demonstrate Stat3 binding to an atypical Stat3-response element in the rat proximal NHE3 promoter, located adjacent to a cluster of Sp cis-elements (SpA/B/C), within -77/-36 nt of the gene. SpB (-58/-55 nt) site was more effective than SpA (-72/-69 nt) site for cooperative binding of Sp1/Sp3. Increasing cell density had no effect on Sp1/Sp3 expression but resulted in their increased binding to the SpA/B/C probe along with Stat3 and concurrently with enhanced nuclear pTyr705-Stat3 level. Immunoprecipitation performed with the nuclear extracts demonstrated physical interaction of Stat3 and Sp1/Sp3 triggered by cell confluency. Stat3 inhibition by overexpression of dominant-negative Stat3-D mutant in MDCK cells or by small interfering RNA-mediated knockdown in Caco-2 cells resulted in inhibition of the cell density-induced NHE3 expression, Sp1/Sp3 binding, and NHE3 promoter activity and in decreased dome formation. Thus, during confluency, ligand-independent Stat3 activation leads to its interaction with Sp1/Sp3, their recruitment to the SpA/B/C cluster in a Stat3 DNA-binding domain-dependent fashion, increased transcription, and expression of NHE3, to coordinate cell density-mediated epithelial dome formation.

  5. Nanophosphor composite scintillators comprising a polymer matrix

    DOEpatents

    Muenchausen, Ross Edward; Mckigney, Edward Allen; Gilbertson, Robert David

    2010-11-16

    An improved nanophosphor composite comprises surface modified nanophosphor particles in a solid matrix. The nanophosphor particle surface is modified with an organic ligand, or by covalently bonding a polymeric or polymeric precursor material. The surface modified nanophosphor particle is essentially charge neutral, thereby preventing agglomeration of the nanophosphor particles during formation of the composite material. The improved nanophosphor composite may be used in any conventional scintillator application, including in a radiation detector.

  6. Sorohalide scintillators, phosphors, and uses thereof

    SciTech Connect

    Yang, Pin; Deng, Haoran; Doty, F. Patrick; Zhou, Xiaowang

    2016-05-10

    The present invention relates to sorohalide compounds having formula A.sub.3B.sub.2X.sub.9, where A is an alkali metal, B is a rare earth metal, and X is a halogen. Optionally, the sorohalide includes a dopant D. Such undoped and doped sorohalides are useful as scintillation materials or phosphors for any number of uses, including for radiation detectors, solid-state light sources, gamma-ray spectroscopy, medical imaging, and drilling applications.

  7. Characteristics of Yerevan High Transparency Scintillators

    SciTech Connect

    Zorn, Carl; Asryan, Gegham; Egiyan, Kim; Tarverdyan, M.; Amaryan, Moscov; Amaryan, Moskov; Demirchyan, Raphael; Stepanyan, Stepan; Burkert, Volker; Sharabian, Youri

    1992-08-01

    Optical transmission, light output and time characteristics are given for long scintillator strips fabricated at the Yerevan Physics Institute using the extrusion method. It is shown that at 45% relative (to anthracene) light output, good transmission (2.5/2.9 m attenuation length with photomultiplier direct readout and 3/3.5 m attenuation length fiber readout) and time characteristics (average decay time 2.8 nsec) were obtained.

  8. Simulating Silicon Photomultiplier Response to Scintillation Light.

    PubMed

    Jha, Abhinav K; van Dam, Herman T; Kupinski, Matthew A; Clarkson, Eric

    2013-02-01

    The response of a Silicon Photomultiplier (SiPM) to optical signals is affected by many factors including photon-detection efficiency, recovery time, gain, optical crosstalk, afterpulsing, dark count, and detector dead time. Many of these parameters vary with overvoltage and temperature. When used to detect scintillation light, there is a complicated non-linear relationship between the incident light and the response of the SiPM. In this paper, we propose a combined discrete-time discrete-event Monte Carlo (MC) model to simulate SiPM response to scintillation light pulses. Our MC model accounts for all relevant aspects of the SiPM response, some of which were not accounted for in the previous models. We also derive and validate analytic expressions for the single-photoelectron response of the SiPM and the voltage drop across the quenching resistance in the SiPM microcell. These analytic expressions consider the effect of all the circuit elements in the SiPM and accurately simulate the time-variation in overvoltage across the microcells of the SiPM. Consequently, our MC model is able to incorporate the variation of the different SiPM parameters with varying overvoltage. The MC model is compared with measurements on SiPM-based scintillation detectors and with some cases for which the response is known a priori. The model is also used to study the variation in SiPM behavior with SiPM-circuit parameter variations and to predict the response of a SiPM-based detector to various scintillators.

  9. a Subminiature Scintillation Detector for Catheter Operation

    NASA Astrophysics Data System (ADS)

    Scafè, R.; Montani, L.; Burgio, N.; Iurlaro, G.; Santagata, A.; Ciavola, C.; Alonge, G.

    2006-04-01

    The feasibility of a subminiature scintillation detector to be inserted in a catheter for lesion localization in nuclear medicine SPECT has been studied. Measurements on a simple laboratory setup have been performed and compared with Monte Carlo results. Further simulations, at 30keV and 140keV, concerning a configuration reproducing severe clinical conditions have shown poor lesion detectability. Several factors affecting the response have to be investigated to improve the capability of lesion localization characterizing such detector.

  10. Improved Neutron Scintillators Based on Nanomaterials

    SciTech Connect

    Dennis Friesel, PhD

    2008-06-30

    The development work conducted in this SBIR has so far not supported the premise that using nano-particles in LiFZnS:Ag foils improves their transparency to 420 (or other frequency) light. This conclusion is based solely on the light absorption properties of LiFZnS foils fabricated from nano- and from micro-particles. Furthermore, even for the case of the Gd{sub 2}O{sub 3} foils, the transmission of 420 nm light gained by using nano-particles all but disappears as the foil thickness is increased beyond about 0.2 mm, a practical scintillator thickness. This was not immediately apparent from the preliminary study since no foils thicker than about 0.04 mm were produced. Initially it was believed that the failure to see an improvement by using nano-particles for the LiFZnS foils was caused by the clumping of the particles in Toluene due to the polarity of the ZnS particles. However, we found, much to our surprise, that nano-particle ZnS alone in polystyrene, and in Epoxy, had worse light transmission properties than the micro-particle foils for equivalent thickness and density foils. The neutron detection measurements, while disappointing, are attributable to our inability to procure or fabricate Bulk Doped ZnS nanoparticles. The cause for the failure of nano-particles to improve the scintillation light, and hence improved neutron detection efficiency, is a fundamental one of light scattering within the scintillator. A consequence of PartTec's documentation of this is that several concepts for the fabrication of improved {sup 6}LiFZnS scintillators were formulated that will be the subject of a future SBIR submission.

  11. Studies of NICADD Extruded Scintillator Strips

    SciTech Connect

    Dychkant, Alexandre; et al.

    2005-03-01

    About four hundred one meter long, 10 cm wide and 5 mm thick extruded scintillating strips were measured at four different points. The results of measurements strip responses to a radioactive source {sup 90}Sr are provided, and details of strip choice, preparation, and method of measurement are included. This work was essential for prototyping a tail catcher and muon tracker for a future international electron positron linear collider detector.

