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Sample records for reduces surface expression

  1. Introduced Amino Terminal Epitopes Can Reduce Surface Expression of Neuronal Nicotinic Receptors

    PubMed Central

    Bracamontes, John R.; Akk, Gustav; Steinbach, Joe Henry

    2016-01-01

    Epitopes accessible on the surface of intact cells are extremely valuable in studies of membrane proteins, allowing quantification and determination of the distribution of proteins as well as identification of cells expressing large numbers of proteins. However for many membrane proteins there are no suitable antibodies to native sequences, due to lack of availability, low affinity or lack of specificity. In these cases the use of an introduced epitope at specific sites in the protein of interest can often provide a suitable tool for studies. However, the introduction of the epitope sequence has the potential to affect protein expression, the assembly of multisubunit proteins or transport to the surface membrane. We find that surface expression of heteromeric neuronal nicotinic receptors containing the α4 and β4 subunits can be affected by introduced epitopes when inserted near the amino terminus of a subunit. The FLAG epitope greatly reduces surface expression when introduced into either α4 or β4 subunits, the V5 epitope has little effect when placed in either, while the Myc epitope reduces expression more when inserted into β4 than α4. These results indicate that the extreme amino terminal region is important for assembly of these receptors, and demonstrate that some widely used introduced epitopes may severely reduce surface expression. PMID:26963253

  2. Rare TREM2 variants associated with Alzheimer's disease display reduced cell surface expression.

    PubMed

    Sirkis, Daniel W; Bonham, Luke W; Aparicio, Renan E; Geier, Ethan G; Ramos, Eliana Marisa; Wang, Qing; Karydas, Anna; Miller, Zachary A; Miller, Bruce L; Coppola, Giovanni; Yokoyama, Jennifer S

    2016-09-02

    Rare variation in TREM2 has been associated with greater risk for Alzheimer's disease (AD). TREM2 encodes a cell surface receptor expressed on microglia and related cells, and the R47H variant associated with AD appears to affect the ability of TREM2 to bind extracellular ligands. In addition, other rare TREM2 mutations causing early-onset neurodegeneration are thought to impair cell surface expression. Using a sequence kernel association (SKAT) analysis in two independent AD cohorts, we found significant enrichment of rare TREM2 variants not previously characterized at the protein level. Heterologous expression of the identified variants showed that novel variants S31F and R47C displayed significantly reduced cell surface expression. In addition, we identified rare variant R136Q in a patient with language-predominant AD that also showed impaired surface expression. The results suggest rare TREM2 variants enriched in AD may be associated with altered TREM2 function and that AD risk may be conferred, in part, from altered TREM2 surface expression.

  3. Sulfonylurea receptors inhibit the epithelial sodium channel (ENaC) by reducing surface expression.

    PubMed

    Konstas, A A; Bielfeld-Ackermann, A; Korbmacher, C

    2001-08-01

    In the kidney the epithelial Na+ channel (ENaC) is co-expressed with the sulfonylurea receptor (SUR), an ABC protein that shares a high degree of homology with the cystic fibrosis transmembrane conductance regulator (CFTR) and reportedly modifies ENaC in various preparations. To investigate a possible regulatory relationship between SUR and ENaC, we performed co-expression studies on Xenopus laevis oocytes, which were assayed for amiloride-sensitive currents (DeltaIami). Moreover, a chemiluminescence assay was used to investigate the surface expression of extracellular hemagglutinin-tagged SUR1 (SUR1-HA) or HA-tagged ENaC (ENaC-HA). In oocytes co-injected with SUR1/ENaC (or SUR2B/ENaC) DeltaIami was reduced by congruent with 53% (or congruent with 45%) compared to DeltaIami measured in matched control oocytes injected with ENaC alone. The inhibitory effect of SUR on DeltaIami was preserved in oocytes expressing ENaC with C-terminally truncated subunits. Co-expression of SURs did not confer sensitivity of DeltaIami to diazoxide, pinacidil, tolbutamide, or glibenclamide. ENaC does not facilitate the surface expression of SUR1-HA, which is known to be retained in the endoplasmatic reticulum (ER) by an ER-retention/retrieval signal. SUR1-HAAAA, a mutant that lacks this signal, still inhibits ENaC currents. Chemiluminescence was reduced by congruent with 49% in oocytes co-expressing ENaC-HA/SUR1 compared to that in control oocytes expressing ENaC-HA alone. We conclude that SUR does not interact with ENaC at the level of the plasma membrane but that it inhibits DeltaIami by reducing surface expression of the channel.

  4. Reduced expression of platelet surface glycoprotein receptor IIb/IIIa at hyperthermic temperatures.

    PubMed

    Pasha, R; Benavides, M; Kottke-Marchant, K; Harasaki, H

    1995-09-01

    Hyperthermic temperatures exist from the heat dissipation of the implantable energy source of an artificial heart. This procedure as well as therapies for cancer and thermal injuries pose a new medical problem. Among many reported effects of heat on biologic systems, platelet functions such as maximal aggregation and adhesion are known to be reduced. Using flow cytometry, we have studied platelet dysfunction at elevated temperatures and have gained a mechanistic comprehension of the loss of platelet function. Platelet rich plasma was incubated at differing temperatures for 1 hour. Immediately after, the platelets were stained using mAb against glycoprotein IIb/IIIa (GPIIb-IIIa) (CD41a) and other platelet surface glycoproteins (GP) involved in aggregation and adhesion. Relative fluorescence intensity was measured using single-labeled, laser flow cytometry to determine changes in GP surface expression. In addition, scanning electron microscopy was used to evaluate morphologic changes. Hyperthermic temperatures between 40 and 44 degrees C significantly lowered the mAb cell surface binding in vitro of GP that participate in aggregation and adhesion. The most dramatic temperature-dependent loss of mAb binding was demonstrated by anti-GPIIb-IIIa, the mAb against the fibrinogen receptor. mAb binding to this receptor at 44 degrees C was decreased to 6.2% of a base-line fluorescence intensity of 654 (arbitrary units). The ADP-induced aggregation of platelets incubated at the same temperature also decreased to 2.1% of maximum aggregation. Other mAb, such as those against the von Willebrand factor receptor (GPIb) (CD42b), the thrombospondin receptor (GPIV) (CD36), and GPIIIa (CD61), also showed statistically significant reduction of mAb binding but to a lesser degree. Finally, scanning electron microscopy as well as side-scatter density plots from flow cytometry revealed that platelets became more spherical after incubation at 44 degrees C. The significant reduction in m

  5. SRC Inhibition Reduces NR2B Surface Expression and Synaptic Plasticity in the Amygdala

    ERIC Educational Resources Information Center

    Sinai, Laleh; Duffy, Steven; Roder, John C.

    2010-01-01

    The Src protein tyrosine kinase plays a central role in the regulation of N-methyl-d-aspartate receptor (NMDAR) activity by regulating NMDAR subunit 2B (NR2B) surface expression. In the amygdala, NMDA-dependent synaptic plasticity resulting from convergent somatosensory and auditory inputs contributes to emotional memory; however, the role of Src…

  6. SRC Inhibition Reduces NR2B Surface Expression and Synaptic Plasticity in the Amygdala

    ERIC Educational Resources Information Center

    Sinai, Laleh; Duffy, Steven; Roder, John C.

    2010-01-01

    The Src protein tyrosine kinase plays a central role in the regulation of N-methyl-d-aspartate receptor (NMDAR) activity by regulating NMDAR subunit 2B (NR2B) surface expression. In the amygdala, NMDA-dependent synaptic plasticity resulting from convergent somatosensory and auditory inputs contributes to emotional memory; however, the role of Src…

  7. Chronic Morphine Reduces Surface Expression of δ-Opioid Receptors in Subregions of Rostral Striatum.

    PubMed

    Leah, Paul M; Heath, Emily M L; Balleine, Bernard W; Christie, Macdonald J

    2016-03-01

    The delta opioid receptor (DOPr), whilst not the primary target of clinically used opioids, is involved in development of opioid tolerance and addiction. There is growing evidence that DOPr trafficking is involved in drug addiction, e.g., a range of studies have shown increased plasma membrane DOPr insertion during chronic treatment with opioids. The present study used a transgenic mouse model in which the C-terminal of the DOPr is tagged with enhanced-green fluorescence protein to examine the effects of chronic morphine treatment on surface membrane expression in striatal cholinergic interneurons that are implicated in motivated learning following both chronic morphine and morphine sensitization treatment schedules in male mice. A sex difference was noted throughout the anterior striatum, which was most prominent in the nucleus accumbens core region. Incontrast with previous studies in other neurons, chronic exposure to a high dose of morphine for 6 days had no effect, or slightly decreased (anterior dorsolateral striatum) surface DOPr expression. A morphine sensitization schedule produced similar results with a significant decrease in surface DOPr expression in nucleus accumbens shell. These results suggest that chronic morphine and morphine sensitisation treatment may have effects on instrumental reward-seeking behaviours and learning processes related to drug addiction, via effects on striatal DOPr function.

  8. Alpha-1 proteinase inhibitor M358R reduces thrombin generation when displayed on the surface of cells expressing tissue factor.

    PubMed

    Gierczak, Richard F; Pepler, Laura; Bhagirath, Vinai; Liaw, Patricia C; Sheffield, William P

    2014-11-01

    The M358R variant of alpha-1-proteinase inhibitor (API) is a potent soluble inhibitor of thrombin. Previously we engineered AR-API M358R, a membrane-bound form of this protein and showed that it inhibited exogenous thrombin when expressed on transfected cells lacking tissue factor (TF). To determine the suitability of AR-API M358R for gene transfer to vascular cells to limit thrombogenicity, we tested the ability of AR-API M358R to inhibit endogenous thrombin generated in plasma via co-expression co-expressing it on the surface of cells expressing TF. Transfected AR-API M358R formed inhibitory complexes with thrombin following exposure of recalcified, defibrinated plasma to TF on T24/83 cells, but discontinuously monitored thrombin generation was unaffected. Similarly, AR-API M358R expression did not reduce continuously monitored thrombin generation by T24/83 cell suspensions exposed to recalcified normal plasma in a Thrombogram-Thrombinoscope-type thrombin generation assay (TGA); in contrast, 1 μM hirudin variant 3 or soluble API M358R abolished thrombin generation. Gene transfer of TF to HEK 293 conferred the ability to support TF-dependent thrombin generation on HEK 293 cells. Co-transfection of HEK 293 cells with a 9:1 excess of DNA encoding AR-API M358R to that encoding TF reduced peak thrombin generation approximately 3-fold compared to controls. These in vitro results suggest that surface display of API M358R inhibits thrombin generation when the tethered serpin is expressed in excess of TF, and suggest its potential to limit thrombosis in appropriate vascular beds in animal models. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. Reduced cell surface expression of processed human immunodeficiency virus type 1 envelope glycoprotein in the presence of Nef.

    PubMed Central

    Schwartz, O; Rivière, Y; Heard, J M; Danos, O

    1993-01-01

    nef genes from two laboratory grown human immunodeficiency virus type 1 (HIV-1) strains and from two proviruses that had not been propagated in vitro were introduced into CD4+ lymphoblastoid CEM cells. The stable expression of all four Nef proteins was associated with an almost complete abrogation of CD4 cell surface localization. The consequences of the presence of Nef on gp160 cleavage, gp120 surface localization, and envelope-induced cytopathic effect were examined in CEM cells in which the HIV-1 env gene was expressed from a vaccinia virus vector. The presence of Nef did not modify the processing of gp160 into its subunits but resulted in a significant decrease of cell surface levels of gp120, associated with a dramatic reduction of the fusion-mediated cell death. Surface levels of mutant envelope glycoproteins unable to bind CD4 were not altered in Nef-expressing cells, suggesting that the phenomenon was CD4 dependent. The intracellular accumulation of fully processed envelope glycoproteins could significantly delay the cytopathic effect associated with envelope surface expression in HIV-infected cells and may be relevant to the selective advantage associated with Nef during the in vivo infectious process. Images PMID:8497051

  10. Potential immunosuppressive and antiinflammatory activities of Malaysian medicinal plants characterized by reduced cell surface expression of cell adhesion molecules.

    PubMed

    Tanaka, S; Yoichi, S; Ao, L; Matumoto, M; Morimoto, K; Akimoto, N; Honda, G; Tabata, M; Oshima, T; Masuda, T; bin Asmawi, M Z; Ismail, Z; Yusof, S M; Din, L B; Said, I M

    2001-12-01

    In the search for agents effective against immune-mediated disorders and inflammation, we have screened Malaysian medicinal plants for the ability to inhibit the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) on the surface of murine endothelial cells (F-2), and mouse myeloid leukaemia cells (M1), respectively. Of 41 kinds (29 species, 24 genera, 16 families) of Malaysian plants tested, 10 and 19 plant samples significantly downregulated the expression of ICAM-1 and VCAM-1, respectively. Bioassay-directed fractionation of an extract prepared from the bark of Goniothalamus andersonii showed that its ingredients, goniothalamin (1) and goniodiol (2) inhibited the cell surface expression of both ICAM-1 and VCAM-1. The present results suggest that Malaysian medicinal plants may be abundant natural resources for immunosuppressive and antiinflammatory substances. Copyright 2001 John Wiley & Sons, Ltd.

  11. Ascorbate inhibits platelet-endothelial adhesion in an in-vitro model of sepsis via reduced endothelial surface P-selectin expression.

    PubMed

    Secor, Dan; Swarbreck, Scott; Ellis, Christopher G; Sharpe, Michael D; Feng, Qingping; Tyml, Karel

    2017-01-01

    Plugging of the capillary bed can lead to organ failure and mortality in sepsis. We have reported that intravenous ascorbate injection reduces platelet adhesion to the capillary wall and capillary plugging in septic mice. Both platelet adhesion and capillary plugging require P-selectin, a key adhesion molecule. To elucidate the beneficial effect of ascorbate, we hypothesized that ascorbate reduces platelet-endothelial adhesion by reducing P-selectin surface expression in endothelial cells. We used mouse platelets, and monolayers of cultured microvascular endothelial cells (mouse skeletal muscle origin) stimulated with lipopolysaccharide, to examine platelet-endothelial adhesion. P-selectin mRNA expression in endothelial cells was determined by real-time PCR and P-selectin protein expression at the surface of these cells by immunofluorescence. Secretion of von Willebrand factor from cells into the supernatant (a measure of P-selectin-containing granule exocytosis) was determined by ELISA. Lipopolysaccharide (10 μg/ml, 1 h) increased platelet-endothelial adhesion. P-selectin-blocking antibody inhibited this adhesion. Lipopolysaccharide also increased P-selectin mRNA in endothelial cells, P-selectin expression at the endothelial surface, and von Willebrand factor secretion. Ascorbate pretreatment (100 μmol/l, 4 h) inhibited the increased platelet adhesion, surface expression of P-selectin, and von Willebrand factor secretion, but not the increase in P-selectin mRNA. The lipopolysaccharide-induced increase in platelet-endothelial adhesion requires P-selectin presence at the endothelial surface. Ascorbate's ability to reduce this presence could be important in reducing both platelet adhesion to the capillary wall and capillary plugging in sepsis.

  12. Heterologous expression of rab4 reduces glucose transport and GLUT4 abundance at the cell surface in oocytes.

    PubMed Central

    Mora, S; Monden, I; Zorzano, A; Keller, K

    1997-01-01

    To evaluate the role of the small rab GTP-binding proteins in glucose transporter trafficking, we have heterologously co-expressed rab4 or rab5 and GLUT4 or GLUT1 glucose transporters in Xenopus oocytes. Co-injection of rab4 and GLUT4 cRNAs resulted in a dose-dependent decrease in glucose transport; this effect was specific for rab4, since co-injection of an inactive rab4 mutant or rab5 cRNA did not have any effect on glucose transport. The effect of rab4 was selective for GLUT4, since no effect was detected in GLUT1-expressing oocytes. The inhibitory effect of rab4 on GLUT4-induced glucose transport was not the result of a change in overall cellular levels of GLUT4 glucose transporters. However, rab4 expression caused a marked decrease in the abundance of GLUT4 transporters present at the cell surface. Finally, rab4 and inhibitors of PtdIns 3-kinase showed additive effects in decreasing glucose transport in GLUT4-expressing oocytes. We conclude that rab4 plays an important role in the regulation of the intracellular GLUT4 trafficking pathway, by contributing to the intracellular retention of GLUT4 through a PtdIns 3-kinase-independent mechanism. PMID:9182703

  13. Domain deletion in the extracellular portion of the EGF-receptor reduces ligand binding and impairs cell surface expression.

    PubMed Central

    Lax, I; Bellot, F; Honegger, A M; Schmidt, A; Ullrich, A; Givol, D; Schlessinger, J

    1990-01-01

    Cultured NIH-3T3 cells were transfected with cDNA constructs encoding human epidermal growth factor-receptor (EGF-R)* and two deletion mutants in the extracellular portion of the receptor molecule. One mutant is devoid of 124 amino-terminal amino acids, and the other lacks 76 residues. Mutant receptors were not delivered to the cell surface unless the transfected cells contained also endogenous EGF-Rs, suggesting that receptor interaction complements the mutation and allows surface display of mutant receptors. Immunoprecipitation experiments revealed an association between mutant and endogenous EGF-Rs when both proteins were expressed in the same cell. Hence, receptor-oligomers may exist in the plane of the membrane even in the absence of ligand binding, and oligomerization may play a role in normal trafficking of EGF-Rs to the cell surface. Mutant receptors retained partial ligand binding activity as 125I-labeled EGF was covalently cross-linked to both mutant receptors, and EGF stimulated, albeit weakly, their protein tyrosine kinase activity. Both mutant EGF-Rs bind EGF with a 10-fold lower affinity than that of the solubilized wild type EGF-R. These results provide further evidence that the region flanked by the two cysteine-rich domains plays a crucial role in defining ligand-binding specificity of EGF-R. Images PMID:2100196

  14. Reduced surface expression of TLR4 by a V254I point mutation accounts for the low lipopolysaccharide responder phenotype of BALB/c B cells.

    PubMed

    Tsukamoto, Hiroki; Fukudome, Kenji; Takao, Shoko; Tsuneyoshi, Naoko; Ohta, Shoichiro; Nagai, Yoshinori; Ihara, Hideyuki; Miyake, Kensuke; Ikeda, Yoshitaka; Kimoto, Masao

    2013-01-01

    LPS is recognized by TLR4 and radioprotective 105 kDa in B cells. Susceptibility to LPS in murine B cells is most closely linked to the locus containing the TLR4 gene. However, the molecular mechanism underlying genetic control of LPS sensitivity by this locus has not been fully elucidated. In this study, we revealed that C57BL/6 (B6) B cells respond to mAb-induced, TLR4-specific signals stronger than BALB/c (BALB) B cells, as assessed by proliferation and upregulation of CD69 and CD86. In contrast, BALB B cells were not hyporesponsive to agonistic anti-radioprotective 105 kDa mAb or the TLR9 agonist CpG. Although the level of TLR4 mRNA in BALB B cells was comparable with that in B6 B cells, surface TLR4 expression in BALB B cells was lower than that in B6 B cells. This lower surface expression of BALB TLR4 was also observed when HEK293 and Ba/F3 cells were transfected with a BALB TLR4 expression construct. We identified a V254I mutation as the responsible single nucleotide polymorphism for lower surface expression of BALB TLR4. Furthermore, cotransfection of myeloid differentiation factor-2 increased BALB TLR4 expression, although it was still lower than B6 TLR4 expression. In concordance with reduced expression, Ba/F3 cells transfected with BALB TLR4 and myeloid differentiation factor-2 were hyporesponsive compared with those with B6 TLR4, as assessed by LPS-induced NF-κB activation. In conclusion, we revealed that LPS sensitivity is genetically controlled by the level of surface TLR4 expression on B cells. A V254I mutation accounts for the LPS hyporesponsive phenotype of BALB B cells.

  15. Blockage of Core Fucosylation Reduces Cell-Surface Expression of PD-1 and Promotes Anti-tumor Immune Responses of T Cells.

    PubMed

    Okada, Masahiro; Chikuma, Shunsuke; Kondo, Taisuke; Hibino, Sana; Machiyama, Hiroaki; Yokosuka, Tadashi; Nakano, Miyako; Yoshimura, Akihiko

    2017-08-01

    Programmed cell death 1 (PD-1) is highly expressed on exhausted T cells and inhibits T cell activation. Antibodies that block the interaction between PD-1 and its ligand prevent this inhibitory signal and reverse T cell dysfunction, providing beneficial anti-tumor responses in a substantial number of patients. Mechanisms for the induction and maintenance of high PD-1 expression on exhausted T cells have not been fully understood. Utilizing a genome-wide loss-of-function screening method based on the CRISPR-Cas9 system, we identified genes involved in the core fucosylation pathway as positive regulators of cell-surface PD-1 expression. Inhibition of Fut8, a core fucosyltransferase, by genetic ablation or pharmacologic inhibition reduced cell-surface expression of PD-1 and enhanced T cell activation, leading to more efficient tumor eradication. Taken together, our findings suggest that blocking core fucosylation of PD-1 can be a promising strategy for improving anti-tumor immune responses. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  16. Expression of Arabidopsis SHN1 in Indian Mulberry (Morus indica L.) Increases Leaf Surface Wax Content and Reduces Post-harvest Water Loss

    PubMed Central

    Sajeevan, R. S.; Nataraja, Karaba N.; Shivashankara, K. S.; Pallavi, N.; Gurumurthy, D. S.; Shivanna, M. B.

    2017-01-01

    Mulberry (Morus species) leaf is the sole food for monophagous silkworms, Bombyx mori L. Abiotic stresses such as drought, salinity, and high temperature, significantly decrease mulberry productivity and post-harvest water loss from leaves influence silkworm growth and cocoon yield. Leaf surface properties regulate direct water loss through the cuticular layer. Leaf surface waxes, contribute for cuticular resistance and protect mesophyll cells from desiccation. In this study we attempted to overexpress AtSHN1, a transcription factor associated with epicuticular wax biosynthesis to increase leaf surface wax load in mulberry. Agrobacterium mediated in vitro transformation was carried out using hypocotyl and cotyledonary explants of Indian mulberry (cv. M5). Mulberry transgenic plants expressing AtSHN1 displayed dark green shiny appearance with increased leaf surface wax content. Scanning electron microscopy (SEM) and gas chromatograph–mass spectrometry (GC-MS) analysis showed change in pattern of surface wax deposition and significant change in wax composition in AtSHN1 overexpressors. Increased wax content altered leaf surface properties as there was significant difference in water droplet contact angle and diameter between transgenic and wild type plants. The transgenic plants showed significant improvement in leaf moisture retention capacity even 5 h after harvest and there was slow degradation of total buffer soluble protein in detached leaves compared to wild type. Silkworm bioassay did not indicate any undesirable effects on larval growth and cocoon yield. This study demonstrated that expression of AtSHN1, can increase surface wax load and reduce the post-harvest water loss in mulberry. PMID:28421085

  17. Expression of Arabidopsis SHN1 in Indian Mulberry (Morus indica L.) Increases Leaf Surface Wax Content and Reduces Post-harvest Water Loss.

    PubMed

    Sajeevan, R S; Nataraja, Karaba N; Shivashankara, K S; Pallavi, N; Gurumurthy, D S; Shivanna, M B

    2017-01-01

    Mulberry (Morus species) leaf is the sole food for monophagous silkworms, Bombyx mori L. Abiotic stresses such as drought, salinity, and high temperature, significantly decrease mulberry productivity and post-harvest water loss from leaves influence silkworm growth and cocoon yield. Leaf surface properties regulate direct water loss through the cuticular layer. Leaf surface waxes, contribute for cuticular resistance and protect mesophyll cells from desiccation. In this study we attempted to overexpress AtSHN1, a transcription factor associated with epicuticular wax biosynthesis to increase leaf surface wax load in mulberry. Agrobacterium mediated in vitro transformation was carried out using hypocotyl and cotyledonary explants of Indian mulberry (cv. M5). Mulberry transgenic plants expressing AtSHN1 displayed dark green shiny appearance with increased leaf surface wax content. Scanning electron microscopy (SEM) and gas chromatograph-mass spectrometry (GC-MS) analysis showed change in pattern of surface wax deposition and significant change in wax composition in AtSHN1 overexpressors. Increased wax content altered leaf surface properties as there was significant difference in water droplet contact angle and diameter between transgenic and wild type plants. The transgenic plants showed significant improvement in leaf moisture retention capacity even 5 h after harvest and there was slow degradation of total buffer soluble protein in detached leaves compared to wild type. Silkworm bioassay did not indicate any undesirable effects on larval growth and cocoon yield. This study demonstrated that expression of AtSHN1, can increase surface wax load and reduce the post-harvest water loss in mulberry.

  18. Swimming Motility Reduces Deposition to Silica Surfaces

    SciTech Connect

    Lu, Nanxi; Massoudieh, Arash; Liang, Xiaomeng; Hu, Dehong; Kamai, Tamir; Ginn, Timothy R.; Zilles, Julie L.; Nguyen, Thanh H.

    2015-01-01

    The role of swimming motility on bacterial transport and fate in porous media was evaluated. We present microscopic evidence showing that strong swimming motility reduces attachment of Azotobacter vinelandii cells to silica surfaces. Applying global and cluster statistical analyses to microscopic videos taken under non-flow conditions, wild type, flagellated A. vinelandii strain DJ showed strong swimming ability with an average speed of 13.1 μm/s, DJ77 showed impaired swimming averaged at 8.7 μm/s, and both the non-flagellated JZ52 and chemically treated DJ cells were non-motile. Quantitative analyses of trajectories observed at different distances above the collector of a radial stagnation point flow cell (RSPF) revealed that both swimming and non-swimming cells moved with the flow when at a distance of at least 20 μm from the collector surface. Near the surface, DJ cells showed both horizontal and vertical movement diverging them from reaching surfaces, while chemically treated DJ cells moved with the flow to reach surfaces, suggesting that strong swimming reduced attachment. In agreement with the RSPF results, the deposition rates obtained for two-dimensional multiple-collector micromodels were also lowest for DJ, while DJ77 and JZ52 showed similar values. Strong swimming specifically reduced deposition on the upstream surfaces of the micromodel collectors.

  19. Mutation Linked to Autosomal Dominant Nocturnal Frontal Lobe Epilepsy Reduces Low-Sensitivity α4β2, and Increases α5α4β2, Nicotinic Receptor Surface Expression

    PubMed Central

    Nichols, Weston A.; Henderson, Brandon J.; Marotta, Christopher B.; Yu, Caroline Y.; Richards, Chris; Dougherty, Dennis A.; Lester, Henry A.

    2016-01-01

    A number of mutations in α4β2-containing (α4β2*) nicotinic acetylcholine (ACh) receptors (nAChRs) are linked to autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE), including one in the β2 subunit called β2V287L. Two α4β2* subtypes with different subunit stoichiometries and ACh sensitivities co-exist in the brain, a high-sensitivity subtype with (α4)2(β2)3 subunit stoichiometry and a low-sensitivity subtype with (α4)3(β2)2 stoichiometry. The α5 nicotinic subunit also co-assembles with α4β2 to form a high-sensitivity α5α4β2 nAChR. Previous studies suggest that the β2V287L mutation suppresses low-sensitivity α4β2* nAChR expression in a knock-in mouse model and also that α5 co-expression improves the surface expression of ADNFLE mutant nAChRs in a cell line. To test these hypotheses further, we expressed mutant and wild-type (WT) nAChRs in oocytes and mammalian cell lines, and measured the effects of the β2V287L mutation on surface receptor expression and the ACh response using electrophysiology, a voltage-sensitive fluorescent dye, and superecliptic pHluorin (SEP). The β2V287L mutation reduced the EC50 values of high- and low-sensitivity α4β2 nAChRs expressed in Xenopus oocytes for ACh by a similar factor and suppressed low-sensitivity α4β2 expression. In contrast, it did not affect the EC50 of α5α4β2 nAChRs for ACh. Measurements of the ACh responses of WT and mutant nAChRs expressed in mammalian cell lines using a voltage-sensitive fluorescent dye and whole-cell patch-clamping confirm the oocyte data. They also show that, despite reducing the maximum response, β2V287L increased the α4β2 response to a sub-saturating ACh concentration (1 μM). Finally, imaging SEP-tagged α5, α4, β2, and β2V287L subunits showed that β2V287L reduced total α4β2 nAChR surface expression, increased the number of β2 subunits per α4β2 receptor, and increased surface α5α4β2 nAChR expression. Thus, the β2V287L mutation alters the subunit

  20. Increased susceptibility of CD4+ T cells from elderly individuals to HIV-1 infection and apoptosis is associated with reduced CD4 and enhanced CXCR4 and FAS surface expression levels.

    PubMed

    Heigele, Anke; Joas, Simone; Regensburger, Kerstin; Kirchhoff, Frank

    2015-10-09

    Elderly HIV-1 infected individuals progress to AIDS more frequently and rapidly than people becoming infected at a young age. To identify possible reasons for these differences in clinical progression, we performed comprehensive phenotypic analyses of CD4+ T cells from uninfected young and elderly individuals, and examined their susceptibility to HIV-1 infection and programmed death. Peripheral blood mononuclear cells (PBMCs) from older people contain an increased percentage of central memory and Th17 CD4+ T cells that are main target cells of HIV-1 and strongly reduced proportions of naïve T cells that are poorly susceptible to HIV-1. Unstimulated T cells from elderly individuals expressed higher levels of activation markers, death receptors, and the viral CXCR4 co-receptor than those from young individuals but responded poorly to stimulation. CD4+ T cells from older individuals were highly susceptible to CXCR4- and CCR5-tropic HIV-1 infection but produced significantly lower quantities of infectious virus than cells from young individuals because they were highly prone to apoptosis and thus presumably had a very short life span. The increased susceptibility of T cells from the elderly to HIV-1 infection correlated directly with CXCR4 and inversely with CD4 expression. The levels of apoptosis correlated with the cell surface expression of FAS but not with the expression of programmed death receptor 1 (PD1) or tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Increased levels of activated and highly susceptible HIV-1 target cells, reduced CD4 and enhanced CXCR4 cell surface expression, together with the high susceptibility to FAS-induced programmed cell death may contribute to the rapid CD4+ T cell depletion and accelerated clinical course of infection in elderly HIV-1-infected individuals.

  1. Developmental Hypothyroidism Reduces the Expression of ...

    EPA Pesticide Factsheets

    Disruption of thyroid hormone (TH) is a known effect of environmental contaminants. Neurotrophins including brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) have been implicated in brain dysfunction resulting from severe developmental TH insufficiency. Neurotrophins are also implicated in activity-dependent plasticity, a process critical for appropriate use-dependent connectivity in the developing brain and for memory formation in the adult. This study examined activity-induced expression of neurotrophin gene products in the hippocampus using the long-term potentiation (LTP) after developmental hypothyroidism induced by propylthiouracil (PTU). Pregnant rats were exposed to PTU (0 or I0ppm) via the drinking water from early gestation to weaning. Adult male offspring were anesthetized with urethane and implanted with electrodes in the dentate gyrus (00) and perforant path (PP). LTP was induced by PP stimulation and responses from 00 were monitored at 15m intervals until sacrifice of the animals 5 h later. The 00 was dissected from the stimulated and nonstimulated hemispheres for rtPCR analysis of the neurotrophins Bdnf, Ngf, Ntf3 and related genes Egrl, Arc, Klf9. We found no PTU-induced difference in basal levels of expression of any of these genes in the nonstimulated 00. LTP increased expression of Bdnf, Ngf, Arc and Klj9 in the control DG, and reduced expression of Ntf3. LTP in DG from PTU animals failed to increase expression of Bdnf,

  2. Solid Surface Combustion at Reduced Gravity

    NASA Technical Reports Server (NTRS)

    Altenkirch, R. A.

    1985-01-01

    The spread of a flame in the gas over the surface of a solid combustible involves in an essential way the transfer of heat from the flame to the solid fuel immediately ahead of it. This heat transfer is affected by the character of the gas phase flame, and so the phenomenon of flame spreading under reduced gravity, in which the flow is generated by gasification of the solid combustible, is apt to be different from what occurs under the Earth's normal gravitational acceleration where the flow is largely buoyancy driven. An experiment is being designed for the Middeck of the Space Shuttle to aid us in understanding the process of flame spreading in the absence of a buoyancy driven flow. A chamber approximately 0.35 cu.m. in volume is to contain either a thin sample of a cellulosic material or a thick sample of polymethyl-methacrylate and an oxidizing environment of O2 and N2. Samples will be ignited at one end, and the ensuing flame spread will be filmed. The spread rate can be determined from the films, and surface and gas-phase temperatures just above the surface will also be recorded. These data will help to clarify the mechanism of forward heat transfer in the low gravity flames.

  3. DNA vaccines expressing the duck hepatitis B virus surface proteins lead to reduced numbers of infected hepatocytes and protect ducks against the development of chronic infection in a virus dose-dependent manner.

    PubMed

    Miller, Darren S; Kotlarski, Ieva; Jilbert, Allison R

    2006-07-20

    We tested the efficacy of DNA vaccines expressing the duck hepatitis B virus (DHBV) pre-surface (pre-S/S) and surface (S) proteins in modifying the outcome of infection in 14-day-old ducks. In two experiments, Pekin Aylesbury ducks were vaccinated on days 4 and 14 of age with plasmid DNA vaccines expressing either the DHBV pre-S/S or S proteins, or the control plasmid vector, pcDNA1.1Amp. All ducks were then challenged intravenously on day 14 of age with 5 x 10(7) or 5 x 10(8) DHBV genomes. Levels of initial DHBV infection were assessed using liver biopsy tissue collected at day 4 post-challenge (p.c.) followed and immunostained for DHBV surface antigen to determine the percentage of infected hepatocytes. All vector vaccinated ducks challenged with 5 x 10(7) and 5 x 10(8) DHBV genomes had an average of 3.21% and 20.1% of DHBV-positive hepatocytes respectively at day 4 p.c. and 16 out of 16 ducks developed chronic DHBV infection. In contrast, pre-S/S and S vaccinated ducks challenged with 5 x 10(7) DHBV genomes had reduced levels of initial infection with an average of 1.38% and 1.93% of DHBV-positive hepatocytes at day 4 p.c. respectively and 10 of 18 ducks were protected against chronic infection. The pre-S/S and the S DNA vaccinated ducks challenged with 5 x 10(8) DHBV genomes had an average of 31.5% and 9.2% of DHBV-positive hepatocytes on day 4 p.c. respectively and only 4 of the 18 vaccinated ducks were protected against chronic infection. There was no statistically significant difference in the efficacy of the DHBV pre-S/S or S DNA vaccines. In conclusion, vaccination of young ducks with DNA vaccines expressing the DHBV pre-S/S and S proteins induced rapid immune responses that reduced the extent of initial DHBV infection in the liver and prevented the development of chronic infection in a virus dose-dependent manner.

  4. Structural features affecting variant surface glycoprotein expression in Trypanosoma brucei.

    PubMed

    Wang, Jun; Böhme, Ulrike; Cross, George A M

    2003-05-01

    The glycosylphosphatidylinositol (GPI)-anchored variant surface glycoprotein (VSG) of Trypanosoma brucei is the most abundant GPI-anchored protein expressed on any cell, and is an essential virulence factor. To determine what structural features affect efficient expression of VSG, we made a series of mutations in two VSGs. Inserting 18 amino acids, between the amino- and carboxy-terminal domains, reduced the expression of VSG 221 to about 3% of the wild-type level. When this insertion was combined with deletion of the single carboxy-terminal subdomain, expression was reduced a further three-fold. In VSG 117, which contains two carboxy-terminal subdomains, point mutation of the intervening N-glycosylation site reduced expression about 15-fold. Deleting the most carboxy-terminal subdomain and intervening region, including the N-glycosylation site, reduced expression to 15-20% of wild type VSG, and deletion of both subdomains reduced expression to <1%. Despite their low abundance, all VSG mutants were GPI anchored on the cell surface. Our results suggest that, for a protein to be efficiently displayed on the surface of bloodstream-form T. brucei, it is essential that it contains the conserved structural motifs of a T. brucei VSG. Serum resistance-associated protein (SRA), which confers human infectivity on T. brucei, strongly resembles a VSG deletion mutant. Expression of three epitope-tagged versions of SRA in T. brucei conferred total resistance to human serum. SRA possesses a canonical GPI signal sequence, but we were unable to obtain unequivocal evidence for the presence of a GPI anchor. SRA was not released during osmotic lysis, indicating that it is not GPI anchored on the cell surface.

  5. Absence of an N-Linked Glycosylation Motif in the Glycoprotein of the Live-Attenuated Argentine Hemorrhagic Fever Vaccine, Candid #1, Results in Its Improper Processing, and Reduced Surface Expression

    PubMed Central

    Manning, John T.; Seregin, Alexey V.; Yun, Nadezhda E.; Koma, Takaaki; Huang, Cheng; Barral, José; de la Torre, Juan C.; Paessler, Slobodan

    2017-01-01

    Junin virus (JUNV), a highly pathogenic New World arenavirus, is the causative agent of Argentine hemorrhagic fever (AHF). The live-attenuated Candid #1 (Can) strain currently serves as a vaccine for at-risk populations. We have previously shown that the Can glycoprotein (GPC) gene is the primary gene responsible for attenuation in a guinea pig model of AHF. However, the mechanisms through which the GPC contributes to the attenuation of the Can strain remain unknown. A more complete understanding of the mechanisms underlying the attenuation and immunogenicity of the Can strain will potentially allow for the rational design of additional safe and novel vaccines. Here, we provide a detailed comparison of both RNA and protein expression profiles between both inter- and intra-segment chimeric JUNV recombinant clones expressing combinations of genes from the Can strain and the pathogenic Romero (Rom) strain. The recombinant viruses that express Can GPC, which were shown to be attenuated in guinea pigs, displayed different RNA levels and GPC processing patterns as determined by Northern and Western blot analyses, respectively. Analysis of recombinant viruses containing amino acid substitutions selected at different mouse brain passages during the generation of Can revealed that altered Can GPC processing was primarily due to the T168A substitution within G1, which eliminates an N-linked glycosylation motif. Incorporation of the T168A substitution in the Rom GPC resulted in a Can-like processing pattern of Rom GPC. In addition, JUNV GPCs containing T168A substitution were retained within the endoplasmic reticulum (ER) and displayed significantly lower cell surface expression than wild-type Rom GPC. Interestingly, the reversion A168T in Can GPC significantly increased GPC expression at the cell surface. Our results demonstrate that recombinant JUNV (rJUNV) expressing Can GPC display markedly different protein expression and elevated genomic RNA expression when compared to

  6. Nanotexturing of surfaces to reduce melting point.

    SciTech Connect

    Garcia, Ernest J.; Zubia, David; Mireles, Jose; Marquez, Noel; Quinones, Stella

    2011-11-01

    This investigation examined the use of nano-patterned structures on Silicon-on-Insulator (SOI) material to reduce the bulk material melting point (1414 C). It has been found that sharp-tipped and other similar structures have a propensity to move to the lower energy states of spherical structures and as a result exhibit lower melting points than the bulk material. Such a reduction of the melting point would offer a number of interesting opportunities for bonding in microsystems packaging applications. Nano patterning process capabilities were developed to create the required structures for the investigation. One of the technical challenges of the project was understanding and creating the specialized conditions required to observe the melting and reshaping phenomena. Through systematic experimentation and review of the literature these conditions were determined and used to conduct phase change experiments. Melting temperatures as low as 1030 C were observed.

  7. Reducing Sliding Friction with Liquid-Impregnated Surfaces

    NASA Astrophysics Data System (ADS)

    Habibi, Mohammad; Collier, C. Patrick; Boreyko, Jonathan; Nature Inspired Fluids; Interfaces Team; CenterNanophase Materials Sciences Team

    2015-11-01

    Liquid-impregnated surfaces are fabricated by infusing a lubricating liquid into the micro/nano roughness of a textured substrate, such that the surface is slippery for any deposited liquid immiscible with the lubricant. To date, liquid-impregnated surfaces have almost exclusively focused on repelling liquids by minimizing the contact angle hysteresis. Here, we demonstrate that liquid-impregnated surfaces are also capable of reducing sliding friction for solid objects. Ordered arrays of silicon micropillars were infused with lubricating liquids varying in viscosity by two orders of magnitude. Five test surfaces were used: two different micropillared surfaces with and without liquid infusion and a smooth, dry control surface. The static and kinetic coefficients of friction were measured using a polished aluminum cube as the sliding object. Compared to the smooth control surface, the sliding friction was reduced by at least a factor of two on the liquid-impregnated surfaces.

  8. Hemodynamic aspects of reduced platelet adhesion on bioinspired microstructured surfaces.

    PubMed

    Pham, Tam Thanh; Wiedemeier, Stefan; Maenz, Stefan; Gastrock, Gunter; Settmacher, Utz; Jandt, Klaus D; Zanow, Jürgen; Lüdecke, Claudia; Bossert, Jörg

    2016-09-01

    Occlusion by thrombosis due to the absence of the endothelial cell layer is one of the most frequent causes of failure of artificial vascular grafts. Bioinspired surface structures may have a potential to reduce the adhesion of platelets contributing to hemostasis. The aim of this study was to investigate the hemodynamic aspects of platelet adhesion, the main cause of thrombosis, on bioinspired microstructured surfaces mimicking the endothelial cell morphology. We tested the hypothesis that platelet adhesion is statistically significantly reduced on bioinspired microstructured surfaces compared to unstructured surfaces. Platelet adhesion as a function of the microstructure dimensions was investigated under flow conditions on polydimethylsiloxane (PDMS) surfaces by a combined experimental and theoretical approach. Platelet adhesion was statistically significantly reduced (by up to 78%; p≤0.05) on the microstructured PDMS surfaces compared to that on the unstructured control surface. Finite element method (FEM) simulations of blood flow dynamic revealed a micro shear gradient on the microstructure surfaces which plays a pivotal role in reducing platelet adhesion. On the surfaces with the highest differences of the shear stress between the top of the microstructures and the ground areas, platelet adhesion was reduced most. In addition, the microstructures help to reduce the interaction strength between fluid and surfaces, resulting in a larger water contact angle but no higher resistance to flow compared to the unstructured surface. These findings provide new insight into the fundamental mechanisms of reducing platelet adhesion on microstructured bioinspired surfaces and may lay the basis for the development of innovative next generation artificial vascular grafts with reduced risk of thrombosis.

  9. Possible rainfall reduction through reduced surface temperatures due to overgrazing

    NASA Technical Reports Server (NTRS)

    Otterman, J.

    1975-01-01

    Surface temperature reduction in terrain denuded of vegetation (as by overgrazing) is postulated to decrease air convection, reducing cloudiness and rainfall probability during weak meteorological disturbances. By reducing land-sea daytime temperature differences, the surface temperature reduction decreases daytime circulation of thermally driven local winds. The described desertification mechanism, even when limited to arid regions, high albedo soils, and weak meteorological disturbances, can be an effective rainfall reducing process in many areas including most of the Mediterranean lands.

  10. The Arabidopsis Abiotic Stress-Induced TSPO-Related Protein Reduces Cell-Surface Expression of the Aquaporin PIP2;7 through Protein-Protein Interactions and Autophagic Degradation[C][W][OPEN

    PubMed Central

    Hachez, Charles; Veljanovski, Vasko; Reinhardt, Hagen; Guillaumot, Damien; Vanhee, Celine; Chaumont, François

    2014-01-01

    The Arabidopsis thaliana multi-stress regulator TSPO is transiently induced by abiotic stresses. The final destination of this polytopic membrane protein is the Golgi apparatus, where its accumulation is strictly regulated, and TSPO is downregulated through a selective autophagic pathway. TSPO-related proteins regulate the physiology of the cell by generating functional protein complexes. A split-ubiquitin screen for potential TSPO interacting partners uncovered a plasma membrane aquaporin, PIP2;7. Pull-down assays and fluorescence imaging approaches revealed that TSPO physically interacts with PIP2;7 at the endoplasmic reticulum and Golgi membranes in planta. Intriguingly, constitutive expression of fluorescently tagged PIP2;7 in TSPO-overexpressing transgenic lines resulted in patchy distribution of the fluorescence, reminiscent of the pattern of constitutively expressed yellow fluorescent protein-TSPO in Arabidopsis. Mutational stabilization of TSPO or pharmacological inhibition of the autophagic pathway affected concomitantly the detected levels of PIP2;7, suggesting that the complex containing both proteins is degraded through the autophagic pathway. Coexpression of TSPO and PIP2;7 resulted in decreased levels of PIP2;7 in the plasma membrane and abolished the membrane water permeability mediated by transgenic PIP2;7. Taken together, these data support a physiological role for TSPO in regulating the cell-surface expression of PIP2;7 during abiotic stress conditions through protein-protein interaction and demonstrate an aquaporin regulatory mechanism involving TSPO. PMID:25538184

  11. Aircraft surface coatings reduce drag, may protect against corrosion

    SciTech Connect

    Kreitinger, R.L.; Middleton, D.B.

    1982-02-01

    The aerodynamic drag on an airplane is a very important design parameter. However, after exposure to the environment and accidental spills the surface of the airplane may become corroded or erode; thus the drag may change. Researchers at Boeing Commercial Airplane Co. and the NASA-Langley Research Center have been studying the possibility of using smooth surface coatings to help reduce drag and protect the surface of the airplane. Elastomeric polyurethanes on portions of a test airplane have reduced total drag by 0.2% (as compared to a bare surface) at cruise Reynolds number.