  12. Boron-Loaded Silicone Rubber Scintillators

    SciTech Connect

    Bell, Z.W.; Maya, L.; Brown, G.M.; Sloop, F.V.Jr

    2003-05-12

    Silicone rubber received attention as an alternative to polyvinyltoluene in applications in which the scintillator is exposed to high doses because of the increased resistance of the rubber to the formation of blue-absorbing color centers. Work by Bowen, et al., and Harmon, et al., demonstrated their properties under gamma/x-ray irradiation, and Bell, et al. have shown their response to thermal neutrons. This last work, however, provided an example of a silicone in which both the boron and the scintillator were contained in the rubber as solutes, a formulation which led to the precipitation of solids and sublimation of the boron component. In the present work we describe a scintillator in which the boron is chemically bonded to the siloxane and so avoids the problem of precipitation and loss of boron to sublimation. Material containing up to 18% boron, by weight, was prepared, mounted on photomultipliers, and exposed to both neutron and gamma fluxes. Pulse height spectra showing the neutron and photon response were obtained, and although the light output was found to be much poorer than from samples in which boron was dissolved, the higher boron concentrations enabled essentially 100% neutron absorption in only a few millimeters' thickness of rubber.

  13. Cerium fluoride, a new fast, heavy scintillator

    SciTech Connect

    Moses, W.W.; Derenzo, S.E.

    1988-11-01

    We describe the scintillation properties of Cerium Fluoride (CeF/sub 3/), a newly discovered, heavy (6.16 g/cm/sup 3/), inorganic scintillator. Its fluorescence decay lifetime, measured with the delayed coincidence method, is described by a single exponential with a 27 /+-/ ns time constant. The emission spectrum peaks at a wavelength of 340 nm, and drops to less than 10% of its peak value at 315 nm and 460 nm. When a 1 cm optical quality cube of CeF/sub 3/ is excited with 511 keV photons, a photopeak with a 20% full width at half maximum is observed at approximately half the light output of a Bismuth Germanate (BGO) crystal with similar geometry. We also present measurements of the decay time and light output of CeF/sub 3/ doped with three rare-earth elements (Dy, Er, and Pr). The short fluorescence lifetime, high density, and reasonable light output of this new scintillator suggest that it would be useful for applications where high counting rates, good stopping power, and nanosecond timing are important, such as medical imaging and nuclear science. 5 refs., 6 figs., 1 tab.

  14. Detecting dark matter with scintillating bubble chambers

    NASA Astrophysics Data System (ADS)

    Zhang, Jianjie; Dahl, C. Eric; Jin, Miaotianzi; Baxter, Daniel

    2016-03-01

    Threshold based direct WIMP dark matter detectors such as the superheated bubble chambers developed by the PICO experiment have demonstrated excellent electron-recoil and alpha discrimination, excellent scalability, ease of change of target fluid, and low cost. However, the nuclear-recoil like backgrounds have been a limiting factor in their dark matter sensitivity. We present a new type of detector, the scintillating bubble chamber, which reads out the scintillation pulse of the scattering events as well as the pressure, temperature, acoustic traces, and bubble images as a conventional bubble chamber does. The event energy provides additional handle to discriminate against the nuclear-recoil like backgrounds. Liquid xenon is chosen as the target fluid in our prototyping detector for its high scintillation yield and suitable vapor pressure which simplifies detector complexity. The detector can be used as an R&D tool to study the backgrounds present in the current PICO bubble chambers or as a prototype for standalone dark matter detectors in the future. Supported by DOE Grant DE-SC0012161.

  15. Chloride, bromide and iodide scintillators with europium

    DOEpatents

    Zhuravleva, Mariya; Yang, Kan

    2016-09-27

    A halide scintillator material is disclosed where the halide may comprise chloride, bromide or iodide. The material is single-crystalline and has a composition of the general formula ABX.sub.3 where A is an alkali, B is an alkali earth and X is a halide which general composition was investigated. In particular, crystals of the formula ACa.sub.1-yEu.sub.yI.sub.3 where A=K, Rb and Cs were formed as well as crystals of the formula CsA.sub.1-yEu.sub.yX.sub.3 (where A=Ca, Sr, Ba, or a combination thereof and X=Cl, Br or I or a combination thereof) with divalent Europium doping where 0.ltoreq.y.ltoreq.1, and more particularly Eu doping has been studied at one to ten mol %. The disclosed scintillator materials are suitable for making scintillation detectors used in applications such as medical imaging and homeland security.

  16. Pulse height model for deuterated scintillation detectors

    NASA Astrophysics Data System (ADS)

    Wang, Haitang; Enqvist, Andreas

    2015-12-01

    An analytical model of light pulse height distribution for finite deuterated scintillation detectors is created using the impulse approximation. Particularly, the energy distribution of a scattered neutron is calculated based on an existing collision probability scheme for general cylindrical shaped detectors considering double differential cross-sections. The light pulse height distribution is analytically and numerically calculated by convoluting collision sequences with the light output function for an EJ-315 detector from our measurements completed at Ohio University. The model provides a good description of collision histories capturing transferred neutron energy in deuterium-based scintillation materials. The resulting light pulse height distribution details pulse compositions and their corresponding contributions. It shows that probabilities of neutron collision with carbon and deuterium nuclei are comparable, however the light pulse amplitude due to collisions with carbon nuclei is small and mainly located at the lower region of the light pulse distribution axis. The model can explore those neutron interaction events that generate pulses near or below a threshold that would be imposed in measurements. A comparison is made between the light pulse height distributions given by the analytical model and measurements. It reveals a significant probability of a neutron generating a small light pulse due to collisions with carbon nuclei when compared to larger light pulse generated by collisions involving deuterium nuclei. This model is beneficial to understand responses of scintillation materials and pulse compositions, as well as nuclei information extraction from recorded pulses.

  17. A large Scintillating Fibre Tracker for LHCb

    NASA Astrophysics Data System (ADS)

    Greim, R.

    2017-02-01

    The LHCb experiment will be upgraded during LHC Long Shutdown 2 to be able to record data at a higher instantaneous luminosity. The readout rate is currently limited to 1 MHz by the Level 1 trigger. In order to achieve the target integrated luminosity of 50 fb‑1 during LHC Run 3, all subdetectors have to be read out by a 40 MHz trigger-less readout system. Especially, the current tracking detectors downstream of the LHCb dipole magnet suffer from large detector dead times and a small granularity in the Outer Tracker, which consists of proportional straw tubes. Therefore, the Downstream Tracker will be replaced by a Scintillating Fibre Tracker with Silicon Photomultiplier readout. The total sensitive area of 340 m2 is made up of 2.5 m long fibre mats consisting of six staggered layers of 250 μm thin scintillating fibres. The scintillation light created by the charged particles traversing the fibre mats is transported to the fibre ends via total internal reflection and detected by state-of-the-art multi-channel SiPM arrays. This paper presents the detector concept, design, challenges, custom-made readout chips, as well as laboratory and beam test results.

  18. New Scintillating Crystals for PET Scanners

    NASA Astrophysics Data System (ADS)

    Lecoq, Paul

    2002-01-01

    Systematic R&D on basic mechanism in inorganic scintillators, initiated by the Crystal Clear Collaboration at CERN 10 years ago, has contributed not to a small amount, to the development of new materials for a new generation of medical imaging devices with increased resolution and sensitivity. The first important requirement for a scintillator to be used in medical imaging devices is the stopping power for the given energy range of X and γ rays to be considered, and more precisely the conversion efficiency. A high light yield is also mandatory to improve the energy resolution, which is essentially limited by the photostatistics and the electronic noise at these energies. A short scintillation decay time allows to reduce the dead time and therefore to increase the limiting counting rate. When all these requirements are fulfilled the sensitivity and image contrast are increased for a given patient dose, or the dose can be reduced. Examples of new materials under development by the Crystal Clear Collaboration will be given with an emphasis on the major breakthrough they can bring in medical imaging, as compared to present equipments.

  19. Telmisartan, a possible PPAR-δ agonist, reduces TNF-α-stimulated VEGF-C production by inhibiting the p38MAPK/HSP27 pathway in human proximal renal tubular cells

    SciTech Connect

    Kimura, Hideki; Mikami, Daisuke; Kamiyama, Kazuko; Sugimoto, Hidehiro; Kasuno, Kenji; Takahashi, Naoki; Yoshida, Haruyoshi; Iwano, Masayuki

    2014-11-14

    Highlights: • TNF-α increased VEGF-C expression by enhancing phosphorylation of p38MAPK and HSP27. • Telmisartan decreased TNF-α-stimulated expression of VEGF-C. • Telmisartan suppressed TNF-α-induced phosphorylation of p38MAPK and HSP27. • Telmisartan activated endogenous PPAR-δ protein. • Telmisartan suppressed p38MAPK phosphorylation in a PPAR-δ-dependent manner. - Abstract: Vascular endothelial growth factor-C (VEGF-C) is a main inducer of inflammation-associated lymphangiogenesis in various inflammatory disorders including chronic progressive kidney diseases, for which angiotensin II receptor type 1 blockers (ARBs) are widely used as the main treatment. Although proximal renal tubular cells may affect the formation of lymphatic vessels in the interstitial area by producing VEGF-C, the molecular mechanisms of VEGF-C production and its manipulation by ARB have not yet been examined in human proximal renal tubular epithelial cells (HPTECs). In the present study, TNF-α dose-dependently induced the production of VEGF-C in HPTECs. The TNF-α-induced production of VEGF-C was mediated by the phosphorylation of p38MAPK and HSP27, but not by that of ERK or NFkB. Telmisartan, an ARB that can activate the peroxisome proliferator-activated receptor (PPAR), served as a PPAR-δ activator and reduced the TNF-α-stimulated production of VEGF-C. This reduction was partially attributed to a PPAR-δ-dependent decrease in p38MAPK phosphorylation. Our results indicate that TNF-α induced the production of VEGF-C in HPTECs by activating p38MAPK/HSP27, and this was partially inhibited by telmisartan in a PPAR-δ dependent manner. These results provide a novel insight into inflammation-associated lymphangiogenesis.

  20. Metal-loaded organic scintillators for neutrino physics

    NASA Astrophysics Data System (ADS)

    Buck, Christian; Yeh, Minfang

    2016-09-01

    Organic liquid scintillators are used in many neutrino physics experiments of the past and present. In particular for low energy neutrinos when realtime and energy information are required, liquid scintillators have several advantages compared to other technologies. In many cases the organic liquid needs to be loaded with metal to enhance the neutrino signal over background events. Several metal loaded scintillators of the past suffered from chemical and optical instabilities, limiting the performance of these neutrino detectors. Different ways of metal loading are described in the article with a focus on recent techniques providing metal loaded scintillators that can be used under stable conditions for many years even in ton scale experiments. Applications of metal loaded scintillators in neutrino experiments are reviewed and the performance as well as the prospects of different scintillator types are compared.

  1. Metal-loaded organic scintillators for neutrino physics

    DOE PAGES

    Buck, Christian; Yeh, Minfang

    2016-08-03

    Organic liquid scintillators are used in many neutrino physics experiments of the past and present. In particular for low energy neutrinos when realtime and energy information are required, liquid scintillators have several advantages compared to other technologies. In many cases the organic liquid needs to be loaded with metal to enhance the neutrino signal over background events. Several metal loaded scintillators of the past suffered from chemical and optical instabilities, limiting the performance of these neutrino detectors. Different ways of metal loading are described in the article with a focus on recent techniques providing metal loaded scintillators that can bemore » used under stable conditions for many years even in ton scale experiments. Lastly, we review applications of metal loaded scintillators in neutrino experiments and compare the performance as well as the prospects of different scintillator types.« less

  2. Metal-loaded organic scintillators for neutrino physics

    SciTech Connect

    Buck, Christian; Yeh, Minfang

    2016-08-03

    Organic liquid scintillators are used in many neutrino physics experiments of the past and present. In particular for low energy neutrinos when realtime and energy information are required, liquid scintillators have several advantages compared to other technologies. In many cases the organic liquid needs to be loaded with metal to enhance the neutrino signal over background events. Several metal loaded scintillators of the past suffered from chemical and optical instabilities, limiting the performance of these neutrino detectors. Different ways of metal loading are described in the article with a focus on recent techniques providing metal loaded scintillators that can be used under stable conditions for many years even in ton scale experiments. Lastly, we review applications of metal loaded scintillators in neutrino experiments and compare the performance as well as the prospects of different scintillator types.

  3. Separation of scintillation and Cherenkov lights in linear alkyl benzene

    SciTech Connect

    Li, Mohan; Guo, Ziyi; Yeh, Minfang; Wang, Zhe; Chen, Shaomin

    2016-09-11

    To separate scintillation and Cherenkov lights in water-based liquid scintillator detectors is a desired feature for future neutrino and proton decay experiments. Linear alkyl benzene (LAB) is one important ingredient of a water-based liquid scintillator currently under development. In this paper we report on the separation of scintillation and Cherenkov lights observed in an LAB sample. The rise and decay times of the scintillation light are measured to be (7.7±3.0)ns and (36.6±2.4)ns, respectively, while the full width [–3σ, 3σ] of the Cherenkov light is 12 ns and is dominated by the time resolution of the photomultiplier tubes. Here, the scintillation light yield was measured to be(1.01±0.12)×103photons/MeV.

  4. Methods for the continuous production of plastic scintillator materials

    DOEpatents

    Bross, Alan; Pla-Dalmau, Anna; Mellott, Kerry

    1999-10-19

    Methods for producing plastic scintillating material employing either two major steps (tumble-mix) or a single major step (inline-coloring or inline-doping). Using the two step method, the polymer pellets are mixed with silicone oil, and the mixture is then tumble mixed with the dopants necessary to yield the proper response from the scintillator material. The mixture is then placed in a compounder and compounded in an inert gas atmosphere. The resultant scintillator material is then extruded and pelletized or formed. When only a single step is employed, the polymer pellets and dopants are metered into an inline-coloring extruding system. The mixture is then processed under a inert gas atmosphere, usually argon or nitrogen, to form plastic scintillator material in the form of either scintillator pellets, for subsequent processing, or as material in the direct formation of the final scintillator shape or form.