  12. Development of antifouling surfaces to reduce bacterial attachment

    NASA Astrophysics Data System (ADS)

    Graham, Mary Viola

    Bacteria are exceptionally good at adhering to surfaces and forming complex structures known as biofilms. This process, known as biofouling, can cause problems for infrastructure (eg, clogging and damaging pipes), for the food industry (eg, contamination of processing surfaces and equipment, and for the medical industry (eg, contamination of indwelling medical devices). Accordingly, multiple strategies have been explored to combat biofouling, including chemical modification of surfaces, development of antibiotic coatings, and more recently, the use of engineered surface topography. When designed properly, engineered surface topographies can significantly reduce bacterial surface attachment, ultimately limiting surface colonization. In this work, we hypothesized that the morphology, size, spacing, and surface pre-treatment of topographical features should directly correlate with the size and shape of target organisms, in order to reduce biofouling. Topographical features with size and spacing from 0.25 to 2 mum were fabricated in silicone elastomer and tested against rod shaped bacteria with an average size of 0.5 x 2 mum and spherical bacteria (cocci) ranging from 0.5 - 1 μm in diameter. Antifouling properties of the different topographical features were tested in both static and flow-based assays, and under oxygen plasma-treated (hydrophilic) and untreated (hydrophobic) surface conditions. We found that surface pre-treatment universally affects the ability bacteria to attach to surfaces, while surface topography limits attachment in a manner dependent on the bacterial size/shape and the size/spacing of the topography.

  13. Aerobic Exercise Program Reduces Anger Expression Among Overweight Children

    PubMed Central

    Tkacz, Joseph; Young-Hyman, Deborah; Boyle, Colleen A.; Davis, Catherine L.

    2009-01-01

    This study tested the effect of a structured aerobic exercise program on anger expression in healthy overweight children. Overweight, sedentary children were randomly assigned to an aerobic exercise program or a no-exercise control condition. All children completed the Pediatric Anger Expression Scale at baseline and posttest. Anger Out and Anger Expression scores were lower for the exercise condition at posttest. Fitness improvements contributed significantly to final models, and points earned for adherence correlated negatively with posttest Anger Out. An aerobic exercise program might be an effective strategy to reduce anger expression, including reduction of aggressive behavior, in overweight children. PMID:19168916

  14. Aerobic exercise program reduces anger expression among overweight children.

    PubMed

    Tkacz, Joseph; Young-Hyman, Deborah; Boyle, Collen A; Davis, Catherine L

    2008-11-01

    This study tested the effect of a structured aerobic exercise program on anger expression in healthy overweight children. Overweight sedentary children were randomly assigned to an aerobic exercise program or a no-exercise control condition. All children completed the Pediatric Anger Expression Scale at baseline and posttest. Anger Out and Anger Expression scores were lower for the exercise condition at posttest. Fitness improvements contributed significantly to final models, and points earned for adherence correlated negatively with posttest Anger Out. An aerobic exercise program might be an effective strategy to reduce anger expression, including reduction of aggressive behavior, in overweight children.

  15. Reduced facial expressiveness in Parkinson's disease: A pure motor disorder?

    PubMed

    Ricciardi, Lucia; Bologna, Matteo; Morgante, Francesca; Ricciardi, Diego; Morabito, Bruno; Volpe, Daniele; Martino, Davide; Tessitore, Alessandro; Pomponi, Massimiliano; Bentivoglio, Anna Rita; Bernabei, Roberto; Fasano, Alfonso

    2015-11-15

    Impaired emotional facial expressiveness is an important feature in Parkinson's disease (PD). Although there is evidence of a possible relationship between reduced facial expressiveness and altered emotion recognition or imagery in PD, it is unknown whether other aspects of the emotional processing, such as subjective emotional experience (alexithymia), might influence hypomimia in this condition. In this study wee aimed to investigate possible relationship between reduced facial expressiveness and altered emotion processing (including facial recognition and alexithymia) in patients with PD. Forty PD patients and seventeen healthy controls were evaluated. Facial expressiveness was rated on video recordings, according to the UPDRS-III item 19 and using an ad hoc scale assessing static and dynamic facial expression and posed emotions. Six blind raters evaluated the patients' videos. Emotion facial recognition was tested using the Ekman Test; alexithymia was assessed using Toronto Alexithymia Scale (TAS-20). PD patients had a significantly reduced static and dynamic facial expressiveness and a deficit in posing happiness and surprise. They performed significantly worse than healthy controls in recognizing surprise (p=0.03). The Ekman total score positively correlated with the global expressiveness (R^2=0.39, p=0.01) and with the expressiveness of disgust (R^2=0.32, p=0.01). The occurrence of alexithymia was not different between PD patients and HC; however, a significant negative correlation between the expressiveness of disgust was found for a subscore of TAS (R^2=-.447, p=0.007). Reduced facial expressiveness in PD may be in part related to difficulties with emotional recognition in a context of an unimpaired subjective emotional experience. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Emotional facial expressions reduce neural adaptation to face identity.

    PubMed

    Gerlicher, Anna M V; van Loon, Anouk M; Scholte, H Steven; Lamme, Victor A F; van der Leij, Andries R

    2014-05-01

    In human social interactions, facial emotional expressions are a crucial source of information. Repeatedly presented information typically leads to an adaptation of neural responses. However, processing seems sustained with emotional facial expressions. Therefore, we tested whether sustained processing of emotional expressions, especially threat-related expressions, would attenuate neural adaptation. Neutral and emotional expressions (happy, mixed and fearful) of same and different identity were presented at 3 Hz. We used electroencephalography to record the evoked steady-state visual potentials (ssVEP) and tested to what extent the ssVEP amplitude adapts to the same when compared with different face identities. We found adaptation to the identity of a neutral face. However, for emotional faces, adaptation was reduced, decreasing linearly with negative valence, with the least adaptation to fearful expressions. This short and straightforward method may prove to be a valuable new tool in the study of emotional processing.

  17. Surface expression of the Chicxulub crater

    PubMed

    Pope, K O; Ocampo, A C; Kinsland, G L; Smith, R

    1996-06-01

    Analyses of geomorphic, soil, and topographic data from the northern Yucatan Peninsula, Mexico, confirm that the buried Chicxulub impact crater has a distinct surface expression and that carbonate sedimentation throughout the Cenozoic has been influenced by the crater. Late Tertiary sedimentation was mostly restricted to the region within the buried crater, and a semicircular moat existed until at least Pliocene time. The topographic expression of the crater is a series of features concentric with the crater. The most prominent is an approximately 83-km-radius trough or moat containing sinkholes (the Cenote ring). Early Tertiary surfaces rise abruptly outside the moat and form a stepped topography with an outer trough and ridge crest at radii of approximately 103 and approximately 129 km, respectively. Two discontinuous troughs lie within the moat at radii of approximately 41 and approximately 62 km. The low ridge between the inner troughs corresponds to the buried peak ring. The moat corresponds to the outer edge of the crater floor demarcated by a major ring fault. The outer trough and the approximately 62-km-radius inner trough also mark buried ring faults. The ridge crest corresponds to the topographic rim of the crater as modified by postimpact processes. These interpretations support previous findings that the principal impact basin has a diameter of approximately 180 km, but concentric, low-relief slumping extends well beyond this diameter and the eroded crater rim may extend to a diameter of approximately 260 km.

  18. LIN28A expression reduces sickling of cultured human erythrocytes.

    PubMed

    de Vasconcellos, Jaira F; Fasano, Ross M; Lee, Y Terry; Kaushal, Megha; Byrnes, Colleen; Meier, Emily R; Anderson, Molly; Rabel, Antoinette; Braylan, Raul; Stroncek, David F; Miller, Jeffery L

    2014-01-01

    Induction of fetal hemoglobin (HbF) has therapeutic importance for patients with sickle cell disease (SCD) and the beta-thalassemias. It was recently reported that increased expression of LIN28 proteins or decreased expression of its target let-7 miRNAs enhances HbF levels in cultured primary human erythroblasts from adult healthy donors. Here LIN28A effects were studied further using erythrocytes cultured from peripheral blood progenitor cells of pediatric subjects with SCD. Transgenic expression of LIN28A was accomplished by lentiviral transduction in CD34(+) sickle cells cultivated ex vivo in serum-free medium. LIN28A over-expression (LIN28A-OE) increased HbF, reduced beta (sickle)-globin, and strongly suppressed all members of the let-7 family of miRNAs. LIN28A-OE did not affect erythroblast differentiation or prevent enucleation, but it significantly reduced or ameliorated the sickling morphologies of the enucleated erythrocytes.

  19. The processing of coreference for reduced expressions in discourse integration.

    PubMed

    Yang, C L; Gordon, P C; Hendrick, R; Wu, J T; Chou, T L

    2001-01-01

    Three reading-time experiments in Chinese are reported that test contrasting views of how pronominal coreference is achieved. On the one hand, studies of reading time and eye tracking suggest that reduced expressions, such as the pronoun he, serve as critical links to integrate separate utterances into a coherent model of discourse. On the other hand, probe-word recognition studies indicate that full anaphoric expressions, such as a repeated name, are more readily interpreted than reduced expressions due to their rich lexical information, which provides effective cues to match the representation of the appropriate referent in memory. The results indicate that the ease of integrating the critical referent into a model of discourse is a function of the congruence of lexical, semantic, and discourse features conveyed by a syntactically prominent reduced expression within linguistic input. This pattern supports the view that a reduced expression is interpreted on-line and indeed plays a critical role in promoting discourse coherence by facilitating the semantic integration of separate utterances.

  20. Surface nanocrystallization of stainless steel for reduced biofilm adherence

    NASA Astrophysics Data System (ADS)

    Yu, Bin; Davis, Elisabeth M.; Hodges, Robert S.; Irvin, Randall T.; Li, D. Y.

    2008-08-01

    Stainless steel is one of the most common metallic biomedical materials. For medical applications, its resistance to the adherence of biofilms is of importance to the elimination or minimization of bacterial infections. In this study, we demonstrate the effectiveness of a process combining surface nanocrystallization and thermal oxidation (or a recovery heat treatment in air) for reducing the biofilm's adherence to stainless steel. During this treatment, a target surface was sandblasted and the resultant dislocation cells in the surface layer were turned into nanosized grains by a subsequent recovery treatment in air. This process generated a more protective oxide film that blocked the electron exchange or reduced the surface activity more effectively. As a result, the biofilm's adherence to the treated surface was markedly minimized. A synthetic peptide was utilized as a substitute of biofilms to evaluate the adhesion between a treated steel surface and biofilms using an atomic force microscope (AFM) through measuring the adhesive force between the target surface and a peptide-coated AFM tip. It was shown that the adhesive force decreased with a decrease in the grain size of the steel. The corresponding surface electron work function (EWF) of the steel was also measured, which showed a trend of variation in EWF with the grain size, consistent with corresponding changes in the adhesive force.

  1. Zwitteration: Coating Surfaces with Zwitterionic Functionality to Reduce Nonspecific Adsorption

    PubMed Central

    2015-01-01

    Coating surfaces with thin or thick films of zwitterionic material is an effective way to reduce or eliminate nonspecific adsorption to the solid/liquid interface. This review tracks the various approaches to zwitteration, such as monolayer assemblies and polymeric brush coatings, on micro- to macroscopic surfaces. A critical summary of the mechanisms responsible for antifouling shows how zwitterions are ideally suited to this task. PMID:24754399

  2. Mars Express radar collects first surface data

    NASA Astrophysics Data System (ADS)

    2005-08-01

    the middle of August, when the night-time portion of the observations will have almost ended. After that, observation priority will be given to other Mars Express instruments that are best suited to operating in daytime, such as the HRSC camera and Omega mapping spectrometer. However, Marsis will continue its surface and ionospheric investigations in daytime, with ionospheric sounding being reserved for more than 20% of all Mars Express orbits, under all possible Sun illumination conditions. In December, the Mars Express orbit pericentre will enter night-time again. By then, the pericentre will have moved closer to the south pole, allowing Marsis to carry out optimal probing of the subsurface once again, this time in the southern hemisphere. Note to editors The first commissioning phase was given over to testing the Marsis electronics and software and the two 20m-long antennas (dipole). The second commissioning phase, lasting about ten days, will be spent calibrating the 7m ‘monopole’ antenna. This antenna is to be used in conjunction with the Marsis dipole to correct any surface roughness effects caused by the radio waves emitted by the dipole and reflected by an irregular surface. The monopole will find its best use during investigations of areas where surface roughness is greater. The Marsis instrument was developed within the framework of a Memorandum of Understanding between the Italian Space Agency (ASI) and NASA. It was developed by Alenia Spazio under ASI management and the scientific supervision of University of Rome ‘La Sapienza’, in partnership with the Jet Propulsion Laboratory (JPL) and the University of Iowa. JPL provided the antenna manufactured by Astro Aerospace. It is the first instrument designed to actually look below the surface of Mars. Its major goals are to characterise the subsurface layers of sediments and possibly detect underground water or ice, conduct large-scale altimetry mapping and provide data on the planet’s ionosphere. For

  3. Reduced Ang2 expression in aging endothelial cells

    SciTech Connect

    Hohensinner, P.J.; Ebenbauer, B.; Kaun, C.; Maurer, G.; Huber, K.; Wojta, J.

    2016-06-03

    Aging endothelial cells are characterized by increased cell size, reduced telomere length and increased expression of proinflammatory cytokines. In addition, we describe here that aging reduces the migratory distance of endothelial cells. Furthermore, we observe an increase of the quiescence protein Ang1 and a decrease of the endothelial activation protein Ang2 upon aging. Supplementing Ang2 to aged endothelial cells restored their migratory capacity. We conclude that aging shifts the balance of the Ang1/Ang2 network favouring a quiescent state. Activation of endothelial cells in aging might be necessary to enhance wound healing capacities. -- Highlights: •Endothelial cells display signs of aging before reaching proliferative senescence. •Aging endothelial cells express more angiopoietin 1 and less angiopoietin 2 than young endothelial cells. •Migratory capacity is reduced in aging endothelial cells.

  4. Ni doping on Cu surfaces: Reduced copper resistivity

    SciTech Connect

    Zheng, P. Y.; Deng, R. P.; Gall, D.

    2014-09-29

    The resistivity of 9.3-nm-thick epitaxial and polycrystalline Cu is reduced by 11%–13% when coated with 0.75 nm Ni. Sequential in situ and ex situ transport measurements show that this is due to electron surface scattering which exhibits a specularity p = 0.7 for the Cu-vacuum interface that transitions to completely diffuse (p = 0) when exposed to air. In contrast, Ni-coated surfaces exhibit partial specularity with p = 0.3 in vacuum and p = 0.15 in air, as Cu{sub 2}O formation is suppressed, leading to a smaller surface potential perturbation and a lower density of localized surface states, yielding less diffuse electron scattering.

  5. Textured bearing surface in artificial joints to reduce macrophage activation

    NASA Astrophysics Data System (ADS)

    Nakanishi, Yoshitaka; Nishi, Naoki; Chikaura, Hiroto; Nakashima, Yuta; Miura, Hiromasa; Higaki, Hidehiko; Mizuta, Hiroshi; Iwamoto, Yukihide; Fujiwara, Yukio; Komohara, Yoshihiro; Takeya, Motohiro

    2015-12-01

    Micro slurry-jet erosion has been proposed as a precision machining technique for the bearing surfaces of artificial joints in order to reduce the total amount of polyethylene wear and to enlarge the size of the wear debris. The micro slurry-jet erosion method is a wet blasting technique which uses alumina particles as the abrasive medium along with compressed air and water to create an ideal surface. Pin-on-disc wear tests with multidirectional sliding motion on the textured surface of a \\text{Co}-\\text{Cr}-\\text{Mo} alloy counterface for polyethylene resulted in both a reduction of wear as well as enlargement of the polyethylene debris size. In this study, primary human peripheral blood mononuclear phagocytes were incubated with the debris, and it was elucidated that the wear debris generated on the textured surface regulated secretion of the proinflammatory cytokines IL-6 and TNF-α, indicating a reduction in the induced tissue reaction and joint loosening.

  6. Domestication reduces alternative splicing expression variations in sorghum.

    PubMed

    Ranwez, Vincent; Serra, Audrey; Pot, David; Chantret, Nathalie

    2017-01-01

    Domestication is known to strongly reduce genomic diversity through population bottlenecks. The resulting loss of polymorphism has been thoroughly documented in numerous cultivated species. Here we investigate the impact of domestication on the diversity of alternative transcript expressions using RNAseq data obtained on cultivated and wild sorghum accessions (ten accessions for each pool). In that aim, we focus on genes expressing two isoforms in sorghum and estimate the ratio between expression levels of those isoforms in each accession. Noticeably, for a given gene, one isoform can either be overexpressed or underexpressed in some wild accessions, whereas in the cultivated accessions, the balance between the two isoforms of the same gene appears to be much more homogenous. Indeed, we observe in sorghum significantly more variation in isoform expression balance among wild accessions than among domesticated accessions. The possibility exists that the loss of nucleotide diversity due to domestication could affect regulatory elements, controlling transcription or degradation of these isoforms. Impact on the isoform expression balance is discussed. As far as we know, this is the first time that the impact of domestication on transcript isoform balance has been studied at the genomic scale. This could pave the way towards the identification of key domestication genes with finely tuned isoform expressions in domesticated accessions while being highly variable in their wild relatives.

  7. Domestication reduces alternative splicing expression variations in sorghum

    PubMed Central

    Ranwez, Vincent; Serra, Audrey; Pot, David

    2017-01-01

    Domestication is known to strongly reduce genomic diversity through population bottlenecks. The resulting loss of polymorphism has been thoroughly documented in numerous cultivated species. Here we investigate the impact of domestication on the diversity of alternative transcript expressions using RNAseq data obtained on cultivated and wild sorghum accessions (ten accessions for each pool). In that aim, we focus on genes expressing two isoforms in sorghum and estimate the ratio between expression levels of those isoforms in each accession. Noticeably, for a given gene, one isoform can either be overexpressed or underexpressed in some wild accessions, whereas in the cultivated accessions, the balance between the two isoforms of the same gene appears to be much more homogenous. Indeed, we observe in sorghum significantly more variation in isoform expression balance among wild accessions than among domesticated accessions. The possibility exists that the loss of nucleotide diversity due to domestication could affect regulatory elements, controlling transcription or degradation of these isoforms. Impact on the isoform expression balance is discussed. As far as we know, this is the first time that the impact of domestication on transcript isoform balance has been studied at the genomic scale. This could pave the way towards the identification of key domestication genes with finely tuned isoform expressions in domesticated accessions while being highly variable in their wild relatives. PMID:28886042

  8. Reduced Ang2 expression in aging endothelial cells.

    PubMed

    Hohensinner, P J; Ebenbauer, B; Kaun, C; Maurer, G; Huber, K; Wojta, J

    2016-06-03

    Aging endothelial cells are characterized by increased cell size, reduced telomere length and increased expression of proinflammatory cytokines. In addition, we describe here that aging reduces the migratory distance of endothelial cells. Furthermore, we observe an increase of the quiescence protein Ang1 and a decrease of the endothelial activation protein Ang2 upon aging. Supplementing Ang2 to aged endothelial cells restored their migratory capacity. We conclude that aging shifts the balance of the Ang1/Ang2 network favouring a quiescent state. Activation of endothelial cells in aging might be necessary to enhance wound healing capacities. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. LIN28A Expression Reduces Sickling of Cultured Human Erythrocytes

    PubMed Central

    de Vasconcellos, Jaira F.; Fasano, Ross M.; Lee, Y. Terry; Kaushal, Megha; Byrnes, Colleen; Meier, Emily R.; Anderson, Molly; Rabel, Antoinette; Braylan, Raul; Stroncek, David F.; Miller, Jeffery L.

    2014-01-01

    Induction of fetal hemoglobin (HbF) has therapeutic importance for patients with sickle cell disease (SCD) and the beta-thalassemias. It was recently reported that increased expression of LIN28 proteins or decreased expression of its target let-7 miRNAs enhances HbF levels in cultured primary human erythroblasts from adult healthy donors. Here LIN28A effects were studied further using erythrocytes cultured from peripheral blood progenitor cells of pediatric subjects with SCD. Transgenic expression of LIN28A was accomplished by lentiviral transduction in CD34(+) sickle cells cultivated ex vivo in serum-free medium. LIN28A over-expression (LIN28A-OE) increased HbF, reduced beta (sickle)-globin, and strongly suppressed all members of the let-7 family of miRNAs. LIN28A-OE did not affect erythroblast differentiation or prevent enucleation, but it significantly reduced or ameliorated the sickling morphologies of the enucleated erythrocytes. PMID:25188417

  10. Preconditioning reduces myocardial complement gene expression in vivo.

    PubMed

    Tanhehco, E J; Yasojima, K; McGeer, P L; McGeer, E G; Lucchesi, B R

    2000-09-01

    This investigation examined the effect of preconditioning in an in vivo model of ischemia-reperfusion injury. Anesthetized New Zealand White rabbits underwent 30 min of regional myocardial ischemia followed by 2 h of reperfusion. Hearts preconditioned with two cycles of 5 min ischemia-10 min reperfusion (IPC) or with the ATP-sensitive K (K(ATP)) channel opener, diazoxide (10 mg/kg), exhibited significantly (P < 0.05) smaller infarcts compared with control. These treatments also significantly (P < 0.001 to P < 0.05) reduced C1q, C1r, C3, C8, and C9 mRNA in the areas at risk (AAR). The K(ATP) channel blocker 5-hydroxydecanoate (5-HD; 10 mg/kg) attenuated infarct size reduction elicited by IPC and diazoxide treatment. 5-HD partially reversed the decrease in complement expression caused by IPC but not diazoxide. There were no significant differences in complement gene expression in the nonrisk regions and livers of all groups. Western blot analysis revealed that IPC also reduced membrane attack complex expression in the AAR. The data demonstrate that preconditioning significantly decreases reperfusion-induced myocardial complement expression in vivo.

  11. Reducing measurement scale mismatch to improve surface energy flux estimation

    NASA Astrophysics Data System (ADS)

    Iwema, Joost; Rosolem, Rafael; Rahman, Mostaquimur; Blyth, Eleanor; Wagener, Thorsten

    2016-04-01

    Soil moisture importantly controls land surface processes such as energy and water partitioning. A good understanding of these controls is needed especially when recognizing the challenges in providing accurate hyper-resolution hydrometeorological simulations at sub-kilometre scales. Soil moisture controlling factors can, however, differ at distinct scales. In addition, some parameters in land surface models are still often prescribed based on observations obtained at another scale not necessarily employed by such models (e.g., soil properties obtained from lab samples used in regional simulations). To minimize such effects, parameters can be constrained with local data from Eddy-Covariance (EC) towers (i.e., latent and sensible heat fluxes) and Point Scale (PS) soil moisture observations (e.g., TDR). However, measurement scales represented by EC and PS still differ substantially. Here we use the fact that Cosmic-Ray Neutron Sensors (CRNS) estimate soil moisture at horizontal footprint similar to that of EC fluxes to help answer the following question: Does reduced observation scale mismatch yield better soil moisture - surface fluxes representation in land surface models? To answer this question we analysed soil moisture and surface fluxes measurements from twelve COSMOS-Ameriflux sites in the USA characterized by distinct climate, soils and vegetation types. We calibrated model parameters of the Joint UK Land Environment Simulator (JULES) against PS and CRNS soil moisture data, respectively. We analysed the improvement in soil moisture estimation compared to uncalibrated model simulations and then evaluated the degree of improvement in surface fluxes before and after calibration experiments. Preliminary results suggest that a more accurate representation of soil moisture dynamics is achieved when calibrating against observed soil moisture and further improvement obtained with CRNS relative to PS. However, our results also suggest that a more accurate

  12. Reducing extrinsic damping of surface acoustic waves at gigahertz frequencies

    NASA Astrophysics Data System (ADS)

    Gelda, Dhruv; Sadhu, Jyothi; Ghossoub, Marc G.; Ertekin, Elif; Sinha, Sanjiv

    2016-04-01

    High-frequency surface acoustic waves (SAWs) in the gigahertz range can be generated using absorption from an ultrafast laser in a patterned metallic grating on a substrate. Reducing the attenuation at these frequencies can yield better sensors as well as enable them to better probe phonon and electron-phonon interactions near surfaces. It is not clear from existing experiments which mechanisms dominate damping at high frequencies. We calculate damping times of SAWs due to various mechanisms in the 1-100 GHz range to find that mechanical loading of the grating on the substrate dominates dissipation by radiating energy from the surface into the bulk. To overcome this and enable future measurements to probe intrinsic damping, we propose incorporating distributed acoustic Bragg reflectors in the experimental structure. Layers of alternating materials with contrasting acoustic impedances embedded a wavelength away from the surface serve to reflect energy back to the surface. Using numerical simulations, we show that a single Bragg reflector is sufficient to increase the energy density at the surface by more than five times. We quantify the resulting damping time to find that it is longer than the intrinsic damping time. The proposed structure can enable future measurements of intrinsic damping in SAWs at ˜100 GHz.

  13. Reducing extrinsic damping of surface acoustic waves at gigahertz frequencies

    SciTech Connect

    Gelda, Dhruv Sadhu, Jyothi; Ghossoub, Marc G.; Ertekin, Elif; Sinha, Sanjiv

    2016-04-28

    High-frequency surface acoustic waves (SAWs) in the gigahertz range can be generated using absorption from an ultrafast laser in a patterned metallic grating on a substrate. Reducing the attenuation at these frequencies can yield better sensors as well as enable them to better probe phonon and electron-phonon interactions near surfaces. It is not clear from existing experiments which mechanisms dominate damping at high frequencies. We calculate damping times of SAWs due to various mechanisms in the 1–100 GHz range to find that mechanical loading of the grating on the substrate dominates dissipation by radiating energy from the surface into the bulk. To overcome this and enable future measurements to probe intrinsic damping, we propose incorporating distributed acoustic Bragg reflectors in the experimental structure. Layers of alternating materials with contrasting acoustic impedances embedded a wavelength away from the surface serve to reflect energy back to the surface. Using numerical simulations, we show that a single Bragg reflector is sufficient to increase the energy density at the surface by more than five times. We quantify the resulting damping time to find that it is longer than the intrinsic damping time. The proposed structure can enable future measurements of intrinsic damping in SAWs at ∼100 GHz.

  14. Reduced cortical surface area in adolescents with conduct disorder.

    PubMed

    Sarkar, Sagari; Daly, Eileen; Feng, Yue; Ecker, Christine; Craig, Michael C; Harding, Duncan; Deeley, Quinton; Murphy, Declan G M

    2015-08-01

    Children with conduct disorder (CD) are at increased risk of developing antisocial personality disorder and psychopathy in adulthood. Neuroimaging research has identified abnormal cortical volume (CV) in CD. However, CV comprises two genetically and developmentally separable components: cortical thickness (CT) and surface area (SA). Aim of this study is to explore the relationship between the cortical constituents of CV in boys with CD. We applied FreeSurfer software to structural MRI data to derive measures of CV, CT, and SA in 21 boys with CD and 19 controls. Relationships between these cortical measures were investigated. Boys with CD had significantly reduced CV and SA compared to non-CD boys in ventromedial and dorsolateral prefrontal cortex. We found no significant between-group differences in CT. Reduced prefrontal CV in boys with CD is associated with significantly reduced SA in the same regions. This finding may help to identify specific neurodevelopmental mechanisms underlying cortical deficits observed in CD.

  15. Reducing Surface Clutter in Cloud Profiling Radar Data

    NASA Technical Reports Server (NTRS)

    Tanelli, Simone; Pak, Kyung; Durden, Stephen; Im, Eastwood

    2008-01-01

    An algorithm has been devised to reduce ground clutter in the data products of the CloudSat Cloud Profiling Radar (CPR), which is a nadir-looking radar instrument, in orbit around the Earth, that measures power backscattered by clouds as a function of distance from the instrument. Ground clutter contaminates the CPR data in the lowest 1 km of the atmospheric profile, heretofore making it impossible to use CPR data to satisfy the scientific interest in studying clouds and light rainfall at low altitude. The algorithm is based partly on the fact that the CloudSat orbit is such that the geodetic altitude of the CPR varies continuously over a range of approximately 25 km. As the geodetic altitude changes, the radar timing parameters are changed at intervals defined by flight software in order to keep the troposphere inside a data-collection time window. However, within each interval, the surface of the Earth continuously "scans through" (that is, it moves across) a few range bins of the data time window. For each radar profile, only few samples [one for every range-bin increment ((Delta)r = 240 m)] of the surface-clutter signature are available around the range bin in which the peak of surface return is observed, but samples in consecutive radar profiles are offset slightly (by amounts much less than (Delta)r) with respect to each other according to the relative change in geodetic altitude. As a consequence, in a case in which the surface area under examination is homogenous (e.g., an ocean surface), a sequence of consecutive radar profiles of the surface in that area contains samples of the surface response with range resolution (Delta)p much finer than the range-bin increment ((Delta)p << r). Once the high-resolution surface response has thus become available, the profile of surface clutter can be accurately estimated by use of a conventional maximum-correlation scheme: A translated and scaled version of the high-resolution surface response is fitted to the observed

  16. Placental Dysferlin Expression is Reduced in Severe Preeclampsia

    PubMed Central

    Lang, Christopher T.; Markham, Kara B.; Behrendt, Nicholas J.; Suarez, Adrian A.; Samuels, Philip; Vandre, Dale D.; Robinson, John M.; Ackerman, William E.

    2009-01-01

    Dysferlin (DYSF) and myoferlin (MYOF), members of the ferlin family of membrane proteins, are co-expressed in human placental syncytiotrophoblast (STB). Although the role of these ferlin proteins in the placenta has yet to be established, it has been suggested that DYSF and MYOF may contribute to the stability of the apical STB plasma membrane. The release of STB-derived cellular debris increases in the setting of preeclampsia (PE), suggesting relative destabilization of the hemochorial interface. To test whether PE was associated with alterations in placental expression of DYSF and/or MYOF, a cross-sectional study was performed using specimens of villous placenta collected form women with severe PE (n = 10) and normotensive controls (n = 10). DYSF and MYOF expression were examined using quantitative real-time RT-PCR, immunoblotting, and immunofluorescence labeling of tissue specimens. Placental DYSF expression was 57% lower at the mRNA level (p = 0.03) and 38% lower at the protein level (p = 0.026) in severe PE as compared to normotensive subjects. There were no differences in placental MYOF protein or mRNA expression between these groups. No appreciable changes in the distribution of DYSF or MYOF within placental villli were observed in PE relative to control specimens. We conclude that DYSF expression is reduced in severe PE relative to gestational age-matched controls. As DYSF has a role in membrane repair, these data suggest a role for DYSF in the stability of the apical STB plasma membrane and may account, at least in part, for the increased shedding of microparticles from this membrane in PE. PMID:19545895

  17. Transdermal 17-beta estradiol replacement therapy reduces megakaryocyte GPVI expression.

    PubMed

    Geng, Hongquan; Zhang, Hui; Zhang, Wei; Nieswandt, Bernhard; Bray, Paul F; Leng, Xinghong

    2008-01-01

    The platelet-collagen interaction is a critical early event in arterial thrombus formation, and platelet GPVI is the major activating receptor for collagen. We have previously used a mouse model to demonstrate that the estrogen effects on platelets depend upon the agonist, estrogen formulation and route of administration. In the current study we used a model of transdermal estradiol (E2) administration to ovariectomized mice to address the potential inhibitory effects of E2 on platelet GPVI. Platelet GPVI expression was reduced after transdermal E2 replacement therapy (p reduced GPVI-mediated fibrinogen binding and aggregation were observed in platelets from mice subjected to 9 days or longer of in vivo E2 treatment, but not in platelets from mice treated for 3 days or shorter, suggesting an indirect pathway. Studies with mouse bone marrow revealed that E2 replacement in ovariectomized mice reduces megakaryocyte GPVI expression. This data suggest that transdermal E2 is able to affect centrally on megakaryocyte GPVI to regulate platelet GPVI and function.

  18. Reducing ZnO nanoparticle cytotoxicity by surface modification

    NASA Astrophysics Data System (ADS)

    Luo, Mingdeng; Shen, Cenchao; Feltis, Bryce N.; Martin, Lisandra L.; Hughes, Anthony E.; Wright, Paul F. A.; Turney, Terence W.

    2014-05-01

    Nanoparticulate zinc oxide (ZnO) is one of the most widely used engineered nanomaterials and its toxicology has gained considerable recent attention. A key aspect for controlling biological interactions at the nanoscale is understanding the relevant nanoparticle surface chemistry. In this study, we have determined the disposition of ZnO nanoparticles within human immune cells by measurement of total Zn, as well as the proportions of extra- and intracellular dissolved Zn as a function of dose and surface coating. From this mass balance, the intracellular soluble Zn levels showed little difference in regard to dose above a certain minimal level or to different surface coatings. PEGylation of ZnO NPs reduced their cytotoxicity as a result of decreased cellular uptake arising from a minimal protein corona. We conclude that the key role of the surface properties of ZnO NPs in controlling cytotoxicity is to regulate cellular nanoparticle uptake rather than altering either intracellular or extracellular Zn dissolution.Nanoparticulate zinc oxide (ZnO) is one of the most widely used engineered nanomaterials and its toxicology has gained considerable recent attention. A key aspect for controlling biological interactions at the nanoscale is understanding the relevant nanoparticle surface chemistry. In this study, we have determined the disposition of ZnO nanoparticles within human immune cells by measurement of total Zn, as well as the proportions of extra- and intracellular dissolved Zn as a function of dose and surface coating. From this mass balance, the intracellular soluble Zn levels showed little difference in regard to dose above a certain minimal level or to different surface coatings. PEGylation of ZnO NPs reduced their cytotoxicity as a result of decreased cellular uptake arising from a minimal protein corona. We conclude that the key role of the surface properties of ZnO NPs in controlling cytotoxicity is to regulate cellular nanoparticle uptake rather than

  19. Modulated surface nanostructures for enhanced light trapping and reduced surface reflection of crystalline silicon solar cells

    NASA Astrophysics Data System (ADS)

    Tayagaki, Takeshi; Hoshi, Yusuke; Hirai, Yuji; Matsuo, Yasutaka; Usami, Noritaka

    2016-05-01

    We demonstrated the fabrication of modulated surface nanostructures as a new surface texture design for thin wafer solar cells. Using a combination of conventional alkali etching and colloidal lithography, we fabricated surface textures with micrometer and nanometre scales on a Si substrate. These modulated surface nanostructures exhibit reduced surface reflection in a broad spectral range, compared with conventional micrometer textures. We investigated optical absorption using a rigorous coupled wave analysis simulation, which revealed a significant reduction in surface reflection over a broad spectral range and efficient light trapping (comparable to that of conventional micrometer-scale textures) for the modulated nanostructures. We found that the modulated surface nanostructures have a high potential of improving the performance of thin wafer crystalline Si solar cells.

  20. Dimethylmercury Formation Mediated by Inorganic and Organic Reduced Sulfur Surfaces

    NASA Astrophysics Data System (ADS)

    Jonsson, Sofi; Mazrui, Nashaat M.; Mason, Robert P.

    2016-06-01

    Underlying formation pathways of dimethylmercury ((CH3)2Hg) in the ocean are unknown. Early work proposed reactions of inorganic Hg (HgII) with methyl cobalamin or of dissolved monomethylmercury (CH3Hg) with hydrogen sulfide as possible bacterial mediated or abiotic pathways. A significant fraction (up to 90%) of CH3Hg in natural waters is however adsorbed to reduced sulfur groups on mineral or organic surfaces. We show that binding of CH3Hg to such reactive sites facilitates the formation of (CH3)2Hg by degradation of the adsorbed CH3Hg. We demonstrate that the reaction can be mediated by different sulfide minerals, as well as by dithiols suggesting that e.g. reduced sulfur groups on mineral particles or on protein surfaces could mediate the reaction. The observed fraction of CH3Hg methylated on sulfide mineral surfaces exceeded previously observed methylation rates of CH3Hg to (CH3)2Hg in seawaters and we suggest the pathway demonstrated here could account for much of the (CH3)2Hg found in the ocean.

  1. Thermomechanical Mechanisms of Reducing Ice Adhesion on Superhydrophobic Surfaces.

    PubMed

    Cohen, N; Dotan, A; Dodiuk, H; Kenig, S

    2016-09-20

    Superhydrophobic (SH) coatings have been shown to reduce freezing and ice nucleation rates, by means of low surface energy chemistry tailored with nano/micro roughness. Durability enhancement of SH surfaces is a crucial issue. Consequently, the present research on reducing ice adhesion is based on radiation-induced radical reaction for covalently bonding SiO2 nanoparticles to polymer coatings to obtain durable roughness. Results indicated that the proposed approach resulted in SH surfaces having high contact angles (>155°) and low sliding angles (<5°) with improved durability and transparency. In a subsequent stage, the synthesized SH coating was investigated for its icephobic characteristics using a variety of substrates. Results indicated that supercooled water drops bounced back when impinging on SH polycarbonate substrate and froze on SH copper substrate held at -10 to -30 °C and were easily peeled off when coated by ice formed during exposure to air/supercooled water drops at -20 °C. The ice shear adhesion investigation (at -20 °C) demonstrated reduction of shear adhesion to a variety of SH treated substrates having low thermal expansion coefficient (copper and aluminum) and high thermal expansion coefficient (polycarbonate and poly(methyl methacrylate)). It was concluded that the thermal mismatch between the adhering ice and the various substrates and its resultant interfacial thermal stresses affect the adhesion strength of the ice to the respective substrate.

  2. Dimethylmercury Formation Mediated by Inorganic and Organic Reduced Sulfur Surfaces

    PubMed Central

    Jonsson, Sofi; Mazrui, Nashaat M.; Mason, Robert P.

    2016-01-01

    Underlying formation pathways of dimethylmercury ((CH3)2Hg) in the ocean are unknown. Early work proposed reactions of inorganic Hg (HgII) with methyl cobalamin or of dissolved monomethylmercury (CH3Hg) with hydrogen sulfide as possible bacterial mediated or abiotic pathways. A significant fraction (up to 90%) of CH3Hg in natural waters is however adsorbed to reduced sulfur groups on mineral or organic surfaces. We show that binding of CH3Hg to such reactive sites facilitates the formation of (CH3)2Hg by degradation of the adsorbed CH3Hg. We demonstrate that the reaction can be mediated by different sulfide minerals, as well as by dithiols suggesting that e.g. reduced sulfur groups on mineral particles or on protein surfaces could mediate the reaction. The observed fraction of CH3Hg methylated on sulfide mineral surfaces exceeded previously observed methylation rates of CH3Hg to (CH3)2Hg in seawaters and we suggest the pathway demonstrated here could account for much of the (CH3)2Hg found in the ocean. PMID:27302323

  3. Deep Coherent Vortices and Their Sea Surface Expressions

    NASA Astrophysics Data System (ADS)

    Ienna, Federico; Bashmachnikov, Igor; Dias, Joaquim; Peliz, Alvaro

    2017-04-01

    Mediterranean Water eddies, known as Meddies, are an important dynamic process occurring at depths of 1000-meters in the Northeast Atlantic Ocean. Meddies occur as a direct result of the Mediterranean Outflow exiting through the Gibraltar Strait, and represent a prevalent mechanism that can be found extensively throughout the ocean. Moreover, Meddy cores are known to produce measurable expressions at the sea surface in the form of rotating coherent vortices, not only affecting the sea surface from beneath, but also allowing for the possibility to remotely study these deep phenomena through data gathered at the sea surface. While many past studies have focused on the properties of Meddy cores, only a handful of studies focus on the physical characteristics and behavior of the surface expressions produced. Are Meddy surface expressions different from other like vortices that dominate the physical ocean surface? What are the relationships between deep and surface mechanisms, and do any feedbacks exist? To shed light on these questions, we investigate the relationship between Meddies and their sea-surface expressions through observations using in-situ float and drifter profiles and satellite altimetry. A total of 782 Meddy cores were examined in the Northeast Atlantic using temperature and salinity data obtained by CTD and Argo during the Mecanismos de transporte e de dispersão da Água Mediterrânica no Atlântico Nordeste (MEDTRANS) project, and their corresponding sea-level expressions were geo-temporally matched in satellite altimetry data. We report several statistical properties of the sea-surface expressions of Meddies, including their mean diameter and vertical magnitude, and compare the properties of their surface features to the underlying Meddy cores. We investigate how the deep core affects the surface, and whether surface expressions may in return yield information about the underlying cores. Additionally, we examine the variability of the surface

  4. Containment of a silicone fluid free surface in reduced gravity

    NASA Technical Reports Server (NTRS)

    Pline, A.; Jacobson, T.

    1988-01-01

    In support of the surface tension driven convection experiment planned for flight aboard the Space Shuttle, tests were conducted under reduced gravity in the 2.2-sec drop tower and the 5.0-sec Zero-G facility at the Lewis Research Center. The dynamics of controlling the test fluid, a 10-centistoke viscosity silicone fluid, in a low-gravity environment were investigated using different container designs and barrier coatings. Three container edge designs were tested without a barrier coating: a square edge, a sharp edge with a 45-deg slope, and a saw-tooth edge. All three edge designs were successful in containing the fluid below the edge.

  5. Calreticulin: Roles in Cell-Surface Protein Expression

    PubMed Central

    Jiang, Yue; Dey, Sandeepa; Matsunami, Hiroaki

    2014-01-01

    In order to perform their designated functions, proteins require precise subcellular localizations. For cell-surface proteins, such as receptors and channels, they are able to transduce signals only when properly targeted to the cell membrane. Calreticulin is a multi-functional chaperone protein involved in protein folding, maturation, and trafficking. However, evidence has been accumulating that calreticulin can also negatively regulate the surface expression of certain receptors and channels. In these instances, depletion of calreticulin enhances cell-surface expression and function. In this review, we discuss the role of calreticulin with a focus on its negative effects on the expression of cell-surface proteins. PMID:25230046

  6. Towards Reducing the Data Exfiltration Surface for the Insider Threat

    SciTech Connect

    Schlicher, Bob G; MacIntyre, Lawrence Paul; Abercrombie, Robert K

    2016-01-01

    Unauthorized data exfiltrations from both insiders and outsiders are costly and damaging. Network communication resources can be used for transporting data illicitly out of the enterprise or cloud. Combined with built-in malware copying utilities, we define this as comprising the Data Exfiltration Surface (DXS). For securing valuable data, it is desirable to reduce the DXS and maintain controls on the egress points. Our approach is to host the data in a protected enclave that includes novel software Data Diode (SDD) installed on a secured, border gateway. The SDD allows copying data into the enclave systems but denies data from being copied out. Simultaneously, it permits remote access with remote desktop and console applications. Our tests demonstrate that we are able to effectively reduce the DXS and we are able to protect data from being exfiltrated through the use of the SDD.

  7. Peripherally expressed neprilysin reduces brain amyloid burden: A novel approach for treating Alzheimer’s disease

    PubMed Central

    Guan, Hanjun; Liu, Yinxing; Daily, Abigail; Police, Sara; Kim, Myung-Hee; Oddo, Salvatore; LaFerla, Frank M.; Pauly, James R.; Murphy, M. Paul; Hersh, Louis B.

    2009-01-01

    A number of therapeutic strategies for treating Alzheimer’s disease have focused on reducing amyloid burden in the brain. Amongst these approaches, the expression of amyloid β peptide (Aβ)-degrading enzymes in the brain has been shown to be effective, but to date not practical for treating patients. We report here a novel strategy for lowering amyloid burden in the brain by peripherally expressing the Aβ-degrading enzyme neprilysin on leukocytes in the 3×Tg-AD mouse model of Alzheimer’s disease. Through transplantation of lentivirus transduced bone marrow cells, the Aβ-degrading protease neprilysin was expressed on the surface of leukocytes. This peripheral neprilysin reduced soluble brain amyloid β peptide levels by ~30% and lowered the accumulation of amyloid β peptides by 50–60% when transplantation was performed at both young and early adult age. In addition, peripheral neprilysin expression reduced amyloid dependent performance deficits as measured by the Morris Water Maze. Unlike other methods designed to lower amyloid β peptide levels in blood, which cause a net increase in peptide, neprilysin expression results in the catabolism of the amyloid β peptide to small innocuous peptide fragments. These findings demonstrate that peripherally expressed neprilysin, and likely other amyloid β peptide degrading enzymes, has the potential for being utilized as a therapeutic approach to prevent and treat Alzheimer’s disease and suggest this approach should be further explored. PMID:19021293

  8. Recovery from Friend disease in mice with reduced major histocompatibility complex class I expression.

    PubMed Central

    Hasenkrug, K J; Sprangrude, G J; Nishio, J; Brooks, D M; Chesebro, B

    1994-01-01

    Mice homozygous for the b allele of the MHC gene, H-2D, have a high incidence of recovery from Friend virus infections, while mice heterozygous for the b allele at H-2D have a very low incidence of recovery. Previous experiments indicated that the low recovery rates associated with heterozygosity at H-2D might be related to a gene dosage effect requiring the expression of two H-2Db alleles for high recovery. We investigated the effects of reduced H-2Db expression on recovery from Friend disease by using H-2b homozygous mice carrying a single beta 2-microglobulin gene disruption. These mice had reductions in cell surface H-2Db expression comparable to those of H-2Da/b heterozygotes. Numerous cell types with various levels of H-2Db expression were examined, and in each case, the expression levels in the beta 2-microglobulin mutants closely reflected those observed in the H-2Da/b heterozygotes. We found, however, that reduced expression did not affect recovery from Friend disease, indicating that heterozygous levels of H-2Db expression are sufficient for the high-recovery phenotype previously associated only with H-2Db homozygotes. PMID:8138991

  9. Nitrate reducing activity pervades surface waters during upwelling

    NASA Astrophysics Data System (ADS)

    Fernandes, Sheryl Oliveira; Halarnekar, Reena; Malik, Ashish; Vijayan, Vijitha; Varik, Sandesh; Kumari, Ritu; V. K., Jineesh; Gauns, Manguesh U.; Nair, Shanta; LokaBharathi, P. A.