  5. Optical scintillation measurements in a desert environment IV: simulated effects of scintillation on communications links

    NASA Astrophysics Data System (ADS)

    Suite, Michele; Rabinovich, W. S.; Mahon, Rita; Moore, Christopher; Ferraro, Mike; Burris, H. R., Jr.; Thomas, L. M.

    2011-09-01

    Optical scintillation is an effect that limits the performance of many optical systems including imagers and free space optical communication links. The Naval Research Laboratory is undertaking a series of measurement campaigns of optical scintillation in a variety of environments. In December of 2010 measurements were made over a one week period in the desert at China Lake, CA. The NRL TATS system was used to measure time resolved scintillation over a variety of different ranges and terrains. This data has been used to determine fade rate and duration as a function of weather and link margin. Temporal correlation of fades has also been calculated. This data allows simulation of a variety of communication protocols and the effects of those protocols on link throughput. In this paper we present a comparison of different protocols for both direct and retroreflector links.

  6. Cresst-II: dark matter search with scintillating absorbers

    NASA Astrophysics Data System (ADS)

    Angloher, G.; Bucci, C.; Cozzini, C.; von Feilitzsch, F.; Frank, T.; Hauff, D.; Henry, S.; Jagemann, Th.; Jochum, J.; Kraus, H.; Majorovits, B.; Ninkovic, J.; Petricca, F.; Pröbst, F.; Ramachers, Y.; Rau, W.; Seidel, W.; Stark, M.; Uchaikin, S.; Stodolsky, L.; Wulandari, H.

    2004-03-01

    In the CRESST-II experiment, scintillating CaWO4 crystals are used as absorbers for direct weakly interacting massive particles (WIMP) detection. Nuclear recoils can be discriminated against electron recoils by measuring phonons and scintillation light simultaneously. The absorber crystal and the silicon light detector are read out by tungsten superconducting phase transition thermometers. Results on the sensitivity of the phonon and the light channel, radiopurity, the scintillation properties of CaWO4, and on the WIMP sensitivity are presented.

  7. The reconstruction software for the MICE scintillating fibre trackers

    NASA Astrophysics Data System (ADS)

    Dobbs, A.; Hunt, C.; Long, K.; Santos, E.; Uchida, M. A.; Kyberd, P.; Heidt, C.; Blot, S.; Overton, E.

    2016-12-01

    The Muon Ionization Cooling Experiment (MICE) will demonstrate the principle of muon beam phase-space reduction via ionization cooling. Muon beam cooling will be required for the proposed Neutrino Factory or Muon Collider. The phase-space before and after the cooling cell must be measured precisely. This is achieved using two scintillating-fibre trackers, each placed in a solenoidal magnetic field. This paper describes the software reconstruction for the fibre trackers: the GEANT4 based simulation; the implementation of the geometry; digitisation; space-point reconstruction; pattern recognition; and the final track fit based on a Kalman filter. The performance of the software is evaluated by means of Monte Carlo studies and the precision of the final track reconstruction is evaluated.

  8. Composite solid-state scintillators for neutron detection

    DOEpatents

    Dai, Sheng; Im, Hee-Jung; Pawel, Michelle D.

    2006-09-12

    Applicant's present invention is a composite scintillator for neutron detection comprising a matrix material fabricated from an inorganic sol-gel precursor solution homogeneously doped with a liquid scintillating material and a neutron absorbing material. The neutron absorbing material yields at least one of an electron, a proton, a triton, an alpha particle or a fission fragment when the neutron absorbing material absorbs a neutron. The composite scintillator further comprises a liquid scintillating material in a self-assembled micelle formation homogeneously doped in the matrix material through the formation of surfactant-silica composites. The scintillating material is provided to scintillate when traversed by at least one of an electron, a proton, a triton, an alpha particle or a fission fragment. The scintillating material is configured such that the matrix material surrounds the micelle formation of the scintillating material. The composite scintillator is fabricated and applied as a thin film on substrate surfaces, a coating on optical fibers or as a glass material.

  9. Ionospheric scintillation observations over Kenyan region - Preliminary results

    NASA Astrophysics Data System (ADS)

    Olwendo, O. J.; Xiao, Yu; Ming, Ou

    2016-11-01

    Ionospheric scintillation refers to the rapid fluctuations in the amplitude and phase of a satellite signal as it passes through small-scale plasma density irregularities in the ionosphere. By analyzing ionospheric scintillation observation datasets from satellite signals such as GPS signals we can study the morphology of ionospheric bubbles. At low latitudes, the diurnal behavior of scintillation is driven by the formation of large-scale equatorial density depletions which form one to two hours after sunset via the Rayleigh-Taylor instability mechanism near the magnetic equator. In this work we present ionospheric scintillation activity over Kenya using data derived from a newly installed scintillation monitor developed by CRIRP at Pwani University (39.78°E, 3.24°S) during the period August to December, 2014. The results reveal the scintillation activity mainly occurs from post-sunset to post-midnight hours, and ceases around 04:00 LT. We also found that the ionospheric scintillation tends to appear at the southwest and northwest of the station. These locations coincide with the southern part of the Equatorial Ionization Anomaly crest over Kenya region. The occurrence of post-midnight L-band scintillation events which are not linked to pre-midnight scintillation observations raises fundamental question on the mechanism and source of electric fields driving the plasma depletion under conditions of very low background electron density.

  10. Scintillation of Un-doped ZnO Single Crystals

    SciTech Connect

    Colosimo, A. M.; Ji, Jianfeng; Stepanov, P. S.; Boatner, L. A.; Selim, F. A.

    2016-01-07

    In this paper, scintillation properties are often studied by photo-luminescence (PL) and scintillation measurements. In this work, we combine X-ray-induced luminescence (XRIL) spectroscopy [Review of Scientific Instruments 83, 103112 (2012)] with PL and standard scintillation measurements to give insight into the scintillation properties of un-doped ZnO single crystals. XRIL revealed that ZnO luminescence proportionally increases with X-ray power and exhibits excellent linearity - indicating the possibility of developing radiation detectors with good energy resolution. Finally, by coupling ZnO crystals to fast photomultiplier tubes and monitoring the anode signal, rise times as fast as 0.9 ns were measured.

  11. Development of polystyrene-based scintillation materials and its mechanisms

    NASA Astrophysics Data System (ADS)

    Nakamura, Hidehito; Kitamura, Hisashi; Shinji, Osamu; Saito, Katashi; Shirakawa, Yoshiyuki; Takahashi, Sentaro

    2012-12-01

    Scintillation materials based on polystyrene (PS) have been investigated. Para-terphenyl was employed as a fluorescent molecule (fluor) that functions as a wavelength shifter. A clear increase in photon yield of the scintillation materials relative to the pure PS was observed, which cannot be explained by the conventional theory of scintillation mechanism. Furthermore, the photon yield increased with flour concentration in accordance with a power-law. Here we reveal the emergence of a luminescence of PS-based scintillation materials and demonstrate that their photon yields can be controlled by the fluor concentration.