    2014-09-01

    Nitrate reducing activity (NRA) is known to be mediated by microaerophilic to anaerobic bacteria and generally occurs in the sub-surface waters. However, we hypothesize that NRA could become prominent in the surface waters during upwelling. Hence, we examined nitrification and nitrate reduction along with hydrographic and environmental parameters off Trivandrum and Kochi, south-west-India in June 2010. Shoaling isolines of temperature, density, and nutrients revealed the onset of upwelling off Trivandrum. Shoaling of these signatures was absent in the northern transect off Kochi. The degree of nutrient consumption (DNC) was low emphasizing the presence of newly upwelled water off Trivandrum. A significant increase in NRA (df = 1, p < 0.05) was observed off Trivandrum than at Kochi. Moreover, as hypothesized, NRA at Trivandrum was pronounced at the surface with a maximum rate of 0.85 (± 0.02) μmol L1 h- 1 nearshore which was ~ 29 × higher than that at Kochi. Further, an inverse relationship between NRA and NO3- concentration (n = 34, r = - 0.415, p < 0.01) suggested transformation of the upwelled nutrient. Nitrification/NRA was ~ 10 × lower at 0.28 off Trivandrum indicating a discernible shift towards reduction. Such contribution from bacterial activity could be a response towards restoration of homeostasis.

  10. Surface Expression of ω-Transaminase in Escherichia coli

    PubMed Central

    Gustavsson, Martin; Muraleedharan, Madhu Nair

    2014-01-01

    Chiral amines are important for the chemical and pharmaceutical industries, and there is rapidly growing interest to use transaminases for their synthesis. Since the cost of the enzyme is an important factor for process economy, the use of whole-cell biocatalysts is attractive, since expensive purification and immobilization steps can be avoided. Display of the protein on the cell surface provides a possible way to reduce the mass transfer limitations of such biocatalysts. However, transaminases need to dimerize in order to become active, and furthermore, they require the cofactor pyridoxal phosphate; consequently, successful transaminase surface expression has not been reported thus far. In this work, we produced an Arthrobacter citreus ω-transaminase in Escherichia coli using a surface display vector based on the autotransporter adhesin involved in diffuse adherence (AIDA-I), which has previously been used for display of dimeric proteins. The correct localization of the transaminase in the E. coli outer membrane and its orientation toward the cell exterior were verified. Furthermore, transaminase activity was detected exclusively in the outer membrane protein fraction, showing that successful dimerization had occurred. The transaminase was found to be present in both full-length and proteolytically degraded forms. The removal of this proteolysis is considered to be the main obstacle to achieving sufficient whole-cell transaminase activity. PMID:24487538

  11. Sulfur passivation of GaAs surfaces: A model for reduced surface recombination without band flattening

    NASA Astrophysics Data System (ADS)

    Spindt, C. J.; Spicer, W. E.

    1989-10-01

    It has been shown by several workers that the passivation of GaAs surfaces using sulfides results in a large reduction in the surface recombination velocity accompanied by an increase in the band bending on n-type samples. This apparently contradictory pair of results leads to the suggestion that the responsible electronic states are a midgap donor compensated by an acceptor near the valence-band maximum. We explore the consequences of such a model, particularly when the midgap state is assumed to be a double donor. In the double donor case, simple qualitative arguments indicate that the surface recombination velocity can be reduced by a factor much greater than the reduction in surface-state density. The model is consistent with observations made using a variety of experimental techniques. A correlation between the electronic states and surface chemistry is made, and the As and Ga antisite defects are discussed as candidates for the donor and acceptor states.

  12. Astaxanthin reduces matrix metalloproteinase expression in human chondrocytes.

    PubMed

    Chen, Wei-Ping; Xiong, Yan; Shi, Yong-Xiang; Hu, Peng-Fei; Bao, Jia-Peng; Wu, Li-Dong

    2014-03-01

    Astaxanthin is a red carotenoid pigment which exerts multiple biological activities. However, little is known about the effects of astaxanthin on matrix metalloproteinases (MMPs) in OA. The present study investigated the effects of astaxanthin on MMPs in human chondrocytes. Human chondrocytes were pretreated with astaxanthin at 1, 10 or 50μM, then, cells were stimulated with IL-1β (10ng/ml) for 24h. MMP-1, MMP-3 and MMP-13 were observed. We found that astaxanthin reduced the expression of MMP-1, MMP-3 and MMP-13 as well as the phosphorylation of two mitogen-activated protein kinases (MAPK) (p38 and ERK1/2) in IL-1β-stimulated chondrocytes. Astaxanthin also blocked the IκB-α degradation. These results suggest that astaxanthin may be beneficial in the treatment of OA. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. Thrombospondin-4 reduces binding affinity of [3H]-gabapentin to calcium-channel α2δ-1-subunit but does not interact with α2δ-1 on the cell-surface when co-expressed

    PubMed Central

    Lana, Beatrice; Page, Karen M.; Kadurin, Ivan; Ho, Shuxian; Nieto-Rostro, Manuela; Dolphin, Annette C.

    2016-01-01

    The α2δ proteins are auxiliary subunits of voltage-gated calcium channels, and influence their trafficking and biophysical properties. The α2δ ligand gabapentin interacts with α2δ-1, and inhibits calcium channel trafficking. However, α2-1 has also been proposed to play a synaptogenic role, independent of calcium channel function. In this regard, α2δ-1 was identified as a ligand of thrombospondins, with the interaction involving the thrombospondin synaptogenic domain and the α2δ-1 von-Willebrand-factor domain. Co-immunoprecipitation between α2δ-1 and the synaptogenic domain of thrombospondin-2 was prevented by gabapentin. We therefore examined whether interaction of thrombospondin with α2δ-1 might reciprocally influence 3H-gabapentin binding. We concentrated on thrombospondin-4, because, like α2δ-1, it is upregulated in neuropathic pain models. We found that in membranes from cells co-transfected with α2δ-1 and thrombospondin-4, there was a Mg2+ -dependent reduction in affinity of 3H-gabapentin binding to α2δ-1. This effect was lost for α2δ-1 with mutations in the von-Willebrand-factor-A domain. However, the effect on 3H-gabapentin binding was not reproduced by the synaptogenic EGF-domain of thrombospondin-4. Partial co-immunoprecipitation could be demonstrated between thrombospondin-4 and α2δ-1 when co-transfected, but there was no co-immunoprecipitation with thrombospondin-4-EGF domain. Furthermore, we could not detect any association between these two proteins on the cell-surface, indicating the demonstrated interaction occurs intracellularly. PMID:27076051

  14. Expressive writing as a brief intervention for reducing drinking intentions.

    PubMed

    Young, Chelsie M; Rodriguez, Lindsey M; Neighbors, Clayton

    2013-12-01

    The present study examined the effectiveness of expressive writing in reducing drinking behavior. We expected that students prompted to write about negative drinking experiences would show greater decreases in future drinking intentions compared to the neutral and the positive writing conditions. We also expected that decreases in drinking intentions following the writing prompts might differ based on current drinking and AUDIT scores. Participants included 200 (76% female) undergraduates who completed measures of their current drinking behavior. They were then randomly assigned to either write about: a time when they had a lot to drink that was a good time (Positive); a time when they had a lot to drink that was a bad time (Negative); or their first day of college (Neutral), followed by measures assessing intended drinking over the next three months. Results revealed that participants intended to drink significantly fewer drinks per week and engage in marginally fewer heavy drinking occasions after writing about a negative drinking occasion when compared to control. Interactions provided mixed findings suggesting that writing about a positive event was associated with higher drinking intentions for heavier drinkers. Writing about a negative event was associated with higher intentions among heavier drinkers, but lower intentions among those with higher AUDIT scores. This research builds on previous expressive writing interventions by applying this technique to undergraduate drinkers. Preliminary results provide some support for this innovative strategy but also suggest the need for further refinement, especially with heavier drinkers. © 2013.

  15. Expressive Writing as a Brief Intervention for Reducing Drinking Intentions

    PubMed Central

    Young, Chelsie M.; Rodriguez, Lindsey M.; Neighbors, Clayton

    2013-01-01

    The present study examined the effectiveness of expressive writing in reducing drinking behavior. We expected that students prompted to write about negative drinking experiences would show greater decreases in future drinking intentions compared to the neutral and the positive writing conditions. We also expected that decreases in drinking intentions following the writing prompts might differ based on current drinking and AUDIT scores. Participants included 200 (76% female) undergraduates who completed measures of their current drinking behavior. They were then randomly assigned to either write about: a time when they had a lot to drink that was a good time (Positive); a time when they had a lot to drink that was a bad time (Negative); or their first day of college (Neutral), followed by measures assessing intended drinking over the next three months. Results revealed that participants intended to drink significantly fewer drinks per week and engage in marginally fewer heavy drinking occasions after writing about a negative drinking occasion when compared to control. Interactions provided mixed findings suggesting that writing about a positive event was associated with higher drinking intentions for heavier drinkers. Writing about a negative event was associated with higher intentions among heavier drinkers, but lower intentions among those with higher AUDIT scores. This research builds on previous expressive writing interventions by applying this technique to undergraduate drinkers. Preliminary results provide some support for this innovative strategy but also suggest the need for further refinement, especially with heavier drinkers. PMID:24064189

  16. Reduced expression of TANGO in colon and hepatocellular carcinomas.

    PubMed

    Arndt, Stephanie; Bosserhoff, Anja K

    2007-10-01

    The TANGO gene was originally identified as a new family member of the MIA gene family. The gene codes for a 14-kDa protein of so far unknown function. Recently, we identified TANGO as a tumor suppressor in malignant melanoma. In this study we evaluated TANGO transcription in different colon and hepatocellular carcinoma cell lines and tissue samples, to analyze whether loss of TANGO expression is a more general process in tumor development. TANGO was down-regulated or lost in all hepatocellular and colon cell lines compared to primary human hepatocytes or normal colon epithelial cells, respectively, and in most of the tumor samples compared to non-tumorous tissue. These results were confirmed in situ by immunohistochemistry on paraffin-embedded sections of colon and hepatocellular tumors. Functional assays with exogenous TANGO treatment of colon and hepatoma cell lines revealed reduced motility and invasion capacity. Our studies present for the first time the down-regulation of TANGO in colon and hepatocellular carcinoma and provide the first indications for a tumor suppressor role of the TANGO gene in human colon and hepatocellular carcinoma. Thus, functional relevant loss of TANGO expression may contribute to general tumor development and progression, and may provide a new target for therapeutic strategies.

  17. Grape seed extract inhibits VEGF expression via reducing HIF-1alpha protein expression.

    PubMed

    Lu, Jianming; Zhang, Keqiang; Chen, Shiuan; Wen, Wei

    2009-04-01

    Grape seed extract (GSE) is a widely consumed dietary supplement that has antitumor activity. Here, we have investigated the inhibitory effect of GSE on the expression of vascular endothelial growth factor (VEGF) and the mechanism underlying this action. We found that GSE inhibited VEGF messenger RNA (mRNA) and protein expression in U251 human glioma cells and MDA-MB-231 human breast cancer cells. GSE inhibited transcriptional activation of the VEGF gene through reducing protein but not mRNA expression of hypoxia-inducible factor (HIF) 1alpha. The inhibitory effect of GSE on HIF-1alpha expression was mainly through inhibiting HIF-1alpha protein synthesis rather than promoting protein degradation. Consistent with this result, GSE-suppressed phosphorylation of several important components involved in HIF-1alpha protein synthesis, such as Akt, S6 kinase and S6 protein. Furthermore, in the MDA-MB-231 tumor, we found that GSE treatment inhibited the expression of VEGF and HIF-1alpha and the phosphorylation of S6 kinase without altering the subcellular localization of HIF-1alpha, correlating with reduced vessel density and tumor size. Depletion of polyphenol with polyvinylpyrrolidone abolished the inhibitory activity of GSE, suggesting a water-soluble fraction of polyphenol in GSE is responsible for the inhibitory activity. Taken together, our results indicate that GSE inhibits VEGF expression by reducing HIF-1alpha protein synthesis through blocking Akt activation. This finding provides new insight into the mechanisms of anticancer activity of GSE and reveals a novel molecular mechanism underlying the antiangiogenic action of GSE.

  18. Grape seed extract inhibits VEGF expression via reducing HIF-1α protein expression

    PubMed Central

    Lu, Jianming; Zhang, Keqiang; Chen, Shiuan; Wen, Wei

    2009-01-01

    Grape seed extract (GSE) is a widely consumed dietary supplement that has antitumor activity. Here, we have investigated the inhibitory effect of GSE on the expression of vascular endothelial growth factor (VEGF) and the mechanism underlying this action. We found that GSE inhibited VEGF messenger RNA (mRNA) and protein expression in U251 human glioma cells and MDA-MB-231 human breast cancer cells. GSE inhibited transcriptional activation of the VEGF gene through reducing protein but not mRNA expression of hypoxia-inducible factor (HIF) 1α. The inhibitory effect of GSE on HIF-1α expression was mainly through inhibiting HIF-1α protein synthesis rather than promoting protein degradation. Consistent with this result, GSE-suppressed phosphorylation of several important components involved in HIF-1α protein synthesis, such as Akt, S6 kinase and S6 protein. Furthermore, in the MDA-MB-231 tumor, we found that GSE treatment inhibited the expression of VEGF and HIF-1α and the phosphorylation of S6 kinase without altering the subcellular localization of HIF-1α, correlating with reduced vessel density and tumor size. Depletion of polyphenol with polyvinylpyrrolidone abolished the inhibitory activity of GSE, suggesting a water-soluble fraction of polyphenol in GSE is responsible for the inhibitory activity. Taken together, our results indicate that GSE inhibits VEGF expression by reducing HIF-1α protein synthesis through blocking Akt activation. This finding provides new insight into the mechanisms of anticancer activity of GSE and reveals a novel molecular mechanism underlying the antiangiogenic action of GSE. PMID:19131542

  19. Reduced expression of sarcospan in muscles of Fukuyama congenital muscular dystrophy.

    PubMed

    Wakayama, Yoshihiro; Inoue, Masahiko; Kojima, Hiroko; Yamashita, Sumimasa; Shibuya, Seiji; Jimi, Takahiro; Hara, Hajime; Matsuzaki, Yoko; Oniki, Hiroaki; Kanagawa, Motoi; Kobayashi, Kazuhiro; Toda, Tatsushi

    2008-12-01

    Expression profiles of sarcospan in muscles with muscular dystrophies are scarcely reported. To examine this, we studied five Fukuyama congenital muscular dystrophy (FCMD) muscles, five Duchenne muscular dystrophy (DMD) muscles, five disease control and five normal control muscles. Immunoblot showed reactions of sarcospan markedly decreased in FCMD and DMD muscle extracts. Immunohistochemistry of FCMD muscles showed that most large diameter myofibers expressed sarcospan discontinuously at their surface membranes. Immature small diameter FCMD myofibers usually did not express sarcospan. Immunoreactivity of sarcospan in DMD muscles was similarly reduced. With regard to dystroglycans and sarcoglycans, immunohistochemistry of FCMD muscles showed selective deficiency of glycosylated alpha-dystroglycan, together with reduced expression of beta-dystroglycan and alpha-, beta-, gamma-, delta-sarcoglycans. Although the expression of glycosylated alpha-dystroglycan was lost, scattered FCMD myofibers showed positive immunoreaction with an antibody against the core protein of alpha-dystroglycan. The group mean ratios of sarcospan mRNA copy number versus GAPDH mRNA copy number by real-time RT-PCR showed that the ratios between FCMD and normal control groups were not significantly different (P>0.1 by the two-tailed t test). This study implied either O-linked glycosylation defects of alpha-dystroglycan in the Golgi apparatus of FCMD muscles may lead to decreased expression of sarcoglycan and sarcospan molecules, or selective deficiency of glycosylated alpha-dystroglycan due to impaired glycosylation in FCMD muscles may affect the molecular integrity of the basal lamina of myofibers. This, in turn, leads to decreased expression of sarcoglycans, and finally of sarcospan at the FCMD myofiber surfaces.

  20. Reduced and Surface-Modified Graphene Oxide with Nonlinear Resistivity.

    PubMed

    Wåhlander, Martin; Nilsson, Fritjof; Andersson, Richard L; Carlmark, Anna; Hillborg, Henrik; Malmström, Eva

    2017-08-01

    Field-grading materials (FGMs) are used to reduce the probability for electrical breakdowns in critical regions of electrical components and are therefore of great importance. Usually, FGMs are heavily filled (40 vol.%) with semi-conducting or conducting particles. Here, polymer-grafted reduced graphene oxide (rGO) is used as a filler to accomplish percolated networks at very low filling ratios (<2 vol.%) in a semi-crystalline polymer matrix: poly(ethylene-co-butyl acrylate) (EBA). Various simulation models are used to predict the percolation threshold and the flake-to-flake distances, to complement the experimental results. A substantial increase in thermal stability of rGO is observed after surface modification, either by silanization or subsequent polymerizations. The non-linear DC resistivity of neat and silanized rGO and its trapping of charge-carriers in semi-crystalline EBA are demonstrated for the first time. It is shown that the polymer-grafted rGO improve the dispersibility in the EBA-matrix and that the graft length controls the inter-flake distances (i.e. charge-carrier hopping distances). By the appropriate selection of graft lengths, both highly resistive materials at 10 kV mm(-1) and FGMs with a large and distinct drop in resistivity (six decades) are obtained, followed by saturation. The nonlinear drop in resistivity is attributed to narrow inter-flake distance distributions of grafted rGO. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Enhancement of G Protein-Coupled Receptor Surface Expression

    PubMed Central

    Dunham, Jill H.; Hall, Randy A.

    2009-01-01

    G protein-coupled receptors (GPCRs) mediate physiological responses to a diverse array of stimuli and are the molecular targets for numerous therapeutic drugs. GPCRs primarily signal from the plasma membrane, but when expressed in heterologous cells many GPCRs exhibit poor trafficking to the cell surface. Multiple approaches have been taken to enhance GPCR surface expression in heterologous cells, including addition/deletion of receptor sequences, co-expression with interacting proteins, and treatment with pharmacological chaperones. In addition to allowing for enhanced surface expression of certain GPCRs in heterologous cells, these approaches have also shed light on the control of GPCR trafficking in vivo and in some cases have led to new therapeutic approaches for treating human diseases that result from defects in GPCR trafficking. PMID:19679364

  2. Botulinum toxin type A reduces hyperalgesia and TRPV1 expression in rats with neuropathic pain.

    PubMed

    Xiao, Lizu; Cheng, Jianguo; Zhuang, Yu; Qu, Wenchun; Muir, Jeffery; Liang, Haowen; Zhang, Deren

    2013-02-01

    We aim to determine the effects of botulinum toxin type A (BTX-A) on the thresholds of pain and the expression of transient receptor potential vanilloid type 1 (TRPV1) in the dorsal root ganglion (DRG) in rats with neuropathic pain induced by selective ventral root transection (VRT). Neuropathic pain was induced by transection of the lumbar 5 ventral root in male Sprague-Dawley rats. BTX-A or saline was administered to the plantar surface by subcutaneous injection. SB366791 (an inhibitor of TRPV1) was administered intraperitoneally. Behavioral tests were conducted preoperatively and at predefined postoperative days. The expression of TRPV1 was detected and quantified by immunohistochemistry and Western blotting at postoperative days 3, 7, 14, and 21. TRPV1 expression increased significantly in the L4 ∼5 dorsal root ganglia 7 days after L5 VRT compared with the sham-operated control (P < 0.05). This increase persisted for at least 21 days. The thresholds of foot withdrawal to mechanical and thermal stimulation decreased significantly as well. Subcutaneous injection of BTX-A significantly and dose-dependently reduced the expression of TRPV1 (P < 0.05) and partially reversed the pain thresholds. Upregulation of TRPV1 expression in the DRG is an important mechanism of neuropathic pain induced by the VRT. The analgesic effect of BTX-A is most likely mediated through reduction of TRPV1 expression in the nociceptors. Wiley Periodicals, Inc.

  3. Arterial Shear Stress Reduces Eph-B4 Expression in Adult Human Veins

    PubMed Central

    Model, Lynn S.; Hall, Michael R.; Wong, Daniel J.; Muto, Akihito; Kondo, Yuka; Ziegler, Kenneth R.; Feigel, Amanda; Quint, Clay; Niklason, Laura; Dardik, Alan

    2014-01-01

    Vein graft adaptation to the arterial environment is characterized by loss of venous identity, with reduced Ephrin type-B receptor 4 (Eph-B4) expression but without increased Ephrin-B2 expression. We examined changes of vessel identity of human saphenous veins in a flow circuit in which shear stress could be precisely controlled. Medium circulated at arterial or venous magnitudes of laminar shear stress for 24 hours; histologic, protein, and RNA analyses of vein segments were performed. Vein endothelium remained viable and functional, with platelet endothelial cell adhesion molecule (PECAM)-expressing cells on the luminal surface. Venous Eph-B4 expression diminished (p = .002), Ephrin-B2 expression was not induced (p = .268), and expression of osteopontin (p = .002) was increased with exposure to arterial magnitudes of shear stress. Similar changes were not found in veins placed under venous flow or static conditions. These data show that human saphenous veins remain viable during ex vivo application of shear stress in a bioreactor, without loss of the venous endothelium. Arterial magnitudes of shear stress cause loss of venous identity without gain of arterial identity in human veins perfused ex vivo. Shear stress alone, without immunologic or hormonal influence, is capable of inducing changes in vessel identity and, specifically, loss of venous identity. PMID:25191151

  4. Reduced CD73 Expression by IL-1β Programmed Th17 Cells Improves Tumor Control

    PubMed Central

    Chatterjee, Shilpak; Thyagarajan, Krishnamurthy; Kesarwani, Pravin; Song, Jin H.; Soloshchenko, Myroslawa; Fu, Jianing; Bailey, Stefanie; Vasu, Chenthamarkshan; Kraft, Andrew; Paulos, Chrystal M.; Yu, Xue-Zhong; Mehrotra, Shikhar

    2014-01-01

    T helper (Th)-17 subsets hold promise in adoptive T cell transfer therapy for cancer. However, ex vivo programming of Th17 cells in presence of TGF-β increases cell surface expression of ectonucleotidases CD39 and CD73, that in turn increases susceptibility to immunosuppression and reduces effector functions. Our data shows that ATP mediated suppression of IFN-γ production by Th17 cells can be overcome either by genetic ablation of CD73 or by generating TGF-β independent Th17 in presence of IL-1β. Th17 cells cultured in IL-1β are also highly polyfunctional, express high level of effector molecules and exhibit better short-term control of B16-F10 murine melanoma, despite reduced stem cell like properties. Adding TGF-β at low dose that does not up regulate CD73 expression, but induces stemness, drastically improves anti-tumor function of IL-1β cultured Th17 cells. It is likely that effector property of IL-1β dependent Th17 is due to their high glycolytic capacity, since generating IL-1β dependent Th17 cells in pyruvate containing media impaired glycolysis and its anti-tumor potential. Thus, our data suggests that due to induction of ectonucleotidase expression by TGF-β, ex vivo culture conditions for generating Th17 cells need to be reconsidered for exploiting their full potential in adoptive T cell therapy. PMID:25205101

  5. MULTIPLE IMAGING TECHNIQUES DEMONSTRATE THE MANIPULATION OF SURFACES TO REDUCE BACTERIAL CONTAMINATION

    EPA Science Inventory

    Surface imaging techniques were combined to determine appropriate manipulation of technologically important surfaces for commercial applications. Stainless steel surfaces were engineered to reduce bacterial contamination, biofilm formation, and corrosion during product processing...

  6. MULTIPLE IMAGING TECHNIQUES DEMONSTRATE THE MANIPULATION OF SURFACES TO REDUCE BACTERIAL CONTAMINATION

    EPA Science Inventory

    Surface imaging techniques were combined to determine appropriate manipulation of technologically important surfaces for commercial applications. Stainless steel surfaces were engineered to reduce bacterial contamination, biofilm formation, and corrosion during product processing...

  7. Cell surface expression of hepatitis B surface and core antigens in transfected rat fibroblast cell lines.

    PubMed

    Gholson, C F; Siddiqui, A; Vierling, J M

    1990-04-01

    Hepatocellular necrosis during hepatitis B virus infection is hypothesized to result from host immune responses against either hepatitis B surface antigen or hepatitis B core antigen expressed on the surface membrane of infected hepatocytes. To study the capacity of hepatitis B deoxyribonucleic acid to induce membrane expression of either hepatitis B surface antigen or hepatitis B core antigen in vitro, we assessed transfected rat fibroblast cell lines by indirect immunofluorescence. Rat fibroblasts were transfected with plasmid vectors containing the natural promoters, native enhancer, and uninterrupted sequences of either the Pre S/S gene or core gene. Resulting cell lines produced hepatitis B surface antigen and hepatitis B core antigen/hepatitis B e antigen, respectively. Immunofluorescence microscopy or flow cytometry showed that hepatitis B surface antigen and hepatitis B core antigen were expressed in a granular pattern in the surface membrane of transfected cells. We conclude that surface membrane expression of both hepatitis B surface antigen and hepatitis B core antigen is an intrinsic consequence of expression of either the Pre S/S or core gene.

  8. Landfill disposal of unused medicines reduces surface water releases.

    PubMed

    Tischler, Lial; Buzby, Mary; Finan, Douglas S; Cunningham, Virginia L

    2013-01-01

    The pharmaceutical industry is conducting research to evaluate the pathways and fate of active pharmaceutical ingredients from the consumer to surface waters. One potential pathway identified by the researchers is the disposal of unused pharmaceutical products that are discarded by consumers in household trash and disposed of in municipal solid waste landfills. This study was designed to evaluate relative amounts of surface water exposures through the landfill disposal pathway compared to patient use and flushing of unused medicine pathways. The estimated releases to surface water of 24 example active pharmaceutical ingredients (APIs) in landfill leachate were calculated for 3 assumed disposal scenarios: 5%, 10%, and 15% of the total annual quantity of API sold is discarded and unused. The estimated releases from landfills to surface waters, after treatment of the leachate, were compared to the total amount of each example API that would be released to surface waters from publicly owned treatment works, generated by patient use and excretion. This study indicates that the disposal of unused medications in municipal solid waste landfills effectively eliminates the unused medicine contribution of APIs to surface waters; greater than 99.9% of APIs disposed of in a landfill are permanently retained.

  9. Reduced Expression of SARM in Mouse Spleen during Polymicrobial Sepsis.

    PubMed

    Gong, Yu; Zou, Lin; Cen, Dongzhi; Chao, Wei; Chen, Dunjin

    2016-12-01

    Objective Immune dysfunction, including prominent apoptosis of immune cells and decreased functioning of the remaining immune cells, plays a central role in the pathogenesis of sepsis. Sterile α and HEAT/armadillo motif-containing protein (SARM) is implicated in the regulation of immune cell apoptosis. This study aimed to elucidate SARM contributes to sepsis-induced immune cell death and immunosuppression. Methods A mouse model of polymicrobial sepsis was generated by cecum ligation and puncture (CLP). SARM gene and protein expression, caspase 3 cleavage and intracellular ATP production were measured in the mouse spleens. Results CLP-induced polymicrobial sepsis specifically attenuated both the gene and protein expression of SARM in the spleens. Moreover, the attenuation of SARM expression synchronized with splenocyte apoptosis, as evidenced by increased caspase 3 cleavage and ATP depletion. Conclusions These findings suggest that SARM is a potential regulator of sepsis-induced splenocyte apoptosis.

  10. Reduced fouling of ultrafiltration membranes via surface fluorination

    SciTech Connect

    Sedath, R.H.; Yates, S.F.; Li, N.N.

    1993-03-01

    Surface fluorination can affect significantly the performance of an ultrafiltration membrane used to concentrate a food-related stream. Membranes fluorinated and tested as flat sheets exhibit higher initial fluxes, and do not foul as rapidly as untreated membranes. This improvement is linked to increased surface hydrophilicity, as shown in decreased contact angle with water. This increased hydrophilicity, in turn, is linked to the addition of fluorine and oxygen to the surface. The pilot plant study did-not show the difference in membrane flux and fouling observed in the flat sheet study. Instead, fluorinated and unfluorinated modules behaved similarly. Fouling by potato waste feed was severe and resulted in formation of an extensive gel layer within the module on the membrane surface. XPS, SEM and FTIR indicate that buildup of organic material occurred on both fluorinated and unfluorinated membranes, but SEM indicates that a fibrous mat of material was observed only on the nonfluorinated membrane. We conclude that in the pilot study, membrane fouling and gel formation were so extensive that the surface interaction effect was overwhelmed.

  11. Reduced fouling of ultrafiltration membranes via surface fluorination

    SciTech Connect

    Sedath, R.H.; Yates, S.F.; Li, N.N.

    1993-03-01

    Surface fluorination can affect significantly the performance of an ultrafiltration membrane used to concentrate a food-related stream. Membranes fluorinated and tested as flat sheets exhibit higher initial fluxes, and do not foul as rapidly as untreated membranes. This improvement is linked to increased surface hydrophilicity, as shown in decreased contact angle with water. This increased hydrophilicity, in turn, is linked to the addition of fluorine and oxygen to the surface. The pilot plant study did-not show the difference in membrane flux and fouling observed in the flat sheet study. Instead, fluorinated and unfluorinated modules behaved similarly. Fouling by potato waste feed was severe and resulted in formation of an extensive gel layer within the module on the membrane surface. XPS, SEM and FTIR indicate that buildup of organic material occurred on both fluorinated and unfluorinated membranes, but SEM indicates that a fibrous mat of material was observed only on the nonfluorinated membrane. We conclude that in the pilot study, membrane fouling and gel formation were so extensive that the surface interaction effect was overwhelmed.

  12. Reducing Motional Decoherence in Ion Traps with Surface Science Methods

    NASA Astrophysics Data System (ADS)

    Haeffner, Hartmut

    2014-03-01

    Many trapped ions experiments ask for low motional heating rates while trapping the ions close to trapping electrodes. However, in practice small ion-electrode distances lead to unexpected high heating rates. While the mechanisms for the heating is still unclear, it is now evident that surface contamination of the metallic electrodes is at least partially responsible for the elevated heating rates. I will discuss heating rate measurements in a microfabricated surface trap complemented with basic surface science studies. We monitor the elemental surface composition of the Cu-Al alloy trap with an Auger spectrometer. After bake-out, we find a strong Carbon and Oxygen contamination and heating rates of 200 quanta/s at 1 MHz trap frequency. After removing most of the Carbon and Oxygen with Ar-Ion sputtering, the heating rates drop to 4 quanta/s. Interestingly, we still measure the decreased heating rate even after the surface oxidized from the background gas throughout a 40-day waiting time in UHV.

  13. Reduced immunohistochemical expression of adhesion molecules in vitiligo skin biopsies.

    PubMed

    Reichert Faria, Adriane; Jung, Juliana Elizabeth; Silva de Castro, Caio César; de Noronha, Lucia

    2017-03-01

    Because defects in adhesion impairment seem to be involved in the etiopathogenesis of vitiligo, this study aimed to compare the immunohistochemical expression of several adhesion molecules in the epidermis of vitiligo and non lesional vitiligo skin. Sixty-six specimens of lesional and non lesional skin from 33 volunteers with vitiligo were evaluated by immunohistochemistry using anti-beta-catenin, anti-E-cadherin, anti-laminin, anti-beta1 integrin, anti-collagen IV, anti-ICAM-1 and anti-VCAM-1 antibodies. Biopsies of vitiligo skin demonstrated a significant reduction in the expression of laminin and integrin. The average value of the immunohistochemically positive reaction area of the vitiligo specimens was 3053.2μm(2), compared with the observed value of 3431.8μm(2) in non vitiligo skin (p=0.003) for laminin. The immuno-positive area was 7174.6μm(2) (vitiligo) and 8966.7μm(2) (non lesional skin) for integrin (p=0.042). A reduction in ICAM-1 and VCAM-1 expression in the basal layer of the epidermis in vitiligo samples was also observed (p=0.001 and p<0.001, respectively). However, no significant differences were observed with respect to the expression of beta-catenin, E-cadherin, and collagen IV between vitiligo and non lesional skin. Our results suggest that an impairment in adhesion exists in vitiligo skin, which is supported by the diminished immunohistochemical expression of laminin, beta1 integrin, ICAM-1 and VCAM-1. Copyright © 2017 Elsevier GmbH. All rights reserved.

  14. Downregulation of transferrin receptor surface expression by intracellular antibody

    SciTech Connect

    Peng Jilin; Wu Sha; Zhao Xiaoping; Wang Min; Li Wenhan; Shen Xin; Liu Jing; Lei Ping; Zhu Huifen; Shen Guanxin . E-mail: guanxin_shen@yahoo.com.cn

    2007-03-23

    To deplete cellular iron uptake, and consequently inhibit the proliferation of tumor cells, we attempt to block surface expression of transferrin receptor (TfR) by intracellular antibody technology. We constructed two expression plasmids (scFv-HAK and scFv-HA) coding for intracellular single-chain antibody against TfR with or without endoplasmic reticulum (ER) retention signal, respectively. Then they were transfected tumor cells MCF-7 by liposome. Applying RT-PCR, Western blotting, immunofluorescence microscopy and immunoelectron microscope experiments, we insure that scFv-HAK intrabody was successfully expressed and retained in ER contrasted to the secreted expression of scFv-HA. Flow cytometric analysis confirmed that the TfR surface expression was markedly decreased approximately 83.4 {+-} 2.5% in scFv-HAK transfected cells, while there was not significantly decrease in scFv-HA transfected cells. Further cell growth and apoptosis characteristics were evaluated by cell cycle analysis, nuclei staining and MTT assay. Results indicated that expression of scFv-HAK can dramatically induce cell cycle G1 phase arrest and apoptosis of tumor cells, and consequently significantly suppress proliferation of tumor cells compared with other control groups. For First time this study demonstrates the potential usage of anti-TfR scFv-intrabody as a growth inhibitor of TfR overexpressing tumors.

  15. Riparian shrub buffers reduce surface water pollutant loads

    Treesearch

    W. A. Geyer; C. Barden; K. Mankin; D. Devlin

    2003-01-01

    Surface water resources in Kansas often contain concentrations of pesticides, nutrients, and sediments that are of concern to local citizens. The United States Geological Survey reported in 1999 that 97 percent of streams and 82 percent of lakes in Kansas would not fully support all uses as designated by state statutes (U.S. Geological Survey 1999). Bacteria and...

  16. Saccharomyces cerevisiae expressing bacteriophage endolysins reduce Lactobacillus contamination during fermentation

    USDA-ARS?s Scientific Manuscript database

    One of the challenges facing the fuel ethanol industry is the management of bacterial contamination during fermentation. Lactobacillus species are the predominant contaminants that decrease the profitability of biofuel production by reducing ethanol yields and causing “stuck” fermentations, which i...

  17. Expression of hepatitis B surface antigen in transgenic banana plants.

    PubMed

    Kumar, G B Sunil; Ganapathi, T R; Revathi, C J; Srinivas, L; Bapat, V A

    2005-10-01

    Embryogenic cells of bananan cv. Rasthali (AAB) have been transformed with the 's' gene of hepatitis B surface antigen (HBsAg) using Agrobacterium mediated transformation. Four different expression cassettes (pHBS, pHER, pEFEHBS and pEFEHER) were utilized to optimize the expression of HBsAg in banana. The transgenic nature of the plants and expression of the antigen was confirmed by PCR, Southern hybridization and reverse transcription (RT)-PCR. The expression levels of the antigen in the plants grown under in vitro conditions as well as the green house hardened plants were estimated by ELISA for all the four constructs. Maximum expression level of 38 ng/g F.W. of leaves was noted in plants transformed with pEFEHBS grown under in vitro conditions, whereas pHER transformed plants grown in the green house showed the maximum expression level of 19.92 ng/g F.W. of leaves. Higher monoclonal antibody binding of 67.87% of the antigen was observed when it was expressed with a C-terminal ER retention signal. The buoyant density in CsCl of HBsAg derived from transgenic banana leaves was determined and found to be 1.146 g/ml. HBsAg obtained from transgenic banana plants is similar to human serum derived one in buoyant density properties. The transgenic plants were grown up to maturity in the green house and the expression of HBsAg in the fruits was confirmed by RT-PCR. These transgenic plants were multiplied under in vitro using floral apex cultures. Attempts were also made to enhance the expression of HBsAg in the leaves of transgenic banana plants by wounding and/or treatment with plant growth regulators. This is the first report on the expression of HBsAg in transgenic banana fruits.

  18. Venus surface investigation based on VIRTIS measurements on Venus Express

    NASA Astrophysics Data System (ADS)

    Arnold, Gabriele; Haus, Rainer; Döhler, Wolfgang; Kappel, David; Piccioni, Giuseppe; Drossart, Pierre

    The dense atmosphere of Venus prevented systematic studies of its surface at optical wavelengths in the past. The discovery of near infrared nightside atmospheric windows has opened a new challenge for detailed surface studies. The Visible and Infrared Thermal Imaging Spectrometer (VIRTIS) on Venus Express is the first experiment collecting continuously nightside surface emission data from the planet. The observed high variability of measured signatures is mainly due to spatial variations of cloud optical depth and surface elevation. The investigation of surface properties requires a convergent approach of radiative transfer simulations and VIR- TIS data analyses. Therefore, a selection of orbits with well calibrated data over the northern hemisphere was performed for footprints that cover a maximum range of surface elevation variations. Radiative transfer calculations demonstrate that the conservative character of cloud multiple scattering below 2 µm and a strong dependence of radiance ratios on surface elevation in this spectral region allow the mapping of surface topography and a retrieval of the surface temperature. To the first order, the surface temperature is a function of ground elevation. Small deviations from this first order dependence have been identified that are possibly due to different surface materials. 1 Institut f¨r Planetologie, Westf¨lische Wilhelms-Universit¨t M¨nster, Wilhelm-Klemm-Str.10, u a a u 48129 M¨nster, Germany u 2 German PlaceNameAerospace PlaceTypeCenter (DLR), Remote Sensing Technology Institute, Dpt. Marine Remote Sensing, Rutherfordstrasse 2, 12489 CityplaceBerlin, countryregionGermany 3 German PlaceNameAerospace PlaceTypeCenter (DLR), Institute for Planetary Research, Rutherfordstrasse 2, 12489 CityplaceBerlin, country-regionGermany 4 LESIA, Observatoire de Paris, CNRS, UPMC, Université Paris-Diderot, 5 place Jules Janssen, e 92195 Meudon, France 5 INAF-IASF (Instituto di Astrofisica Spaziale e Fisica Cosmica), via

  19. Paired Expression Analysis of Tumor Cell Surface Antigens

    PubMed Central

    Orentas, Rimas J.; Sindiri, Sivasish; Duris, Christine; Wen, Xinyu; He, Jianbin; Wei, Jun S.; Jarzembowski, Jason; Khan, Javed

    2017-01-01

    Adoptive immunotherapy with antibody-based therapy or with T cells transduced to express chimeric antigen receptors (CARs) is useful to the extent that the cell surface membrane protein being targeted is not expressed on normal tissues. The most successful CAR-based (anti-CD19) or antibody-based therapy (anti-CD20) in hematologic malignancies has the side effect of eliminating the normal B cell compartment. Targeting solid tumors may not provide a similar expendable marker. Beyond antibody to Her2/NEU and EGFR, very few antibody-based and no CAR-based therapies have seen broad clinical application for solid tumors. To expand the way in which the surfaceome of solid tumors can be analyzed, we created an algorithm that defines the pairwise relative overexpression of surface antigens. This enables the development of specific immunotherapies that require the expression of two discrete antigens on the surface of the tumor target. This dyad analysis was facilitated by employing the Hotelling’s T-squared test (Hotelling–Lawley multivariate analysis of variance) for two independent variables in comparison to a third constant entity (i.e., gene expression levels in normal tissues). We also present a unique consensus scoring mechanism for identifying transcripts that encode cell surface proteins. The unique application of our bioinformatics processing pipeline and statistical tools allowed us to compare the expression of two membrane protein targets as a pair, and to propose a new strategy based on implementing immunotherapies that require both antigens to be expressed on the tumor cell surface to trigger therapeutic effector mechanisms. Specifically, we found that, for MYCN amplified neuroblastoma, pairwise expression of ACVR2B or anaplastic lymphoma kinase (ALK) with GFRA3, GFRA2, Cadherin 24, or with one another provided the strongest hits. For MYCN, non-amplified stage 4 neuroblastoma, neurotrophic tyrosine kinase 1, or ALK paired with GFRA2, GFRA3, SSK1, GPR173, or

  20. Physiological capillary regression is not dependent on reducing VEGF expression

    PubMed Central

    Olfert, I. Mark

    2015-01-01

    Investigations into physiologically-controlled capillary regression report the provocative finding that microvessel regression occurs in the face of persistent elevation of skeletal muscle vascular endothelial growth factor-A (VEGF) expression. Thrombospondin-1 (TSP-1), a negative angiogenic regulator, is increasingly being observed to temporally correlate with capillary regression, suggesting that increased TSP-1 (and not reduction in VEGF per se) is needed to initiate, and likely regulate, capillary regression. Based on evidence being gleaned from physiologically-mediated regression of capillaries, it needs to be recognized that capillary regression (and perhaps capillary rarefaction with disease) is not simply the reversal of factors used to stimulate angiogenesis. Rather, the conceptual understanding that angiogenesis and capillary regression each have specific and unique requirements that are biologically constrained to opposite sides of the balance between positive and negative angioregulatory factors may shed light on why anti-VEGF therapies have not lived up to the promise in reversing angiogenesis and providing the cure that many had hoped toward fighting cancer. Emerging evidence from physiological controlled angiogenesis suggest that cases involving excessive or uncontrolled capillary expansion may be best treated by therapies designed to increase expression of negative angiogenic regulators, whereas those involving capillary rarefaction may benefit from inhibiting negative regulators (like TSP-1). PMID:26660949

  1. On the Use of Surface Porosity to Reduce Unsteady Lift

    NASA Technical Reports Server (NTRS)

    Tinetti, Ana F.; Kelly, Jeffrey J.; Bauer, Steven X. S.; Thomas, Russell H.

    2001-01-01

    An innovative application of existing technology is proposed for attenuating the effects of transient phenomena, such as rotor-stator and rotor-strut interactions, linked to noise and fatigue failure in turbomachinery environments. A computational study was designed to assess the potential of passive porosity technology as a mechanism for alleviating interaction effects by reducing the unsteady lift developed on a stator airfoil subject to wake impingement. The study involved a typical high bypass fan Stator airfoil (solid baseline and several porous configurations), immersed in a free field and exposed to the effects of a transversely moving wake. It was found that, for the airfoil under consideration, the magnitude of the unsteady lift could be reduced more than 18% without incurring significant performance losses.

  2. Surface Polysaccharide Mutants Reveal that Absence of O Antigen Reduces Biofilm Formation of Actinobacillus pleuropneumoniae

    PubMed Central

    Hathroubi, S.; Hancock, M. A.; Langford, P. R.; Tremblay, Y. D. N.; Labrie, J.

    2015-01-01

    Actinobacillus pleuropneumoniae is a Gram-negative bacterium belonging to the Pasteurellaceae family and the causative agent of porcine pleuropneumonia, a highly contagious lung disease causing important economic losses. Surface polysaccharides, including lipopolysaccharides (LPS) and capsular polysaccharides (CPS), are implicated in the adhesion and virulence of A. pleuropneumoniae, but their role in biofilm formation is still unclear. In this study, we investigated the requirement for these surface polysaccharides in biofilm formation by A. pleuropneumoniae serotype 1. Well-characterized mutants were used: an O-antigen LPS mutant, a truncated core LPS mutant with an intact O antigen, a capsule mutant, and a poly-N-acetylglucosamine (PGA) mutant. We compared the amount of biofilm produced by the parental strain and the isogenic mutants using static and dynamic systems. Compared to the findings for the biofilm of the parental or other strains, the biofilm of the O antigen and the PGA mutants was dramatically reduced, and it had less cell-associated PGA. Real-time PCR analyses revealed a significant reduction in the level of pgaA, cpxR, and cpxA mRNA in the biofilm cells of the O-antigen mutant compared to that in the biofilm cells of the parental strain. Specific binding between PGA and LPS was consistently detected by surface plasmon resonance, but the lack of O antigen did not abolish these interactions. In conclusion, the absence of the O antigen reduces the ability of A. pleuropneumoniae to form a biofilm, and this is associated with the reduced expression and production of PGA. PMID:26483403

  3. Surface Polysaccharide Mutants Reveal that Absence of O Antigen Reduces Biofilm Formation of Actinobacillus pleuropneumoniae.

    PubMed

    Hathroubi, S; Hancock, M A; Bossé, J T; Langford, P R; Tremblay, Y D N; Labrie, J; Jacques, M

    2015-10-19

    Actinobacillus pleuropneumoniae is a Gram-negative bacterium belonging to the Pasteurellaceae family and the causative agent of porcine pleuropneumonia, a highly contagious lung disease causing important economic losses. Surface polysaccharides, including lipopolysaccharides (LPS) and capsular polysaccharides (CPS), are implicated in the adhesion and virulence of A. pleuropneumoniae, but their role in biofilm formation is still unclear. In this study, we investigated the requirement for these surface polysaccharides in biofilm formation by A. pleuropneumoniae serotype 1. Well-characterized mutants were used: an O-antigen LPS mutant, a truncated core LPS mutant with an intact O antigen, a capsule mutant, and a poly-N-acetylglucosamine (PGA) mutant. We compared the amount of biofilm produced by the parental strain and the isogenic mutants using static and dynamic systems. Compared to the findings for the biofilm of the parental or other strains, the biofilm of the O antigen and the PGA mutants was dramatically reduced, and it had less cell-associated PGA. Real-time PCR analyses revealed a significant reduction in the level of pgaA, cpxR, and cpxA mRNA in the biofilm cells of the O-antigen mutant compared to that in the biofilm cells of the parental strain. Specific binding between PGA and LPS was consistently detected by surface plasmon resonance, but the lack of O antigen did not abolish these interactions. In conclusion, the absence of the O antigen reduces the ability of A. pleuropneumoniae to form a biofilm, and this is associated with the reduced expression and production of PGA. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  4. Real-time absorption reduced surface fluorescence imaging

    NASA Astrophysics Data System (ADS)

    Yang, Bin; Tunnell, James W.

    2014-09-01

    We introduce a technique that limits absorption effects in fluorescence imaging and does not require extensive imaging processing, thus allowing for video rate imaging. The absorption minimization is achieved using spatial frequency domain imaging at a single high spatial frequency with standard three-phase demodulation. At a spatial frequency f=0.5 mm-1, we demonstrated in both in-vitro phantoms and ex-vivo tissue that the absorption can be significantly reduced. In the real-time implementation, we achieved a video rate of 19 frames/s. This technique has potential in cancer visualization and tumor margin detection.

  5. Real-time absorption reduced surface fluorescence imaging.

    PubMed

    Yang, Bin; Tunnell, James W

    2014-09-01

    We introduce a technique that limits absorption effects in fluorescence imaging and does not require extensive imaging processing, thus allowing for video rate imaging. The absorption minimization is achieved using spatial frequency domain imaging at a single high spatial frequency with standard three-phase demodulation. At a spatial frequency f ¼ 0.5 mm−1, we demonstrated in both in-vitro phantoms and ex-vivo tissue that the absorption can be significantly reduced. In the real-time implementation, we achieved a video rate of 19 frames∕s. This technique has potential in cancer visualization and tumor margin detection.

  6. Competition-based transfer of carbohydrate expression information from a cell-adhered surface to a secondary surface.

    PubMed

    Ding, Lin; Xiao, Xirui; Chen, Yunlong; Qian, Ruocan; Bao, Lei; Ju, Huangxian

    2011-04-07

    An information transfer strategy was developed for the visualization of carbohydrate expression by the competition of a primary cell-adhered solid surface with a carbohydrate assembled surface as an artificial secondary surface for one species. The strategy could be effectively utilized for in situ monitoring of dynamic carbohydrate expression on an adhesive cell surface.

  7. Leukocyte Cell Surface Proteinases: Regulation of Expression, Functions, and Mechanisms of Surface Localization

    PubMed Central

    Owen, Caroline A.

    2008-01-01

    A number of proteinases are expressed on the surface of leukocytes including members of the serine, metallo-, and cysteine proteinase superfamilies. Some proteinases are anchored to the plasma membrane of leukocytes by a transmembrane domain or a glycosyl phosphatidyl inositol (GPI) anchor. Other proteinases bind with high affinity to classical receptors, or with lower affinity to integrins, proteoglycans, or other leukocyte surface molecules. Leukocyte surface levels of proteinases are regulated by: 1) cytokines, chemokines, bacterial products, and growth factors which stimulate synthesis and/or release of proteinase by cells; 2) the availability of surface binding sites for proteinases; and/or 3) internalization or shedding of surface-bound proteinases. The binding of proteinases to leukocyte surfaces serves many functions including: 1) concentrating the activity of proteinases to the immediate pericellular environment; 2) facilitating pro-enzyme activation; 3) increasing proteinase stability and retention in the extracellular space; 4) regulating leukocyte function by proteinases signaling through cell surface binding sites or other surface proteins; and 5) protecting proteinases from inhibition by extracellular proteinase inhibitors. There is strong evidence that membrane-associated proteinases on leukocytes play critical roles in wound healing, inflammation, extracellular matrix remodeling, fibrinolysis, and coagulation. This review will outline the biology of membrane-associated proteinases expressed by leukocytes and their roles in physiologic and pathologic processes. PMID:18329945

  8. Leukocyte cell surface proteinases: regulation of expression, functions, and mechanisms of surface localization.

    PubMed

    Owen, Caroline A

    2008-01-01

    A number of proteinases are expressed on the surface of leukocytes including members of the serine, metallo-, and cysteine proteinase superfamilies. Some proteinases are anchored to the plasma membrane of leukocytes by a transmembrane domain or a glycosyl phosphatidyl inositol (GPI) anchor. Other proteinases bind with high affinity to classical receptors, or with lower affinity to integrins, proteoglycans, or other leukocyte surface molecules. Leukocyte surface levels of proteinases are regulated by: (1) cytokines, chemokines, bacterial products, and growth factors which stimulate synthesis and/or release of proteinases by cells; (2) the availability of surface binding sites for proteinases; and/or (3) internalization or shedding of surface-bound proteinases. The binding of proteinases to leukocyte surfaces serves many functions including: (1) concentrating the activity of proteinases to the immediate pericellular environment; (2) facilitating pro-enzyme activation; (3) increasing proteinase stability and retention in the extracellular space; (4) regulating leukocyte function by proteinases signaling through cell surface binding sites or other surface proteins; and (5) protecting proteinases from inhibition by extracellular proteinase inhibitors. There is strong evidence that membrane-associated proteinases on leukocytes play critical roles in wound healing, inflammation, extracellular matrix remodeling, fibrinolysis, and coagulation. This review will outline the biology of membrane-associated proteinases expressed by leukocytes and their roles in physiologic and pathologic processes.

  9. Reduced Protein Expression in a Virus Attenuated by Codon Deoptimization

    PubMed Central

    Jack, Benjamin R.; Boutz, Daniel R.; Paff, Matthew L.; Smith, Bartram L.; Bull, James J.; Wilke, Claus O.

    2017-01-01

    A general means of viral attenuation involves the extensive recoding of synonymous codons in the viral genome. The mechanistic underpinnings of this approach remain unclear, however. Using quantitative proteomics and RNA sequencing, we explore the molecular basis of attenuation in a strain of bacteriophage T7 whose major capsid gene was engineered to carry 182 suboptimal codons. We do not detect transcriptional effects from recoding. Proteomic observations reveal that translation is halved for the recoded major capsid gene, and a more modest reduction applies to several coexpressed downstream genes. We observe no changes in protein abundances of other coexpressed genes that are encoded upstream. Viral burst size, like capsid protein abundance, is also decreased by half. Together, these observations suggest that, in this virus, reduced translation of an essential polycistronic transcript and diminished virion assembly form the molecular basis of attenuation. PMID:28698233

  10. Expression of the RNase III enzyme DROSHA is reduced during progression of human cutaneous melanoma

    PubMed Central

    Jafarnejad, Seyed Mehdi; Sjoestroem, Cecilia; Martinka, Magdalena; Li, Gang

    2016-01-01

    Aberrant expression of miRNAs and their biogenesis factors has been frequently observed in different types of cancer. We recently reported that expression of DICER1 is reduced in metastatic melanoma. Nevertheless, so far very little is known about the expression pattern of other miRNA biogenesis factors in this type of malignancy. Here, we investigated the expression pattern of DROSHA in a large set of melanocytic lesions by tissue microarray and immunohistochemistry (n = 409). We found that nuclear expression of DROSHA is markedly reduced in the early stages of melanoma progression (P = 0.0001) and is inversely correlated with melanoma thickness (P = 0.0001), AJCC stages (P = 0.0001), and ulceration status (P = 0.002). We also confirmed the reduced expression of nuclear DROSHA by a second specific antibody raised against a different region of the DROSHA protein. In addition, we observed that the reduced nuclear expression of DROSHA during melanoma progression is accompanied by an increased cytoplasmic expression of this protein (P = 0.0001). Finally, we found that expression pattern of DROSHA varies from that of DICER1 and concomitant loss of expression of both DICER1 and DROSHA confers the worse outcome for melanoma patients. Our results demonstrate a reduced nuclear expression of DROSHA which further highlights a perturbed miRNA biogenesis pathway in melanoma. In addition, the aberrant subcellular localization of DROSHA indicates possible deregulation in the mechanisms responsible for its proper localization in the nucleus. PMID:23370771

  11. Muscle cell depolarization induces a gain in surface GLUT4 via reduced endocytosis independently of AMPK.

    PubMed

    Wijesekara, Nadeeja; Tung, Amanda; Thong, Farah; Klip, Amira

    2006-06-01

    Contracting skeletal muscle increases glucose uptake to sustain energy demand. This is achieved through a gain in GLUT4 at the membrane, but the traffic mechanisms and regulatory signals involved are unknown. Muscle contraction is elicited by membrane depolarization followed by a rise in cytosolic Ca2+ and actomyosin activation, drawing on ATP stores. It is unknown whether one or more of these events triggers the rise in surface GLUT4. Here, we investigate the effect of membrane depolarization on GLUT4 cycling using GLUT4myc-expressing L6 myotubes devoid of sarcomeres and thus unable to contract. K+-induced membrane depolarization elevated surface GLUT4myc, and this effect was additive to that of insulin, was not prevented by inhibiting phosphatidylinositol 3-kinase (PI3K) or actin polymerization, and did not involve Akt activation. Instead, depolarization elevated cytosolic Ca2+, and the surface GLUT4myc elevation was prevented by dantrolene (an inhibitor of Ca2+ release from sarcoplasmic reticulum) and by extracellular Ca2+ chelation. Ca2+-calmodulin-dependent protein kinase-II (CaMKII) was not phosphorylated after 10 min of K+ depolarization, and the CaMK inhibitor KN62 did not prevent the gain in surface GLUT4myc. Interestingly, although 5'-AMP-activated protein kinase (AMPK) was phosphorylated upon depolarization, lowering AMPKalpha via siRNA did not alter the surface GLUT4myc gain. Conversely, the latter response was abolished by the PKC inhibitors bisindolylmaleimide I and calphostin C. Unlike insulin, K+ depolarization caused only a small increase in GLUT4myc exocytosis and a major reduction in its endocytosis. We propose that K+ depolarization reduces GLUT4 internalization through signals and mechanisms distinct from those engaged by insulin. Such a pathway(s) is largely independent of PI3K, Akt, AMPK, and CaMKII but may involve PKC.

  12. Cell surface expression of biologically active influenza C virus HEF glycoprotein expressed from cDNA.

    PubMed

    Pekosz, A; Lamb, R A

    1999-10-01

    The hemagglutinin, esterase, and fusion (HEF) glycoprotein of influenza C virus possesses receptor binding, receptor destroying, and membrane fusion activities. The HEF cDNAs from influenza C/Ann Arbor/1/50 (HEF-AA) and influenza C/Taylor/1223/47 (HEF-Tay) viruses were cloned and expressed, and transport of HEF to the cell surface was monitored by susceptibility to cleavage by exogenous trypsin, indirect immunofluorescence microscopy, and flow cytometry. Previously it has been found in studies with the C/Johannesburg/1/66 strain of influenza C virus (HEF-JHB) that transport of HEF to the cell surface is severely inhibited, and it is thought that the short cytoplasmic tail, Arg-Thr-Lys, is involved in blocking HEF cell surface expression (F. Oeffner, H.-D. Klenk, and G. Herrler, J. Gen. Virol. 80:363-369, 1999). As the cytoplasmic tail amino acid sequences of HEF-AA and HEF-Tay are identical to that of HEF-JHB, the data indicate that cell surface expression of HEF-AA and HEF-Tay is not inhibited by this amino acid sequence. Furthermore, the abundant cell surface transport of HEF-AA and HEF-Tay indicates that their cell surface expression does not require coexpression of another viral protein. The HEF-AA and HEF-Tay HEF glycoproteins bound human erythrocytes, promoted membrane fusion in a low-pH and trypsin-dependent manner, and displayed esterase activity, indicating that the HEF glycoprotein alone mediates all three known functions at the cell surface.

  13. Forced heat loss from body surface reduces heat flow to body surface.

    PubMed

    Berman, A

    2010-01-01

    Heat stress is commonly relieved by forced evaporation from body surfaces. The mode of heat stress relief by heat extraction from the periphery is not clear, although it reduces rectal temperature. Radiant surface temperature (Ts) of the right half of the body surface was examined by thermovision in 4 lactating Holstein cows (30 kg of milk/d) during 7 repeated cycles of forced evaporation created by 30s of wetting followed by 4.5 min of forced airflow. Wetting was performed by an array of sprinklers (0.76 m(3)/h), and forced airflow (>3m/s velocity) over the right side of the body surface was produced by fans mounted at a height of 3m above the ground. Sprinkling wetted the hind legs, rump, and chest, but not the lower abdomen side, front legs, or neck. The animals were maintained in shade at an air temperature of 28 degrees C and relative humidity of 47%. Coat thickness was 1 to 2mm, so Ts closely represented skin temperature. Mean Ts of 5 x 20cm areas on the upper and lower hind and front legs, rump, chest, abdomen side, and neck were obtained by converting to temperature their respective gray intensity in single frames obtained at 10-s intervals. Little change occurred in Ts during the first wetting (0.1+/-0.6 degrees C), but it decreased rapidly thereafter (1.6+/-0.6 degrees C in the fifth wetting). The Ts also decreased, to a smaller extent, in areas that remained dry (0.7+/-1.0 degrees C). In all body sites, a plateau in Ts was reached by 2 min after wetting. The difference between dry and wet areas in the first cooling cycle was approximately 1.2 degrees C. The Ts of different body areas decreased during consecutive cooling cycles and reached a plateau by 3 cooling cycles in dry sites (front leg, neck, abdomen side), by 5 cooling cycles in the hind leg, and 7 cooling cycles in the rump and chest. The reduction in mean Ts produced by 7 cycles was 4.0 to 6.0 degrees C in wetted areas and 1.6 to 3.7 degrees C in sites that were not wetted. Initial rectal

  14. Biochar-amended filter socks reduce herbicide losses via tile line surface inlets

    USDA-ARS?s Scientific Manuscript database

    Standing water in depressions and behind terraces in fields with subsurface drainage systems can result in reduced crop yields. This concern can be partially alleviated by installing surface inlets that reduce the duration of ponding. Unfortunately, these inlets provide an open conduit for surface w...

  15. Analysis of the limitations of hepatitis B surface antigen expression in soybean cell suspension cultures.

    PubMed

    Ganapathi, T R; Sunil Kumar, G B; Srinivas, L; Revathi, C J; Bapat, V A

    2007-09-01

    Soybean cell suspension cultures were transformed using Agrobacterium tumefaciens harboring pHBS/pHER constructs to express hepatitis B surface antigen (HBsAg). The transformed colonies were selected and analyzed for the expression of HBsAg by PCR, reverse transcription (RT) PCR, Western blot and ELISA analysis. The maximum expression of 700 ng/g F.W. was noted in pHER transformed cells. The highest expressing colonies were used to initiate the cell suspension cultures and the expression of HBsAg was estimated periodically. The expression levels were reduced drastically in cell suspension cultures compared to the colonies maintained on semi-solid medium. Various parameters were studied to maximize the cell growth and to retain the expression levels. The supplementation of culture medium with a protease inhibitor, leupeptin hemisulfate could restore up to 50% of HBsAg expression in cell suspension cultures. This is the first report to investigate the possible cause and solution to the loss of recombinant protein expression levels in plant cell suspension cultures.

  16. Effects of space flight on surface marker expression

    NASA Astrophysics Data System (ADS)

    Sonnenfeld, G.

    1999-01-01

    Space flight has been shown to affect expression of several cell surface markers. These markers play important roles in regulation of immune responses, including CD4 and CD8. The studies have involved flight of experimental animals and humans followed by analysis of tissue samples (blood in humans, rats and monkeys, spleen, thymus, lymph nodes and bone marrow in rodents). The degree and direction of the changes induced by space flight have been determined by the conditions of the flight. Also, there may be compartmentalization of the response of surface markers to space flight, with differences in the response of cells isolated from blood and local immune tissue. The same type of compartmentalization was also observed with cell adhesion molecules (integrins). In this case, the expression of integrins from lymph node cells differed from that of splenocytes isolated from rats immediately after space flight. Cell culture studies have indicated that there may be an inhibition in conversion of a precursor cell line to cells exhibiting mature macrophage characteristics after space flight, however, these experiments were limited as a result of technical difficulties. In general, it is clear that space flight results in alterations of cell surface markers. The biological significance of these changes remains to be established.

  17. Reduction of CD147 surface expression on primary T cells leads to enhanced cell proliferation.

    PubMed

    Biegler, Brian; Kasinrerk, Watchara

    2012-12-01

    CD147 is a ubiquitously expressed membrane glycoprotein that has numerous functional associations in health and disease. However, the molecular mechanisms by which CD147 participates in these processes are unclear. Establishing physiologically relevant silencing of CD147 in primary T cells could provide clues essential for elucidating some aspects of CD147 biology. To date, achieving the knockdown of CD147 in primary T cells has remained elusive. Utilizing RNA interference and the Nucleofector transfection system, we were able to reduce the expression of CD147 in primary T cells. Comparison of basic functions, such as proliferation and CD25 expression, were then made between control populations and populations with reduced expression. Up-regulation of CD147 was found upon T-cell activation, indicating a role in T-cell responses. To better understand the possible importance of this up-regulation, we knocked down the expression of CD147 using RNA interference. When compared to control populations the CD147 knockdown populations exhibited increased proliferation. This alteration of cell proliferation, however, was not linked to a change in CD25 expression. We achieved reduction of CD147 surface expression in primary T cells by siRNA-mediated gene silencing. Our results point to CD147 having a possible negative regulatory role in T cell-mediated immune responses.

  18. Cell surface expression of CD147/EMMPRIN is regulated by cyclophilin 60.

    PubMed

    Pushkarsky, Tatiana; Yurchenko, Vyacheslav; Vanpouille, Christophe; Brichacek, Beda; Vaisman, Iosif; Hatakeyama, Shigetsugu; Nakayama, Keiichi I; Sherry, Barbara; Bukrinsky, Michael I

    2005-07-29

    CD147, also known as extracellular matrix metalloproteinase inducer, is a regulator of matrix metalloproteinase production and also serves as a signaling receptor for extracellular cyclophilins. Previously, we demonstrated that cell surface expression of CD147 is sensitive to cyclophilin-binding drug cyclosporin A, suggesting involvement of a cyclophilin in the regulation of intracellular transport of CD147. In this report, we identify this cyclophilin as cyclophilin 60 (Cyp60), a distinct member of the cyclophilin family of proteins. CD147 co-immunoprecipitated with Cyp60, and confocal immunofluorescent microscopy revealed intracellular co-localization of Cyp60 and CD147. This interaction with Cyp60 involved proline 211 of CD147, which was shown previously to be critical for interaction between CD147 and another cyclophilin, cyclophilin A, in solution. Mutation of this proline residue abrogated co-immunoprecipitation of CD147 and Cyp60 and reduced surface expression of CD147 on the plasma membrane. Suppression of Cyp60 expression using RNA interference had an effect similar to that of cyclosporin A: reduction of cell surface expression of CD147. These results suggest that Cyp60 plays an important role in the translocation of CD147 to the cell surface. Therefore, Cyp60 may present a novel target for therapeutic interventions in diseases where CD147 functions as a pathogenic factor, such as cancer, human immunodeficiency virus infection, or rheumatoid arthritis.

  19. Erbin interacts with TARP γ-2 for surface expression of AMPA receptors in cortical interneurons.

    PubMed

    Tao, Yanmei; Chen, Yong-Jun; Shen, Chengyong; Luo, Zhengyi; Bates, C Ryan; Lee, Daehoon; Marchetto, Sylvie; Gao, Tian-Ming; Borg, Jean-Paul; Xiong, Wen-Cheng; Mei, Lin

    2013-03-01

    Inhibitory neurons control the firing of glutamatergic neurons and synchronize brain activity. However, little is known about mechanisms of excitatory synapse formation in inhibitory neurons. Here we demonstrate that Erbin is specifically expressed in cortical inhibitory neurons. It localizes at excitatory synapses and regulates AMPA receptor (AMPAR) surface expression. Erbin mutation reduced mEPSCs and AMPAR currents specifically in parvalbumin (PV)-positive interneurons but not in pyramidal neurons. We found that the AMPAR auxiliary protein TARP γ-2 was specifically expressed in cortical interneurons. Erbin interacts with TARP γ-2 and is crucial for its stability. Deletion of the γ-2-interacting domain in Erbin attenuated surface AMPAR and excitatory transmission in PV-positive interneurons. Furthermore, we observed behavioral deficits in Erbin-null mice and in mice expressing an Erbin truncation mutant that is unable to interact with TARP γ-2. These observations demonstrate a crucial function for Erbin in AMPAR surface expression in cortical PV-positive interneurons and may contribute to a better understanding of psychiatric disorders.

  20. Bromelain treatment reduces CD25 expression on activated CD4+ T cells in vitro✩

    PubMed Central

    Secor, Eric R.; Singh, Anurag; Guernsey, Linda A.; McNamara, Jeff T.; Zhan, Lijun; Maulik, Nilanjana; Thrall, Roger S.

    2009-01-01

    Bromelain (Br), an extract from pineapple stem with cysteine protease activity, exerts anti-inflammatory effects in a number of inflammatory models. We have previously shown that Br treatment decreased activated CD4+ T cells and has a therapeutic role in an ovalbumin-induced murine model of allergic airway disease. The current study was designed to determine the effect of Br on CD4+ T cell activation, specifically the expression of CD25 in vitro. CD25 is up regulated upon T cell activation, found as a soluble fraction (sCD25) and is a therapeutic target in inflammation, autoimmunity and allergy. Br treatment of anti-CD3 stimulated CD4+ T cells reduced CD25 expression in a dose and time dependent manner. This reduction of CD25 was dependent on the proteolytic action of Br as the addition of E64 (a cysteine protease inhibitor) abrogated this response. The concentration of sCD25 was increased in supernatants of Br treated activated CD4+ T cells as compared to control cells, suggesting that Br proteolytically cleaved cell-surface CD25. This novel mechanism of action identifies how Br may exert its therapeutic benefits in inflammatory conditions. PMID:19162239

  1. Insecticides reduce survival and the expression of traits associated with carnivory of carnivorous plants.

    PubMed

    Jennings, David E; Congelosi, Alexandra M; Rohr, Jason R

    2012-03-01

    While agrochemical pollution is thought to be an important conservation threat to carnivorous plants, the effects of insecticides on these taxa have not been quantified previously. Using a combination of lab- and field-based experiments, we tested the effects of commercial and technical grades of three widely used insecticides (carbaryl, lambda-cyhalothrin, and malathion) on survival and the expression of traits associated with carnivory of pink sundews (Drosera capillaris) and Venus flytraps (Dionaea muscipula). Commercial grades were generally more harmful than technical grades under lab and field conditions, but all three insecticides were capable of reducing both survival and the expression of traits associated with carnivory within recommended application rates. However, pink sundews appeared to be more susceptible to insecticides than Venus flytraps, perhaps because of larger numbers of digestive glands on the leaf surfaces. We make several recommendations for future research directions, such as examining the long-term effects of insecticides on carnivorous plant populations, for example in terms of growth rates and fitness. Additionally, future research should include representative species from a wider-range of carnivorous plant growth forms, and explore the mechanism by which insecticides are harming the plants. Given the effects we observed in the present study, we suggest that the use of insecticides should be carefully managed in areas containing vulnerable carnivorous plant species.

  2. Bromelain treatment reduces CD25 expression on activated CD4+ T cells in vitro.

    PubMed

    Secor, Eric R; Singh, Anurag; Guernsey, Linda A; McNamara, Jeff T; Zhan, Lijun; Maulik, Nilanjana; Thrall, Roger S

    2009-03-01

    Bromelain (Br), an extract from pineapple stem with cysteine protease activity, exerts anti-inflammatory effects in a number of inflammatory models. We have previously shown that Br treatment decreased activated CD4(+) T cells and has a therapeutic role in an ovalbumin-induced murine model of allergic airway disease. The current study was designed to determine the effect of Br on CD4(+) T cell activation, specifically the expression of CD25 in vitro. CD25 is up regulated upon T cell activation, found as a soluble fraction (sCD25) and is a therapeutic target in inflammation, autoimmunity and allergy. Br treatment of anti-CD3 stimulated CD4(+) T cells reduced CD25 expression in a dose and time dependent manner. This reduction of CD25 was dependent on the proteolytic action of Br as the addition of E64 (a cysteine protease inhibitor) abrogated this response. The concentration of sCD25 was increased in supernatants of Br treated activated CD4(+) T cells as compared to control cells, suggesting that Br proteolytically cleaved cell-surface CD25. This novel mechanism of action identifies how Br may exert its therapeutic benefits in inflammatory conditions.

  3. Surface-expressed enolases of Plasmodium and other pathogens

    PubMed Central

    Ghosh, Anil Kumar; Jacobs-Lorena, Marcelo

    2014-01-01

    Enolase is the eighth enzyme in the glycolytic pathway, a reaction that generates ATP from phosphoenol pyruvate in cytosolic compartments. Enolase is essential, especially for organisms devoid of the Krebs cycle that depend solely on glycolysis for energy. Interestingly, enolase appears to serve a separate function in some organisms, in that it is also exported to the cell surface via a poorly understood mechanism. In these organisms, surface enolase assists in the invasion of their host cells by binding plasminogen, an abundant plasma protease precursor. Binding is mediated by the interaction between a lysine motif of enolase with Kringle domains of plasminogen. The bound plasminogen is then cleaved by specific proteases to generate active plasmin. Plasmin is a potent serine protease that is thought to function in the degradation of the extracellular matrix surrounding the targeted host cell, thereby facilitating pathogen invasion. Recent work revealed that the malaria parasite Plasmodium also expresses surface enolase, and that this feature may be essential for completion of its life cycle. The therapeutic potential of targeting surface enolases of pathogens is discussed. PMID:21881761

  4. Face in profile view reduces perceived facial expression intensity: an eye-tracking study.

    PubMed

    Guo, Kun; Shaw, Heather

    2015-02-01

    Recent studies measuring the facial expressions of emotion have focused primarily on the perception of frontal face images. As we frequently encounter expressive faces from different viewing angles, having a mechanism which allows invariant expression perception would be advantageous to our social interactions. Although a couple of studies have indicated comparable expression categorization accuracy across viewpoints, it is unknown how perceived expression intensity and associated gaze behaviour change across viewing angles. Differences could arise because diagnostic cues from local facial features for decoding expressions could vary with viewpoints. Here we manipulated orientation of faces (frontal, mid-profile, and profile view) displaying six common facial expressions of emotion, and measured participants' expression categorization accuracy, perceived expression intensity and associated gaze patterns. In comparison with frontal faces, profile faces slightly reduced identification rates for disgust and sad expressions, but significantly decreased perceived intensity for all tested expressions. Although quantitatively viewpoint had expression-specific influence on the proportion of fixations directed at local facial features, the qualitative gaze distribution within facial features (e.g., the eyes tended to attract the highest proportion of fixations, followed by the nose and then the mouth region) was independent of viewpoint and expression type. Our results suggest that the viewpoint-invariant facial expression processing is categorical perception, which could be linked to a viewpoint-invariant holistic gaze strategy for extracting expressive facial cues.

  5. The European Continent : Surface Expression of Upper Mantle Dynamics

    NASA Astrophysics Data System (ADS)

    Tondi, M. R.; Schivardi, R.; Molinari, I.; Morelli, A.

    2012-12-01

    images of the European upper mantle isotropic shear-wave speeds and mass densities, recently recovered by combined inversion of surface-wave information and GRACE satellite gravity data (Tondi et al., 2012) are used to select the regions where the residual topography and the residual mantle gravity anomalies are strongly correlated (correlation coefficient is equal to 1). We assume surface uplift processes with negative density anomalies and downward pull with positive anomalies. Our work shows a strong correlation among the areas where, on the basis of our assumptions, the mantle dynamics have surface expression and the areas of low values of radial anisotropy: (1) the southern margins of the East European Craton, (2) the North-Eastern edges of the Arabian Plateau, (3) the northern edge of the CEVP (Central European Volcanic Province), (4) the North-Eastern part of the Atlantic Ocean, between Greenland and Iceland.

  6. Systemic alteration of cell-surface and secreted glycoprotein expression in malignant breast cancer cell lines.

    PubMed

    Timpe, Leslie C; Yen, Roger; Haste, Nicole V; Litsakos-Cheung, Christina; Yen, Ten-Yang; Macher, Bruce A

    2013-11-01

    Breast cancer cell lines express fewer transmembrane and secreted glycoproteins than nonmalignant ones. The objective of these experiments was to characterize the changes in the expression of several hundred glycoproteins quantitatively. Secreted and cell-surface glycoproteins were isolated using a glycoprotein capture protocol and then identified by tandem mass spectrometry. Glycoproteins expressed by a group of cell lines originating from malignant tumors of the breast were compared with those expressed by a nonmalignant set. The average number of spectral counts (proportional to relative protein abundance) and the total number of glycopeptides in the malignant samples were reduced to about two-thirds of the level in the nonmalignant samples. Most glycoproteins were expressed at a different level in the malignant samples, with nearly as many increasing as decreasing. The glycoproteins with reduced expression accounted for a larger change in spectral counts, and hence for the net loss of spectral counts in the malignant lines. Similar results were found when the glycoproteins were studied via identified glycosylation sites only, or through identified sites together with non-glycopeptides. The overall reduction is largely due to the loss of integrins, laminins and other proteins that form or interact with the basement membrane.

  7. GABAA Receptor α1 Subunit Mutation A322D Associated with Autosomal Dominant Juvenile Myoclonic Epilepsy Reduces the Expression and Alters the Composition of Wild Type GABAA Receptors*

    PubMed Central

    Ding, Li; Feng, Hua-Jun; Macdonald, Robert L.; Botzolakis, Emanuel J.; Hu, Ningning; Gallagher, Martin J.

    2010-01-01

    A GABAA receptor (GABAAR) α1 subunit mutation, A322D (AD), causes an autosomal dominant form of juvenile myoclonic epilepsy (ADJME). Previous studies demonstrated that the mutation caused α1(AD) subunit misfolding and rapid degradation, reducing its total and surface expression substantially. Here, we determined the effects of the residual α1(AD) subunit expression on wild type GABAAR expression to determine whether the AD mutation conferred a dominant negative effect. We found that although the α1(AD) subunit did not substitute for wild type α1 subunits on the cell surface, it reduced the surface expression of α1β2γ2 and α3β2γ2 receptors by associating with the wild type subunits within the endoplasmic reticulum and preventing them from trafficking to the cell surface. The α1(AD) subunit reduced surface expression of α3β2γ2 receptors by a greater amount than α1β2γ2 receptors, thus altering cell surface GABAAR composition. When transfected into cultured cortical neurons, the α1(AD) subunit altered the time course of miniature inhibitory postsynaptic current kinetics and reduced miniature inhibitory postsynaptic current amplitudes. These findings demonstrated that, in addition to causing a heterozygous loss of function of α1(AD) subunits, this epilepsy mutation also elicited a modest dominant negative effect that likely shapes the epilepsy phenotype. PMID:20551311

  8. Tocotrienol is the most effective vitamin E for reducing endothelial expression of adhesion molecules and adhesion to monocytes.

    PubMed

    Theriault, Andre; Chao, Jun-Tzo; Gapor, Abdul; Chao, Jun Tzo; Gapor, Abeli

    2002-01-01

    Alpha-tocopherol and its esterified derivatives have been shown to be effective in reducing monocytic-endothelial cell adhesion. However, the effect of alpha-tocotrienol (alpha-T3) has not been characterized. In the present study, using human umbilical vein endothelial cells (HUVEC) as the model system, we examined the relative inhibitory effects of alpha-T3 and other vitamin E derivatives on cell surface adhesion molecule expression under TNF-alpha stimulation. Using enzyme-linked immunosorbent assay, we demonstrated that alpha-T3 markedly inhibited the surface expression of vascular cell adhesion molecule-1 in TNF-alpha activated HUVEC in a dose- and time-dependent manner. The optimal inhibition was observed at 25 micromol/l alpha-T3 within 24 h (77+/-5%) without cytotoxicity. In addition, the surface expression of intercellular adhesion molecule-1 and E-selectin were also reduced by 40+/-7 and 42+/-5%, respectively. In order to further evaluate the effects of alpha-T3 on the vascular endothelium, we investigated the ability of monocytes to adhere to endothelial cells. Interestingly, a 63+/-3% decrease in monocytic cell adherence was observed. Compared to alpha-tocopherol and alpha-tocopheryl succinate, alpha-T3 displayed a more profound inhibitory effect on adhesion molecule expression and monocytic cell adherence. This inhibitory action by alpha-T3 on TNF-alpha-induced monocyte adhesion was shown to be NF-kappaB dependent and was interestingly reversed with co-incubation with farnesol and geranylgeraniol, suggesting a role for prenylated proteins in the regulation of adhesion molecule expression. In summary, the above results suggest that alpha-T3 is a potent and effective agent in the reduction of cellular adhesion molecule expression and monocytic cell adherence.

  9. Reduced expression of Autographa californica nucleopolyhedrovirus ORF34, an essential gene, enhances heterologous gene expression

    SciTech Connect

    Salem, Tamer Z.; Zhang, Fengrui; Thiem, Suzanne M.

    2013-01-20

    Autographa californica multiple nucleopolyhedrovirus ORF34 is part of a transcriptional unit that includes ORF32, encoding a viral fibroblast growth factor (FGF) and ORF33. We identified ORF34 as a candidate for deletion to improve protein expression in the baculovirus expression system based on enhanced reporter gene expression in an RNAi screen of virus genes. However, ORF34 was shown to be an essential gene. To explore ORF34 function, deletion (KO34) and rescue bacmids were constructed and characterized. Infection did not spread from primary KO34 transfected cells and supernatants from KO34 transfected cells could not infect fresh Sf21 cells whereas the supernatant from the rescue bacmids transfection could recover the infection. In addition, budded viruses were not observed in KO34 transfected cells by electron microscopy, nor were viral proteins detected from the transfection supernatants by western blots. These demonstrate that ORF34 is an essential gene with a possible role in infectious virus production.

  10. Reduced expression of Paternally Expressed Gene-3 enhances somatic cell reprogramming through mitochondrial activity perturbation.

    PubMed

    Theka, Ilda; Sottile, Francesco; Aulicino, Francesco; Garcia, Alvaro Castells; Cosma, Maria Pia

    2017-08-29

    Imprinted genes control several cellular and metabolic processes in embryonic and adult tissues. In particular, paternally expressed gene-3 (Peg3) is active in the adult stem cell population and during muscle and neuronal lineage development. Here we have investigated the role of Peg3 in mouse embryonic stem cells (ESCs) and during the process of somatic cell reprogramming towards pluripotency. Our data show that Peg3 knockdown increases expression of pluripotency genes in ESCs and enhances reprogramming efficiency of both mouse embryonic fibroblasts and neural stem cells. Interestingly, we observed that altered activity of Peg3 correlates with major perturbations of mitochondrial gene expression and mitochondrial function, which drive metabolic changes during somatic cell reprogramming. Overall, our study shows that Peg3 is a regulator of pluripotent stem cells and somatic cell reprogramming.

  11. Flumazenil decreases surface expression of α4β2δ GABAA receptors by increasing the rate of receptor internalization

    PubMed Central

    Kuver, Aarti; Smith, Sheryl S.

    2015-01-01

    Increases in expression of α4βδ GABAA receptors (GABARs), triggered by fluctuations in the neurosteroid THP (3α-OH-5α[β]-pregnan-20-one), are associated with changes in mood and cognition. We tested whether α4βδ trafficking and surface expression would be altered by in vitro exposure to flumazenil, a benzodiazepine ligand which reduces α4βδ expression in vivo. We first determined that flumazenil (100 nM – 100 μM, IC50=~1 μM) acted as a negative modulator, reducing GABA (10 μM)-gated current in the presence of 100 nM THP (to increase receptor efficacy), assessed with whole cell patch clamp recordings of recombinant α4β2δ expressed in HEK-293 cells. Surface expression of recombinant α4β2δ receptors was detected using a 3XFLAG reporter at the C-terminus of α4 (α4F) using confocal immunocytochemical techniques following 48 h exposure of cells to GABA (10 μM) + THP (100 nM). Flumazenil (10 μM) decreased surface expression of α4F by ~60%, while increasing its intracellular accumulation, after 48 h. Reduced surface expression of α4β2δ after flumazenil treatment was confirmed by decreases in the current responses to 100 nM of the GABA agonist gaboxadol. Flumazenil-induced decreases in surface expression of α4β2δ were prevented by the dynamin blocker, dynasore, and by leupeptin, which blocks lysosomal enzymes, suggesting that flumazenil is acting to increase endocytosis and lysosomal degradation of the receptor. Flumazenil increased the rate of receptor removal from the cell surface by 2-fold, assessed using botulinum toxin B to block insertion of new receptors. These findings may suggest new therapeutic strategies for regulation of α4β2δ expression using flumazenil. PMID:26592470

  12. Reduced COX-2 Expression in Aged Mice Is Associated With Impaired Fracture Healing

    PubMed Central

    Naik, Amish A; Xie, Chao; Zuscik, Michael J; Kingsley, Paul; Schwarz, Edward M; Awad, Hani; Guldberg, Robert; Drissi, Hicham; Puzas, J Edward; Boyce, Brendan; Zhang, Xinping; O'Keefe, Regis J

    2009-01-01

    The cellular and molecular events responsible for reduced fracture healing with aging are unknown. Cyclooxygenase 2 (COX-2), the inducible regulator of prostaglandin E2 (PGE2) synthesis, is critical for normal bone repair. A femoral fracture repair model was used in mice at either 7–9 or 52–56 wk of age, and healing was evaluated by imaging, histology, and gene expression studies. Aging was associated with a decreased rate of chondrogenesis, decreased bone formation, reduced callus vascularization, delayed remodeling, and altered expression of genes involved in repair and remodeling. COX-2 expression in young mice peaked at 5 days, coinciding with the transition of mesenchymal progenitors to cartilage and the onset of expression of early cartilage markers. In situ hybridization and immunohistochemistry showed that COX-2 is expressed primarily in early cartilage precursors that co-express col-2. COX-2 expression was reduced by 75% and 65% in fractures from aged mice compared with young mice on days 5 and 7, respectively. Local administration of an EP4 agonist to the fracture repair site in aged mice enhanced the rate of chondrogenesis and bone formation to levels observed in young mice, suggesting that the expression of COX-2 during the early inflammatory phase of repair regulates critical subsequent events including chondrogenesis, bone formation, and remodeling. The findings suggest that COX-2/EP4 agonists may compensate for deficient molecular signals that result in the reduced fracture healing associated with aging. PMID:18847332

  13. The Glycophosphatidylinositol Anchor of the MCMV Evasin, m157, Facilitates Optimal Cell Surface Expression and Ly49 Receptor Recognition

    PubMed Central

    Carlin, Lindsey E.; Guseva, Natalya V.; Shey, Michael R.; Ballas, Zuhair K.; Heusel, Jonathan W.

    2013-01-01

    The murine cytomegalovirus-encoded protein m157 is a cognate ligand for both inhibitory and activating receptors expressed by natural killer cells. Additionally, m157 is expressed on the surface of infected cells by a glycophosphatidylinositol (GPI) anchor. Although endogenous GPI-anchored proteins are known to be ligands for the NK cell receptor, NKG2D, the contribution of the GPI anchor for viral m157 ligand function is unknown. To determine whether the GPI anchor for m157 is dispensable for m157 function, we generated m157 variants expressed as transmembrane fusion proteins and tested cells expressing transmembrane m157 for the capacity to activate cognate Ly49 receptors. We found that the GPI anchor is required for high-level cell surface expression of m157, and that the transmembrane m157 ligand retains the capacity to activate reporter cells and NK cells expressing Ly49H, as well as Ly49I129 reporter cells, but with reduced potency. Importantly, target cells expressing the transmembrane form of m157 were killed less efficiently and failed to mediate Ly49H receptor downregulation on fresh NK cells compared to targets expressing GPI-anchored m157. Taken together, these results show that the GPI anchor for m157 facilitates robust cell surface expression, and that NK cells are sensitive to the altered cell surface expression of this potent viral evasin. PMID:23840655

  14. Thrombin Increases Expression of Fibronectin Antigen on the Platelet Surface

    NASA Astrophysics Data System (ADS)

    Ginsberg, Mark H.; Painter, Richard G.; Forsyth, Jane; Birdwell, Charles; Plow, Edward F.

    1980-02-01

    Fibronectins (fn) are adhesive glycoproteins which bind to collagen and to fibrin and appear to be important in cellular adhesion to other cells or surfaces. Fn-related antigen is present in human platelets, suggesting a possible role for fn in the adhesive properties of platelets. We have studied the localization of fn in resting and thrombin-stimulated platelets by immunofluorescence and quantitative binding of radiolabeled antibody. In resting fixed platelets, variable light surface staining for fn was observed. When these cells were made permeable to antibody with detergent, staining for fn was markedly enhanced and was present in a punctate distribution, suggesting intracellular localization. Stimulation with thrombin, which is associated with increased platelet adhesiveness, resulted in increased staining for fn antigen on intact platelets. These stimulated cells did not leak 51Cr nor did they stain for F-actin, thus documenting that the increased fn staining was not due to loss of plasma membrane integrity. The thrombin-induced increase in accessible platelet fn antigen was confirmed by quantitative antibody binding studies in which thrombin-stimulated platelets specifically bound 15 times as much radiolabeled F(ab')2 anti-fn as did resting cells. Thus, thrombin stimulation results in increased expression of fn antigen on the platelet surface. Here it may participate in interactions with fibrin, connective tissue, or other cells.

  15. Modified expression of surface glyconjugates in stored human platelets

    SciTech Connect

    Dhar, A.; Ganguly, P.

    1987-05-01

    Platelets are anucleated cells which play an important part in blood coagulation and thrombosis. These cells may be stored in the blood bank for only 4/5 days. In order to improve the storage of platelets, it is essential to first understand the changes in these cells due to storage. In this work, human platelets were stored in autologous plasma at 4/sup 0/ or 22/sup 0/ and their surface changes were monitored with three lectins - wheat germ afflutinin (WGA), concanavalin A (Con A) and lentil lectin (LL). Blood was drawn from healthy donors and platelet rich plasma (PRP) was collected by slow speed centrifugation. Platelets stored at either temperature for different times showed increased sensitivity to agglutination by WGA after 34-48 hrs. Lectins, Con A and LL, which were not agglutinating to fresh platelets readily caused agglutination after 48-72 hrs. The platelets stored for 25 hrs or longer period were insensitive to thrombin but showed enhanced aggregation with WGA. Labelling of surface glycoconjugates of stored platelets with /sup 3/H-boro-hydride revealed progressive loss of a glycoprotein of Mr 150,000 (GPIb infinity) together with the appearance of components of Mr 69,000; Mr 60,000; Mr 25,000. New high molecular weight glycoproteins were also detected only in stored platelets. The author studies clearly indicate that modification or altered expression of platelets surface glycoproteins may be one factor of storage related dysfunction of platelets.

  16. Reduced Duodenal Cytochrome P450 3A Protein Expression and Catalytic Activity in Patients with Cirrhosis

    PubMed Central

    McConn, Donavon J.; Lin, Yvonne S.; Mathisen, Terri L.; Blough, David K.; Xu, Yang; Hashizume, Takanori; Taylor, Shari L.; Thummel, Kenneth E.; Shuhart, Margaret C.

    2009-01-01

    The small intestine and liver express high levels of cytochrome P450 3A (CYP3A), an enzyme subfamily contributing significantly to drug metabolism. In patients with cirrhosis, reduced metabolism of drugs is typically attributed to decreased liver function, but it is unclear whether intestinal drug metabolism is also compromised. In this study, we compared CYP3A protein expression and in vitro midazolam hydroxylation in duodenal mucosal biopsies from subjects with normal liver function (controls; n=20) and subjects with varying severity of cirrhosis (n=23). Compared to samples from controls, duodenal CYP3A expression and total midazolam hydroxylation was reduced by 47% and 34%, respectively in samples from subjects with cirrhosis. Greater decreases in CYP3A expression were seen in subjects with increasing severity of cirrhosis. Thus, patients with advanced cirrhosis may have increased drug exposure following oral dosing as a result of both impaired liver function and decreased intestinal CYP3A expression and activity. PMID:19212316

  17. Assessing and reducing sources of gene expression variability in Staphylococcus epidermidis biofilms.

    PubMed

    Sousa, Cármen; França, Angela; Cerca, Nuno

    2014-12-01

    Gene expression quantification can be a useful tool in studying the properties of bacterial biofilms. Unfortunately, techniques such as RNA extraction, cDNA synthesis, and quantitative PCR (qPCR) can introduce variability into mRNA transcript measurements, obscuring biologically relevant results. Here we sought to identify the steps that impair accurate gene expression quantification from Staphylococcus epidermidis biofilm samples. We devised an experimental setup that could be used to determine the contribution of each experimental step to the variability of mRNA transcript measurement. Among factors tested, biofilm growth contributed the most bias to gene expression quantification. Additional experiments demonstrated that pooling biofilms together reduced this variability, resulting in more accurate gene expression analysis results. We therefore recommend pooling in order to reduce the variability associated with gene expression quantification from biofilm samples.

  18. Post-fire logging reduces surface woody fuels up to four decades following wildfire

    Treesearch

    David W. Peterson; Erich Kyle Dodson; Richy J. Harrod

    2015-01-01

    Severe wildfires create pulses of dead trees that influence future fuel loads, fire behavior, and fire effects as they decay and deposit surface woody fuels. Harvesting fire-killed trees may reduce future surface woody fuels and related fire hazards, but the magnitude and timing of post-fire logging effects on woody fuels have not been fully assessed. To address this...