  12. Plasmonic light yield enhancement of a liquid scintillator

    NASA Astrophysics Data System (ADS)

    Bignell, Lindsey J.; Mume, Eskender; Jackson, Timothy W.; Lee, George P.

    2013-05-01

    We demonstrate modifications to the light yield properties of an organic liquid scintillator due to the localization of the tertiary fluorophore component to the surface of Ag-core silica-shell nanoparticles. We attribute this enhancement to the near-field interaction of Ag nanoparticle plasmons with these fluor molecules. The scintillation light yield enhancement is shown to be equal to the fluorescence enhancement within measurement uncertainties. With a suitable choice of plasmon energy and scintillation fluor, this effect may be used to engineer scintillators with enhanced light yields for radiation detection applications.

  13. Energy Transfer Based Nanocomposite Scintillator for Radiation Detection

    NASA Astrophysics Data System (ADS)

    Aslam, Soha; Sahi, Sunil; Chen, Wei; Ma, Lun; Kenarangui, Rasool

    2014-09-01

    Scintillators are the materials that emit light upon irradiation with high energy radiation like X-ray or gamma-ray. Inorganic single crystal and organic (plastic and liquid) are the two most used scintillator types. Both of these scintillator kinds have advantages and disadvantages. Inorganic single crystals are expensive and difficult to grow in desire shape and size. Also, single crystal scintillator such as NaI and CsI are very hygroscopic. On the other hand, organic scintillators have low density which limits their applications in gamma spectroscopy. Due to high quantum yield and size dependent emission, nanoparticles have attracted interested in various field of research. Here, we have studies the nanoparticles for radiation detection. We have synthesized nanoparticles of Cerium fluoride (CeF3), Zinc Oxide (ZnO), Cadmium Telluride (CdTe), Copper complex and Zinc sulfide (ZnS). We have used Fluorescence Resonance Energy Transfer (FRET) principle to enhance the luminescence properties of nanocomposite scintillator. Nanocomposites scintillators are structurally characterized with X-ray diffraction (XRD) and Transmission Electron Microscopy (TEM). Optical properties are studied using Photoluminescence, UV-Visible and X-ray. Enhancements in the luminescence are observed under UV and X-ray excitation. Preliminary studies shows nanocomposite scintillators are promising for radiation detection. Scintillators are the materials that emit light upon irradiation with high energy radiation like X-ray or gamma-ray. Inorganic single crystal and organic (plastic and liquid) are the two most used scintillator types. Both of these scintillator kinds have advantages and disadvantages. Inorganic single crystals are expensive and difficult to grow in desire shape and size. Also, single crystal scintillator such as NaI and CsI are very hygroscopic. On the other hand, organic scintillators have low density which limits their applications in gamma spectroscopy. Due to high quantum

  14. Multifrequency equatorial ionospheric scintillations in American and Indian zones

    NASA Technical Reports Server (NTRS)

    Rastogi, R. G.; Aarons, J.; Whitney, H. E.; Mullen, J. P.; Pantoja, J.; Deshpande, M. R.; Vats, H. O.; Chandra, H.; Davies, K.

    1980-01-01

    Amplitude scintillations of 40/41, 140 and 360 MHz transmissions recorded at Huancayo (phase I) and at Ootacamund (phase II) of the ATS-6 program are compared. The scintillations were found to be strongest between 20 and 24 hr LT with another peak around midday. The daytime scintillations do not show a significant seasonal variation at either of these stations. The nighttime scintillations were maximum during December solstices at Huancayo and during equinoxes at Ootacamund and suggested to be due to non-q type of sporadic E following the occurrence of counter-electrojet.

  15. Lanthanide doped strontium-barium cesium halide scintillators

    DOEpatents

    Bizarri, Gregory; Bourret-Courchesne, Edith; Derenzo, Stephen E.; Borade, Ramesh B.; Gundiah, Gautam; Yan, Zewu; Hanrahan, Stephen M.; Chaudhry, Anurag; Canning, Andrew

    2015-06-09

    The present invention provides for a composition comprising an inorganic scintillator comprising an optionally lanthanide-doped strontium-barium, optionally cesium, halide, useful for detecting nuclear material.

  16. Report on radiation exposure of lead-scintillator stack

    SciTech Connect

    Underwood, D.G.

    1990-11-08

    A stack of lead and scintillator was placed in a neutral beam obtained from targeting 800 GeV protons. Small pieces of film containing radiochromic dye were placed adjacent to the layers of scintillator for the purpose of measuring the radiation dose to the scintillator. Our motivation was to calibrate the radiation dose obtainable in this manner for future tests of scintillator for SSC experiments and to relate dose to flux to check absolute normalization for calculations. We also observed several other radiation effects which should be considered for both damage and compensation in a calorimeter.

  17. Plasmonic light yield enhancement of a liquid scintillator

    SciTech Connect

    Bignell, Lindsey J.; Jackson, Timothy W.; Mume, Eskender; Lee, George P.

    2013-05-27

    We demonstrate modifications to the light yield properties of an organic liquid scintillator due to the localization of the tertiary fluorophore component to the surface of Ag-core silica-shell nanoparticles. We attribute this enhancement to the near-field interaction of Ag nanoparticle plasmons with these fluor molecules. The scintillation light yield enhancement is shown to be equal to the fluorescence enhancement within measurement uncertainties. With a suitable choice of plasmon energy and scintillation fluor, this effect may be used to engineer scintillators with enhanced light yields for radiation detection applications.

  18. SIRT1 overexpression decreases cisplatin-induced acetylation of NF-{kappa}B p65 subunit and cytotoxicity in renal proximal tubule cells

    SciTech Connect

    Jung, Yu Jin; Lee, Jung Eun; Lee, Ae Sin; Kang, Kyung Pyo; Lee, Sik; Park, Sung Kwang; Lee, Sang Yong; Han, Myung Kwan; Kim, Duk Hoon; Kim, Won

    2012-03-09

    Highlights: Black-Right-Pointing-Pointer Cisplatin increases acetylation of NF-{kappa}B p65 subunit in HK2 cells. Black-Right-Pointing-Pointer SIRT1 overexpression decreases cisplatin-induced p65 acetylation and -cytotoxicity. Black-Right-Pointing-Pointer Resveratrol decreased cisplatin-induced cell viability through deacetylation of p65. -- Abstract: As the increased acetylation of p65 is linked to nuclear factor-{kappa}B (NF-{kappa}B) activation, the regulation of p65 acetylation can be a potential target for the treatment of inflammatory injury. Cisplatin-induced nephrotoxicity is an important issue in chemotherapy of cancer patients. SIRT1, nicotinamide adenine dinucleotide (NAD{sup +})-dependent protein deacetylase, has been implicated in a variety of cellular processes such as inflammatory injury and the control of multidrug resistance in cancer. However, there is no report on the effect of SIRT1 overexpression on cisplatin-induced acetylation of p65 subunit of NF-{kappa}B and cell injury. To investigate the effect of SIRT1 in on cisplatin-induced acetylation of p65 subunit of NF-{kappa}B and cell injury, HK2 cells were exposed with SIRT1 overexpression, LacZ adenovirus or dominant negative adenovirus after treatment with cisplatin. While protein expression of SIRT1 was decreased by cisplatin treatment compared with control buffer treatment, acetylation of NF-{kappa}B p65 subunit was significantly increased after treatment with cisplatin. Overexpression of SIRT1 ameliorated the increased acetylation of p65 of NF-{kappa}B during cisplatin treatment and cisplatin-induced cytotoxicity. Further, treatment of cisplatin-treated HK2 cells with resveratrol, a SIRT1 activator, also decreased acetylation of NF-{kappa}B p65 subunit and cisplatin-induced increase of the cell viability in HK2 cells. Our findings suggests that the regulation of acetylation of p65 of NF-{kappa}B through SIRT1 can be a possible target to attenuate cisplatin-induced renal cell damage.