  19. Increased expression of LDL receptor-related protein 1 during human cytomegalovirus infection reduces virion cholesterol and infectivity

    PubMed Central

    Gudleski-O’Regan, Nicole; Greco, Todd M.; Cristea, Ileana M.; Shenk, Thomas

    2012-01-01

    In response to virus infection, cells can alter protein expression to modify cellular functions and limit viral replication. To examine host protein expression during infection with human cytomegalovirus (HCMV), an enveloped DNA virus, we performed a semi-quantitative, temporal analysis of the cell surface proteome in infected fibroblasts. We determined that resident low density lipoprotein related receptor 1 (LRP1), a plasma membrane receptor that regulates lipid metabolism, is elevated early after HCMV infection, resulting in decreased intracellular cholesterol. siRNA knockdown or antibody-mediated inhibition of LRP1 increased intracellular cholesterol, and concomitantly increased the infectious virus yield. Virions produced under these conditions contained elevated cholesterol, resulting in increased infectivity. Depleting cholesterol from virions reduced their infectivity by blocking fusion of the virion envelope with the cell membrane. Thus, LRP1 restricts HCMV infectivity by controlling the availability of cholesterol for the virion envelope and increased LRP1 expression is likely a defense response to infection. PMID:22817990

  20. Palmitoylation of the β4-Subunit Regulates Surface Expression of Large Conductance Calcium-activated Potassium Channel Splice Variants*

    PubMed Central

    Chen, Lie; Bi, Danlei; Tian, Lijun; McClafferty, Heather; Steeb, Franziska; Ruth, Peter; Knaus, Hans Guenther; Shipston, Michael J.

    2013-01-01

    Regulatory β-subunits of large conductance calcium- and voltage-activated potassium (BK) channels play an important role in generating functional diversity and control of cell surface expression of the pore forming α-subunits. However, in contrast to α-subunits, the role of reversible post-translational modification of intracellular residues on β-subunit function is largely unknown. Here we demonstrate that the human β4-subunit is S-acylated (palmitoylated) on a juxtamembrane cysteine residue (Cys-193) in the intracellular C terminus of the regulatory β-subunit. β4-Subunit palmitoylation is important for cell surface expression and endoplasmic reticulum (ER) exit of the β4-subunit alone. Importantly, palmitoylated β4-subunits promote the ER exit and surface expression of the pore-forming α-subunit, whereas β4-subunits that cannot be palmitoylated do not increase ER exit or surface expression of α-subunits. Strikingly, however, this palmitoylation- and β4-dependent enhancement of α-subunit surface expression was only observed in α-subunits that contain a putative trafficking motif (… REVEDEC) at the very C terminus of the α-subunit. Engineering this trafficking motif to other C-terminal α-subunit splice variants results in α-subunits with reduced surface expression that can be rescued by palmitoylated, but not depalmitoylated, β4-subunits. Our data reveal a novel mechanism by which palmitoylated β4-subunit controls surface expression of BK channels through masking of a trafficking motif in the C terminus of the α-subunit. As palmitoylation is dynamic, this mechanism would allow precise control of specific splice variants to the cell surface. Our data provide new insights into how complex interplay between the repertoire of post-transcriptional and post-translational mechanisms controls cell surface expression of BK channels. PMID:23504458

  1. Cyclosporin A reduces expression of adhesion molecules in the kidney of rats with chronic serum sickness

    PubMed Central

    Rincón, J; Parra, G; Quiroz, Y; Benatuil, L; Rodríguez-Iturbe, B

    2000-01-01

    Treatment with cyclosporin A (CsA) improves proteinuria and reduces renal cellular infiltration in chronic serum sickness (CSS). We examined if these effects were associated with a reduced renal expression of CD54 and its ligands, interferon-gamma (IFN-γ), tumour necrosis factor-alpha (TNF-α) and MHC class II molecules. We studied two groups of rats in which CSS was induced by daily injections of ovalbumin (OVA): a group treated with CsA (OVA.CsA group, n = 11) and a group that received no treatment (OVA.CSS group, n = 11). An additional group of five rats (control group) received only phosphate buffer. Immunostaining techniques were used to follow CSS and to study the expression of CD54, CD18, CD11b/c, IFN-γ, TNF-α and MHC class molecules. Proteinuria (mg/24 h) was reduced from 248·2 ± 73·1 (OVA.CCS group) to 14·5 ± 13·1 with CsA treatment (P < 0·0001). The renal expression of CD54 and its ligands (CD18 and CD11b/c) was reduced by 50% to 75%. Correspondingly, there was a 60% to 85% reduction in the number of infiltrating leucocytes. The number of cells expressing TNF-α, IFN-γ and MHC II molecules was also reduced. CsA reduces expression of CD54 and its ligands. This effect is associated with a reduction of cellular infiltration, IFN-γ, TNF-α-producing cells and with MHC II expression in the kidney. These findings suggest that expression of adhesion molecules plays a critical role in CSS and underline the importance of cellular immunity in this experimental model. PMID:10931158

  2. A surface with a biomimetic micropattern reduces colonization of Mycobacterium abscessus.

    PubMed

    Kim, Eun; Kinney, William H; Ovrutsky, Alida R; Vo, Danthy; Bai, Xiyuan; Honda, Jennifer R; Marx, Grace; Peck, Emily; Lindberg, Leslie; Falkinham, Joseph O; May, Rhea M; Chan, Edward D

    2014-11-01

    Nontuberculous mycobacteria (NTM) are ubiquitous organisms found in soil, water, and biofilms. Engineered surface topography has been proposed as a method to reduce microbial biofilm formation. The Sharklet(®) micropattern silicone surface has been shown to reduce biofilm formation of pyogenic bacteria. We hypothesized that this micropattern surface will also reduce colonization by Mycobacterium abscessus, a human pathogen. Smooth and micropattern silicone samples were incubated with 1 × 10(6) M. abscessus mL(-1) for 2 and 4 days. After processing to optimize recovery of adhered mycobacteria, there was a 75% and 50% reduction in the number of viable M. abscessus recovered from the micropattern surfaces compared to the smooth surfaces at 2 and 4 days after inoculation, respectively. Ziehl-Neelsen staining after measures to remove the adherent microorganisms revealed fewer residual M. abscessus on the micropattern samples as compared to smooth samples, validating the quantitative culture results. Microscopic observation of 2, 4, and 8 day M. abscessus cultures on micropattern samples showed that the organisms preferentially colonized within the channels between the rectangular features. In summary, a micropattern surface reduces the colonization of a pathogenic NTM. It remains to be seen whether this micropattern can reduce infections in humans. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  3. Mechanisms of Reduced Astrocyte Surface Coverage in Cortical Neuron-Glia Co-cultures on Nanoporous Gold Surfaces.

    PubMed

    Chapman, Christopher A R; Chen, Hao; Stamou, Marianna; Lein, Pamela J; Seker, Erkin

    2016-09-01

    Nanoporous gold (np-Au) is a promising multifunctional material for neural electrodes. We have previously shown that np-Au nanotopography reduces astrocyte surface coverage (linked to undesirable gliosis) while maintaining high neuronal coverage in a cortical primary neuron-glia co-culture model as long as two weeks in vitro. Here, we investigate the potential influence of secreted soluble factors from cells grown on np-Au on the cell type-specific surface coverage of cells grown on conventional tissue culture plastic and test the hypothesis that secretion of factors is responsible for inhibiting astrocyte coverage on np-Au. In order to assess whether factors secreted from cells grown on np-Au surfaces reduced surface coverage by astrocytes, we seeded fresh primary rat neuron-glia co-cultures on conventional polystyrene culture dishes, but maintained the cells in conditioned media from co-cultures grown on np-Au surfaces. After one week in vitro, a preferential reduction in astrocyte surface coverage was not observed, suggesting that soluble factors are not playing a role. In contrast, four hours after cell seeding there were a significant number of non-adhered, yet still viable, cells for the cultures on np-Au surfaces. We hypothesize that the non-adherent cells are mainly astrocytes, because: (i) there was no difference in neuronal cell coverage between np-Au and pl-Au for long culture durations and (ii) neurons are post-mitotic and not expected to increase in number upon attaching to the surface. Overall, the results suggest that the np-Au topography leads to preferential neuronal attachment shortly after cell seeding and limits astrocyte-specific np-Au surface coverage at longer culture durations.

  4. Surface L-type Ca2+ channel expression levels are increased in aged hippocampus

    PubMed Central

    Núñez-Santana, Félix Luis; Oh, Myongsoo Matthew; Antion, Marcia Diana; Lee, Amy; Hell, Johannes Wilhelm; Disterhoft, John Francis

    2014-01-01

    Age-related increase in L-type Ca2+ channel (LTCC) expression in hippocampal pyramidal neurons has been hypothesized to underlie the increased Ca2+ influx and subsequent reduced intrinsic neuronal excitability of these neurons that lead to age-related cognitive deficits. Here, using specific antibodies against Cav1.2 and Cav1.3 subunits of LTCCs, we systematically re-examined the expression of these proteins in the hippocampus from young (3 to 4 month old) and aged (30 to 32 month old) F344xBN rats. Western blot analysis of the total expression levels revealed significant reductions in both Cav1.2 and Cav1.3 subunits from all three major hippocampal regions of aged rats. Despite the decreases in total expression levels, surface biotinylation experiments revealed significantly higher proportion of expression on the plasma membrane of Cav1.2 in the CA1 and CA3 regions and of Cav1.3 in the CA3 region from aged rats. Furthermore, the surface biotinylation results were supported by immunohistochemical analysis that revealed significant increases in Cav1.2 immunoreactivity in the CA1 and CA3 regions of aged hippocampal pyramidal neurons. In addition, we found a significant increase in the level of phosphorylated Cav1.2 on the plasma membrane in the dentate gyrus of aged rats. Taken together, our present findings strongly suggest that age-related cognitive deficits cannot be attributed to a global change in L-type channel expression nor to the level of phosphorylation of Cav1.2 on the plasma membrane of hippocampal neurons. Rather, increased expression and density of LTCCs on the plasma membrane may underlie the age-related increase in L-type Ca2+ channel activity in CA1 pyramidal neurons. PMID:24033980

  5. Reducing the In2O3(111) Surface Results in Ordered Indium Adatoms

    SciTech Connect

    Wagner, Margareta; Seiler, Steffen; Meyer, Bernd; Boatner, Lynn A; Schmid, M.; Diebold, U.

    2014-01-01

    The In2O3(111) surface can be transformed from an oxidized bulk termination to one that is covered by single In adatoms. As each adatom sits at one specific site within the surface unit cell they form a well-ordered (1 1) superstructure. Annealing at 500 C in O2 or in ultrahigh vacuum results in a fully reversible conversion between these two surface terminations; this transformation and intermediate stages were followed with Scanning Tunneling Microscopy (STM). Formation of this novel surface structure under reducing conditions is corroborated by Density Functional Theory (DFT). The reduced adatom-covered and the oxidized In2O3(111) surfaces are expected to exhibit different chemical and electronic properties, which can easily be exploited by the facile and reversible switching between the two terminations.

  6. Improved chemical and electrochemical stability of perovskite oxides with less reducible cations at the surface

    SciTech Connect

    Tsvetkov, Nikolai; Lu, Qiyang; Sun, Lixin; Crumlin, Ethan J.; Yildiz, Bilge

    2016-06-13

    Segregation and phase separation of aliovalent dopants on perovskite oxide (ABO 3 ) surfaces are detrimental to the performance of energy conversion systems such as solid oxide fuel/electrolysis cells and catalysts for thermochemical H 2 O and CO 2 splitting. One key reason behind the instability of perovskite oxide surfaces is the electrostatic attraction of the negatively charged A-site dopants (for example, Sr La ') by the positively charged oxygen vacancies (V $••\\atop{o}$) enriched at the surface. Here we show that reducing the surface V $••\\atop{o}$ concentration improves the oxygen surface exchange kinetics and stability significantly, albeit contrary to the well-established understanding that surface oxygen vacancies facilitate reactions with O 2 molecules. We take La 0.8 Sr 0.2 CoO 3 (LSC) as a model perovskite oxide, and modify its surface with additive cations that are more and less reducible than Co on the B-site of LSC. By using ambient-pressure X-ray absorption and photoelectron spectroscopy, we proved that the dominant role of the less reducible cations is to suppress the enrichment and phase separation of Sr while reducing the concentration of V $••\\atop{o}$ and making the LSC more oxidized at its surface. Consequently, we found that these less reducible cations significantly improve stability, with up to 30 times faster oxygen exchange kinetics after 54 h in air at 530 °C achieved by Hf addition onto LSC. Finally, the results revealed a 'volcano' relation between the oxygen exchange kinetics and the oxygen vacancy formation enthalpy of the binary oxides of the additive cations. This volcano relation highlights the existence of an optimum surface oxygen vacancy concentration that balances the gain in oxygen exchange kinetics and the chemical stability loss.

  7. Improved chemical and electrochemical stability of perovskite oxides with less reducible cations at the surface

    NASA Astrophysics Data System (ADS)

    Tsvetkov, Nikolai; Lu, Qiyang; Sun, Lixin; Crumlin, Ethan J.; Yildiz, Bilge

    2016-09-01

    Segregation and phase separation of aliovalent dopants on perovskite oxide (ABO3) surfaces are detrimental to the performance of energy conversion systems such as solid oxide fuel/electrolysis cells and catalysts for thermochemical H2O and CO2 splitting. One key reason behind the instability of perovskite oxide surfaces is the electrostatic attraction of the negatively charged A-site dopants (for example, ) by the positively charged oxygen vacancies () enriched at the surface. Here we show that reducing the surface concentration improves the oxygen surface exchange kinetics and stability significantly, albeit contrary to the well-established understanding that surface oxygen vacancies facilitate reactions with O2 molecules. We take La0.8Sr0.2CoO3 (LSC) as a model perovskite oxide, and modify its surface with additive cations that are more and less reducible than Co on the B-site of LSC. By using ambient-pressure X-ray absorption and photoelectron spectroscopy, we proved that the dominant role of the less reducible cations is to suppress the enrichment and phase separation of Sr while reducing the concentration of and making the LSC more oxidized at its surface. Consequently, we found that these less reducible cations significantly improve stability, with up to 30 times faster oxygen exchange kinetics after 54 h in air at 530 °C achieved by Hf addition onto LSC. Finally, the results revealed a `volcano' relation between the oxygen exchange kinetics and the oxygen vacancy formation enthalpy of the binary oxides of the additive cations. This volcano relation highlights the existence of an optimum surface oxygen vacancy concentration that balances the gain in oxygen exchange kinetics and the chemical stability loss.

  8. Improved chemical and electrochemical stability of perovskite oxides with less reducible cations at the surface.

    PubMed

    Tsvetkov, Nikolai; Lu, Qiyang; Sun, Lixin; Crumlin, Ethan J; Yildiz, Bilge

    2016-09-01

    Segregation and phase separation of aliovalent dopants on perovskite oxide (ABO3) surfaces are detrimental to the performance of energy conversion systems such as solid oxide fuel/electrolysis cells and catalysts for thermochemical H2O and CO2 splitting. One key reason behind the instability of perovskite oxide surfaces is the electrostatic attraction of the negatively charged A-site dopants (for example, ) by the positively charged oxygen vacancies () enriched at the surface. Here we show that reducing the surface concentration improves the oxygen surface exchange kinetics and stability significantly, albeit contrary to the well-established understanding that surface oxygen vacancies facilitate reactions with O2 molecules. We take La0.8Sr0.2CoO3 (LSC) as a model perovskite oxide, and modify its surface with additive cations that are more and less reducible than Co on the B-site of LSC. By using ambient-pressure X-ray absorption and photoelectron spectroscopy, we proved that the dominant role of the less reducible cations is to suppress the enrichment and phase separation of Sr while reducing the concentration of and making the LSC more oxidized at its surface. Consequently, we found that these less reducible cations significantly improve stability, with up to 30 times faster oxygen exchange kinetics after 54 h in air at 530 °C achieved by Hf addition onto LSC. Finally, the results revealed a 'volcano' relation between the oxygen exchange kinetics and the oxygen vacancy formation enthalpy of the binary oxides of the additive cations. This volcano relation highlights the existence of an optimum surface oxygen vacancy concentration that balances the gain in oxygen exchange kinetics and the chemical stability loss.

  9. Improved chemical and electrochemical stability of perovskite oxides with less reducible cations at the surface

    DOE PAGES

    Tsvetkov, Nikolai; Lu, Qiyang; Sun, Lixin; ...

    2016-06-13

    Segregation and phase separation of aliovalent dopants on perovskite oxide (ABO 3 ) surfaces are detrimental to the performance of energy conversion systems such as solid oxide fuel/electrolysis cells and catalysts for thermochemical H 2 O and CO 2 splitting. One key reason behind the instability of perovskite oxide surfaces is the electrostatic attraction of the negatively charged A-site dopants (for example, Sr La ') by the positively charged oxygen vacancies (Vmore » $$••\\atop{o}$$) enriched at the surface. Here we show that reducing the surface V $$••\\atop{o}$$ concentration improves the oxygen surface exchange kinetics and stability significantly, albeit contrary to the well-established understanding that surface oxygen vacancies facilitate reactions with O 2 molecules. We take La 0.8 Sr 0.2 CoO 3 (LSC) as a model perovskite oxide, and modify its surface with additive cations that are more and less reducible than Co on the B-site of LSC. By using ambient-pressure X-ray absorption and photoelectron spectroscopy, we proved that the dominant role of the less reducible cations is to suppress the enrichment and phase separation of Sr while reducing the concentration of V $$••\\atop{o}$$ and making the LSC more oxidized at its surface. Consequently, we found that these less reducible cations significantly improve stability, with up to 30 times faster oxygen exchange kinetics after 54 h in air at 530 °C achieved by Hf addition onto LSC. Finally, the results revealed a 'volcano' relation between the oxygen exchange kinetics and the oxygen vacancy formation enthalpy of the binary oxides of the additive cations. This volcano relation highlights the existence of an optimum surface oxygen vacancy concentration that balances the gain in oxygen exchange kinetics and the chemical stability loss.« less

  10. Characterization of Staphylococcus aureus mutants expressing reduced susceptibility to common house-cleaners

    PubMed Central

    Davis, A.O.; O’Leary, J.O.; Muthaiyan, A.; Langevin, M.J.; Delgado, A.; Abalos, A.T.; Fajardo, A.R.; Marek, J.; Wilkinson, B.J.; Gustafson, J.E.

    2013-01-01

    Aims To characterize mutants of Staphylococcus aureus expressing reduced susceptibility to house cleaners (HC), assess the impact of the alternative sigma factor SigB on HC susceptibility, and determine the MIC of clinical methicillin-resistant S. aureus (MRSA) to a HC. Methods and Results Susceptibility to HC, HC components, H2O2, vancomycin and oxacillin and physiological parameters were determined for HC-reduced susceptibility (HCRS) mutants, parent strain COL and COLsigB::kan. HCRS mutants selected with three HC expressed reduced susceptibility to multiple HC, HC components, H2O2 and vancomycin. Two unique HCRS mutants also lost the methicillin resistance determinant. In addition, all HCRS mutants exhibited better growth at two temperatures, and one HCRS mutant expressed reduced carotenoid production. COLsigB::kan demonstrated increased susceptibility to all HC and many HC components. sigB operon mutations were not detected in one HCRS mutant background. Of 76 clinical MRSA, 20 exhibited reduced susceptibility to a HC. Conclusions HCRS mutants demonstrate altered susceptibility to multiple antimicrobials. While sigB is required for full HC resistance, one HCRS mechanism does not involve sigB operon mutations. Clinical MRSA expressing reduced susceptibility to a common HC were detected. Significance and Impact of the Study This study suggests that HCRS mutants are not protected against, nor selected by, practical HC concentrations. PMID:15659191

  11. Increased NY-ESO-1 Expression and Reduced Infiltrating CD3+ T Cells in Cutaneous Melanoma

    PubMed Central

    Giavina-Bianchi, Mara; Giavina-Bianchi, Pedro; Sotto, Mirian Nacagami; Muzikansky, Alona; Kalil, Jorge; Festa-Neto, Cyro; Duncan, Lyn M.

    2015-01-01

    NY-ESO-1 is a cancer-testis antigen aberrantly expressed in melanomas, which may serve as a robust and specific target in immunotherapy. NY-ESO-1 antigen expression, tumor features, and the immune profile of tumor infiltrating lymphocytes were assessed in primary cutaneous melanoma. NY-ESO-1 protein was detected in 20% of invasive melanomas (16/79), rarely in in situ melanoma (1/10) and not in benign nevi (0/20). Marked intratumoral heterogeneity of NY-ESO-1 protein expression was observed. NY-ESO-1 expression was associated with increased primary tumor thickness (P = 0.007) and inversely correlated with superficial spreading melanoma (P < 0.02). NY-ESO-1 expression was also associated with reduced numbers and density of CD3+ tumor infiltrating lymphocytes (P = 0.017). When NY-ESO-1 protein was expressed, CD3+ T cells were less diffusely infiltrating the tumor and were more often arranged in small clusters (P = 0.010) or as isolated cells (P = 0.002) than in large clusters of more than five lymphocytes. No correlation of NY-ESO-1 expression with gender, age, tumor site, ulceration, lymph node sentinel status, or survival was observed. NY-ESO-1 expression in melanoma was associated with tumor progression, including increased tumor thickness, and with reduced tumor infiltrating lymphocytes. PMID:25954764

  12. Increased NY-ESO-1 expression and reduced infiltrating CD3+ T cells in cutaneous melanoma.

    PubMed

    Giavina-Bianchi, Mara; Giavina-Bianchi, Pedro; Sotto, Mirian Nacagami; Muzikansky, Alona; Kalil, Jorge; Festa-Neto, Cyro; Duncan, Lyn M

    2015-01-01

    NY-ESO-1 is a cancer-testis antigen aberrantly expressed in melanomas, which may serve as a robust and specific target in immunotherapy. NY-ESO-1 antigen expression, tumor features, and the immune profile of tumor infiltrating lymphocytes were assessed in primary cutaneous melanoma. NY-ESO-1 protein was detected in 20% of invasive melanomas (16/79), rarely in in situ melanoma (1/10) and not in benign nevi (0/20). Marked intratumoral heterogeneity of NY-ESO-1 protein expression was observed. NY-ESO-1 expression was associated with increased primary tumor thickness (P = 0.007) and inversely correlated with superficial spreading melanoma (P < 0.02). NY-ESO-1 expression was also associated with reduced numbers and density of CD3+ tumor infiltrating lymphocytes (P = 0.017). When NY-ESO-1 protein was expressed, CD3+ T cells were less diffusely infiltrating the tumor and were more often arranged in small clusters (P = 0.010) or as isolated cells (P = 0.002) than in large clusters of more than five lymphocytes. No correlation of NY-ESO-1 expression with gender, age, tumor site, ulceration, lymph node sentinel status, or survival was observed. NY-ESO-1 expression in melanoma was associated with tumor progression, including increased tumor thickness, and with reduced tumor infiltrating lymphocytes.

  13. Electrophysiological and behavioral responses to ketamine in mice with reduced Akt1 expression

    PubMed Central

    Featherstone, Robert E.; Tatard-Leitman, Valerie M.; Suh, Jimmy D.; Lin, Robert; Lucki, Irwin; Siegel, Steven J.

    2013-01-01

    Rationale A number of studies have associated reduced Akt1 expression with vulnerability for schizophrenia. Although mice with deletion of a single copy of the Akt1 gene (Akt1+/−) show reduced Akt1 expression relative to wild-type (WT) animals, the extent to which these mice show schizophrenia-like phenotypic changes and/or increased susceptibility to epigenetic or non-genetic factors related to schizophrenia is unknown. Objectives Mutant mice were assessed on electroencephalographic/event related potential (EEG/ERP) and behavioral (acoustic startle and pre-pulse inhibition) measures relevant to schizophrenia. Mice were also assessed following exposure to the NMDA receptor antagonist ketamine, a potent psychotomimetic drug, in order to assess the role of reduced Akt1 expression as a vulnerability factor for schizophrenia. Methods Akt1+/−, Akt1−/− and WT mice received a series of paired-click, white noise stimuli, following ketamine (50 mg/kg) and saline injections. EEG was analyzed for ERPs and event-related power. Akt1+/− and WT mice were also assessed on PPI following ketamine (50 mg/kg) or saline injection. Results Akt1+/− and Akt1−/− mice displayed reduced amplitude of the P20 component of the ERP to the first click of a paired click stimulus, as well as reduced S1-S2 difference for P20 and N40 components, following ketamine. Mutant mice also showed increased reduction in gamma synchrony and theta suppression following ketamine. Akt1+/− mice displayed reduced pre-pulse inhibition. Conclusions Reduced genetic expression of Akt1 facilitated ketamine-induced changes of EEG and behavior in mice, suggesting that reduced Akt1 expression can serve as a vulnerability factor for schizophrenia. PMID:23392353

  14. Surface modification of platelet concentrate bags to reduce biofilm formation and transfusion sepsis.

    PubMed

    Wilson-Nieuwenhuis, Joels S T; Dempsey-Hibbert, Nina; Liauw, Christopher M; Whitehead, Kathryn A

    2017-09-07

    Bacterial contamination of blood products poses a major risk in transfusion medicine, including transfusions involving platelet products. Although testing systems are in place for routine screening of platelet units, the formation of bacterial biofilms in such units may decrease the likelihood that bacteria will be detected. This work determined the surface properties of p-PVC platelet concentrate bags and investigated how these characteristics influenced biofilm formation. Serratia marcescens and Staphylococcus epidermidis, two species commonly implicated in platelet contamination, were used to study biofilm growth. The platelet concentrate bags were physically flattened to determine if reducing the surface roughness altered biofilm formation. The results demonstrated that the flattening process of the platelet bags affected the chemistry of the surface and reduced the surface hydrophobicity. Flattening of the surfaces resulted in a reduction in biofilm formation for both species after 5 days, with S. marcescens demonstrating a greater reduction. However, there was no significant difference between the smooth and flat surfaces following 7 days' incubation for S. marcescens and no significant differences between any of the surfaces following 7 days' incubation for S. epidermidis. The results suggest that flattening the p-PVC surfaces may limit potential biofilm formation for the current duration of platelet storage time of 5 days. It is hoped that this work will enhance the understanding of how surface properties influence the development of microbial biofilms in platelet concentrate bags in order to devise a solution to discourage biofilm formation. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Influence of hydrophobic and superhydrophobic surfaces on reducing aerodynamic insect residues

    NASA Astrophysics Data System (ADS)

    Krishnan, K. Ghokulla; Milionis, Athanasios; Loth, Eric; Farrell, Thomas E.; Crouch, Jeffrey D.; Berry, Douglas H.

    2017-01-01

    Insect fouling during takeoff, climb and landing can result in increased drag and fuel consumption for aircrafts with laminar-flow surfaces. This study investigates the effectiveness of various hydrophobic and superhydrophobic surfaces in reducing residue of insects on an aerodynamic surface at relatively high impact speeds (about 45 m/s). An experimental setup consisting of a wind tunnel and a method to inject live flightless fruit flies was used to test the effectiveness of various surfaces against insect fouling. Insect fouling was analyzed based on residue area and height from multiple impacts. In general most of the residue area was due to the hemolymph spreading while most of the residue height was due to adhesion of exoskeleton parts. Hydrophobic and especially superhydrophobic surfaces performed better than a hydrophilic aluminum surface in terms of minimizing the residue area of various insect components (exoskeleton, hemolymph, and red fluid). Surfaces with reduced wettability and short lateral length scales tended to have the smallest residue area. Residue height was not as strongly influenced by surface wettability since even a single exoskeleton adhered to the surface upon impact was enough to produce a residue height of the order of one mm. In general, the results indicate that hemolymph spread needs to be avoided (e.g. by having reduced wettability and short lateral correlation lengths) in order to minimize the residue area, while exoskeleton adherence needs to be avoided (e.g. by having oleophobic properties and micro/nano roughness) in order to minimize the residue height. In particular, two of the superhydrophobic coatings produced substantial reduction in residue height and area, relative to the baseline surface of aluminum. However, the surfaces also showed poor mechanical durability on the high-speed insect impact location. This suggests that although low wettability materials show great insect anti-fouling behavior, their durability needs to

  16. Endothelial microparticles reduce ICAM-1 expression in a microRNA-222-dependent mechanism

    PubMed Central

    Jansen, Felix; Yang, Xiaoyan; Baumann, Katharina; Przybilla, David; Schmitz, Theresa; Flender, Anna; Paul, Kathrin; Alhusseiny, Adil; Nickenig, Georg; Werner, Nikos

    2015-01-01

    Endothelial microparticles (EMP) are released from activated or apoptotic endothelial cells (ECs) and can be taken up by adjacent ECs, but their effect on vascular inflammation after engulfment is largely unknown. We sought to determine the role of EMP in EC inflammation. In vitro, EMP treatment significantly reduced tumour necrosis factor-α-induced endothelial intercellular adhesion molecule (ICAM)-1 expression on mRNA and protein level, whereas there was no effect on vascular cell adhesion molecule-1 expression. Reduced ICAM-1 expression after EMP treatment resulted in diminished monocyte adhesion in vitro. In vivo, systemic treatment of ApoE−/− mice with EMP significantly reduced murine endothelial ICAM-1 expression. To explore the underlying mechanisms, Taqman microRNA array was performed and microRNA (miR)-222 was identified as the strongest regulated miR between EMP and ECs. Following experiments demonstrated that miR-222 was transported into recipient ECs by EMP and functionally regulated expression of its target protein ICAM-1 in vitro and in vivo. After simulating diabetic conditions, EMP derived from glucose-treated ECs contained significantly lower amounts of miR-222 and showed reduced anti-inflammatory capacity in vitro and in vivo. Finally, circulating miR-222 level was diminished in patients with coronary artery disease (CAD) compared to patients without CAD. EMPs promote anti-inflammatory effects in vitro and in vivo by reducing endothelial ICAM-1 expression via the transfer of functional miR-222 into recipient cells. In pathological hyperglycaemic conditions, EMP-mediated miR-222-dependent anti-inflammatory effects are reduced. PMID:26081516

  17. Endothelial microparticles reduce ICAM-1 expression in a microRNA-222-dependent mechanism.

    PubMed

    Jansen, Felix; Yang, Xiaoyan; Baumann, Katharina; Przybilla, David; Schmitz, Theresa; Flender, Anna; Paul, Kathrin; Alhusseiny, Adil; Nickenig, Georg; Werner, Nikos

    2015-09-01

    Endothelial microparticles (EMP) are released from activated or apoptotic endothelial cells (ECs) and can be taken up by adjacent ECs, but their effect on vascular inflammation after engulfment is largely unknown. We sought to determine the role of EMP in EC inflammation. In vitro, EMP treatment significantly reduced tumour necrosis factor-α-induced endothelial intercellular adhesion molecule (ICAM)-1 expression on mRNA and protein level, whereas there was no effect on vascular cell adhesion molecule-1 expression. Reduced ICAM-1 expression after EMP treatment resulted in diminished monocyte adhesion in vitro. In vivo, systemic treatment of ApoE-/- mice with EMP significantly reduced murine endothelial ICAM-1 expression. To explore the underlying mechanisms, Taqman microRNA array was performed and microRNA (miR)-222 was identified as the strongest regulated miR between EMP and ECs. Following experiments demonstrated that miR-222 was transported into recipient ECs by EMP and functionally regulated expression of its target protein ICAM-1 in vitro and in vivo. After simulating diabetic conditions, EMP derived from glucose-treated ECs contained significantly lower amounts of miR-222 and showed reduced anti-inflammatory capacity in vitro and in vivo. Finally, circulating miR-222 level was diminished in patients with coronary artery disease (CAD) compared to patients without CAD. EMPs promote anti-inflammatory effects in vitro and in vivo by reducing endothelial ICAM-1 expression via the transfer of functional miR-222 into recipient cells. In pathological hyperglycaemic conditions, EMP-mediated miR-222-dependent anti-inflammatory effects are reduced.

  18. PROX1 gene is differentially expressed in oral cancer and reduces cellular proliferation.

    PubMed

    Rodrigues, Maria F S D; de Oliveira Rodini, Camila; de Aquino Xavier, Flávia C; Paiva, Katiúcia B; Severino, Patrícia; Moyses, Raquel A; López, Rossana M; DeCicco, Rafael; Rocha, Lília A; Carvalho, Marcos B; Tajara, Eloiza H; Nunes, Fabio D

    2014-12-01

    Homeobox genes are a family of transcription factors that play a pivotal role in embryogenesis. Prospero homeobox 1 (PROX1) has been shown to function as a tumor suppressor gene or oncogene in various types of cancer, including oral squamous cell carcinoma (OSCC). We have previously identified PROX1 as a downregulated gene in OSCC. The aim of this study is to clarify the underlying mechanism by which PROX1 regulates tumorigenicity of OSCC cells. PROX1 mRNA and protein expression levels were first investigated in 40 samples of OSCC and in nontumor margins. Methylation and amplification analysis was also performed to assess the epigenetic and genetic mechanisms involved in controlling PROX1 expression. OSCC cell line SCC9 was also transfected to stably express the PROX1 gene. Next, SCC9-PROX1-overexpressing cells and controls were subjected to proliferation, differentiation, apoptosis, migration, and invasion assays in vitro. OSCC samples showed reduced PROX1 expression levels compared with nontumor margins. PROX1 amplification was associated with better overall survival. PROX1 overexpression reduces cell proliferation and downregulates cyclin D1. PROX1-overexpressing cells also exhibited reduced CK18 and CK19 expression and transcriptionally altered the expression of WISP3, GATA3, NOTCH1, and E2F1. Our results suggest that PROX1 functions as a tumor suppressor gene in oral carcinogenesis.

  19. PROX1 Gene is Differentially Expressed in Oral Cancer and Reduces Cellular Proliferation

    PubMed Central

    Rodrigues, Maria F.S.D.; de Oliveira Rodini, Camila; de Aquino Xavier, Flávia C.; Paiva, Katiúcia B.; Severino, Patrícia; Moyses, Raquel A.; López, Rossana M.; DeCicco, Rafael; Rocha, Lília A.; Carvalho, Marcos B.; Tajara, Eloiza H.; Nunes, Fabio D.

    2014-01-01

    Abstract Homeobox genes are a family of transcription factors that play a pivotal role in embryogenesis. Prospero homeobox 1 (PROX1) has been shown to function as a tumor suppressor gene or oncogene in various types of cancer, including oral squamous cell carcinoma (OSCC). We have previously identified PROX1 as a downregulated gene in OSCC. The aim of this study is to clarify the underlying mechanism by which PROX1 regulates tumorigenicity of OSCC cells. PROX1 mRNA and protein expression levels were first investigated in 40 samples of OSCC and in nontumor margins. Methylation and amplification analysis was also performed to assess the epigenetic and genetic mechanisms involved in controlling PROX1 expression. OSCC cell line SCC9 was also transfected to stably express the PROX1 gene. Next, SCC9-PROX1-overexpressing cells and controls were subjected to proliferation, differentiation, apoptosis, migration, and invasion assays in vitro. OSCC samples showed reduced PROX1 expression levels compared with nontumor margins. PROX1 amplification was associated with better overall survival. PROX1 overexpression reduces cell proliferation and downregulates cyclin D1. PROX1-overexpressing cells also exhibited reduced CK18 and CK19 expression and transcriptionally altered the expression of WISP3, GATA3, NOTCH1, and E2F1. Our results suggest that PROX1 functions as a tumor suppressor gene in oral carcinogenesis. PMID:25526434

  20. STAT3 Knockdown Reduces Pancreatic Cancer Cell Invasiveness and Matrix Metalloproteinase-7 Expression in Nude Mice

    PubMed Central

    Huang, Ke jian; Wu, Wei dong; Jiang, Tao; Cao, Jun; Feng, Zhen zhong; Qiu, Zheng jun

    2011-01-01

    Aims Transducer and activator of transcription-3 (STAT3) plays an important role in tumor cell invasion and metastasis. The aim of the present study was to investigate the effects of STAT3 knockdown in nude mouse xenografts of pancreatic cancer cells and underlying gene expression. Methods A STAT3 shRNA lentiviral vector was constructed and infected into SW1990 cells. qRT-PCR and western immunoblot were performed to detect gene expression. Nude mouse xenograft assays were used to assess changes in phenotypes of these stable cells in vivo. HE staining was utilized to evaluate tumor cell invasion and immunohistochemistry was performed to analyze gene expression. Results STAT3 shRNA successfully silenced expression of STAT3 mRNA and protein in SW1990 cells compared to control cells. Growth rate of the STAT3-silenced tumor cells in nude mice was significantly reduced compared to in the control vector tumors and parental cells-generated tumors. Tumor invasion into the vessel and muscle were also suppressed in the STAT3-silenced tumors compared to controls. Collagen IV expression was complete and continuous surrounding the tumors of STAT3-silenced SW1990 cells, whereas collagen IV expression was incomplete and discontinuous surrounding the control tumors. Moreover, microvessel density was significantly lower in STAT3-silenced tumors than parental or control tumors of SW1990 cells. In addition, MMP-7 expression was reduced in STAT3-silenced tumors compared to parental SW1990 xenografts and controls. In contrast, expression of IL-1β and IgT7α was not altered. Conclusion These data clearly demonstrate that STAT3 plays an important role in regulation of tumor growth, invasion, and angiogenesis, which could be act by reducing MMP-7 expression in pancreatic cancer cells. PMID:21991388

  1. Nanorough titanium surfaces reduce adhesion of Escherichia coli and Staphylococcus aureus via nano adhesion points.

    PubMed

    Lüdecke, Claudia; Roth, Martin; Yu, Wenqi; Horn, Uwe; Bossert, Jörg; Jandt, Klaus D

    2016-09-01

    Microbial adhesion to natural and synthetic materials surfaces is a key issue e.g. in food industry, sewage treatment and most importantly in the biomedical field. The current development and progress in nanoscale structuring of materials surfaces to control microbial adhesion requires an advanced understanding of the microbe-material-interaction. This study aimed to investigate the nanostructure of the microbe-material-interface and link it to microbial adhesion kinetics as function of titanium surface nanoroughness to gain new insight into controlling microbial adhesion via materials' surface nanoroughness. Adhesion of Escherichia coli and Staphylococcus aureus was statistically significantly reduced (p≤0.05) by 55.6 % and 40.5 %, respectively, on physical vapor deposited titanium thin films with a nanoroughness of 6nm and the lowest surface peak density compared to 2nm with the highest surface peak density. Cross-sectioning of the microbial cells with a focused ion beam (FIB) and SEM imaging provided for the first time direct insight into the titanium-microbe-interface. High resolution SEM micrographs gave evidence that the surface peaks are the loci of initial contact between the microbial cells and the material's surface. In a qualitative model we propose that the initial microbial adhesion on nanorough surfaces is controlled by the titanium surface peak density via nano adhesion points. This new understanding will help towards the design of materials surfaces for controlling microbial adhesion.

  2. Macrotextured spoked surfaces reduce the residence time of a bouncing Leidenfrost drop.

    PubMed

    Patterson, Colin J; Shiri, Samira; Bird, James C

    2017-02-15

    Liquid drops can bounce when they impact non-wetting surfaces. Recently, studies have demonstrated that the time that the bouncing drop contacts a superhydrophobic surface can be reduced by incorporating ridged macrotextures on the surface. Yet the existing models aimed at explaining this phenomenon offer incompatible predictions of the contact time when a drop impacts multiple intersecting macrotextures, or spokes. Furthermore, it is unclear whether the effects of the macrotexture on the drop hydrodynamics extend to non-wetting surfaces in which direct contact is avoided by a thin vapor layer. Here we demonstrate that the phenomenon observed for macrotextured, superhydrophobic surfaces extends to macrotextured, wettable surfaces above the Leidenfrost temperature. We show that the number of droplets and overall residence time both depend on the number of intersecting spokes. Finally, we compare and contrast our results with mechanistic models to rationalize various elements of the phenomenon.

  3. Sub-micrometer scale surface roughness of titanium reduces fibroblasts function.

    PubMed

    Migita, Satoshi; Okuyama, So; Araki, Kunitaka

    2016-04-06

    Titanium and its alloys are conventionally used to produce medical devices, but their biocompatibility has not yet been optimized. Surface modification, especially control of the surface roughness of titanium, is one strategy for improving biocompatibility and providing effective binding to hard tissue. However, the soft tissue compatibility of metallic materials is currently poorly understood, and effective techniques for tight binding between metal surfaces and soft tissue are still under development. Therefore, we here investigated whether the surface roughness of titanium affects fibroblast adhesion and proliferation. Our results showed that a surface roughness of ~100 nm reduces fibroblast function. On such surfaces, distinct focal adhesion was not observed. These findings improve the general understanding of the binding compatibility between soft tissues and metallic materials.

  4. Macrotextured spoked surfaces reduce the residence time of a bouncing Leidenfrost drop

    NASA Astrophysics Data System (ADS)

    Patterson, Colin J.; Shiri, Samira; Bird, James C.

    2017-02-01

    Liquid drops can bounce when they impact non-wetting surfaces. Recently, studies have demonstrated that the time that the bouncing drop contacts a superhydrophobic surface can be reduced by incorporating ridged macrotextures on the surface. Yet the existing models aimed at explaining this phenomenon offer incompatible predictions of the contact time when a drop impacts multiple intersecting macrotextures, or spokes. Furthermore, it is unclear whether the effects of the macrotexture on the drop hydrodynamics extend to non-wetting surfaces in which direct contact is avoided by a thin vapor layer. Here we demonstrate that the phenomenon observed for macrotextured, superhydrophobic surfaces extends to macrotextured, wettable surfaces above the Leidenfrost temperature. We show that the number of droplets and overall residence time both depend on the number of intersecting spokes. Finally, we compare and contrast our results with mechanistic models to rationalize various elements of the phenomenon.

  5. Expression of surface platelet receptors (CD62P and CD41/61) in horses with recurrent airway obstruction (RAO).

    PubMed

    Iwaszko-Simonik, Alicja; Niedzwiedz, Artur; Graczyk, Stanislaw; Slowikowska, Malwina; Pliszczak-Krol, Aleksandra

    2015-03-15

    Recurrent airway obstruction (RAO) is an allergic disease of horses similar to human asthma, which is characterized by airway inflammation and activation of neutrophils, lymphocytes and platelets. Platelet activation and an increase in circulating platelet-leukocyte aggregates may lead to airway remodeling. The aim of this study was to investigate platelet status in RAO-affected horses based on the platelet morphology and platelet surface expression of CD41/61 and CD62P. Ten RAO-affected horses and ten healthy horses were included in this study. Blood samples were obtained to determine the platelet count (PLT), mean platelet volume (MPV) and platelet large cell ratio (P-LCR). Expression of CD62P and CD41/61 was detected by flow cytometry on activated platelets. The median PLT was significantly reduced in horses with RAO compared to the controls. The MPV and the P-LCR values were significantly higher in RAO horses than controls. Expression of CD41/61 on platelets was increased in RAO horses, while CD62P expression was reduced. This study demonstrated the morphological changes in platelets and expression of platelet surface receptors. Despite the decrease of CD62P expression, the observed increased surface expression of CD41/61 on platelets in horses with RAO may contribute to the formation of platelet aggregates in their respiratory system.

  6. Metabolite and gene expression responses in juvenile flounder Paralichthys olivaceus exposed to reduced salinities.

    PubMed

    Wu, Huifeng; Liu, Jialin; Lu, Zhen; Xu, Lanlan; Ji, Chenglong; Wang, Qing; Zhao, Jianmin

    2017-04-01

    Seawater salinity is one of the most important changeable environmental factors influencing the behavior, survival, growth and production of marine organisms. In this work, metabolite and gene expression profiles were used to elucidate the biological effects of reduced salinities in juvenile flounder Paralichthys olivaceus. Metabolic profiling indicated that both reduced salinities (23.3‰ and 15.6‰) enhanced proteolysis and disturbed osmotic regulation and energy metabolism in juvenile flounder P. olivaceus. Furthermore, the low salinity (15.6‰) enhanced anaerobic metabolism indicated by the elevated lactate in flounder tissue extracts. Gene expression profiles exhibited that reduced salinities could induce immune stress and oxidative stress and disturb energy metabolism in juvenile flounder P. olivaceus. In addition, reduced salinities might promote the growth and gonadal differentiation in juvenile flounder P. olivaceus.

  7. Overt expression of AP-1 reduces alpha myosin heavy chain expression and contributes to heart failure from chronic volume overload.

    PubMed

    Freire, Grace; Ocampo, Catherina; Ilbawi, Nadim; Griffin, Andrew J; Gupta, Madhu

    2007-10-01

    Reduced expression of alpha-MHC plays a significant role in cardiac contractile dysfunction from hemodynamic overload. Previously, Pur proteins and YY1 have been shown to play a role in alpha-MHC repression during heart failure induced by pressure overload and by spontaneous hypertension, respectively. This was not observed in volume-overload-induced heart failure, suggesting additional regulatory mechanisms for alpha-MHC repression. The present study was performed to identify volume overload responsive transcription factors involved in alpha-MHC gene regulation. DNA binding activity of several transcription factors was evaluated in a functionally characterized rat model of heart failure induced by aorto-caval shunt. After 10 weeks of shunt, severe LV dilatation and reduced LV function were accompanied by increased expression of ANF and beta-MHC, and decreased expression of alpha-MHC. This was associated with dramatic (10-fold) activation of AP-1 together with increased expression of c-fos and c-jun. AP-1 activation was not observed following 4 weeks of shunt when cardiac function was preserved. In cultured cardiomyocytes, induction of AP-1 by PMA attenuated alpha-MHC mRNA by 60%. Transient transfection assays mapped PMA responsive sequence to -582 to -588 bp of alpha-MHC promoter. Deletion or mutation of these nucleotides had minimal effect on basal promoter activity but played a dominant role in PMA-mediated repression of alpha-MHC promoter activity. Over-expression of c-fos and c-jun in cardiomyocytes inhibited alpha-MHC promoter activity in a concentration dependent manner. Data suggest a repressive role of AP-1 in alpha-MHC expression and its possible involvement in the transition from compensatory hypertrophy to heart failure in chronic volume overload.