  19. Cubesat Proximity Operations Demonstration (CPOD)

    NASA Technical Reports Server (NTRS)

    Villa, Marco; Martinez, Andres; Petro, Andrew

    2015-01-01

    The CubeSat Proximity Operations Demonstration (CPOD) project will demonstrate rendezvous, proximity operations and docking (RPOD) using two 3-unit (3U) CubeSats. Each CubeSat is a satellite with the dimensions 4 inches x 4 inches x 13 inches (10 centimeters x 10 centimeters x 33 centimeters) and weighing approximately 11 pounds (5 kilograms). This flight demonstration will validate and characterize many new miniature low-power proximity operations technologies applicable to future missions. This mission will advance the state of the art in nanosatellite attitude determination,navigation and control systems, in addition to demonstrating relative navigation capabilities.The two CPOD satellites are scheduled to be launched together to low-Earth orbit no earlier than Dec. 1, 2015.

  20. Role of H{sub 2}O{sub 2} on the kinetics of low-affinity high-capacity Na{sup +}-dependent alanine transport in SHR proximal tubular epithelial cells

    SciTech Connect

    Pinto, Vanda; Pinho, Maria Joao; Jose, Pedro A.; Soares-da-Silva, Patricio

    2010-07-30

    Research highlights: {yields} H{sub 2}O{sub 2} in excess is required for the presence of a low-affinity high-capacity component for the Na{sup +}-dependent [{sup 14}C]-L-alanine uptake in SHR PTE cells only. {yields} It is suggested that Na{sup +} binding in renal ASCT2 may be regulated by ROS in SHR PTE cells. -- Abstract: The presence of high and low sodium affinity states for the Na{sup +}-dependent [{sup 14}C]-L-alanine uptake in immortalized renal proximal tubular epithelial (PTE) cells was previously reported (Am. J. Physiol. 293 (2007) R538-R547). This study evaluated the role of H{sub 2}O{sub 2} on the Na{sup +}-dependent [{sup 14}C]-L-alanine uptake of ASCT2 in immortalized renal PTE cells from Wistar Kyoto rat (WKY) and spontaneously hypertensive rat (SHR). Na{sup +} dependence of [{sup 14}C]-L-alanine uptake was investigated replacing NaCl with an equimolar concentration of choline chloride in vehicle- and apocynin-treated cells. Na{sup +} removal from the uptake solution abolished transport activity in both WKY and SHR PTE cells. Decreases in H{sub 2}O{sub 2} levels in the extracellular medium significantly reduced Na{sup +}-K{sub m} and V{sub max} values of the low-affinity high-capacity component in SHR PTE cells, with no effect on the high-affinity low-capacity state of the Na{sup +}-dependent [{sup 14}C]-L-alanine uptake. After removal of apocynin from the culture medium, H{sub 2}O{sub 2} levels returned to basal values within 1 to 3 h in both WKY and SHR PTE cells and these were found stable for the next 24 h. Under these experimental conditions, the Na{sup +}-K{sub m} and V{sub max} of the high-affinity low-capacity state were unaffected and the low-affinity high-capacity component remained significantly decreased 1 day but not 4 days after apocynin removal. In conclusion, H{sub 2}O{sub 2} in excess is required for the presence of a low-affinity high-capacity component for the Na{sup +}-dependent [{sup 14}C]-L-alanine uptake in SHR PTE cells only

  1. In vitro studies on the stability in the proximal gastrointestinal tract and bioaccessibility in Caco-2 cells of chlorogenic acids from spent coffee grounds.

    PubMed

    Monente, Carmen; Ludwig, Iziar A; Stalmach, Angelique; de Peña, Maria Paz; Cid, Concepción; Crozier, Alan

    2015-01-01

    Spent coffee grounds are a potential commercial source of substantial amounts of chlorogenic acids (CGAs). The aim of this study was to evaluate the stability of spent coffee CGAs using in vitro simulated gastroduodenal digestion and to investigate their potential absorption using an in vitro Caco-2 model of human small intestinal epithelium. During in vitro digestion, lactones were partially degraded while caffeoylquinic and feruloylquinic acids were much more stable. Transport and metabolism studies showed that 1% of the total CGAs were absorbed and transported from the apical to the basolateral side of a Caco-2 cell monolayer after 1 h. Lactones and coumaroylquinic acids showed the rate of highest absorption. Caco-2 cells possessed low metabolic activity. In conclusion, spent coffee extracts contain large amounts of CGAs, which remained bioaccessible across the intestinal barrier, albeit to a relatively low degree.

  2. Integrative genomic analysis in K562 chronic myelogenous leukemia cells reveals that proximal NCOR1 binding positively regulates genes that govern erythroid differentiation and Imatinib sensitivity.

    PubMed

    Long, Mark D; van den Berg, Patrick R; Russell, James L; Singh, Prashant K; Battaglia, Sebastiano; Campbell, Moray J

    2015-09-03

    To define the functions of NCOR1 we developed an integrative analysis that combined ENCODE and NCI-60 data, followed by in vitro validation. NCOR1 and H3K9me3 ChIP-Seq, FAIRE-seq and DNA CpG methylation interactions were related to gene expression using bootstrapping approaches. Most NCOR1 combinations (24/44) were associated with significantly elevated level expression of protein coding genes and only very few combinations related to gene repression. DAVID's biological process annotation revealed that elevated gene expression was uniquely associated with acetylation and ETS binding. A matrix of gene and drug interactions built on NCI-60 data identified that Imatinib significantly targeted the NCOR1 governed transcriptome. Stable knockdown of NCOR1 in K562 cells slowed growth and significantly repressed genes associated with NCOR1 cistrome, again, with the GO terms acetylation and ETS binding, and significantly dampened sensitivity to Imatinib-induced erythroid differentiation. Mining public microarray data revealed that NCOR1-targeted genes were significantly enriched in Imatinib response gene signatures in cell lines and chronic myelogenous leukemia (CML) patients. These approaches integrated cistrome, transcriptome and drug sensitivity relationships to reveal that NCOR1 function is surprisingly most associated with elevated gene expression, and that these targets, both in CML cell lines and patients, associate with sensitivity to Imatinib.

  3. Driven shielding capacitive proximity sensor

    NASA Technical Reports Server (NTRS)

    Vranish, John M. (Inventor); McConnell, Robert L. (Inventor)

    2000-01-01

    A capacitive proximity sensing element, backed by a reflector driven at the same voltage as and in phase with the sensor, is used to reflect the field lines away from a grounded robot arm towards an intruding object, thus dramatically increasing the sensor's range and sensitivity.

  4. Proximity Sensors Make Robot Dexterous

    NASA Technical Reports Server (NTRS)

    Hess, Cliff; Li, Larry C. H.

    1990-01-01

    Control system enables robot hand to grasp objects of varied shapes. Key features of system: reflective proximity sensors furnishing data on position, orientation, and distance of object and software protocol controlling sequence of operations in approaching and grasping objects. Reflected-beam sensing concept applied to simple opposed-jaw industrial grippers as well as to dexterous robot hands.

  5. Comparing the response of PSD-capable plastic scintillator to standard liquid scintillator

    NASA Astrophysics Data System (ADS)

    Woolf, Richard S.; Hutcheson, Anthony L.; Gwon, Chul; Phlips, Bernard F.; Wulf, Eric A.

    2015-06-01

    This work discusses a test campaign to characterize the response of the recently developed plastic scintillator with pulse shape discrimination (PSD) capabilities (EJ-299-33). PSD is a property exhibited by certain types of scintillating material in which incident stimuli (fast neutrons or γ rays) can be separated by exploiting differences in the scintillation light pulse tail. Detector geometries used were: a 10 cm×10 cm×10 cm cube and a 10-cm diameter×10-cm long cylinder. EJ-301 and EJ-309 liquid scintillators with well-known responses were also tested. The work was conducted at the University of Massachusetts Lowell Van De Graaff accelerator. The facility accelerated protons on a thin Li target to yield quasi-monoenergetic neutrons from the 7Li(p,n)7Be reaction (Q-value: -1.644 MeV). Collimated fast neutrons were obtained by placing detectors behind a neutron spectrometer. Rotating the spectrometer, and thus changing the neutron energy, allowed us to achieve 0.5-3.2 MeV neutrons in 200-300 keV steps. Data were acquired through a flash analog-to-digital converter (ADC) capable of performing digital PSD measurements. By using the PSD technique to separate the neutron events from unwanted γ background, we constructed a pulse height spectrum at each energy. Obtaining a relationship of the relative light output versus energy allowed us to construct the response function for the EJ-299-33 and liquid scintillator. The EJ-299-33 response in terms of electron equivalent energy (Ee.e.) vs. proton equivalent energy (Ep.e.), how it compared with the standard xylene-based EJ-301 (or, NE-213/BC-501 A equivalent) and EJ-309 liquid scintillator response, and how the EJ-301 and EJ-309 compared, are presented. We find that the EJ-299-33 demonstrated a lower light output by up to 40% for <1.0 MeV neutrons; and ranging between a 5-35% reduction for 2.5-3.0 MeV neutrons compared to the EJ-301/309, depending on the scintillator and geometry. Monte Carlo modeling techniques were

  6. Fundamental Limits of Scintillation Detector Timing Precision

    PubMed Central

    Derenzo, Stephen E.; Choong, Woon-Seng; Moses, William W.

    2014-01-01

    In this paper we review the primary factors that affect the timing precision of a scintillation detector. Monte Carlo calculations were performed to explore the dependence of the timing precision on the number of photoelectrons, the scintillator decay and rise times, the depth of interaction uncertainty, the time dispersion of the optical photons (modeled as an exponential decay), the photodetector rise time and transit time jitter, the leading-edge trigger level, and electronic noise. The Monte Carlo code was used to estimate the practical limits on the timing precision for an energy deposition of 511 keV in 3 mm × 3 mm × 30 mm Lu2SiO5:Ce and LaBr3:Ce crystals. The calculated timing precisions are consistent with the best experimental literature values. We then calculated the timing precision for 820 cases that sampled scintillator rise times from 0 to 1.0 ns, photon dispersion times from 0 to 0.2 ns, photodetector time jitters from 0 to 0.5 ns fwhm, and A from 10 to 10,000 photoelectrons per ns decay time. Since the timing precision R was found to depend on A−1/2 more than any other factor, we tabulated the parameter B, where R = BA−1/2. An empirical analytical formula was found that fit the tabulated values of B with an rms deviation of 2.2% of the value of B. The theoretical lower bound of the timing precision was calculated for the example of 0.5 ns rise time, 0.1 ns photon dispersion, and 0.2 ns fwhm photodetector time jitter. The lower bound was at most 15% lower than leading-edge timing discrimination for A from 10 to 10,000 photoelectrons/ns. A timing precision of 8 ps fwhm should be possible for an energy deposition of 511 keV using currently available photodetectors if a theoretically possible scintillator were developed that could produce 10,000 photoelectrons/ns. PMID:24874216

  7. Global Morphology of Ionospheric Scintillations II

    DTIC Science & Technology

    1975-03-11

    Fading at Huancayo , Peru 3. Scintillation Contours at 136 MHz for 6 Years Data from Huancayo , Peru 4. Fading on 254 MHz as Observed at Kwajalein 5...Flight Center, NASA Z861-71-239. 8 3.2 Dependence on Magnetic Activity Data from Huancayo , Peru, when analyzed over a long period has shown as a gross...a second increase, that is, after midnight during magnetically disturbed days. 60 HUANCAYO , PERU LES - 6, 254 MHz S1 > 60 -- Kp = 0 - 3 ---Kp=4-9

  8. Thin GSO scintillator for neutron detection

    SciTech Connect

    Reeder, P.L.

    1994-05-01

    The new scintillator cerium-doped gadolinium orthosilicate (GSO -- Gd{sub 2}SiO{sub 5}:Ce) has a light output that is about 20% that of NaI(T1). The enormous cross section of Gd for capture of.thermal neutrons makes GSO a candidate for novel types of neutron.detectors. The characteristic radiations from neutron capture in Gd can be stopped in about 75 {mu}m of GSO. Data obtained from a GSO detector that was about 0.6-mm thick demonstrated that thermal neutrons could easily be detected and that higher energy gamma rays caused minimal interference.

  9. Cherenkov and Scintillation Properties of Cubic Zirconium

    NASA Technical Reports Server (NTRS)

    Christl, M.J.; Adams, J.H.; Parnell, T.A.; Kuznetsov, E.N.

    2008-01-01

    Cubic zirconium (CZ) is a high index of refraction (n =2.17) material that we have investigated for Cherenkov counter applications. Laboratory and proton accelerator tests of an 18cc sample of CZ show that the expected fast Cherenkov response is accompanied by a longer scintillation component that can be separated by pulse shaping. This presents the possibility of novel particle spectrometers which exploits both properties of CZ. Other high index materials being examined for Cherenkov applications will be discussed. Results from laboratory tests and an accelerator exposure will be presented and a potential application in solar energetic particle instruments will be discussed

  10. Changes in gene expression in human renal proximal tubule cells exposed to low concentrations of S-(1,2-dichlorovinyl)-L-cysteine, a metabolite of trichloroethylene

    SciTech Connect

    Lock, Edward A. . E-mail: e.lock@ljmu.ac.uk; Barth, Jeremy L.; Argraves, Scott W.; Schnellmann, Rick G.