  8. Synthesis and Characterization of a Lubricin Mimic (mLub) To Reduce Friction and Adhesion on the Articular Cartilage Surface

    PubMed Central

    Lawrence, Alexandra; Xu, Xin; Bible, Melissa D.; Calve, Sarah; Neu, Corey P.; Panitch, Alyssa

    2015-01-01

    The lubricating proteoglycan, lubricin, facilitates the remarkable low friction and wear properties of articular cartilage in the synovial joints of the body. Lubricin lines the joint surfaces and plays a protective role as a boundary lubricant in sliding contact; decreased expression of lubricin is associated with cartilage degradation and the pathogenesis of osteoarthritis. An unmet need for early osteoarthritis treatment is the development of therapeutic molecules that mimic lubricin function and yet are also resistant to enzymatic degradation common in the damaged joint. Here, we engineered a lubricin mimic (mLub) that is less susceptible to enzymatic degradation and binds to the articular surface to reduce friction. mLub was synthesized using a chondroitin sulfate backbone with type II collagen and hyaluronic acid (HA) binding peptides to promote interaction with the articular surface and synovial fluid constituents. In vitro and in vivo characterization confirmed the binding ability of mLub to isolated type II collagen and HA, and to the cartilage surface. Following trypsin treatment to the cartilage surface, application of mLub, in combination with purified or commercially available hyaluronan, reduced the coefficient of friction, and adhesion, to control levels as assessed over macro- to micro-scales by rheometry and atomic force microscopy. In vivo studies demonstrate an mLub residency time of less than 1 week. Enhanced lubrication by mLub reduces surface friction and adhesion, which may suppress the progression of degradation and cartilage loss in the joint. mLub therefore shows potential for treatment in early osteoarthritis following injury. PMID:26398308

  9. Reduced tissue osmolarity increases TRPV4 expression and pro-inflammatory cytokines in intervertebral disc cells.

    PubMed

    Walter, B A; Purmessur, D; Moon, A; Occhiogrosso, J; Laudier, D M; Hecht, A C; Iatridis, J C

    2016-07-19

    The mechanical behaviour and cellular metabolism of intervertebral discs (IVDs) and articular cartilage are strongly influenced by their proteoglycan content and associated osmotic properties. This osmotic environment is a biophysical signal that changes with disease and may contribute to the elevated matrix breakdown and altered biologic response to loading observed in IVD degeneration and osteoarthritis. This study tested the hypothesis that changes in osmo-sensation by the transient receptor potential vallinoid-4 (TRPV4) ion channel occur with disease and contribute to the inflammatory environment found during degeneration. Immunohistochemistry on bovine IVDs from an inflammatory organ culture model were used to investigate if TRPV4 is expressed in the IVD and how expression changes with degeneration. Western blot, live-cell calcium imaging, and qRT-PCR were used to investigate whether osmolarity changes or tumour necrosis factor α (TNFα) regulate TRPV4 expression, and how altered TRPV4 expression influences calcium signalling and pro-inflammatory cytokine expression. TRPV4 expression correlated with TNFα expression, and was increased when cultured in reduced medium osmolarity and unaltered with TNFα-stimulation. Increased TRPV4 expression increased the calcium flux following TRPV4 activation and increased interleukin-1β (IL-1β) and IL-6 gene expression in IVD cells. TRPV4 expression was qualitatively elevated in regions of aggrecan depletion in degenerated human IVDs. Collectively, results suggest that reduced tissue osmolarity, likely following proteoglycan degradation, can increase TRPV4 signalling and enhance pro-inflammatory cytokine production, suggesting changes in TRPV4 mediated osmo-sensation may contribute to the progressive matrix breakdown in disease.

  10. REDUCED TISSUE OSMOLARITY INCREASES TRPV4 EXPRESSION AND PRO-INFLAMMATORY CYTOKINES IN INTERVERTEBRAL DISC CELLS

    PubMed Central

    Walter, B.A.; Purmessur, D; Moon, A.; Occhiogrosso, J.; Laudier, D.M.; Hecht, A.C.; Iatridis, J.C.

    2016-01-01

    The mechanical behaviour and cellular metabolism of intervertebral discs (IVDs) and articular cartilage are strongly influenced by their proteoglycan content and associated osmotic properties. This osmotic environment is a biophysical signal that changes with disease and may contribute to the elevated matrix breakdown and altered biologic response to loading observed in IVD degeneration and osteoarthritis. This study tested the hypothesis that changes in osmo-sensation by the transient receptor potential vallinoid-4 (TRPV4) ion channel occur with disease and contribute to the inflammatory environment found during degeneration. Immunohistochemistry on bovine IVDs from an inflammatory organ culture model were used to investigate if TRPV4 is expressed in the IVD and how expression changes with degeneration. Western blot, live-cell calcium imaging, and qRT-PCR were used to investigate whether osmolarity changes or tumour necrosis factor α (TNFα) regulate TRPV4 expression, and how altered TRPV4 expression influences calcium signalling and pro-inflammatory cytokine expression. TRPV4 expression correlated with TNFα expression, and was increased when cultured in reduced medium osmolarity and unaltered with TNFα-stimulation. Increased TRPV4 expression increased the calcium flux following TRPV4 activation and increased interleukin-1β (IL-1β) and IL-6 gene expression in IVD cells. TRPV4 expression was qualitatively elevated in regions of aggrecan depletion in degenerated human IVDs. Collectively, results suggest that reduced tissue osmolarity, likely following proteoglycan degradation, can increase TRPV4 signalling and enhance pro-inflammatory cytokine production, suggesting changes in TRPV4 mediated osmo-sensation may contribute to the progressive matrix breakdown in disease. PMID:27434269

  11. Plasmodium vivax merozoite surface protein 8 cloning, expression, and characterisation.

    PubMed

    Perez-Leal, Oscar; Sierra, Adriana Y; Barrero, Carlos A; Moncada, Camilo; Martinez, Pilar; Cortes, Jimena; Lopez, Yolanda; Torres, Elizabeth; Salazar, Luz M; Patarroyo, Manuel A

    2004-11-26

    Plasmodium vivax, one of the four parasite species causing malaria in humans, is the most widespread throughout the world, leading to nearly 80 million cases per year, mainly in Latin-America and Asia. An open reading frame encoding the Plasmodium falciparum merozoite surface protein 8 P. vivax homologue has been identified in the present study by screening the current data obtained from this parasite's partially sequenced genome. This new protein contains 487 amino-acids, two epidermal growth factor like domains, hydrophobic regions at the N- and C-termini compatible with a signal peptide, and a glycosylphosphatidylinositol anchor site, respectively. This gene's transcription and its encoded protein expression have been assessed, as well as its recognition by P. vivax-infected patients' sera. Based on this recognition, and a previous study showing that mice immunised with the Plasmodium yoelii homologous protein were protected, we consider the PvMSP8 a good candidate to be included in a multi-stage multi-antigen P. vivax vaccine.

  12. Standing on a declining surface reduces transient prolonged standing induced low back pain development.

    PubMed

    Gallagher, Kaitlin M; Callaghan, Jack P

    2016-09-01

    While alternating standing position on a sloped surface has proven successful at reducing low back pain during standing, the purpose of this study was to evaluate standing solely on a declining surface to isolate the influence of the postural change. Seventeen participants performed two 75-min prolonged standing occupational simulations- level ground and declining surface. Fifty-three percent of participants (9/17) were categorized as pain developers during the level ground standing condition. For these same pain developers, their average maximum pain scores were 58% lower during sloped standing. All participants showed greater hip flexion, trunk-to-thigh angle flexion, and posterior translation of the trunk center of gravity when standing on the sloped surface. These postural changes could cause the muscles crossing the hip posteriorly to increase passive stiffness and assist with stabilizing the pelvis. This study stresses the importance of hip kinematics, not just lumbar spine posture, in reducing prolonged standing induced low back pain.

  13. Abiotic surface sensing and biofilm-dependent regulation of gene expression in Escherichia coli.

    PubMed

    Prigent-Combaret, C; Vidal, O; Dorel, C; Lejeune, P

    1999-10-01

    To get further information on bacterial surface sensing and biofilm-dependent regulation of gene expression in Escherichia coli K-12, random insertion mutagenesis with Mu dX, a mini-Mu carrying the promoterless lacZ gene, was performed with an ompR234 adherent strain, and a simple screen was developed to assess changes in gene expression in biofilm cells versus planktonic cells. This screen revealed that major changes in the pattern of gene expression occur during biofilm development: the transcription of 38% of the genes was affected within biofilms. Different cell functions were more expressed in sessile bacteria: the OmpC porin, the high-affinity transport system of glycine betaine (encoded by the proU operon), the colanic acid exopolysaccharide (wca locus, formerly called cps), tripeptidase T (pepT), and the nickel high-affinity transport system (nikA). On the other hand, the syntheses of flagellin (fliC) and of a putative protein of 92 amino acids (f92) were both reduced in biofilms. Such a genetic reprogramming of gene expression in biofilms seems to result from changes in multiple environmental physicochemical conditions. In this work, we show that bacteria within biofilms encounter higher-osmolarity conditions, greater oxygen limitation, and higher cell density than in the liquid phase.

  14. fREDUCE: Detection of degenerate regulatory elements using correlation with expression

    PubMed Central

    Wu, Randy Z; Chaivorapol, Christina; Zheng, Jiashun; Li, Hao; Liang, Shoudan

    2007-01-01

    Background The precision of transcriptional regulation is made possible by the specificity of physical interactions between transcription factors and their cognate binding sites on DNA. A major challenge is to decipher transcription factor binding sites from sequence and functional genomic data using computational means. While current methods can detect strong binding sites, they are less sensitive to degenerate motifs. Results We present fREDUCE, a computational method specialized for the detection of weak or degenerate binding motifs from gene expression or ChIP-chip data. fREDUCE is built upon the widely applied program REDUCE, which elicits motifs by global statistical correlation of motif counts with expression data. fREDUCE introduces several algorithmic refinements that allow efficient exhaustive searches of oligonucleotides with a specified number of degenerate IUPAC symbols. On yeast ChIP-chip benchmarks, fREDUCE correctly identified motifs and their degeneracies with accuracies greater than its predecessor REDUCE as well as other known motif-finding programs. We have also used fREDUCE to make novel motif predictions for transcription factors with poorly characterized binding sites. Conclusion We demonstrate that fREDUCE is a valuable tool for the prediction of degenerate transcription factor binding sites, especially from array datasets with weak signals that may elude other motif detection methods. PMID:17941998

  15. Neurotrophin-3 significantly reduces sodium channel expression linked to neuropathic pain states

    PubMed Central

    Wilson-Gerwing, Tracy D.; Stucky, Cheryl L.; McComb, Geoffrey W.; Verge, Valerie M. K.

    2009-01-01

    Neuropathic pain resulting from chronic constriction injury (CCI) is critically linked to sensitization of peripheral nociceptors. Voltage gated sodium channels are major contributors to this state and their expression can be upregulated by nerve growth factor (NGF). We have previously demonstrated that neurotrophin-3 (NT-3) acts antagonistically to NGF in modulation of aspects of CCI-induced changes in trkA-associated nociceptor phenotype and thermal hyperalgesia. Thus, we hypothesized that exposure of neurons to increased levels of NT-3 would reduce expression of Nav1.8 and Nav1.9 in DRG neurons subject to CCI. In adult male rats, Nav1.8 and Nav1.9 mRNAs are expressed at high levels in predominantly small to medium size neurons. One week following CCI, there is reduced incidence of neurons expressing detectable Nav1.8 and Nav1.9 mRNA, but without a significant decline in mean level of neuronal expression, and similar findings observed immunohistochemically. There is also increased accumulation/redistribution of channel protein in the nerve most apparent proximal to the first constriction site. Intrathecal infusion of NT-3 significantly attenuates neuronal expression of Nav1.8 and Nav1.9 mRNA contralateral and most notably, ipsilateral to CCI, with a similar impact on relative protein expression at the level of the neuron and constricted nerve. We also observe reduced expression of the common neurotrophin receptor p75 in response to CCI that is not reversed by NT-3 in small to medium sized neurons and may confer an enhanced ability of NT-3 to signal via trkA, as has been previously shown in other cell types. These findings are consistent with an analgesic role for NT-3. PMID:18601922

  16. Reducing workplace cytotoxic surface contamination using a closed-system drug transfer device.

    PubMed

    Siderov, Jim; Kirsa, Sue; McLauchlan, Robert

    2010-03-01

    The potential for staff exposure to antineoplastic agents exists in the workplace despite current recommended safe handling procedures. Reliance on cytotoxic drug safety cabinets (CDSC) to provide total protection from exposure to hazardous drugs is insufficient. Preventing workplace contamination is the best strategy to minimise exposure. PhaSeal is a commercially available system for ensuring the leak-free transfer of hazardous drugs, fitting both the NIOSH and ISOPP definitions of a closed system. To date, there have been no published studies examining the use of a closed system drug transfer device (PhaSeal) under Australian conditions.The purpose of this study is to determine the impact of a closed system drug transfer device on cytotoxic surface contamination in the cytotoxic preparation areas of two Australian metropolitan public hospitals. This was a pre- and post-intervention study in which chemical contamination was tested at baseline then at five and 12 months after the introduction of the a closed system drug transfer device. Cyclophosphamide was used as a surrogate marker for all cytotoxic drugs. Surface wipe sampling was performed at specified sites within the cytotoxic suite using a standardized technique. Commercial products of cyclophosphamide were also sampled. After five months, contamination was reduced in 13 of the 22 sites sampled (59%), with four of these samples showing undetectable levels of contamination. Two other site samples (9%) remained unchanged. The total contamination of surfaces tested was reduced by 24%. After five months hospital 1 withdrew from the study. After 12 months, surface contamination was reduced in 75% of sample sites. The total contamination of surfaces tested was reduced by 68%. The wipes of the external surface of commercial products detected cyclophosphamide contamination. When used inside a CDSC, the closed system drug transfer device PhaSeal further reduces surface contamination, in some instances to

  17. Contributions of feature shapes and surface cues to the recognition of facial expressions.

    PubMed

    Sormaz, Mladen; Young, Andrew W; Andrews, Timothy J

    2016-10-01

    Theoretical accounts of face processing often emphasise feature shapes as the primary visual cue to the recognition of facial expressions. However, changes in facial expression also affect the surface properties of the face. In this study, we investigated whether this surface information can also be used in the recognition of facial expression. First, participants identified facial expressions (fear, anger, disgust, sadness, happiness) from images that were manipulated such that they varied mainly in shape or mainly in surface properties. We found that the categorization of facial expression is possible in either type of image, but that different expressions are relatively dependent on surface or shape properties. Next, we investigated the relative contributions of shape and surface information to the categorization of facial expressions. This employed a complementary method that involved combining the surface properties of one expression with the shape properties from a different expression. Our results showed that the categorization of facial expressions in these hybrid images was equally dependent on the surface and shape properties of the image. Together, these findings provide a direct demonstration that both feature shape and surface information make significant contributions to the recognition of facial expressions.

  18. New Variance-Reducing Methods for the PSD Analysis of Large Optical Surfaces

    NASA Technical Reports Server (NTRS)

    Sidick, Erkin

    2010-01-01

    Edge data of a measured surface map of a circular optic result in large variance or "spectral leakage" behavior in the corresponding Power Spectral Density (PSD) data. In this paper we present two new, alternative methods for reducing such variance in the PSD data by replacing the zeros outside the circular area of a surface map by non-zero values either obtained from a PSD fit (method 1) or taken from the inside of the circular area (method 2).

  19. A process to enhance the specific surface area and capacitance of hydrothermally reduced graphene oxide.

    PubMed

    Alazmi, Amira; El Tall, Omar; Rasul, Shahid; Hedhili, Mohamed N; Patole, Shashikant P; Costa, Pedro M F J

    2016-10-20

    The impact of post-synthesis processing in reduced graphene oxide materials for supercapacitor electrodes has been analyzed. A comparative study of vacuum, freeze and critical point drying was carried out for hydrothermally reduced graphene oxide demonstrating that the optimization of the specific surface area and preservation of the porous network are critical to maximize its supercapacitance performance. As described below, using a supercritical fluid as the drying medium, unprecedented values of the specific surface area (364 m(2) g(-1)) and supercapacitance (441 F g(-1)) for this class of materials have been achieved.

  20. Modulation of Mcl-1 expression reduces age-related cochlear degeneration.

    PubMed

    Yang, Wei Ping; Xu, Yang; Guo, Wei Wei; Liu, Hui Zhan; Hu, Bo Hua

    2013-11-01

    Mcl-1 is an anti-apoptotic member of the Bcl-2 family that modulates apoptosis-related signaling pathways and promotes cell survival. We have previously demonstrated a reduction of Mcl-1 expression in aging cochleae. To investigate whether restoring Mcl-1 expression would reduce aging-related cochlear degeneration, we developed a rat model of Mcl-1 overexpression. A plasmid encoding human Mcl-1/enhanced green fluorescent protein was applied to the round window of the cochlea. This in vivo treatment transfected both the sensory and supporting cells of the cochlear sensory epithelium and enhanced Mcl-1 expression at both the mRNA and the protein level. The upregulation of Mcl-1 expression reduced the progression of age-related cochlear dysfunction and sensory cell death. Furthermore, the transfection of Mcl-1 exerted its protective effect by suppressing cochlear apoptosis at the mitochondrial level. This study demonstrates that the genetic modulation of Mcl-1 expression reduces the progression of age-related cochlear degeneration.

  1. Application of bacteriophages to reduce Salmonella attachment and biofilms on hard surfaces.

    PubMed

    Gong, Chao; Jiang, Xiuping

    2017-02-23

    This study evaluated the effectiveness of bacteriophage treatment for reducing Salmonella attachment and biofilms on hard surfaces. Bacteriophages (n = 6) were selected for bacteriophage treatment based on host ranges against Salmonella isolates (n = 10) obtained from rendering plants. The effectiveness of bacteriophage treatment (104-108 PFU/mL) was initially evaluated against strong Salmonella biofilm formers in 96-well microplate. Then, the bacteriophage treatment (109 PFU/mL) was applied for 7 d to reduce Salmonella attached to the stainless steel surfaces under laboratory and greenhouse conditions. The inhibition of biofilm formation and reduction of pre-formed biofilm in 96-well microplate with bacteriophage treatment reached up to 90 and 66%, respectively. Under laboratory conditions, bacteriophage treatment reduced up to 2.9 and 3.0 log CFU/cm2 of attachment and slightly formed biofilm of selected top 10 Salmonella strains and an avirulent Salmonella Typhimurium strain 8243, respectively, as compared with reductions of 3.4, 1.4, and 3.0 log CFU/cm2 of S. Typhimurium strain 8243 in summer, fall/winter, and spring seasons under greenhouse conditions, respectively. Clearly, bacteriophages were effective on reducing Salmonella attachment and biofilms formed on hard surfaces under both laboratory and greenhouse conditions. The use of bacteriophages on hard surfaces may have merits in reducing the likelihood of finished rendered products being recontaminated with Salmonella in rendering plants.

  2. An Acceptance-Based Psychoeducation Intervention to Reduce Expressed Emotion in Relatives of Bipolar Patients

    ERIC Educational Resources Information Center

    Eisner, Lori R.; Johnson, Sheri L.

    2008-01-01

    Expressed emotion (EE) is a robust predictor of outcome in bipolar disorder. Despite decades of research, interventions to reduce EE levels have had only modest effects. This study used an expanded model of EE to develop an intervention. Research has demonstrated a strong link between attributions and EE in families of patients with psychiatric…

  3. 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.

  4. An Acceptance-Based Psychoeducation Intervention to Reduce Expressed Emotion in Relatives of Bipolar Patients

    ERIC Educational Resources Information Center

    Eisner, Lori R.; Johnson, Sheri L.

    2008-01-01

    Expressed emotion (EE) is a robust predictor of outcome in bipolar disorder. Despite decades of research, interventions to reduce EE levels have had only modest effects. This study used an expanded model of EE to develop an intervention. Research has demonstrated a strong link between attributions and EE in families of patients with psychiatric…

  5. Lovastatin inhibits human B lymphoma cell proliferation by reducing intracellular ROS and TRPC6 expression

    PubMed Central

    Song, Xiang; Liu, Bing-Chen; Lu, Xiao-Yu; Yang, Li-Li; Zhai, Yu-Jia; Eaton, Amity F.; Thai, Tiffany L.; Eaton, Douglas C.; Ma, He-Ping; Shen, Bao-Zhong

    2014-01-01

    Clinical evidence suggests that statins reduce cancer incidence and mortality. However, there is lack of in vitro data to show the mechanism by which statins can reduce the malignancies of cancer cells. We used a human B lymphoma Daudi cells as a model and found that lovastatin inhibited, whereas exogenous cholesterol (Cho) stimulated, proliferation cell cycle progression in control Daudi cells, but not in the cells when transient receptor potential canonical 6 (TRPC6) channel was knocked down. Lovastatin decreased, whereas Cho increased, the levels of intracellular reactive oxygen species (ROS) respectively by decreasing or increasing the expression of p47-phox and gp91-phox (NOX2). Reducing intracellular ROS with either a mimetic superoxide dismutase (TEMPOL) or a NADPH oxidase inhibitor (apocynin) inhibited cell proliferation, particularly in Cho-treated cells. The effects of TEMPOL or apocynin were mimicked by inhibition of TRPC6 with SKF-96365. Lovastatin decreased TRPC6 expression and activity via a Cho-dependent mechanism, whereas Cho increased TRPC6 expression and activity via an ROS-dependent mechanism. Consistent with the fact that TRPC6 is a Ca2+-permeable channel, lovastatin decreased, but Cho increased, intracellular Ca2+ also via ROS. These data suggest that lovastatin inhibits malignant B cell proliferation by reducing membrane Cho, intracellular ROS, TRPC6 expression and activity, and intracellular Ca2+. PMID:24518247

  6. Lovastatin inhibits human B lymphoma cell proliferation by reducing intracellular ROS and TRPC6 expression.

    PubMed

    Song, Xiang; Liu, Bing-Chen; Lu, Xiao-Yu; Yang, Li-Li; Zhai, Yu-Jia; Eaton, Amity F; Thai, Tiffany L; Eaton, Douglas C; Ma, He-Ping; Shen, Bao-Zhong

    2014-05-01

    Clinical evidence suggests that statins reduce cancer incidence and mortality. However, there is lack of in vitro data to show the mechanism by which statins can reduce the malignancies of cancer cells. We used a human B lymphoma Daudi cells as a model and found that lovastatin inhibited, whereas exogenous cholesterol (Cho) stimulated, proliferation cell cycle progression in control Daudi cells, but not in the cells when transient receptor potential canonical 6 (TRPC6) channel was knocked down. Lovastatin decreased, whereas Cho increased, the levels of intracellular reactive oxygen species (ROS) respectively by decreasing or increasing the expression of p47-phox and gp91-phox (NOX2). Reducing intracellular ROS with either a mimetic superoxide dismutase (TEMPOL) or an NADPH oxidase inhibitor (apocynin) inhibited cell proliferation, particularly in Cho-treated cells. The effects of TEMPOL or apocynin were mimicked by inhibition of TRPC6 with SKF-96365. Lovastatin decreased TRPC6 expression and activity via a Cho-dependent mechanism, whereas Cho increased TRPC6 expression and activity via an ROS-dependent mechanism. Consistent with the fact that TRPC6 is a Ca(2+)-permeable channel, lovastatin decreased, but Cho increased, intracellular Ca(2+) also via ROS. These data suggest that lovastatin inhibits malignant B cell proliferation by reducing membrane Cho, intracellular ROS, TRPC6 expression and activity, and intracellular Ca(2+). Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Reduced expression of pain mediators and pain sensitivity in amyloid precursor protein over-expressing CRND8 transgenic mice.

    PubMed

    Shukla, M; Quirion, R; Ma, W

    2013-10-10

    β-Amyloid (Aβ) peptides are derived from the sequential cleavage of the amyloid precursor protein (APP). They are enriched in plaques present in Alzheimer's brains and thus play important roles in the pathogenesis of this disease. APP is also known to be expressed in the neurons of dorsal root ganglion (DRG) and contributes to neuronal survival and axonal growth during development. However, whether APP and Aβ peptides are involved in nociception and pathological pain states is mostly unknown. In the present study, we have used behavioral, biochemical and morphological approaches to address this issue in both adult rats and APP over-expressing CRND8 transgenic mice. We observed that the Aβ peptide (17-24) was predominantly expressed in small-sized DRG neurons of rats. Following intraplantar (i.pl.) injection of complete Freud's adjuvant (CFA), the levels of APP and Aβ peptides were significantly reduced in the ipsilateral lumbar 4-6 rat DRG. In 3-, 12- and 24-month-old CRND8 mice, pain sensitivity in response to heat and mechanical stimulation was significantly dampened compared to their age-matched wild-type littermates. In parallel with reduced pain sensitivity, the expression of pain mediators such as substance P, calcitonin gene-related peptide and transient receptor potential vanilloid-1 was significantly reduced in L4-6 DRG of CRND8 mice. Although i.pl. injection of CFA induced a rather similar pattern of inflammatory pain in 3-month-old CRND8 mice and their wild-type littermates, recovery from inflammatory pain seemed faster in 12-month-old CRND8 mice than wild-type mice. These findings suggest that APP and Aβ peptides suppress both nociception and inflammatory pain and are likely involved in blunt pain perception of Alzheimer's patients in clinical settings.

  8. Reduced capacity in automatic processing of facial expression in restrictive anorexia nervosa and obesity.

    PubMed

    Cserjési, Renáta; Vermeulen, Nicolas; Lénárd, László; Luminet, Olivier

    2011-07-30

    There is growing evidence that disordered eating is associated with facial expression recognition and emotion processing problems. In this study, we investigated the question of whether anorexia and obesity occur on a continuum of attention bias towards negative facial expressions in comparison with healthy individuals of normal weight. Thirty-three patients with restrictive anorexia nervosa (AN-R), 30 patients with obesity (OB) and 63 healthy age and social-economic status matched controls were recruited. Our results indicated that AN-R patients were more attentive to angry faces and had difficulties in being attentive to positive expressions, whilst OB patients had problems in looking for or being attentive to negative expressions independently of self-reported depression and anxiety. Our findings did not support the idea that AN-R and OB occur on a continuum. We found that AN-R was associated with a reduced capacity in positive facial expression processing, whereas OB was associated with a reduced capacity in negative facial expressions processing. The social relevance of our findings and a possible explanation based upon neuroscience are discussed. Copyright © 2010 Elsevier Ltd. All rights reserved.

  9. Oxygen impairs oligodendroglial development via oxidative stress and reduced expression of HIF-1α

    PubMed Central

    Brill, Christina; Scheuer, Till; Bührer, Christoph; Endesfelder, Stefanie; Schmitz, Thomas

    2017-01-01

    The premature increase of oxygen tension may contribute to oligodendroglial precursor cell (OPC) damage in preterm infants. Fetal OPCs are exposed to low oxygen tissue tensions not matched when cells are cultured in room air. Maturation (A2B5, O4, O1, MBP, CNP, arborization), oxidative stress (nitrotyrosine Western blot, NRF2 and SOD2 expression), apoptosis (TUNEL), proliferation (Ki67), and expression of transcription factors regulated by Hypoxia-Inducible-Factor-1-alpha (Hif-1α) expressed in OPCs (Olig1, Olig2, Sox9, Sox10) were assessed in rat OPCs and OLN93 cells cultured at 5% O2 and 21% O2. Influences of Hif-1α were investigated by Hif-1α luciferase reporter assays and Hif-1α-knockdown experiments. At 21% O2, cell proliferation was decreased and process arborization of OPCs was reduced. Expression of MBP, CNP, Olig1, Sox9 and Sox10 was lower at 21% O2, while Nrf2, SOD2, nitrotyrosine were increased. Apoptosis was unchanged. Luciferease reporter assay in OLN93 cells indicated increased Hif-1α activity at 5% O2. In OLN93 cells at 5% O2, Hif-1α knockdown decreased the expression of MBP and CNP, similar to that observed at 21% O2. These data indicate that culturing OPCs at 21% O2 negatively affects development and maturation. Both enhanced oxidative stress and reduced expression of Hif-1α-regulated genes contribute to these hyperoxia-induced changes. PMID:28230075

  10. Reduced expression of monocyte CD200R is associated with enhanced proinflammatory cytokine production in sarcoidosis

    PubMed Central

    Fraser, Simon D.; Sadofsky, Laura R.; Kaye, Paul M.; Hart, Simon P.

    2016-01-01

    In sarcoidosis, the proinflammatory cytokines interferon gamma, tumour necrosis factor and interleukin-6 are released by monocyte-derived macrophages and lymphocytes in the lungs and other affected tissues. Regulatory receptors expressed on monocytes and macrophages act to suppress cytokine production, and reduced expression of regulatory receptors may thus promote tissue inflammation. The aim of this study was to characterise the role of regulatory receptors on blood monocytes in patients with sarcoidosis. Cytokine release in response to stimulation of whole blood was measured in healthy controls and Caucasian non-smoking patients with sarcoidosis who were not taking disease modifying therapy. Expression of the regulatory molecules IL-10R, SIRP-α/β, CD47, CD200R, and CD200L was measured by flow cytometry, and functional activity was assessed using blocking antibodies. Stimulated whole blood and monocytes from patients with sarcoidosis produced more TNF and IL-6 compared with healthy controls. 52.9% of sarcoidosis patients had monocytes characterised by low expression of CD200R, compared with 11.7% of controls (p < 0.0001). Patients with low monocyte CD200R expression produced higher levels of proinflammatory cytokines. In functional studies, blocking the CD200 axis increased production of TNF and IL-6. Reduced expression of CD200R on monocytes may be a mechanism contributing to monocyte and macrophage hyper-activation in sarcoidosis. PMID:27929051

  11. Micro-structuring of polycarbonate-urethane surfaces in order to reduce platelet activation and adhesion.

    PubMed

    Clauser, Johanna; Gester, Kathrin; Roggenkamp, Jan; Mager, Ilona; Maas, Judith; Jansen, Sebastian V; Steinseifer, Ulrich

    2014-01-01

    In the development of new hemocompatible biomaterials, surface modification appears to be a suitable method in order to reduce the thrombogenetic potential of such materials. In this study, polycarbonate-urethane (PCU) tubes with different surface microstructures to be used for aortic heart valve models were investigated with regard to the thrombogenicity. The surface structures were produced by using a centrifugal casting process for manufacturing PCU tubes with defined casting mold surfaces which are conferred to the PCU surface during the process. Tubes with different structures defined by altering groove widths were cut into films and investigated under dynamic flow conditions in contact with porcine blood. The analysis was carried out by laser scanning microscopy which allowed for counting various morphological types of platelets with regard to the grade of activation. The comparison between plain and shaped PCU samples showed that the surface topography led to a decline of the activation of the coagulation cascade and thus to the reduction of the fibrin synthesis. Comparing different types of structures revealed that smooth structures with a small groove width (d ~ 3 μm) showed less platelet activation as well as less adhesion in contrast to a distinct wave structure (d ~ 90 μm). These results prove surface modification of polymer biomaterials to be a suitable method for reducing thrombogenicity and hence give reason for further alterations and improvements.

  12. Reducing Ice Adhesion on Nonsmooth Metallic Surfaces: Wettability and Topography Effects.

    PubMed

    Ling, Edwin Jee Yang; Uong, Victor; Renault-Crispo, Jean-Sébastien; Kietzig, Anne-Marie; Servio, Phillip

    2016-04-06

    The effects of ice formation and accretion on external surfaces range from being mildly annoying to potentially life-threatening. Ice-shedding materials, which lower the adhesion strength of ice to its surface, have recently received renewed research attention as a means to circumvent the problem of icing. In this work, we investigate how surface wettability and surface topography influence the ice adhesion strength on three different surfaces: (i) superhydrophobic laser-inscribed square pillars on copper, (ii) stainless steel 316 Dutch-weave meshes, and (iii) multiwalled carbon nanotube-covered steel meshes. The finest stainless steel mesh displayed the best performance with a 93% decrease in ice adhesion relative to polished stainless steel, while the superhydrophobic square pillars exhibited an increase in ice adhesion by up to 67% relative to polished copper. Comparisons of dynamic contact angles revealed little correlation between surface wettability and ice adhesion. On the other hand, by considering the ice formation process and the fracture mechanics at the ice-substrate interface, we found that two competing mechanisms governing ice adhesion strength arise on nonplanar surfaces: (i) mechanical interlocking of the ice within the surface features that enhances adhesion, and (ii) formation of microcracks that act as interfacial stress concentrators, which reduce adhesion. Our analysis provides insight toward new approaches for the design of ice-releasing materials through the use of surface topographies that promote interfacial crack propagation.

  13. Changes in Gene Expression of E. coli under Conditions of Modeled Reduced Gravity

    NASA Astrophysics Data System (ADS)

    Vukanti, Raja; Mintz, Eric; Leff, Laura

    2008-06-01

    Relatively few studies have examined bacterial responses to the reduced gravity conditions that are experienced by bacteria grown in space. In this study, whole genome expression of Escherichia coli K12 under clinorotation (which models some of the conditions found under reduced gravity) was analyzed. We hypothesized that phenotypic differences at cellular and population levels under clinorotation (hereafter referred to as modeled reduced gravity) are directly coupled to changes in gene expression. Further, we hypothesized that these responses may be due to indirect effects of these environmental conditions on nutrient accessibility for bacteria. Overall, 430 genes were identified as significantly different between modeled reduced gravity conditions and controls. Up-regulated genes included those involved in the starvation response ( csiD, cspD, ygaF, gabDTP, ygiG, fliY, cysK) and redirecting metabolism under starvation ( ddpX, acs, actP, gdhA); responses to multiple stresses, such as acid stress ( asr, yhiW), osmotic stress ( yehZYW), oxidative stress ( katE, btuDE); biofilm formation ( lldR, lamB, yneA, fadB, ydeY); curli biosynthesis ( csgDEF), and lipid biosynthesis ( yfbEFG). Our results support the previously proposed hypothesis that under conditions of modeled reduced gravity, zones of nutrient depletion develop around bacteria eliciting responses similar to entrance into stationary phase which is generally characterized by expression of starvation inducible genes and genes associated with multiple stress responses.

  14. Aquaporin-4 expression is severely reduced in human sarcoglycanopathies and dysferlinopathies.

    PubMed

    Assereto, Stefania; Mastrototaro, Mauro; Stringara, Silvia; Gazzerro, Elisabetta; Broda, Paolo; Nicchia, Grazia Paola; Svelto, Maria; Bruno, Claudio; Nigro, Vincenzo; Lisanti, M P; Frigeri, Antonio; Minetti, Carlo

    2008-07-15

    Aquaporin-4 (AQP4) is the major water channel expressed in fast-twitch skeletal muscle fibers. AQP4 is reduced in Duchenne and Becker Muscular Dystrophies, but not in caveolinopathies, thus suggesting an interaction with dystrophin or with members of the dystrophin-glycoprotein complex (DGC) rather than a nonspecific effect due to muscle membrane damage. To establish the role of sarcoglycans in AQP4 decrease occurring in muscular dystrophy, AQP4 expression was analyzed in muscle biopsies from patients affected by Limb Girdle Muscular Dystrophies (LGMDs) 2C-F genetically confirmed. In all the LGMD 2C-F (2alpha-, 1beta-, 2gamma-, 1delta-deficiency), AQP4 was severely decreased. This effect was associated to a marked reduction in alpha1-syntrophin levels. In control muscle AQP4 did not show a direct interaction with any of the four sarcoglycans but, it co-immunoprecipitated with alpha1-syntrophin, indicating that this modular protein may link AQP4 levels with the DGC complex. To determine whether AQP4 expression could be affected in other LGMDs due to the defect of a membrane protein not associated to the dystrophin complex, we examined AQP4 expression in 6 patients affected by dysferlin deficiency genetically confirmed. All the patients displayed a reduction of the water channel, and AQP4 expression appeared to correlate with the severity of the muscle histopathological lesions. However, differently from what observed in the sarcoglycans, alpha1-syntrophin expression was normal or just slightly reduced. These results seem to indicate an additional mechanism of regulation of AQP4 levels in muscle cells. In accordance with a specific effect of membrane muscle disorders, AQP4 protein levels were not changed in 3 mitochondrial and 3 metabolic myopathies. In conclusion, AQP4 expression and membrane localization are markedly reduced in LGMD 2B-2F. The role of AQP4 in the degenerative mechanism occurring in these diseases will be the object of our future research.

  15. Arsenic transport between surface and groundwater in a moderately reducing zone: Geochemical approach

    NASA Astrophysics Data System (ADS)

    Khaska, Mahmoud; Le Gal La Salle, Corinne; Verdoux, Patrick

    2015-04-01

    Arsenic contamination represents a major risk to human health as one of the most prominent environmental causes of cancer mortality. Mining activities, particularly those involving arsenic rich ores have an impact on the environment and on human health that may persist for many decades after mine closure. The relationships between As released from alluvial aquifer in the vicinity of the sulfide-rich mine dumps was demonstrated with geochemical and isotopic tracers (major and traces elements, 87Sr/86Sr, 18O, 2H). Strontium isotopes were used to trace the transport of As downstream from a As rich tailing dam. Increasing As and Fe concentrations in surface water are explained by As release associated with alluvial groundwater discharge to the stream. This process occurs in a moderately reduced section of the stream downgradient from the sulfide-rich tailing dam. High As, total Fe and low Eh in groundwater confirm the discharge of alluvial groundwater and explain its impact on surface water. Transport of As between surface and groundwater can be described as follows: 1- Subsurface moderately reducing conditions prevail in groundwater downgradient from the tailing dams. This suggests a flux of reduced water from sulfide-rich tailing dams which is characterized by its high As and Fe content resulting from the reduction of Fe-sulfides. 2- Upon mixing with surface water, oxidizing conditions prevails and precipitate as Fe hydroxide on the stream bed. As and Sr subsequently adsorbed on the Fe -oxyhydroxide surface. This process contributes to the immobilization of As in surface water. Remaining dissolved As in surface water can be re-introduced in alluvial groundwater downstream of the reducing zone.

  16. Regulation of collagen fibrillogenesis by cell-surface expression of kinase dead DDR2.

    PubMed

    Blissett, Angela R; Garbellini, Derek; Calomeni, Edward P; Mihai, Cosmin; Elton, Terry S; Agarwal, Gunjan

    2009-01-23

    The assembly of collagen fibers, the major component of the extracellular matrix (ECM), governs a variety of physiological processes. Collagen fibrillogenesis is a tightly controlled process in which several factors, including collagen binding proteins, have a crucial role. Discoidin domain receptors (DDR1 and DDR2) are receptor tyrosine kinases that bind to and are phosphorylated upon collagen binding. The phosphorylation of DDRs is known to activate matrix metalloproteases, which in turn cleave the ECM. In our earlier studies, we established a novel mechanism of collagen regulation by DDRs; that is, the extracellular domain (ECD) of DDR2, when used as a purified, soluble protein, inhibits collagen fibrillogenesis in-vitro. To extend this novel observation, the current study investigates how the DDR2-ECD, when expressed as a membrane-anchored, cell-surface protein, affects collagen fibrillogenesis by cells. We generated a mouse osteoblast cell line that stably expresses a kinase-deficient form of DDR2, termed DDR2/-KD, on its cell surface. Transmission electron microscopy, fluorescence microscopy, and hydroxyproline assays demonstrated that the expression of DDR2/-KD reduced the rate and abundance of collagen deposition and induced significant morphological changes in the resulting fibers. Taken together, our observations extend the functional roles that DDR2 and possibly other membrane-anchored, collagen-binding proteins can play in the regulation of cell adhesion, migration, proliferation and in the remodeling of the extracellular matrix.

  17. How to reduce resistance to movement of alkane liquid drops across tilted surfaces without relying on surface roughening and perfluorination.

    PubMed

    Urata, Chihiro; Masheder, Benjamin; Cheng, Dalton F; Hozumi, Atsushi

    2012-12-21

    Alkylsilane-derived monolayer-covered surfaces generally display a reasonably good level of hydrophobicity but poor oleophobicity. Here, we demonstrate that the physical attributes of alkylsilane-derived surfaces (liquid-like or solid-like) are dependent on the alkyl chain length and density, and these factors subsequently have significant influence upon the dynamic dewetting behavior toward alkanes (C(n)H(2n+2), where n = 7-16). In this study, we prepared and characterized hybrid films through a simple sol-gel process based on the cohydrolysis and co-condensation of a mixture of a range of alkyltriethoxysilanes (C(n)H(2n+1)Si(OEt)(3), where n = 3, 6, 8, 10, 12, 14, 16, and 18) and tetramethoxysilane (TMOS). Surprisingly, when the carbon number (C(n)) of alkyl chain was 10 and below, the produced hybrid films were all smooth, highly transparent, and showed negligible contact angle (CA) hysteresis. On these hybrid surfaces, 5 μL drops of alkanes (n-hexadecane, n-dodecane, and n-decane) could move easily at low tilt angles (<5°) without pinning. On the other hand, when the C(n) exceeded 12, both transparency and mobility of probe liquids significantly worsened. In the former case, TMOS molecules played key roles in both forming continuous films (as a binder) and improving flexibility of alkyl chains (as a molecular spacer), resulting in the smooth liquid-like surfaces. Silylation of the hybrid film and subsequent dynamic CA measurements proved the presence of silanol groups on the outermost surfaces and demonstrated that the dynamic dewettability of hybrid films worsened as packing densities increased. Additionally, solvent effects (high affinity) between the alkyl chains and alkane liquids imparted a more liquid-like character to the surface. Thanks to these simple physical effects, the resistance to the alkane droplet motion across tilted surfaces was markedly reduced. With the longer carbon chains, the chain mobility was strictly inhibited by mutual interactions

  18. Norlichexanthone Reduces Virulence Gene Expression and Biofilm Formation in Staphylococcus aureus

    PubMed Central

    Bojer, Martin S.; Zhao, Yu; Friberg, Cathrine; Ifrah, Dan; Glasser Heede, Nina; Larsen, Thomas O.; Frøkiær, Hanne; Frees, Dorte; Zhang, Lixin; Dai, Huanqin

    2016-01-01

    Staphylococcus aureus is a serious human pathogen and antibiotic resistant, community-associated strains, such as the methicillin resistant S. aureus (MRSA) strain USA300, continue to spread. To avoid resistance, anti-virulence therapy has been proposed where toxicity is targeted rather than viability. Previously we have shown that norlichexanthone, a small non-reduced tricyclic polyketide produced by fungi and lichens, reduces expression of hla encoding α-hemolysin as well as the regulatory RNAIII of the agr quorum sensing system in S. aureus 8325–4. The aim of the present study was to further characterise the mode of action of norlichexanthone and its effect on biofilm formation. We find that norlichexanthone reduces expression of both hla and RNAIII also in strain USA300. Structurally, norlichexanthone resembles ω-hydroxyemodin that recently was shown to bind the agr two component response regulator, AgrA, which controls expression of RNAIII and the phenol soluble modulins responsible for human neutrophil killing. We show that norlichexanthone reduces S. aureus toxicity towards human neutrophils and interferes directly with AgrA binding to its DNA target. In contrast to ω-hydroxyemodin however, norlichexanthone reduces staphylococcal biofilm formation. Transcriptomic analysis revealed that genes regulated by the SaeRS two-component system are repressed by norlichexanthone when compared to untreated cells, an effect that was mitigated in strain Newman carrying a partially constitutive SaeRS system. Our data show that norlichexanthone treatment reduces expression of key virulence factors in CA-MRSA strain USA300 via AgrA binding and represses biofilm formation. PMID:28005941

  19. Method and apparatus for reducing the drag of flows over surfaces

    NASA Technical Reports Server (NTRS)

    Keefe, Laurence R. (Inventor)

    1998-01-01

    An apparatus, and its accompanying method, for reducing the drag of flows over a surface includes arrays of small disks and sensors. The arrays are embedded in the surface and may extend above, or be depressed below, the surface, provided they remain hydraulically smooth either when operating or when inactive. The disks are arranged in arrays of various shapes, and spaced according to the cruising speed of the vehicle on which the arrays are installed. For drag reduction at speeds of the order of 30 meters/second, preferred embodiments include disks that are 0.2 millimeter in diameter and spaced 0.4 millimeter apart. For drag reduction at speeds of the order of 300 meters/second, preferred embodiments include disks that are 0.045 millimeter in diameter and spaced 0.09 millimeter apart. Smaller and larger dimensions for diameter and spacing are also possible. The disks rotate in the plane of the surface, with their rotation axis substantially perpendicular to the surface. The rotating disks produce velocity perturbations parallel to the surface in the overlying boundary layer. The sensors sense the flow at the surface and connect to control circuitry that adjusts the rotation rates and duty cycles of the disks accordingly. Suction and blowing holes can be interspersed among, or made coaxial with, the disks for creating general three-component velocity perturbations in the near-surface region. The surface can be a flat, planar surface or a nonplanar surface, such as a triangular riblet surface. The present apparatus and method have potential applications in the field of aeronautics for improving performance and efficiency of commercial and military aircraft, and in other industries where drag is an obstacle, including gas and oil delivery through long-haul pipelines.

  20. Indoxyl sulfate reduces klotho expression and promotes senescence in the kidneys of hypertensive rats.

    PubMed

    Adijiang, Ayinuer; Shimizu, Hidehisa; Higuchi, Yusuke; Nishijima, Fuyuhiko; Niwa, Toshimitsu

    2011-01-01

    Administration of indoxyl sulfate, a uremic toxin, promotes progression of chronic kidney disease in rats affected by the disease. Klotho, an anti-aging gene, is expressed in the kidneys, and its renal expression is decreased in chronic kidney disease. This study aimed to clarify whether indoxyl sulfate could reduce klotho expression and contribute to cell senescence in the kidneys of hypertensive rats. The rats used for this study were segregated in to the following 4 groups: (1) Dahl salt-resistant normotensive rats (DN), (2) Dahl salt-resistant normotensive indoxyl sulfate-administered rats (DN + IS), (3) Dahl salt-sensitive hypertensive rats (DH), and (4) Dahl salt-sensitive hypertensive indoxyl sulfate-administered rats (DH + IS). After 32 weeks, their kidneys were excised for histological and immunohistochemical analysis for klotho, senescence-associated β-galactosidase, p16(INK4a), p21(WAF1/CIP1), p53, and retinoblastoma protein (Rb). DH + IS rats showed decreased expression of klotho, increased expression of senescence-associated β-galactosidase, p16(INK4a), p21(WAF1/CIP1), p53, and Rb in renal tubular cells, and increased tubulointerstitial fibrosis and mesangial expansion as compared with DH rats. Further, DN + IS rats showed decreased expression of klotho as compared with DN rats. Administration of indoxyl sulfate to hypertensive rats reduced renal expression of klotho and promoted cell senescence with expression of senescence-related proteins, such as p16(INK4a), p21(WAF1/CIP1), p53, and Rb, which was accompanied by renal fibrosis. Copyright © 2011 National Kidney Foundation, Inc. Published by Elsevier Inc. All rights reserved.