    2006-10-15

    Epidemiology studies suggest that there may be a weak association between high level exposure to trichloroethylene (TCE) and renal tubule cell carcinoma. Laboratory animal studies have shown an increased incidence of renal tubule carcinoma in male rats but not mice. TCE can undergo metabolism via glutathione (GSH) conjugation to form metabolites that are known to be nephrotoxic. The GSH conjugate, S-(1,2-dichlorovinyl)glutathione (DCVG), is processed further to the cysteine conjugate, S-(1,2-dichlorovinyl)-L-cysteine (DCVC), which is the penultimate nephrotoxic species. We have cultured human renal tubule cells (HRPTC) in serum-free medium under a variety of different culture conditions and observed growth, respiratory control and glucose transport over a 20 day period in medium containing low glucose. Cell death was time- and concentration-dependent, with the EC{sub 5} for DCVG being about 3 {mu}M and for DCVC about 7.5 {mu}M over 10 days. Exposure of HRPTC to sub-cytotoxic doses of DCVC (0.1 {mu}M and 1 {mu}M for 10 days) led to a small number of changes in gene expression, as determined by transcript profiling with Affymetrix human genome chips. Using the criterion of a mean 2-fold change over control for the four samples examined, 3 genes at 0.1 {mu}M DCVC increased, namely, adenosine kinase, zinc finger protein X-linked and an enzyme with lyase activity. At 1 {mu}M DCVC, two genes showed a >2-fold decrease, N-acetyltransferase 8 and complement factor H. At a lower stringency (1.5-fold change), a total of 63 probe sets were altered at 0.1 {mu}M DCVC and 45 at 1 {mu}M DCVC. Genes associated with stress, apoptosis, cell proliferation and repair and DCVC metabolism were altered, as were a small number of genes that did not appear to be associated with the known mode of action of DCVC. Some of these genes may serve as molecular markers of TCE exposure and effects in the human kidney.

  11. Measurement of Scintillation and Ionization Yield and Scintillation Pulse Shape from Nuclear Recoils in Liquid Argon

    DOE PAGES

    Cao, H.

    2015-05-26

    We have measured the scintillation and ionization yield of recoiling nuclei in liquid argon as a function of applied electric field by exposing a dual-phase liquid argon time projection chamber (LAr-TPC) to a low energy pulsed narrow band neutron beam produced at the Notre Dame Institute for Structure and Nuclear Astrophysics. Liquid scintillation counters were arranged to detect and identify neutrons scattered in the TPC and to select the energy of the recoiling nuclei. We also report measurements of the scintillation yields for nuclear recoils with energies from 10.3 to 57.3 keV and for median applied electric fields from 0more » to 970 V/cm. For the ionization yields, we report measurements from 16.9 to 57.3 keV and for electric fields from 96.4 to 486 V/cm. Furthermore, we report the observation of an anticorrelation between scintillation and ionization from nuclear recoils, which is similar to the anticorrelation between scintillation and ionization from electron recoils. Assuming that the energy loss partitions into excitons and ion pairs from 83mKr internal conversion electrons is comparable to that from 207Bi conversion electrons, we obtained the numbers of excitons (Nex) and ion pairs (Ni) and their ratio (Nex/Ni) produced by nuclear recoils from 16.9 to 57.3 keV. Motivated by arguments suggesting direction sensitivity in LAr-TPC signals due to columnar recombination, a comparison of the light and charge yield of recoils parallel and perpendicular to the applied electric field is presented for the first time.« less

  12. Measurement of scintillation and ionization yield and scintillation pulse shape from nuclear recoils in liquid argon

    NASA Astrophysics Data System (ADS)

    Cao, H.; Alexander, T.; Aprahamian, A.; Avetisyan, R.; Back, H. O.; Cocco, A. G.; Dejongh, F.; Fiorillo, G.; Galbiati, C.; Grandi, L.; Guardincerri, Y.; Kendziora, C.; Lippincott, W. H.; Love, C.; Lyons, S.; Manenti, L.; Martoff, C. J.; Meng, Y.; Montanari, D.; Mosteiro, P.; Olvitt, D.; Pordes, S.; Qian, H.; Rossi, B.; Saldanha, R.; Sangiorgio, S.; Siegl, K.; Strauss, S. Y.; Tan, W.; Tatarowicz, J.; Walker, S.; Wang, H.; Watson, A. W.; Westerdale, S.; Yoo, J.; Scene Collaboration

    2015-05-01

    We have measured the scintillation and ionization yield of recoiling nuclei in liquid argon as a function of applied electric field by exposing a dual-phase liquid argon time projection chamber (LAr-TPC) to a low energy pulsed narrow band neutron beam produced at the Notre Dame Institute for Structure and Nuclear Astrophysics. Liquid scintillation counters were arranged to detect and identify neutrons scattered in the TPC and to select the energy of the recoiling nuclei. We report measurements of the scintillation yields for nuclear recoils with energies from 10.3 to 57.3 keV and for median applied electric fields from 0 to 970 V /cm . For the ionization yields, we report measurements from 16.9 to 57.3 keV and for electric fields from 96.4 to 486 V /cm . We also report the observation of an anticorrelation between scintillation and ionization from nuclear recoils, which is similar to the anticorrelation between scintillation and ionization from electron recoils. Assuming that the energy loss partitions into excitons and ion pairs from Krm83 internal conversion electrons is comparable to that from 207Bi conversion electrons, we obtained the numbers of excitons (Nex) and ion pairs (Ni) and their ratio (Nex/Ni ) produced by nuclear recoils from 16.9 to 57.3 keV. Motivated by arguments suggesting direction sensitivity in LAr-TPC signals due to columnar recombination, a comparison of the light and charge yield of recoils parallel and perpendicular to the applied electric field is presented for the first time.

  13. Measurement of Scintillation and Ionization Yield and Scintillation Pulse Shape from Nuclear Recoils in Liquid Argon

    SciTech Connect

    Cao, H.

    2015-05-26

    We have measured the scintillation and ionization yield of recoiling nuclei in liquid argon as a function of applied electric field by exposing a dual-phase liquid argon time projection chamber (LAr-TPC) to a low energy pulsed narrow band neutron beam produced at the Notre Dame Institute for Structure and Nuclear Astrophysics. Liquid scintillation counters were arranged to detect and identify neutrons scattered in the TPC and to select the energy of the recoiling nuclei. We also report measurements of the scintillation yields for nuclear recoils with energies from 10.3 to 57.3 keV and for median applied electric fields from 0 to 970 V/cm. For the ionization yields, we report measurements from 16.9 to 57.3 keV and for electric fields from 96.4 to 486 V/cm. Furthermore, we report the observation of an anticorrelation between scintillation and ionization from nuclear recoils, which is similar to the anticorrelation between scintillation and ionization from electron recoils. Assuming that the energy loss partitions into excitons and ion pairs from 83mKr internal conversion electrons is comparable to that from 207Bi conversion electrons, we obtained the numbers of excitons (Nex) and ion pairs (Ni) and their ratio (N