  1. Hypothermia reduces VEGF-165 expression, but not osteogenic differentiation of human adipose stem cells under hypoxia

    PubMed Central

    Bakker, Astrid D.; Hogervorst, Jolanda M. A.; Nolte, Peter A.; Klein-Nulend, Jenneke

    2017-01-01

    Cryotherapy is successfully used in the clinic to reduce pain and inflammation after musculoskeletal damage, and might prevent secondary tissue damage under the prevalent hypoxic conditions. Whether cryotherapy reduces mesenchymal stem cell (MSC) number and differentiation under hypoxic conditions, causing impaired callus formation is unknown. We aimed to determine whether hypothermia modulates proliferation, apoptosis, nitric oxide production, VEGF gene and protein expression, and osteogenic/chondrogenic differentiation of human MSCs under hypoxia. Human adipose MSCs were cultured under hypoxia (37°C, 1% O2), hypothermia and hypoxia (30°C, 1% O2), or control conditions (37°C, 20% O2). Total DNA, protein, nitric oxide production, alkaline phosphatase activity, gene expression, and VEGF protein concentration were measured up to day 8. Hypoxia enhanced KI67 expression at day 4. The combination of hypothermia and hypoxia further enhanced KI67 gene expression compared to hypoxia alone, but was unable to prevent the 1.2-fold reduction in DNA amount caused by hypoxia at day 4. Addition of hypothermia to hypoxic cells did not alter the effect of hypoxia alone on BAX-to-BCL-2 ratio, alkaline phosphatase activity, gene expression of SOX9, COL1, or osteocalcin, or nitric oxide production. Hypothermia decreased the stimulating effect of hypoxia on VEGF-165 gene expression by 6-fold at day 4 and by 2-fold at day 8. Hypothermia also decreased VEGF protein expression under hypoxia by 2.9-fold at day 8. In conclusion, hypothermia decreased VEGF-165 gene and protein expression, but did not affect differentiation, or apoptosis of MSCs cultured under hypoxia. These in vitro results implicate that hypothermia treatment in vivo, applied to alleviate pain and inflammation, is not likely to harm early stages of callus formation. PMID:28166273

  2. Evolution of the insect body plan as revealed by the Sex combs reduced expression pattern.

    PubMed

    Rogers, B T; Peterson, M D; Kaufman, T C

    1997-01-01

    The products of the HOM/Hox homeotic genes form a set of evolutionarily conserved transcription factors that control elaborate developmental processes and specify cell fates in many metazoans. We examined the expression of the ortholog of the homeotic gene Sex combs reduced (Scr) of Drosophila melanogaster in insects of three divergent orders: Hemiptera, Orthoptera and Thysanura. Our data reflect how the conservation and variation of Scr expression has affected the morphological evolution of insects. Whereas the anterior epidermal expression of Scr, in a small part of the posterior maxillary and all of the labial segment, is found to be in common among all four insect orders, the posterior (thoracic) expression domains vary. Unlike what is observed in flies, the Scr orthologs of other insects are not expressed broadly over the first thoracic segment, but are restricted to small patches. We show here that Scr is required for suppression of wings on the prothorax of Drosophila. Moreover, Scr expression at the dorsal base of the prothoracic limb in two other winged insects, crickets (Orthoptera) and milkweed bugs (Hemiptera), is consistent with Scr acting as a suppressor of prothoracic wings in these insects. Scr is also expressed in a small patch of cells near the basitarsal-tibial junction of milkweed bugs, precisely where a leg comb develops, suggesting that Scr promotes comb formation, as it does in Drosophila. Surprisingly, the dorsal prothoracic expression of Scr is also present in the primitively wingless firebrat (Thysanura) and the leg patch is seen in crickets, which have no comb. Mapping both gene expression patterns and morphological characters onto the insect phylogenetic tree demonstrates that in the cases of wing suppression and comb formation the appearance of expression of Scr in the prothorax apparently precedes these specific functions.

  3. Inducing a concurrent motor load reduces categorization precision for facial expressions.

    PubMed

    Ipser, Alberta; Cook, Richard

    2016-05-01

    Motor theories of expression perception posit that observers simulate facial expressions within their own motor system, aiding perception and interpretation. Consistent with this view, reports have suggested that blocking facial mimicry induces expression labeling errors and alters patterns of ratings. Crucially, however, it is unclear whether changes in labeling and rating behavior reflect genuine perceptual phenomena (e.g., greater internal noise associated with expression perception or interpretation) or are products of response bias. In an effort to advance this literature, the present study introduces a new psychophysical paradigm for investigating motor contributions to expression perception that overcomes some of the limitations inherent in simple labeling and rating tasks. Observers were asked to judge whether smiles drawn from a morph continuum were sincere or insincere, in the presence or absence of a motor load induced by the concurrent production of vowel sounds. Having confirmed that smile sincerity judgments depend on cues from both eye and mouth regions (Experiment 1), we demonstrated that vowel production reduces the precision with which smiles are categorized (Experiment 2). In Experiment 3, we replicated this effect when observers were required to produce vowels, but not when they passively listened to the same vowel sounds. In Experiments 4 and 5, we found that gender categorizations, equated for difficulty, were unaffected by vowel production, irrespective of the presence of a smiling expression. These findings greatly advance our understanding of motor contributions to expression perception and represent a timely contribution in light of recent high-profile challenges to the existing evidence base.

  4. Defective erythropoiesis in a mouse model of reduced Fbxo7 expression due to decreased p27 expression

    PubMed Central

    Randle, Suzanne J; Nelson, David E; Patel, Shachi P; Laman, Heike

    2015-01-01

    During the final stages of erythropoiesis, lineage-restricted progenitors mature over three to five cell divisions, culminating with withdrawal from the cell cycle and the loss of most organelles, including mitochondria and nuclei. Recent genome-wide association studies in human populations have associated several SNPs near or within FBXO7 with erythrocyte phenotypes. Fbxo7 encodes a multi-functional F-box protein known to bind p27 and participate in selective mitophagy. One SNP causes an amino acid substitution (Met115Ile) and is associated with smaller erythrocytes. We find that the less common IIe115 allele of Fbxo7 binds less efficiently to p27, and cells expressing this allele proliferate faster than cells expressing Met115. We show that an erythroleukaemic cell line with reduced Fbxo7 expression fails to stabilize p27 levels, exit the cell cycle, and produce haemoglobin. In addition, mice deficient in Fbxo7 expression are anaemic due to a reduction in erythrocyte numbers, and this is associated with lower p27 levels, increased numbers of late-stage erythroblasts with greater than 2N DNA content, and delayed mitophagy during terminal differentiation. Collectively, these data support an important physiological, cell cycle regulatory role for Fbxo7 during erythropoiesis. © 2015 Authors. Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland. PMID:26095538

  5. Airfoil-shaped micro-mixers for reducing fouling on membrane surfaces

    DOEpatents

    Ho, Clifford K; Altman, Susan J; Clem, Paul G; Hibbs, Michael; Cook, Adam W

    2012-10-23

    An array of airfoil-shaped micro-mixers that enhances fluid mixing within permeable membrane channels, such as used in reverse-osmosis filtration units, while minimizing additional pressure drop. The enhanced mixing reduces fouling of the membrane surfaces. The airfoil-shaped micro-mixer can also be coated with or comprised of biofouling-resistant (biocidal/germicidal) ingredients.

  6. Soluble calcium amendment: Co-Application with poultry litter to reduce P loss following surface application

    USDA-ARS?s Scientific Manuscript database

    This paper will discuss the utilization of gypsum (CaSO4 .2H2O) to reduce P losses from surface runoff when poultry litter is used as a fertilizer source in agriculture. Utilization of poultry litter as a fertilizer source is common in regions with intense poultry production. While poultry litter ha...

  7. Increased hand washing reduces influenza virus surface contamination in Bangkok households, 2009-2010.

    PubMed

    Levy, Jens W; Suntarattiwong, Piyarat; Simmerman, James M; Jarman, Richard G; Johnson, Kara; Olsen, Sonja J; Chotpitayasunondh, Tawee

    2014-01-01

    Within a hand-washing clinical trial, we evaluated factors associated with fomite contamination in households with an influenza-infected child. Influenza virus RNA contamination was higher in households with low absolute humidity and in control households, suggesting that hand washing reduces surface contamination.

  8. Coupled Phase-Variable Expression and Epitope Masking of Selective Surface Lipoproteins Increase Surface Phenotypic Diversity in Mycoplasma hominis

    PubMed Central

    Zhang, Qijing; Wise, Kim S.

    2001-01-01

    A new mechanism expanding mycoplasmal surface diversity is described. Exposure of surface epitopes on a constitutively expressed membrane protein (P56) of Mycoplasma hominis was subject to high-frequency phase variation due to phase-variable expression of the P120 antigen and its selective masking of P56 epitopes. Phase-variable masking may confer previously unrealized adaptive capabilities on mycoplasmas. PMID:11447202

  9. Reduced responses of macrophages on nanometer surface features of altered alumina crystalline phases.

    PubMed

    Khang, Dongwoo; Liu-Snyder, Peishan; Pareta, Rajesh; Lu, Jing; Webster, Thomas J

    2009-06-01

    Extensive prolonged interactions of inflammatory cells (such as macrophages) at the host-implant interface may lead to implant failure. While previous studies have shown increased in vitro and in vivo bone cell adhesion, proliferation and mineralization on nanophase compared to currently implanted ceramics, few studies have been conducted to elucidate inflammatory cell responses on such nanophase ceramics. Controlling surface feature size and corresponding surface roughness on implants may clearly alter immune cell responses, which would be an extremely important consideration for the use of nanostructured materials as improved biomaterials. In this study, reduced macrophage density was observed on alumina (Al(2)O(3)) compacts with greater nanometer surface roughness accompanied by changes in crystallinity for up to 24 h in culture. Since alumina is a commonly used ceramic in orthopedic applications, this in vitro study continues to support the use of nanophase ceramics as improved orthopedic implants by demonstrating reduced macrophage responses.

  10. Shear stress reduces protease activated receptor-1 expression in human endothelial cells

    NASA Technical Reports Server (NTRS)

    Nguyen, K. T.; Eskin, S. G.; Patterson, C.; Runge, M. S.; McIntire, L. V.

    2001-01-01

    Shear stress has been shown to regulate several genes involved in the thrombotic and proliferative functions of endothelial cells. Thrombin receptor (protease-activated receptor-1: PAR-1) increases at sites of vascular injury, which suggests an important role for PAR-1 in vascular diseases. However, the effect of shear stress on PAR-1 expression has not been previously studied. This work investigates effects of shear stress on PAR-1 gene expression in both human umbilical vein endothelial cells (HUVECs) and microvascular endothelial cells (HMECs). Cells were exposed to different shear stresses using a parallel plate flow system. Northern blot and flow cytometry analysis showed that shear stress down-regulated PAR-1 messenger RNA (mRNA) and protein levels in both HUVECs and HMECs but with different thresholds. Furthermore, shear-reduced PAR-1 mRNA was due to a decrease of transcription rate, not increased mRNA degradation. Postshear stress release of endothelin-1 in response to thrombin was reduced in HUVECs and HMECs. Moreover, inhibitors of potential signaling pathways applied during shear stress indicated mediation of the shear-decreased PAR-1 expression by protein kinases. In conclusion, shear stress exposure reduces PAR-1 gene expression in HMECs and HUVECs through a mechanism dependent in part on protein kinases, leading to altered endothelial cell functional responses to thrombin.

  11. Shear stress reduces protease activated receptor-1 expression in human endothelial cells

    NASA Technical Reports Server (NTRS)

    Nguyen, K. T.; Eskin, S. G.; Patterson, C.; Runge, M. S.; McIntire, L. V.

    2001-01-01

    Shear stress has been shown to regulate several genes involved in the thrombotic and proliferative functions of endothelial cells. Thrombin receptor (protease-activated receptor-1: PAR-1) increases at sites of vascular injury, which suggests an important role for PAR-1 in vascular diseases. However, the effect of shear stress on PAR-1 expression has not been previously studied. This work investigates effects of shear stress on PAR-1 gene expression in both human umbilical vein endothelial cells (HUVECs) and microvascular endothelial cells (HMECs). Cells were exposed to different shear stresses using a parallel plate flow system. Northern blot and flow cytometry analysis showed that shear stress down-regulated PAR-1 messenger RNA (mRNA) and protein levels in both HUVECs and HMECs but with different thresholds. Furthermore, shear-reduced PAR-1 mRNA was due to a decrease of transcription rate, not increased mRNA degradation. Postshear stress release of endothelin-1 in response to thrombin was reduced in HUVECs and HMECs. Moreover, inhibitors of potential signaling pathways applied during shear stress indicated mediation of the shear-decreased PAR-1 expression by protein kinases. In conclusion, shear stress exposure reduces PAR-1 gene expression in HMECs and HUVECs through a mechanism dependent in part on protein kinases, leading to altered endothelial cell functional responses to thrombin.

  12. Reduced seed germination in Arabidopsis over-expressing SWI/SNF2 ATPase genes.

    PubMed

    Leeggangers, Hendrika A C F; Folta, Adam; Muras, Aleksandra; Nap, Jan-Peter; Mlynarova, Ludmila

    2015-02-01

    In the life of flowering plants, seed germination is a critical step to ensure survival into the next generation. Generally the seed prior to germination has been in a dormant state with a low rate of metabolism. In the transition from a dormant seed to a germinating seed, various epigenetic mechanisms play a regulatory role. Here, we demonstrate that the over-expression of chromatin remodeling ATPase genes (AtCHR12 or AtCHR23) reduced the frequency of seed germination in Arabidopsis thaliana up to 30% relative to the wild-type seeds. On the other hand, single loss-of-function mutations of the two genes did not affect seed germination. The reduction of germination in over-expressing mutants was more pronounced in stress conditions (salt or high temperature), showing the impact of the environment. Reduced germinations upon over-expression coincided with increased transcript levels of seed maturation genes and with reduced degradation of their mRNAs stored in dry seeds. Our results indicate that repression of AtCHR12/23 gene expression in germinating wild-type Arabidopsis seeds is required for full germination. This establishes a functional link between chromatin modifiers and regulatory networks towards seed maturation and germination.

  13. Modeling of reduced effective secondary electron emission yield from a velvet surface

    DOE PAGES

    Swanson, Charles; Kaganovich, Igor D.

    2016-12-05

    Complex structures on a material surface can significantly reduce total secondary electron emission from that surface. A velvet is a surface that consists of an array of vertically standing whiskers. The reduction occurs due to the capture of low-energy, true secondary electrons emitted at the bottom of the structure and on the sides of the velvet whiskers. We performed numerical simulations and developed an approximate analytical model that calculates the net secondary electron emission yield from a velvet surface as a function of the velvet whisker length and packing density, and the angle of incidence of primary electrons. We foundmore » that to suppress secondary electrons, the following condition on dimensionless parameters must be met: (π/2) DΑ tan θ >> 1, where theta is the angle of incidence of the primary electron from the normal, D is the fraction of surface area taken up by the velvet whisker bases, and A is the aspect ratio, A = h/r, the ratio of height to radius of the velvet whiskers. We find that velvets available today can reduce the secondary electron yield by 90% from the value of a flat surface. As a result, the values of optimal velvet whisker packing density that maximally suppresses the secondary electron emission yield are determined as a function of velvet aspect ratio and the electron angle of incidence.« less

  14. Impact of a Reducing Agent on the Dynamic Surface Properties of Lysozyme Solutions.

    PubMed

    Tihonov, Michael M; Kim, Viktoria V; Noskov, Boris A

    2016-05-01

    Disulfide bond shuffling in the presence of the reducing agents dithiothreitol (DTT) or β-mercaptoethanol (BME) strongly affects the surface properties of lysozyme solutions. The addition of 0.32 mM DTT substantially alters the kinetic dependencies of the dynamic surface elasticity and surface tension relative to those of pure protein solutions. The significant increase in the dynamic surface elasticity likely relates to the cross-linking between lysozyme molecules and the formation of a dense layer of protein globules stabilized by intermolecular disulfide bonds at the liquid/gas interface. This effect differs from the previously described influence of chaotropic denaturants, such as guanidine hydrochloride (GuHCl) and urea, on the surface properties of lysozyme solutions. If both chaotropic and reducing agents are added to protein solutions simultaneously, their effects become superimposed. In the case of mixed lysozyme/GuHCl/DTT solutions, the dynamic surface elasticity near equilibrium decreases as the GuHCl concentration increases because of the gradual loosening of the cross-linked layer of protein globules but remains much higher than that of lysozyme/GuHCl solutions.

  15. Role of humidity in reducing the friction of graphene layers on textured surfaces

    NASA Astrophysics Data System (ADS)

    Li, Zheng-yang; Yang, Wen-jing; Wu, Yan-ping; Wu, Song-bo; Cai, Zhen-bing

    2017-05-01

    A multiple-layer graphene was prepared on steel surface to reduce friction and wear. A graphene-containing ethanol solution was dripped on the steel surface, and several layers of graphene flakes were deposited on the surface after ethanol evaporated. Tribological performance of graphene-contained surface (GCS) was induced by reciprocating ball against plate contact in different RH (0% (dry nitrogen), 30%, 60%, and 90%). Morphology and wear scar were analyzed by OM, 2D profile, SEM, Raman spectroscopy, and XPS. Results show that GCS can substantially reduce the wear and coefficient of friction (COF) in 60% relative humidity (RH). Low COF occurs due to graphene layer providing a small shear stress on the friction interface. Meanwhile, conditions of high RH and textured surface could make the low COF persist for a longer time. High moisture content can stabilize and protect the graphene C-network from damage due to water dissociative chemisorption with carbon dangling bonds, and the textured surface was attributed to release graphene layer stored in the dimple.

  16. Modeling of reduced effective secondary electron emission yield from a velvet surface

    SciTech Connect

    Swanson, Charles; Kaganovich, Igor D.

    2016-12-05

    Complex structures on a material surface can significantly reduce total secondary electron emission from that surface. A velvet is a surface that consists of an array of vertically standing whiskers. The reduction occurs due to the capture of low-energy, true secondary electrons emitted at the bottom of the structure and on the sides of the velvet whiskers. We performed numerical simulations and developed an approximate analytical model that calculates the net secondary electron emission yield from a velvet surface as a function of the velvet whisker length and packing density, and the angle of incidence of primary electrons. We found that to suppress secondary electrons, the following condition on dimensionless parameters must be met: (π/2) DΑ tan θ >> 1, where theta is the angle of incidence of the primary electron from the normal, D is the fraction of surface area taken up by the velvet whisker bases, and A is the aspect ratio, A = h/r, the ratio of height to radius of the velvet whiskers. We find that velvets available today can reduce the secondary electron yield by 90% from the value of a flat surface. As a result, the values of optimal velvet whisker packing density that maximally suppresses the secondary electron emission yield are determined as a function of velvet aspect ratio and the electron angle of incidence.

  17. Modeling of reduced effective secondary electron emission yield from a velvet surface

    NASA Astrophysics Data System (ADS)

    Swanson, Charles; Kaganovich, Igor D.

    2016-12-01

    Complex structures on a material surface can significantly reduce total secondary electron emission from that surface. A velvet is a surface that consists of an array of vertically standing whiskers. The reduction occurs due to the capture of low-energy, true secondary electrons emitted at the bottom of the structure and on the sides of the velvet whiskers. We performed numerical simulations and developed an approximate analytical model that calculates the net secondary electron emission yield from a velvet surface as a function of the velvet whisker length and packing density, and the angle of incidence of primary electrons. We found that to suppress secondary electrons, the following condition on dimensionless parameters must be met: (π/2 )D A tan θ≫1 , where θ is the angle of incidence of the primary electron from the normal, D is the fraction of surface area taken up by the velvet whisker bases, and A is the aspect ratio, A ≡ h/r, the ratio of height to radius of the velvet whiskers. We find that velvets available today can reduce the secondary electron yield by 90% from the value of a flat surface. The values of optimal velvet whisker packing density that maximally suppresses the secondary electron emission yield are determined as a function of velvet aspect ratio and the electron angle of incidence.

  18. Spirulina and C-phycocyanin reduce cytotoxicity and inflammation-related genes expression of microglial cells.

    PubMed

    Chen, Jin-Cherng; Liu, Kris Sun; Yang, Ting-Ju; Hwang, Juen-Haur; Chan, Yin-Ching; Lee, I-Te

    2012-11-01

    Our aim was to investigate the effects of Spirulina on BV-2 microglial cell cytotoxicity and inflammatory genes expression. BV-2 microglial cells were treated with lipopolysaccharide (LPS) (1 µg/ml) and various concentrations of Spirulina platensis water extract or its active component (C-phycocyanin (C-PC)) for 24 hours. Cytotoxicity (lactate dehydrogenase (LDH) release) and expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) mRNAs were assayed. LPS increased LDH production and up-regulated expression of iNOS, COX-2, TNF-α, and IL-6 by BV-2 microglial cells. However, Spirulina platensis water extract and C-PC significantly reduced LPS-induced LDH release, and expression of iNOS, COX-2, TNF-α, and IL-6 mRNAs. Spirulina can reduce the cytotoxicity and inhibit expression of inflammation-related genes of LPS-stimulated BV-2 microglial cells.

  19. A Complex Interaction Between Reduced Reelin Expression and Prenatal Organophosphate Exposure Alters Neuronal Cell Morphology.

    PubMed

    Mullen, Brian R; Ross, Brennan; Chou, Joan Wang; Khankan, Rana; Khialeeva, Elvira; Bui, Kimberly; Carpenter, Ellen M

    2016-06-01

    Genetic and environmental factors are both likely to contribute to neurodevelopmental disorders including schizophrenia, autism spectrum disorders, and major depressive disorders. Prior studies from our laboratory and others have demonstrated that the combinatorial effect of two factors-reduced expression of reelin protein and prenatal exposure to the organophosphate pesticide chlorpyrifos oxon-gives rise to acute biochemical effects and to morphological and behavioral phenotypes in adolescent and young adult mice. In the current study, we examine the consequences of these factors on reelin protein expression and neuronal cell morphology in adult mice. While the cell populations that express reelin in the adult brain appear unchanged in location and distribution, the levels of full length and cleaved reelin protein show persistent reductions following prenatal exposure to chlorpyrifos oxon. Cell positioning and organization in the hippocampus and cerebellum are largely normal in animals with either reduced reelin expression or prenatal exposure to chlorpyrifos oxon, but cellular complexity and dendritic spine organization is altered, with a skewed distribution of immature dendritic spines in adult animals. Paradoxically, combinatorial exposure to both factors appears to generate a rescue of the dendritic spine phenotypes, similar to the mitigation of behavioral and morphological changes observed in our prior study. Together, our observations support an interaction between reelin expression and chlorpyrifos oxon exposure that is not simply additive, suggesting a complex interplay between genetic and environmental factors in regulating brain morphology. © The Author(s) 2016.

  20. A Complex Interaction Between Reduced Reelin Expression and Prenatal Organophosphate Exposure Alters Neuronal Cell Morphology

    PubMed Central

    Mullen, Brian R.; Ross, Brennan; Chou, Joan Wang; Khankan, Rana; Khialeeva, Elvira; Bui, Kimberly

    2016-01-01

    Genetic and environmental factors are both likely to contribute to neurodevelopmental disorders including schizophrenia, autism spectrum disorders, and major depressive disorders. Prior studies from our laboratory and others have demonstrated that the combinatorial effect of two factors—reduced expression of reelin protein and prenatal exposure to the organophosphate pesticide chlorpyrifos oxon—gives rise to acute biochemical effects and to morphological and behavioral phenotypes in adolescent and young adult mice. In the current study, we examine the consequences of these factors on reelin protein expression and neuronal cell morphology in adult mice. While the cell populations that express reelin in the adult brain appear unchanged in location and distribution, the levels of full length and cleaved reelin protein show persistent reductions following prenatal exposure to chlorpyrifos oxon. Cell positioning and organization in the hippocampus and cerebellum are largely normal in animals with either reduced reelin expression or prenatal exposure to chlorpyrifos oxon, but cellular complexity and dendritic spine organization is altered, with a skewed distribution of immature dendritic spines in adult animals. Paradoxically, combinatorial exposure to both factors appears to generate a rescue of the dendritic spine phenotypes, similar to the mitigation of behavioral and morphological changes observed in our prior study. Together, our observations support an interaction between reelin expression and chlorpyrifos oxon exposure that is not simply additive, suggesting a complex interplay between genetic and environmental factors in regulating brain morphology. PMID:27364165

  1. Reduced WIF-1 expression stimulates skin hyperpigmentation in patients with melasma.

    PubMed

    Kim, Ji-Young; Lee, Tae-Ryong; Lee, Ai-Young

    2013-01-01

    The expression of Wnt inhibitory factor-1 (WIF-1) gene, which was detected by a microarray analysis of hyperpigmented and normally pigmented skin sets of melasma patients, was significantly reduced in the hyperpigmented skin from melasma patients, but not in healthy controls, regardless of UV irradiation. Wnt signals regulate skin pigmentation; however, WIF-1 is expressed in cultured skin keratinocytes and fibroblasts, but not in melanocytes. Therefore, we examined whether WIF-1 knockdown in neighboring keratinocytes and fibroblasts plays a role in melasma. Additionally, the effect of WIF-1 overexpression on the amelioration of hyperpigmentation was examined. WIF-1 knockdown, either in fibroblasts or in keratinocytes, significantly stimulated tyrosinase expression and melanosome transfer, whereas melanocytes with WIF-1 overexpression significantly reduced those parameters. The WIF-1 knockdown decreased glycogen synthase kinase-3β (GSK-3β), β-catenin, and NFATc2 (nuclear factor of activated T cells, cytoplasmic, calcineurin-dependent 2) phosphorylation and increased microphthalmia-associated transcription factor (MITF) expression as in melanocytes with Wnt-1 overexpression, whereas the WIF-1 overexpression reversed the results. Expression of Wnts, both canonical and noncanonical, was increased in the hyperpigmented skin of melasma patients. Collectively, WIF-1 downregulation, which may occur in epidermal keratinocytes and in dermal fibroblasts, is involved in melasma development because of the stimulation of melanogenesis and melanosome transfer through upregulation of the canonical and the noncanonical Wnt signaling pathway.

  2. Reducing CD73 expression by IL1β-Programmed Th17 cells improves immunotherapeutic control of tumors.

    PubMed

    Chatterjee, Shilpak; Thyagarajan, Krishnamurthy; Kesarwani, Pravin; Song, Jin H; Soloshchenko, Myroslawa; Fu, Jianing; Bailey, Stefanie R; Vasu, Chenthamarkshan; Kraft, Andrew S; Paulos, Chrystal M; Yu, Xue-Zhong; Mehrotra, Shikhar

    2014-11-01

    T cells of the T helper (Th)17 subset offer promise in adoptive T-cell therapy for cancer. However, current protocols for ex vivo programming of Th17 cells, which include TGFβ exposure, increase the expression of CD39 and CD73, two cell surface ATP ectonucleotidases that reduce T-cell effector functions and promote immunosuppression. Here, we report that ATP-mediated suppression of IFNγ production by Th17 cells can be overcome by genetic ablation of CD73 or by using IL1β instead of TGFβ to program Th17 cells ex vivo. Th17 cells cultured in IL1β were also highly polyfunctional, expressing high levels of effector molecules and exhibiting superior short-term control of melanoma in mice, despite reduced stem cell-like properties. TGFβ addition at low doses that did not upregulate CD73 expression but induced stemness properties drastically improved the antitumor effects of IL1β-cultured Th17 cells. Effector properties of IL1β-dependent Th17 cells were likely related to their high glycolytic capacity, since ex vivo programming in pyruvate impaired glycolysis and antitumor effects. Overall, we show that including TGFβ in ex vivo cultures used to program Th17 cells blunts their immunotherapeutic potential and demonstrate how this potential can be more fully realized for adoptive T-cell therapy.

  3. The MOC31PE immunotoxin reduces cell migration and induces gene expression and cell death in ovarian cancer cells

    PubMed Central

    2014-01-01

    Background The standard treatment of ovarian cancer with chemotherapy often leads to drug resistance and relapse of the disease, and the need for development of novel therapy alternatives is obvious. The MOC31PE immunotoxin binds to the cell surface antigen EpCAM, which is expressed by the majority of epithelial cancers including ovarian carcinomas, and we studied the cytotoxic effects of MOC31PE in ovarian cancer cells. Methods Investigation of the effects of MOC31PE treatment on protein synthesis, cell viability, proliferation and gene expression of the ovarian cancer cell lines B76 and HOC7. Results MOC31PE treatment for 24 h caused a dose-dependent reduction of protein synthesis with ID50 values of less than 10 ng/ml, followed by reduced cell viability. In a gene expression array monitoring the expression of 84 key genes in cancer pathways, 13 of the genes were differentially expressed by MOC31PE treatment in comparison to untreated cells. By combining MOC31PE and the immune suppressor cyclosporin A (CsA) the MOC31PE effect on protein synthesis inhibition and cell viability increased tenfold. Cell migration was also reduced, both in the individual MOC31PE and CsA treatment, but even more when combining MOC31PE and CsA. In tumor metastasis PCR arrays, 23 of 84 genes were differentially expressed comparing CsA versus MOC31PE + CsA treatment. Increased expression of the tumor suppressor KISS1 and the nuclear receptor NR4A3 was observed, and the differential candidate gene expression was confirmed in complementary qPCR analyses. For NR4A3 this was not accompanied by increased protein expression. However, a subcellular fractionation assay revealed increased mitochondrial NR4A3 in MOC31PE treated cells, suggesting a role for this protein in MOC31PE-induced apoptotic cell death. Conclusion The present study demonstrates that MOC31PE may become a new targeted therapy for ovarian cancer and that the MOC31PE anti-cancer effect is potentiated by CsA. PMID:24528603

  4. Expression and functional properties of the Streptococcus intermedius surface protein antigen I/II.

    PubMed

    Petersen, F C; Pasco, S; Ogier, J; Klein, J P; Assev, S; Scheie, A A

    2001-07-01

    Streptococcus intermedius is associated with deep-seated purulent infections. In this study, we investigated expression and functional activities of antigen I/II in S. intermedius. The S. intermedius antigen I/II appeared to be cell surface associated, with a molecular mass of approximately 160 kDa. Northern blotting indicated that the S. intermedius NCTC 11324 antigen I/II gene was transcribed as a monocistronic message. Maximum expression was seen during the early exponential phase. Insertional inactivation of the antigen I/II gene resulted in reduced hydrophobicity during early exponential phase, whereas no effect was detected during mid- and late exponential phases. Binding to human fibronectin and laminin was reduced in the isogenic mutant, whereas binding to human collagen types I and IV and to rat collagen type I was not significant for either the wild type or the mutant. Compared to the wild type, the capacity of the isogenic mutant to induce interleukin 8 (IL-8) release by THP-1 monocytic cells was significantly reduced. The results indicate that the S. intermedius antigen I/II is involved in adhesion to human receptors and in IL-8 induction.

  5. NR4A1 Knockdown Suppresses Seizure Activity by Regulating Surface Expression of NR2B.

    PubMed

    Zhang, Yanke; Chen, Guojun; Gao, Baobing; Li, Yunlin; Liang, Shuli; Wang, Xiaofei; Wang, Xuefeng; Zhu, Binglin

    2016-11-23

    Nuclear receptor subfamily 4 group A member 1 (NR4A1), a downstream target of CREB that is a key regulator of epileptogenesis, has been implicated in a variety of biological processes and was previously identified as a seizure-associated molecule. However, the relationship between NR4A1 and epileptogenesis remains unclear. Here, we showed that NR4A1 protein was predominantly expressed in neurons and up-regulated in patients with epilepsy as well as pilocarpine-induced mouse epileptic models. NR4A1 knockdown by lentivirus transfection (lenti-shNR4A1) alleviated seizure severity and prolonged onset latency in mouse models. Moreover, reciprocal coimmunoprecipitation of NR4A1 and NR2B demonstrated their interaction. Furthermore, the expression of p-NR2B (Tyr1472) in epileptic mice and the expression of NR2B in the postsynaptic density (PSD) were significantly reduced in the lenti-shNR4A1 group, indicating that NR4A1 knockdown partly decreased surface NR2B by promoting NR2B internalization. These results are the first to indicate that the expression of NR4A1 in epileptic brain tissues may provide new insights into the molecular mechanisms underlying epilepsy.

  6. NR4A1 Knockdown Suppresses Seizure Activity by Regulating Surface Expression of NR2B

    PubMed Central

    Zhang, Yanke; Chen, Guojun; Gao, Baobing; Li, Yunlin; Liang, Shuli; Wang, Xiaofei; Wang, Xuefeng; Zhu, Binglin

    2016-01-01

    Nuclear receptor subfamily 4 group A member 1 (NR4A1), a downstream target of CREB that is a key regulator of epileptogenesis, has been implicated in a variety of biological processes and was previously identified as a seizure-associated molecule. However, the relationship between NR4A1 and epileptogenesis remains unclear. Here, we showed that NR4A1 protein was predominantly expressed in neurons and up-regulated in patients with epilepsy as well as pilocarpine-induced mouse epileptic models. NR4A1 knockdown by lentivirus transfection (lenti-shNR4A1) alleviated seizure severity and prolonged onset latency in mouse models. Moreover, reciprocal coimmunoprecipitation of NR4A1 and NR2B demonstrated their interaction. Furthermore, the expression of p-NR2B (Tyr1472) in epileptic mice and the expression of NR2B in the postsynaptic density (PSD) were significantly reduced in the lenti-shNR4A1 group, indicating that NR4A1 knockdown partly decreased surface NR2B by promoting NR2B internalization. These results are the first to indicate that the expression of NR4A1 in epileptic brain tissues may provide new insights into the molecular mechanisms underlying epilepsy. PMID:27876882

  7. Candida krusei and Candida glabrata reduce the filamentation of Candida albicans by downregulating expression of HWP1 gene.

    PubMed

    de Barros, Patrícia Pimentel; Freire, Fernanda; Rossoni, Rodnei Dennis; Junqueira, Juliana Campos; Jorge, Antonio Olavo Cardoso

    2017-07-01

    Pathogenicity of Candida albicans is associated with its capacity switch from yeast-like to hyphal growth. The hyphal form is capable to penetrate the epithelial surfaces and to damage the host tissues. Therefore, many investigations have focused on mechanisms that control the morphological transitions of C. albicans. Recently, certain studies have showed that non-albicans Candida species can reduce the capacity of C. albicans to form biofilms and to develop candidiasis in animal models. Then, the objective of this study was to evaluate the effects of Candida krusei and Candida glabrata on the morphogenesis of C. albicans. Firstly, the capacity of reference and clinical strains of C. albicans in forming hyphae was tested in vitro. After that, the expression of HWP1 (hyphal wall protein 1) gene was determined by quantitative real-time PCR (polymerase chain reaction) assay. For both reference and clinical strains, a significant inhibition of the hyphae formation was observed when C. albicans was incubated in the presence of C. krusei or C. glabrata compared to the control group composed only by C. albicans. In addition, the culture mixed of C. albicans-C. krusei or C. albicans-C. glabrata reduced significantly the expression of HWP1 gene of C. albicans in relation to single cultures of this specie. In both filamentation and gene expression assays, C. krusei showed the higher inhibitory activity on the morphogenesis of C. albicans compared to C. glabrata. C. krusei and C. glabrata are capable to reduce the filamentation of C. albicans and consequently decrease the expression of the HWP1 gene.

  8. A novel method for monitoring the cell surface expression of heteromeric protein complexes in dispersed neurons and acute hippocampal slices

    PubMed Central

    Holman, David; Henley, Jeremy M.

    2012-01-01

    The subunit composition of multimeric protein complexes is critical in determining their trafficking and functional properties. Despite there being multiple techniques to investigate the trafficking events of individual subunits there are currently limited means to monitor the trafficking properties of heteromeric protein complexes. Here, we combine surface biotinylation with co-immunoprecipitation to monitor the cell surface expression of native, heteromeric AMPA receptor complexes. Using this method, we demonstrate that the surface levels of GluR1/2 and GluR2/3 complexes are reduced following NMDA-evoked long-term depression (NMDA-LTD) in acute hippocampal slices. Finally, we discuss how this method can be adapted to monitor the cell surface expression of other heteromeric protein complexes. PMID:17083981

  9. Carbohydrate Coating Reduces Adhesion of Biofilm-Forming Bacillus subtilis to Gold Surfaces

    PubMed Central

    Kesel, S.; Mader, A.; Seeberger, P. H.; Lieleg, O.

    2014-01-01

    The growth of bacterial biofilms in pipes and food tanks causes severe problems in industry. Biofilms growing on medical implants or catheters are of great concern, as they can cause serious infections and decrease the functionality of the medical device. The prevention of bacterial adhesion—the first step in colonization and biofilm formation—is therefore very important. Current research comprises alterations in surface properties, the prevention of adhesin biosynthesis, inhibition with receptor analogs, or the development of anti-adhesive vaccines. We present a new approach that allows us to study bacterial adhesion with high sensitivity in real-time while testing several different surfaces in parallel. Using the cantilever-array technique we demonstrate that coating of gold surfaces with mono- or disaccharides results in a reduction of the bacterial adhesion of the biofilm-forming bacterium Bacillus subtilis NCIB 3610 to these gold surfaces. This reduction in bacterial adhesion is independent of the studied carbohydrate. Using several mutant strains, we investigate the underlying molecular interactions, and our results suggest that adhesion to gold surfaces is mediated by thiol groups present in proteins of the bacterial cell membrane or biofilm matrix proteins expressed at low levels by the wild-type strain. Furthermore, our data indicate that the adhesion of B. subtilis NCIB 3610 to carbohydrate-coated gold surfaces is facilitated by interactions between carbohydrates installed on the cantilever gold surface and an exopolysaccharide expressed by this strain. Understanding general and specific contributions of molecular interactions mediating bacterial adhesion will enable its prevention in the future. PMID:25038098

  10. Impact of gypsum applied to grass buffer strips on reducing soluble p in surface water runoff.

    PubMed

    Watts, D B; Torbert, H A

    2009-01-01

    The threat of P transport from land applied manure has resulted in water quality concerns. Research was conducted to evaluate gypsum as a soil amendment applied to grass buffer strips for reducing soluble P in surface runoff. A simulated concentrated flow was created in an established tall fescue (Festuca arundinacea Schreb.) pasture. Poultry litter (PL) was applied at a rate of 250 kg N ha(-1) to the upper 3.05 m of each plot, while gypsum was applied at rates of 0, 1, 3.2, and 5.6 Mg ha-1to the lower 1.52 m of the plot functioning as a grass buffer strip. Two 30-min runoff events ( approximately 4 L min(-1)) were conducted, immediately after PL application and 4 wk later to determined soluble P concentration in the surface water samples. The greatest concentration of soluble P was in the runoff event occurring immediately after the PL application. Gypsum applied to grass buffer strips was effective in reducing soluble P concentrations (32-40%) in surface runoff, while the untreated buffer strip was somewhat effective in reducing soluble P (18%). No significant differences were observed between gypsum rates, suggesting that land managers would achieve the greatest benefit from the lowest application rate (1Mgha(-1)). In the second runoff event, although concentrations of soluble P in the surface water runoff were greatly reduced, the effect of gypsum had disappeared. Thus, these results show that gypsum is most effective in reducing the initial P losses from PL application when applied to grass buffer strips. The information obtained from this study may be useful in aiding land managers in developing management practices that reduce soluble P loss at the edge of a field.

  11. Reduced Fhit expression in sporadic and BRCA2-linked breast carcinomas.

    PubMed

    Ingvarsson, S; Agnarsson, B A; Sigbjornsdottir, B I; Kononen, J; Kallioniemi, O P; Barkardottir, R B; Kovatich, A J; Schwarting, R; Hauck, W W; Huebner, K; McCue, P A

    1999-06-01

    Evidence for alteration of the FHIT gene in a significant fraction of breast carcinomas has been reported, in apparent concordance with loss of heterozygosity (LOH) at chromosome region 3p14.2 in breast cancer and benign proliferative breast disease. A significantly higher frequency of LOH at the FHIT locus was reported for BRCA2-/- tumors, possibly due to misrepaired double-strand breaks at this common fragile region. To determine whether such genomic alterations lead to Fhit inactivation, we have assessed the level of Fhit expression by immunohistochemical detection in sporadic tumors and cancers occurring in BRCA2 999del5 carriers. To determine whether Fhit inactivation may have prognostic significance, we have also assessed expression of breast cancer markers and clinical features in sporadic tumors relative to Fhit expression. Of 40 consecutive sporadic breast carcinomas studied for tumor markers, 50% showed reduced Fhit expression. In these sporadic cancers, loss of Fhit expression was not correlated significantly with the presence or absence of other tumor markers. In a study of 58 sporadic and 34 BRCA2 999del5 Icelandic invasive cancers, there was a significant association of LOH at 3p14.2 with reduced expression of Fhit (P = 0.001); also the lower expression of Fhit and higher LOH at 3p14.2 in BRCA2 999del5 tumors relative to sporadic cancers was significant (P = 0.002). Thus, genetic alteration at the fragile site within the FHIT gene leads to loss of Fhit protein in a significant fraction of sporadic breast cancers and a much larger fraction of familial breast cancers with an inherited BRCA2 mutation, consistent with the idea that loss of BRCA2 function affects stability of the FHIT/FRA3B locus.

  12. Betaine reduces the expression of inflammatory adipokines caused by hypoxia in human adipocytes.

    PubMed

    Olli, K; Lahtinen, S; Rautonen, N; Tiihonen, K

    2013-01-14

    Obesity is characterised by a state of chronic low-grade inflammation and the elevated circulating and tissue levels of inflammatory markers, including inflammation-related adipokines, released from white adipose tissue. The expression and release of these adipokines generally rises as the adipose tissue expands and hypoxic conditions start to develop within the tissue. Here, the effect of betaine, a trimethylglycine having a biological role as an osmolyte and a methyl donor, on the expression of inflammation-related markers was tested in human adipocytes under hypoxia. Differentiated adipocytes were cultivated under low (1 %) oxygen tension for 8-20 h. The expression of different adipokines, including IL-6, leptin, PPARγ, TNF-α and adiponectin, was measured by quantitative PCR by determining the relative mRNA level from the adipocytes. Hypoxia, in general, led to a decrease in the expression of PPARγ mRNA in human adipocytes, whereas the expression levels of leptin and IL-6 mRNA were substantially increased by hypoxia. The cultivation of adipocytes under hypoxia also led to a reduction in the expression of TNF-α mRNA. The results showed that hypoxia increased the relative quantification of leptin gene transcription, and that betaine (250 μmol/l) reduced this effect, caused by low oxygen conditions. Under hypoxia, betaine also reduced the mRNA level of the pro-inflammatory markers IL-6 and TNF-α. These results demonstrate that the extensive changes in the expression of inflammation-related adipokines in human adipocytes caused by hypoxia can be diminished by the presence of physiologically relevant concentrations of betaine.

  13. Biodentine Reduces Tumor Necrosis Factor Alpha-induced TRPA1 Expression in Odontoblastlike Cells.

    PubMed

    El Karim, Ikhlas A; McCrudden, Maelíosa T C; McGahon, Mary K; Curtis, Tim M; Jeanneau, Charlotte; Giraud, Thomas; Irwin, Chris R; Linden, Gerard J; Lundy, Fionnuala T; About, Imad

    2016-04-01

    The transient receptor potential (TRP) ion channels have emerged as important cellular sensors in both neuronal and non-neuronal cells, with TRPA1 playing a central role in nociception and neurogenic inflammation. The functionality of TRP channels has been shown to be modulated by inflammatory cytokines. The aim of this study was to investigate the effect of inflammation on odontoblast TRPA1 expression and to determine the effect of Biodentine (Septodent, Paris, France) on inflammatory-induced TRPA1 expression. Immunohistochemistry was used to study TRPA1 expression in pulp tissue from healthy and carious human teeth. Pulp cells were differentiated to odontoblastlike cells in the presence of 2 mmol/L beta-glycerophosphate, and these cells were used in quantitative polymerase chain reaction, Western blotting, calcium imaging, and patch clamp studies. Immunofluorescent staining revealed TRPA1 expression in odontoblast cell bodies and odontoblast processes, which was more intense in carious versus healthy teeth. TRPA1 gene expression was induced in cultured odontoblastlike cells by tumor necrosis factor alpha, and this expression was significantly reduced in the presence of Biodentine. The functionality of the TRPA1 channel was shown by calcium microfluorimetry and patch clamp recording, and our results showed a significant reduction in tumor necrosis factor alpha-induced TRPA1 responses after Biodentine treatment. In conclusion, this study showed TRPA1 to be modulated by caries-induced inflammation and that Biodentine reduced TRPA1 expression and functional responses. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  14. Reduced retinoids and retinoid receptors' expression in pancreatic cancer: A link to patient survival.

    PubMed

    Bleul, Tim; Rühl, Ralph; Bulashevska, Svetlana; Karakhanova, Svetlana; Werner, Jens; Bazhin, Alexandr V

    2015-09-01

    Pancreatic ductal adenocarcinoma (PDAC) represents one of the deadliest cancers in the world. All-trans retinoic acid (ATRA) is the major physiologically active form of vitamin A, regulating expression of many genes. Disturbances of vitamin A metabolism are prevalent in some cancer cells. The main aim of this work was to investigate deeply the components of retinoid signaling in PDAC compared to in the normal pancreas and to prove the clinical importance of retinoid receptor expression. For the study, human tumor tissues obtained from PDAC patients and murine tumors from the orthotopic Panc02 model were used for the analysis of retinoids, using high performance liquid chromatography mass spectrometry and real-time RT-PCR gene expression analysis. Survival probabilities in univariate analysis were estimated using the Kaplan-Meier method and the Cox proportional hazards model was used for the multivariate analysis. In this work, we showed for the first time that the ATRA and all-trans retinol concentration is reduced in PDAC tissue compared to their normal counterparts. The expression of RARα and β as well as RXRα and β are down-regulated in PDAC tissue. This reduced expression of retinoid receptors correlates with the expression of some markers of differentiation and epithelial-to-mesenchymal transition as well as of cancer stem cell markers. Importantly, the expression of RARα and RXRβ is associated with better overall survival of PDAC patients. Thus, reduction of retinoids and their receptors is an important feature of PDAC and is associated with worse patient survival outcomes.

  15. Klotho expression is reduced in COPD airway epithelial cells: effects on inflammation and oxidant injury.

    PubMed

    Gao, Wei; Yuan, Cheng; Zhang, Jingying; Li, Lingling; Yu, Like; Wiegman, Coen H; Barnes, Peter J; Adcock, Ian M; Huang, Mao; Yao, Xin

    2015-12-01

    COPD (chronic obstructive pulmonary disease) is associated with sustained inflammation, excessive injury, and accelerated lung aging. Human Klotho (KL) is an anti-aging protein that protects cells against inflammation and damage. In the present study, we quantified KL expression in the lungs of COPD patients and in an ozone-induced mouse model of COPD, and investigated the mechanisms that control KL expression and function in the airways. KL distribution and levels in human and mouse airways were measured by immunohistochemistry and Western blotting. The effect of CSE (cigarette smoke extract) on KL expression was detected in human bronchial epithelial cells. Moreover, the effect of KL on CSE-mediated inflammation and hydrogen peroxide-induced cellular injury/apoptosis was determined using siRNAs. KL expression was decreased in the lungs of smokers and further reduced in patients with COPD. Similarly, 6 weeks of exposure to ozone decreased KL levels in airway epithelial cells. CSE and TNFα (tumour necrosis factor α) decreased KL expression and release from airway epithelial cells, which was associated with enhanced pro-inflammatory cytokine expression. Moreover, KL depletion increased cell sensitivity to cigarette smoke-induced inflammation and oxidative stress-induced cell damage. These effects involved the NF-κB (nuclear factor κB), MAPK (mitogen-activated protein kinase) and Nrf2 (nuclear factor erythroid 2-related factor 2) pathways. Reduced KL expression in COPD airway epithelial cells was associated with increased oxidative stress, inflammation and apoptosis. These data provide new insights into the mechanisms associated with the accelerated lung aging in COPD development.

  16. Klotho expression is reduced in COPD airway epithelial cells: effects on inflammation and oxidant injury

    PubMed Central

    Gao, Wei; Yuan, Cheng; Zhang, Jingying; Li, Lingling; Yu, Like; Wiegman, Coen H.; Barnes, Peter J.; Adcock, Ian M.; Huang, Mao

    2015-01-01

    COPD (chronic obstructive pulmonary disease) is associated with sustained inflammation, excessive injury, and accelerated lung aging. Human Klotho (KL) is an anti-aging protein that protects cells against inflammation and damage. In the present study, we quantified KL expression in the lungs of COPD patients and in an ozone-induced mouse model of COPD, and investigated the mechanisms that control KL expression and function in the airways. KL distribution and levels in human and mouse airways were measured by immunohistochemistry and Western blotting. The effect of CSE (cigarette smoke extract) on KL expression was detected in human bronchial epithelial cells. Moreover, the effect of KL on CSE-mediated inflammation and hydrogen peroxide-induced cellular injury/apoptosis was determined using siRNAs. KL expression was decreased in the lungs of smokers and further reduced in patients with COPD. Similarly, 6 weeks of exposure to ozone decreased KL levels in airway epithelial cells. CSE and TNFα (tumour necrosis factor α) decreased KL expression and release from airway epithelial cells, which was associated with enhanced pro-inflammatory cytokine expression. Moreover, KL depletion increased cell sensitivity to cigarette smoke-induced inflammation and oxidative stress-induced cell damage. These effects involved the NF-κB (nuclear factor κB), MAPK (mitogen-activated protein kinase) and Nrf2 (nuclear factor erythroid 2-related factor 2) pathways. Reduced KL expression in COPD airway epithelial cells was associated with increased oxidative stress, inflammation and apoptosis. These data provide new insights into the mechanisms associated with the accelerated lung aging in COPD development. PMID:26201096

  17. Endothelial P-selectin expression is reduced in advanced primary melanoma and melanoma metastasis.

    PubMed Central

    Nooijen, P. T.; Westphal, J. R.; Eggermont, A. M.; Schalkwijk, C.; Max, R.; de Waal, R. M.; Ruiter, D. J.

    1998-01-01

    Some malignant tumors induce a cellular immune response that results in the formation of an inflammatory infiltrate and subsequent tumor regression. The infiltrating leukocytes extravasate from the bloodstream after binding to adhesion receptors on the surface of the endothelium. One of these receptors is the P-selectin molecule (CD62P) that is constitutively present on normal capillaries. We observed that P-selectin expression is absent from the microvasculature in advanced primary melanoma and in melanoma metastasis in contrast to benign melanocytic lesions where P-selectin expression was identical to that in normal skin. We suggest that one of the mechanisms by which advanced melanoma lesions evade inflammatory regression operates via a decrease of endothelial P-selectin expression. Images Figure 1 PMID:9502409

  18. Murine pancreatic adenocarcinoma reduces Ikaros expression and disrupts T cell homeostasis.

    PubMed

    Nelson, Nadine; Xiang, Shengyan; Zhang, Xiaohong; Gilvary, Danielle; Djeu, Julie; Husain, Kazim; Malafa, Mokenge; Vohra, Nasreen; Pilon-Thomas, Shari; Ghansah, Tomar

    2015-01-01

    Maintenance of T cell immune homeostasis is critical for adequate anti-tumor immunity. The transcription factor Ikaros is essential for lymphocyte development including T cells. Alterations in Ikaros expression occur in blood malignancies in humans and mice. In this study, we investigated the role of Ikaros in regulating T cell immune balance in pancreatic cancer mouse models. Using our Panc02 tumor-bearing (TB) mouse model, western blot analysis revealed a reduction in Ikaros proteins while qRT-PCR showed no differences in Ikaros mRNA levels in TB splenocytes compared to control. Treatment of naïve splenocytes with the proteasomal inhibitor, MG132, stabilized Ikaros expression and prevented Ikaros downregulation by Panc02 cells, in vitro. Western blot analyses showed a reduction in protein phosphatase 1 (PP1) and protein kinase CK2 expression in TB splenocytes while CK2 activity was increased. Immunofluorescence microscopy revealed altered punctate staining of Ikaros in TB splenocytes. Flow cytometry revealed a significant decrease in effector CD4+ and CD8+ T cell percentages but increased CD4+CD25+ regulatory T cells in TB splenocytes. Similar alterations in T cell percentages, as well as reduced Ikaros and CK2 but not PP1 expression, were observed in a transgenic, triple mutant (TrM) pancreatic cancer model. Ikaros expression was also reduced in enriched TB CD3+ T cells. MG132 treatment of naïve CD3+ T cells stabilized Ikaros expression in the presence of Panc02 cells. Western blots showed reduced PP1 and CK2 expression in TB CD3+ T cells. The results of this study suggest that the pancreatic tumor microenvironment may cause proteasomal degradation of Ikaros, possibly via dysregulation of PP1 and CK2 expression and activity, respectively. This loss of Ikaros expression may contribute to an imbalance in T cell percentages. Ikaros may potentially be a therapeutic target to restore T cell homeostasis in pancreatic cancer hosts, which may be critical for effective

  19. Brain SERT Expression of Male Rats Is Reduced by Aging and Increased by Testosterone Restitution

    PubMed Central

    Herrera-Pérez, José Jaime; Fernández-Guasti, Alonso; Martínez-Mota, Lucía

    2013-01-01

    In preclinical and clinical studies aging has been associated with a deteriorated response to antidepressant treatment. We hypothesize that such impairment is explained by an age-related decrease in brain serotonin transporter (SERT) expression associated with low testosterone (T) levels. The objectives of this study were to establish (1) if brain SERT expression is reduced by aging and (2) if the SERT expression in middle-aged rats is increased by T-restitution. Intact young rats (3–5 months) and gonad-intact middle-aged rats with or without T-restitution were used. The identification of the brain SERT expression was done by immunofluorescence in prefrontal cortex, lateral septum, hippocampus, and raphe nuclei. An age-dependent reduction of SERT expression was observed in all brain regions examined, while T-restitution recovered the SERT expression only in the dorsal raphe of middle-aged rats. This last action seems relevant since dorsal raphe plays an important role in the antidepressant action of selective serotonin reuptake inhibitors. All data suggest that this mechanism accounts for the T-replacement usefulness to improve the response to antidepressants in the aged population. PMID:26317087

  20. Systemic RNAi Delivery to the Muscles of ROSA26 Mice Reduces lacZ Expression

    PubMed Central

    Wei, Jessica; Chamberlain, Joel R.

    2014-01-01

    RNAi has potential for therapeutically downregulating the expression of dominantly inherited genes in a variety of human genetic disorders. Here we used the ROSA26 mouse, which constitutively expresses the bacterial lacZ gene in tissues body wide, as a model to test the ability to downregulate gene expression in striated muscles. Recombinant adeno-associated viral vectors (rAAVs) were generated that express short hairpin RNAs (shRNAs) able to target the lacZ mRNA. Systemic delivery of these rAAV6 vectors led to a decrease of β-galactosidase expression of 30–50-fold in the striated muscles of ROSA26 mice. However, high doses of vectors expressing 21 nucleotide shRNA sequences were associated with significant toxicity in both liver and cardiac muscle. This toxicity was reduced in cardiac muscle using lower vector doses. Furthermore, improved knockdown in the absence of toxicity was obtained by using a shorter (19 nucleotide) shRNA guide sequence. These results support the possibility of using rAAV vectors to deliver RNAi sequences systemically to treat dominantly inherited disorders of striated muscle. PMID:25127128

  1. Asymptotic expressions for the surface currents induced on a cylindrically curved impedance strip

    NASA Astrophysics Data System (ADS)

    Akduman, Ibrahim; Buyukaksoy, Alinur

    1995-05-01

    The present paper is concerned with the derivation of the electric and magnetic surface currents induced on a cylindrically curved impedance strip. By considering the locality of the high-frequency diffraction phenomena the physical (- pi, pi) interval for the usual cylindrical polar angle is replaced by an abstract infinite interval (- infinity, infinity) whereby the related mixed boundary value problem is formulated as a 'modified matrix Hilbert' problem. By using the Debye approximations for the Hankel and Bessel functions involved, the modified matrix Hilbert problem is first decoupled and then reduced to two pairs of simultaneous Fredholm integral equations of the second kind which are solved by iterations. The explicit expressions for the electric and magnetic surface current components attributable to the reflection, edge or surface diffractions of the incident field as well as to the edge reflections of these components themselves are obtained by evaluating the current integrals asymptotically. The results derived in this paper constitute also a rigorous proof for a conjecture made by Idemen on the reflections of the surface currents at the edges.

  2. Atorvastatin suppresses glioma invasion and migration by reducing microglial MT1-MMP expression.

    PubMed

    Yongjun, Yi; Shuyun, Huang; Lei, Chen; Xiangrong, Chen; Zhilin, Yang; Yiquan, Ke

    2013-07-15

    Microglia, the immune cells of the brain, often present in large numbers in gliomas, where they promote tumor growth and invasiveness. This study found that atorvastatin reduced the pro-tumorigenic effects of microglia on glioma migration and invasion by reducing the microglial expression of membrane type 1 metalloproteinase (MT1-MMP). The results suggest that down-regulation of MT1-MMP is controlled by a p38 MAPK pathway in microglia. Taken together, the results support further research on atorvastatin as a candidate for glioma therapy by targeting microglia.

  3. Elimination of Kalrn Expression in POMC Cells Reduces Anxiety-Like Behavior and Contextual Fear Learning

    PubMed Central

    Mandela, Prashant; Yan, Yan; LaRese, Taylor; Eipper, Betty A.; Mains, Richard E.

    2014-01-01

    Kalirin, a Rho GDP/GTP exchange factor for Rac1 and RhoG, is known to play an essential role in the formation and maintenance of excitatory synapses and in the secretion of neuropeptides. Mice unable to express any of the isoforms of Kalrn in cells that produce POMC at any time during development (POMC cells) exhibited reduced anxiety-like behavior and reduced acquisition of passive avoidance behavior, along with sex-specific alteration in the corticosterone response to restraint stress. Strikingly, lack of Kalrn expression in POMC cells closely mimicked the effects of global Kalrn knockout on anxiety-like behavior and passive avoidance conditioning without causing the other deficits noted in Kalrn knockout mice. Our data suggest that deficits in excitatory inputs onto POMC neurons are responsible for the behavioral phenotypes observed. PMID:25014196

  4. Reduced GABAA receptor α6 expression in the trigeminal ganglion alters inflammatory TMJ hypersensitivity

    PubMed Central

    Puri, Jyoti; Vinothini, Priya; Reuben, Jayne; Bellinger, Larry L.; Ailing, Li; Peng, Yuan B.; Kramer, Phillip R.

    2012-01-01

    Trigeminal ganglia neurons express the GABAA receptor subunit alpha 6 (Gabrα6) but the role of this particular subunit in orofacial hypersensitivity is unknown. In this report the function of Gabrα6 was tested by reducing its expression in the trigeminal ganglia and measuring the effect of this reduction on inflammatory temporomandibular joint (TMJ) hypersensitivity. Gabrα6 expression was reduced by infusing the trigeminal ganglia of male Sprague Dawley rats with small interfering RNA (siRNA) having homology to either the Gabrα6 gene (Gabrα6 siRNA) or no known gene (control siRNA). Sixty hours after siRNA infusion the rats received a bilateral TMJ injection of complete Freund’s adjuvant to induce an inflammatory response. Hypersensitivity was then quantitated by measuring meal duration, which lengthens when hypersensitivity increases. Neuronal activity in the trigeminal ganglia was also measured by quantitating the amount of phosphorylated ERK. Rats in a different group that did not have TMJ inflammation had an electrode placed in the spinal cord at the level of C1 sixty hours after siRNA infusion to record extracellular electrical activity of neurons that responded to TMJ stimulation. Our results show that Gabrα6 was expressed in both neurons and satellite glia of the trigeminal ganglia and that Gabrα6 positive neurons within the trigeminal ganglia have afferents in the TMJ. Gabrα6 siRNA infusion reduced Gabrα6 gene expression by 30% and significantly lengthened meal duration in rats with TMJ inflammation. Gabrα6 siRNA infusion also significantly increased p-ERK expression in the trigeminal ganglia of rats with TMJ inflammation and increased electrical activity in the spinal cord of rats without TMJ inflammation. These results suggest that maintaining Gabrα6 expression was necessary to inhibit primary sensory afferents in the trigeminal pathway and reduce inflammatory orofacial nociception. PMID:22521829

  5. Investigation of water washes suitable for very small meat plants to reduce pathogens on beef surfaces.

    PubMed

    Yoder, Sally Flowers; Henning, William R; Mills, Edward W; Doores, Stephanie; Ostiguy, Nancy; Cutter, Catherine N

    2010-05-01

    Water washing with a handheld hose was performed on beef surfaces to ascertain the most effective combination of methods needed to remove potentially harmful microorganisms. For these experiments, beef brisket surfaces were experimentally inoculated with a fecal slurry containing Escherichia coli O157:H7, Salmonella Typhimurium, Campylobacter coli, and Campylobacter jejuni. In a pilot study, surfaces were washed with cold water (15 degrees C) at various water pressures, spray distances, application times, and drip times, and remaining bacterial populations were determined following the enumeration and isolation of pathogens and naturally occurring hygiene indicators (mesophilic aerobic bacteria, coliforms, and E. coli). The most efficacious combinations of these washing conditions were applied subsequently to artificially contaminated beef brisket surfaces in conjunction with hot (77 degrees C), warm (54 degrees C), and additional cold (15 degrees C) water washes. In the cold water washing pilot study, combinations of physical washing conditions significantly reduced all bacterial populations (P < 0.05). Further studies clearly indicated the superior bactericidal effectiveness of hot water washing; E. coli O157:H7 and Salmonella Typhimurium were reduced by 3.8 and 4.1 log CFU/cm(2), respectively. Overall, higher water temperature, longer application times, and shorter spray distances more effectively removed pathogens from inoculated beef surfaces. These findings will be used to formulate water washing recommendations for very small meat processing establishments.

  6. MD2 expression is reduced in large airways of smokers and COPD smokers.

    PubMed

    Pace, Elisabetta; Ferraro, Maria; Chiappara, Giuseppina; Vitulo, Patrizio; Pipitone, Loredana; Di Vincenzo, Serena; Gjomarkaj, Mark

    2015-09-01

    Toll-like receptor 4 (TLR4) signaling requires a number of accessory proteins to initiate a signal. MD-2 is one of the accessory proteins with a relevant role in lipopolysaccharide responses. Although cigarette smoke increases TLR4 expression, TLR4 signaling is altered in smokers and in smokers COPD patients. The main aims of this study were to explore whether MD2 is altered in large and small airways of COPD and of smokers without COPD. The expression of MD2 ex vivo was assessed by immunohistochemistry in surgical specimens from current smokers COPD (s-COPD; n = 14), smokers without COPD (S; n = 7), and from non-smoker non-COPD subjects (C; n = 11. The in vitro effects of cigarette smoke extracts on the MD2 expression in a human bronchial epithelial cell line (16-HBE) were also assessed by flow cytometry. MD2 is reduced in the epithelium and in the submucosa in large airways but not in the epithelium and in the submucosa in small airways of smokers and of s-COPD. The expression of MD2 in the submucosa of the large airways is significantly higher in comparison to the submucosa of the small airways in all the studied groups. In vitro, cigarette smoke is able to increase TLR4 but it reduces MD2 in a dose-dependent manner in bronchial epithelial cells. Cigarette smoke may alter innate immune responses reducing the expression of the MD2, a molecule with an important role in TLR4 signaling.

  7. Reduced mycorrhizal colonization (rmc) tomato mutant lacks expression of SymRK signaling pathway genes

    PubMed Central

    Nair, Aswathy; Bhargava, Sujata

    2012-01-01

    Comparison of the expression of 13 genes involved in arbuscular mycorrhizal (AM) symbiosis was performed in a wild type tomato (Solanum lycopersicum cv 76R) and its reduced mycorrhizal colonization mutant rmc in response to colonization with Glomus fasiculatum. Four defense-related genes were induced to a similar extent in the mutant and wild type AM colonized plants, indicating a systemic response to AM colonization. Genes related to nutrient exchange between the symbiont partners showed higher expression in the AM roots of wild type plants than the mutant plants, which correlated with their arbuscular frequency. A symbiosis receptor kinase that is involved in both nodulation and AM symbiosis was not expressed in the rmc mutant. The fact that some colonization was observed in rmc was suggestive of the existence of an alternate colonization signaling pathway for AM symbiosis in this mutant. PMID:23221680

  8. Nandrolone reduces activation of Notch signaling in denervated muscle associated with increased Numb expression.

    PubMed

    Liu, Xin-Hua; Yao, Shen; Qiao, Rui-Fang; Levine, Alice C; Kirschenbaum, Alexander; Pan, Jiangping; Wu, Yong; Qin, Weiping; Bauman, William A; Cardozo, Christopher P

    2011-10-14

    Nandrolone, an anabolic steroid, slows denervation-atrophy in rat muscle. The molecular mechanisms responsible for this effect are not well understood. Androgens and anabolic steroids activate Notch signaling in animal models of aging and thereby mitigate sarcopenia. To explore the molecular mechanisms by which nandrolone prevents denervation-atrophy, we investigated the effects of nandrolone on Notch signaling in denervated rat gastrocnemius muscle. Denervation significantly increased Notch activity reflected by elevated levels of nuclear Notch intracellular domain (NICD) and expression of Hey1 (a Notch target gene). Activation was greatest at 7 and 35 days after denervation but remained present at 56 days after denervation. Activation of Notch in denervated muscle was prevented by nandrolone associated with upregulated expression of Numb mRNA and protein. These data demonstrate that denervation activates Notch signaling, and that nandrolone abrogates this response associated with increased expression of Numb, suggesting a potential mechanism by which nandrolone reduces denervation-atrophy.

  9. Interleukin-18 deficiency reduces neuropeptide gene expressions in the mouse amygdala related with behavioral change.

    PubMed

    Yamamoto, Yuta; Tanahashi, Toshihito; Katsuura, Sakurako; Kurokawa, Ken; Nishida, Kensei; Kuwano, Yuki; Kawai, Tomoko; Teshima-Kondo, Shigetada; Chikahisa, Sachiko; Tsuruo, Yoshihiro; Sei, Hiroyoshi; Rokutan, Kazuhito

    2010-12-15

    In this study, we examined the effects of IL-18 deficiency on behaviors and gene expression profiles in 6 brain regions. IL-18(-/-) mice reduced depressive-like behavior and changed gene expressions predominantly in the amygdala compared with wild-type mice. Pathway analysis of the differentially expressed genes ranked behavior as the top-scored biological function. Of note, the absence of IL-18 decreased Avp, Hcrt, Oxt, and Pmch mRNA levels and the number of arginine vasopressin- and oxytocin-positive cells in the amygdala, but not in the hypothalamus. Our results suggest that IL-18-dependent vasopressinergic and oxytocinergic circuitry in the amygdala may regulate depressive-like behaviors in mice.

  10. Contactin-1 reduces E-cadherin expression via activating AKT in lung cancer.

    PubMed

    Yan, Judy; Wong, Nicholas; Hung, Claudia; Chen, Wendy Xin-Yi; Tang, Damu

    2013-01-01

    Contactin-1 has been shown to promote cancer metastasis. However, the underlying mechanisms remain unclear. We report here that knockdown of contactin-1 in A549 lung cancer cells reduced A549 cell invasion and the cell's ability to grow in soft agar without affecting cell proliferation. Reduction of contactin-1 resulted in upregulation of E-cadherin, consistent with E-cadherin being inhibitive of cancer cell invasion. In an effort to investigate the mechanisms whereby contactin-1 reduces E-cadherin expression, we observed that contactin-1 plays a role in AKT activation, as knockdown of contactin-1 attenuated AKT activation. Additionally, inhibition of AKT activation significantly enhanced E-cadherin expression, an observation that mimics the situation observed in contactin-1 knockdown, suggesting that activation of AKT plays a role in contactin-1-mediated downregulation of E-cadherin. In addition, we were able to show that knockdown of contactin-1 did not further reduce A549 cell's invasion ability, when AKT activation was inhibited by an AKT inhibitor. To further support our findings, we overexpressed CNTN-1 in two CNTN-1 null breast cancer cell lines expressing E-cadherin. Upon overexpression, CNTN-1 reduced E-cadherin levels in one cell line and increased AKT activation in the other. Furthermore, in our study of 63 primary lung cancers, we observed 65% of primary lung cancers being contactin-1 positive and in these carcinomas, 61% were E-cadherin negative. Collectively, we provide evidence that contactin-1 plays a role in the downregulation of E-cadherin in lung cancer and that AKT activation contributes to this process. In a study of mechanisms responsible for contactin-1 to activate AKT, we demonstrated that knockdown of CNTN-1 in A549 cells did not enhance PTEN expression but upregulated PHLPP2, a phosphatase that dephosphorylates AKT. These observations thus suggest that contactin-1 enhances AKT activation in part by preventing PHLPP2-mediated AKT

  11. Expression of a retroposon-like sequence upstream of the putative Trypanosoma brucei variant surface glycoprotein gene expression site promoter.

    PubMed Central

    Lodes, M J; Smiley, B L; Stadnyk, A W; Bennett, J L; Myler, P J; Stuart, K

    1993-01-01

    We have cloned the region spanning the putative promoter from two variant surface glycoprotein gene expression sites that are at each end of chromosome M4 of Trypanosoma brucei IsTat 7. Both expression sites contain a retroposon-like sequence (ESR) pseudogene whose 3' end is approximately 30 bp upstream of the putative expression site promoter. The ESRs from both expression sites share considerable sequence homology and are related to LINE-like elements, especially the T. brucei ingi retroposon. Other ESRs are located on large, but not intermediate or mini-, chromosomes in the IsTaR 1 serodeme, and the total copy number is 10 to 20, similar to that estimated for variant surface glycoprotein expression sites. No DNA rearrangements in the vicinity of the ESR and putative expression site promoter were detected following antigenic switches in the IsTaR 1 serodeme. ESR transcripts are present in bloodstream, but not procyclic, forms. Variation in transcript size and sequence between bloodstream variant antigenic types implies that only the ESR from the active expression site is transcribed. This pattern of expression reflects that of sequences downstream of the putative expression site promoter, suggesting that the region of coordinately controlled expression extends upstream of this promoter. Images PMID:8413293

  12. Increased Serotonin Transporter Expression Reduces Fear and Recruitment of Parvalbumin Interneurons of the Amygdala.

    PubMed

    Bocchio, Marco; Fucsina, Giulia; Oikonomidis, Lydia; McHugh, Stephen B; Bannerman, David M; Sharp, Trevor; Capogna, Marco

    2015-12-01

    Genetic association studies suggest that variations in the 5-hydroxytryptamine (5-HT; serotonin) transporter (5-HTT) gene are associated with susceptibility to psychiatric disorders such as anxiety or posttraumatic stress disorder. Individuals carrying high 5-HTT-expressing gene variants display low amygdala reactivity to fearful stimuli. Mice overexpressing the 5-HTT (5-HTTOE), an animal model of this human variation, show impaired fear, together with reduced fear-evoked theta oscillations in the basolateral amygdala (BLA). However, it is unclear how variation in 5-HTT gene expression impacts on the microcircuitry of the BLA to change behavior. We addressed this issue by investigating the activity of parvalbumin (PV)-expressing interneurons (PVINs), the biggest IN population in the basal amygdala (BA). We found that increased 5-HTT expression impairs the recruitment of PVINs (measured by their c-Fos immunoreactivity) during fear. Ex vivo patch-clamp recordings demonstrated that the depolarizing effect of 5-HT on PVINs was mediated by 5-HT2A receptor. In 5-HTTOE mice, 5-HT-evoked depolarization of PVINs and synaptic inhibition of principal cells, which provide the major output of the BA, were impaired. This deficit was because of reduced 5-HT2A function and not because of increased 5-HT uptake. Collectively, these findings provide novel cellular mechanisms that are likely to contribute to differences in emotional behaviors linked with genetic variations of the 5-HTT.

  13. Testosterone reduces AGTR1 expression to prevent β-cell and islet apoptosis from glucotoxicity.

    PubMed

    Kooptiwut, Suwattanee; Hanchang, Wanthanee; Semprasert, Namoiy; Junking, Mutita; Limjindaporn, Thawornchai; Yenchitsomanus, Pa-thai

    2015-03-01

    Hypogonadism in men is associated with an increased incidence of type 2 diabetes. Supplementation with testosterone has been shown to protect pancreatic β-cell against apoptosis due to toxic substances including streptozotocin and high glucose. One of the pathological mechanisms of glucose-induced pancreatic β-cell apoptosis is the induction of the local rennin-angiotensin-aldosterone system (RAAS). The role of testosterone in regulation of the pancreatic RAAS is still unknown. This study aims to investigate the protective action of testosterone against glucotoxicity-induced pancreatic β-cell apoptosis via alteration of the pancreatic RAAS pathway. Rat insulinoma cell line (INS-1) cells or isolated male mouse islets were cultured in basal and high-glucose media in the presence or absence of testosterone, losartan, and angiotensin II (Ang II), then cell apoptosis, cleaved caspase 3 expression, oxidative stress, and expression of angiotensin II type 1 receptor (AGTR1) and p47(phox) mRNA and protein were measured. Testosterone and losartan showed similar effects in reducing pancreatic β-cell apoptosis. Testosterone significantly reduced expression of AGTR1 protein in INS-1 cells cultured in high-glucose medium or high-glucose medium with Ang II. Testosterone decreased the expression of AGTR1 and p47(phox) mRNA and protein in comparison with levels in cells cultured in high-glucose medium alone. Furthermore, testosterone attenuated superoxide production when co-cultured with high-glucose medium. In contrast, when cultured in basal glucose, supplementation of testosterone did not have any effect on cell apoptosis, oxidative stress, and expression of AGT1R and p47(phox). In addition, high-glucose medium did not increase cleaved caspase 3 in AGTR1 knockdown experiments. Thus, our results indicated that testosterone prevents pancreatic β-cell apoptosis due to glucotoxicity through reduction of the expression of ATGR1 and its signaling pathway.

  14. Benzylglucosinolate Derived Isothiocyanate from Tropaeolum majus Reduces Gluconeogenic Gene and Protein Expression in Human Cells

    PubMed Central

    Guzmán-Pérez, Valentina; Bumke-Vogt, Christiane; Schreiner, Monika; Mewis, Inga; Borchert, Andrea; Pfeiffer, Andreas F. H.

    2016-01-01

    Nasturtium (Tropaeolum majus L.) contains high concentrations of benzylglcosinolate. We found that a hydrolysis product of benzyl glucosinolate—the benzyl isothiocyanate (BITC)—modulates the intracellular localization of the transcription factor Forkhead box O 1 (FOXO1). FoxO transcription factors can antagonize insulin effects and trigger a variety of cellular processes involved in tumor suppression, longevity, development and metabolism. The current study evaluated the ability of BITC—extracted as intact glucosinolate from nasturtium and hydrolyzed with myrosinase—to modulate i) the insulin-signaling pathway, ii) the intracellular localization of FOXO1 and, iii) the expression of proteins involved in gluconeogenesis, antioxidant response and detoxification. Stably transfected human osteosarcoma cells (U-2 OS) with constitutive expression of FOXO1 protein labeled with GFP (green fluorescent protein) were used to evaluate the effect of BITC on FOXO1. Human hepatoma HepG2 cell cultures were selected to evaluate the effect on gluconeogenic, antioxidant and detoxification genes and protein expression. BITC reduced the phosphorylation of protein kinase B (AKT/PKB) and FOXO1; promoted FOXO1 translocation from cytoplasm into the nucleus antagonizing the insulin effect; was able to down-regulate the gene and protein expression of gluconeogenic enzymes; and induced the gene expression of antioxidant and detoxification enzymes. Knockdown analyses with specific siRNAs showed that the expression of gluconeogenic genes was dependent on nuclear factor (erythroid derived)-like2 (NRF2) and independent of FOXO1, AKT and NAD-dependent deacetylase sirtuin-1 (SIRT1). The current study provides evidence that BITC might have a role in type 2 diabetes T2D by reducing hepatic glucose production and increasing antioxidant resistance. PMID:27622707

  15. The roughness surface expressed by the mathematical model

    NASA Astrophysics Data System (ADS)

    Macurova, Anna

    2010-07-01

    The work investigates the effect of some characteristics of a cut surface and studies roughness of the cutting process. There is elaborated theoretical information and new aspects on calculation of the theoretical values of the roughness of the cut surface for the chosen materials are formulated. In the area of the experimental investigation, results on characteristics of the chosen materials are formulated in this work. Obtained results are fundamental for the mathematical modulation and mathematical analysis for the investigated dependencies for the cut surfaces. The mathematical model also represents the specific dependencies of the technological process. The characteristics of the observed parameters are approximated by characteristics of the quasi-linear models. The solution of this model offers acceptable results. The mathematical models of the roughness of the cut surface are a mathematical description of the dependency of the maximum roughness of the cut surface of the feed represented by the differential equation and by the integral curves.

  16. Reduced ubiquitin C-terminal hydrolase-1 expression levels in dementia with Lewy bodies.

    PubMed

    Barrachina, Marta; Castaño, Esther; Dalfó, Esther; Maes, Tamara; Buesa, Carlos; Ferrer, Isidro

    2006-05-01

    Parkinson disease (PD) and dementia with Lewy bodies (DLB) are characterized by the accumulation of abnormal alpha-synuclein and ubiquitin in protein aggregates conforming Lewy bodies and Lewy neurites. Ubiquitin C-terminal hydrolase-1 (UCHL-1) disassembles polyubiquitin chains to increase the availability of free monomeric ubiquitin to the ubiquitin proteasome system (UPS) thus favoring protein degradation. Since mutations in the UCHL-1 gene, reducing UPS activity by 50%, have been reported in autosomal dominant PD, and UCHL-1 inhibition results in the formation of alpha-synuclein aggregates in mesencephalic cultured neurons, the present study was initiated to test UCHL-1 mRNA and protein levels in post-mortem frontal cortex (area 8) of PD and DLB cases, compared with age-matched controls. TaqMan PCR assays, and Western blots demonstrated down-regulation of UCHL-1 mRNA and UCHL-1 protein in the cerebral cortex in DLB (either in pure forms, not associated with Alzheimer disease: AD, and in common forms, with accompanying AD changes), but not in PD, when compared with age-matched controls. Interestingly, UCHL-1 mRNA and protein expressions were reduced in the medulla oblongata in the same PD cases. Moreover, UCHL-1 protein was decreased in the substantia nigra in cases with Lewy body pathology. UCHL-1 down-regulation was not associated with reduced protein levels of several proteasomal subunits, including 20SX, 20SY, 19S and 11Salpha. Yet UCHL-3 expression was reduced in the cerebral cortex of PD and DLB patients. Together, these observations show reduced UCHL-1 expression as a contributory factor in the abnormal protein aggregation in DLB, and points UCHL-1 as a putative therapeutic target in the treatment of DLB.

  17. Decreased tumorigenicity correlates with expression of altered cell surface carbohydrates in Lec9 CHO cells.

    PubMed Central

    Ripka, J; Shin, S; Stanley, P

    1986-01-01

    To investigate a role for surface carbohydrates in cellular malignancy, 15 different glycosylation-defective CHO cell mutants were examined for their tumorigenic and metastatic capacities after subcutaneous injection into nude mice. Most of the glycosylation mutants displayed similar or slightly decreased tumorigenicity compared with parental CHO cells. Neither parental CHO cells nor any of the mutants were observed to metastasize. However, independent isolates of one mutant type, Lec9, showed a dramatic reduction in tumor formation. The altered carbohydrates expressed at the surface of Lec9 cells appeared to be responsible for their loss of tumorigenicity, because revertants for lectin resistance were able to form tumors, and a double mutant (Lec9.Lec1) that expressed a Lec1 glycosylation phenotype also formed tumors. Finally, Lec9 cells were able to form tumors in gamma-irradiated nude mice, suggesting that recognition by an irradiation-sensitive host cell(s) was responsible for their reduced tumorigenicity in untreated nude mice. PMID:3785164

  18. The brain-specific Beta4 subunit downregulates BK channel cell surface expression.

    PubMed

    Shruti, Sonal; Urban-Ciecko, Joanna; Fitzpatrick, James A; Brenner, Robert; Bruchez, Marcel P; Barth, Alison L

    2012-01-01

    The large-conductance K(+) channel (BK channel) can control neural excitability, and enhanced channel currents facilitate high firing rates in cortical neurons. The brain-specific auxiliary subunit β4 alters channel Ca(++)- and voltage-sensitivity, and β4 knock-out animals exhibit spontaneous seizures. Here we investigate β4's effect on BK channel trafficking to the plasma membrane. Using a novel genetic tag to track the cellular location of the pore-forming BKα subunit in living cells, we find that β4 expression profoundly reduces surface localization of BK channels via a C-terminal ER retention sequence. In hippocampal CA3 neurons from C57BL/6 mice with endogenously high β4 expression, whole-cell BK channel currents display none of the characteristic properties of BKα+β4 channels observed in heterologous cells. Finally, β4 knock-out animals exhibit a 2.5-fold increase in whole-cell BK channel current, indicating that β4 also regulates current magnitude in vivo. Thus, we propose that a major function of the brain-specific β4 subunit in CA3 neurons is control of surface trafficking.

  19. The Brain-Specific Beta4 Subunit Downregulates BK Channel Cell Surface Expression

    PubMed Central

    Shruti, Sonal; Urban-Ciecko, Joanna; Fitzpatrick, James A.; Brenner, Robert; Bruchez, Marcel P.; Barth, Alison L.

    2012-01-01

    The large-conductance K+ channel (BK channel) can control neural excitability, and enhanced channel currents facilitate high firing rates in cortical neurons. The brain-specific auxiliary subunit β4 alters channel Ca++- and voltage-sensitivity, and β4 knock-out animals exhibit spontaneous seizures. Here we investigate β4's effect on BK channel trafficking to the plasma membrane. Using a novel genetic tag to track the cellular location of the pore-forming BKα subunit in living cells, we find that β4 expression profoundly reduces surface localization of BK channels via a C-terminal ER retention sequence. In hippocampal CA3 neurons from C57BL/6 mice with endogenously high β4 expression, whole-cell BK channel currents display none of the characteristic properties of BKα+β4 channels observed in heterologous cells. Finally, β4 knock-out animals exhibit a 2.5-fold increase in whole-cell BK channel current, indicating that β4 also regulates current magnitude in vivo. Thus, we propose that a major function of the brain-specific β4 subunit in CA3 neurons is control of surface trafficking. PMID:22438928

  20. Development of electrically conductive-superoleophobic micropillars for reducing surface adhesion of oil at low temperatures

    NASA Astrophysics Data System (ADS)

    Pan, Zihe; Wang, Tianchang; Zhou, Yikang; Zhao, Boxin

    2016-12-01

    Electrically conductive and superoleophobic micropillars have been developed through the construction of biomimetic micropillars using Ag-filled epoxy composites and the incorporation of FDTS on the micropillar surface. These micropillars are found to be superoleophobic with an oil contact angle of 140°, demonstrating excellent self-cleaning properties. The conductivity of micropillars allows for the Joule-heating effect to actively reduce the adhesion and even unfreeze the frozen oil droplets by passing electrical current. Electrical resistance of the composite micropillars was modulated by two orders of magnitudes by varying the contents of Ag flakes from 45 wt% to 65 wt%. The effectiveness of conductive micropillars for surface un-freezing was investigated by applying DC current to decrease the adhesion strength of frozen oil droplets on surfaces. The results showed a pronounced reduction of frozen oil adhesion force by 60% when the resistance increased from 7.5 Ω to 877 Ω after applying DC current for 2 min. By continuously applying DC current for 3 min, the frozen oil adhesion decreased to 0.05 N, reaching zero when the surface was heated up to -10 °C after applying DC current for 5 min. In contrast, when the droplet was heated up to -5 °C by hot air, there is still a substantial force of adhesion. The research findings demonstrate the use of constructing conductive-superoleophobic composite micropillars at surface for eliminating the frozen oil from surfaces at low temperatures.

  1. Physical origins of ruled surfaces on the reduced density matrices geometry

    NASA Astrophysics Data System (ADS)

    Chen, Ji-Yao; Ji, Zhengfeng; Liu, Zheng-Xin; Qi, Xiaofei; Yu, Nengkun; Zeng, Bei; Zhou, Duanlu

    2017-02-01

    The reduced density matrices (RDMs) of many-body quantum states form a convex set. The boundary of low dimensional projections of this convex set may exhibit nontrivial geometry such as ruled surfaces. In this paper, we study the physical origins of these ruled surfaces for bosonic systems. The emergence of ruled surfaces was recently proposed as signatures of symmetry-breaking phase. We show that, apart from being signatures of symmetry-breaking, ruled surfaces can also be the consequence of gapless quantum systems by demonstrating an explicit example in terms of a two-mode Ising model. Our analysis was largely simplified by the quantum de Finetti's theorem—in the limit of large system size, these RDMs are the convex set of all the symmetric separable states. To distinguish ruled surfaces originated from gapless systems from those caused by symmetry-breaking, we propose to use the finite size scaling method for the corresponding geometry. This method is then applied to the two-mode XY model, successfully identifying a ruled surface as the consequence of gapless systems.

  2. Expression and production of staphylococcal enterotoxin C is substantially reduced in milk.

    PubMed

    Valihrach, Lukas; Alibayov, Babek; Zdenkova, Kamila; Demnerova, Katerina

    2014-12-01

    Staphylococcal food poisoning is a global problem. The gene encoding enterotoxin C (sec) has been reported several times as the most frequent enterotoxin gene identified in food poisoning cases caused by contaminated milk. In this study, the expression of sec was examined during the growth of Staphylococcus aureus in milk compared to routinely used laboratory media. Additionally, expression of several regulatory genes (sarA, saeS, codY, srrA, rot, hld, agrA, sigB) and other five enterotoxin genes (sea, seg, seh, sek, sel) were observed. It has been well established for that S. aureus is able to grow in milk and we found significantly reduced expression of sec in milk compared to the laboratory medium (P < 0.05). Here, we report the first study providing a comprehensive view on the expression of enterotoxin genes and its regulation in milk. The milk environment dramatically changed the expression profiles of several enterotoxin genes although staphylococcal growth was not affected at all. The mechanism of the reduction may be explained by downregulation of the agr system, although other factors are expected to be involved. The constituent of milk causing the inhibitory effect remains unidentified.

  3. Increased expression of the diabetes gene SOX4 reduces insulin secretion by impaired fusion pore expansion

    PubMed Central

    Collins, Stephan C.; Do, Hyun Woong; Hastoy, Benoit; Hugill, Alison; Adam, Julie; Chibalina, Margarita V.; Galvanovskis, Juris; Godazgar, Mahdieh; Lee, Sheena; Goldsworthy, Michelle; Salehi, Albert; Tarasov, Andrei I.; Rosengren, Anders H.; Cox, Roger; Rorsman, Patrik

    2016-01-01

    The transcription factor Sox4 has been proposed to underlie the increased type-2 diabetes risk linked to an intronic SNP in CDKAL1. In a mouse model expressing a mutant form of Sox4, glucose-induced insulin secretion is reduced by 40% despite normal intracellular Ca2+ signalling and depolarization-evoked exocytosis. This paradox is explained by a 4-fold increase in kiss-and-run exocytosis (as determined by single-granule exocytosis measurements), in which the fusion pore connecting the granule lumen to the exterior only expands to a diameter of 2 nm that does not allow the exit of insulin. Microarray analysis indicated that this correlated with an increased expression of the exocytosis-regulating protein Stxbp6. In a large collection of human islet preparations (n=63), STXBP6 expression and GIIS correlated positively and negatively with SOX4 expression, respectively. Overexpression of SOX4 in the human insulin-secreting cell EndoC-βH2 interfered with granule emptying and inhibited hormone release, the latter effect was reversed by silencing of STXBP6. These data suggest that increased SOX4 expression inhibits insulin secretion and increased diabetes risk by upregulation of STXBP6 and an increase in kiss-and-run exocytosis at the expense of full fusion. We propose that pharmacological interventions promoting fusion pore expansion may be effective in diabetes therapy. PMID:26993066

  4. Increased Expression of Follistatin in Breast Cancer Reduces Invasiveness and Clinically Correlates with Better Survival.

    PubMed

    Zabkiewicz, Catherine; Resaul, Jeyna; Hargest, Rachel; Jiang, Wen Guo; Ye, Lin

    2017-01-01

    Activin and its antagonist follistatin (FST) have been implicated in several solid tumours. This study investigated the role of FST in breast cancer. FST expression was examined using reverse transcription polymerase chain reaction (RT-PCR), real-time quantitative polymerase chain reaction (qPCR) and immunohistochemistry in a cohort of breast cancer samples. Expression was correlated to pathological and prognostic parameters in our patient cohort. FST was overexpressed in MCF-7 cells and assays for growth and invasion were performed. FST is expressed in breast tissue, in the cytoplasm of mammary epithelial cells. Expression was decreased in breast cancer tissue in comparison to normal mammary tissue. Over-expression of FST in vitro led to significantly increased growth rate and reduced invasion. Higher FST associates with lower-grade tumours and better survival. Our results suggest a role for FST as a suppressor of invasion and metastasis in breast cancer. Copyright© 2017, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.

  5. Disturbed shear stress reduces Klf2 expression in arterial-venous fistulae in vivo

    PubMed Central

    Yamamoto, Kota; Protack, Clinton D; Kuwahara, Go; Tsuneki, Masayuki; Hashimoto, Takuya; Hall, Michael R; Assi, Roland; Brownson, Kirstyn E; Foster, Trenton R; Bai, Hualong; Wang, Mo; Madri, Joseph A; Dardik, Alan

    2015-01-01

    Laminar shear stress (SS) induces an antiproliferative and anti-inflammatory endothelial phenotype and increases Klf2 expression. We altered the diameter of an arteriovenous fistula (AVF) in the mouse model to determine whether increased fistula diameter produces disturbed SS in vivo and if acutely increased disturbed SS results in decreased Klf2 expression. The mouse aortocaval fistula model was performed with 22, 25, or 28 gauge needles to puncture the aorta and the inferior vena cava. Duplex ultrasound was used to examine the AVF and its arterial inflow and venous outflow, and SS was calculated. Arterial samples were examined with western blot, immunohistochemistry, and immunofluorescence analysis for proteins and qPCR for RNA. Mice with larger diameter fistulae had diminished survival but increased AVF patency. Increased SS magnitudes and range of frequencies were directly proportional to the needle diameter in the arterial limb proximal to the fistula but not in the venous limb distal to the fistula, with 22-gauge needles producing the most disturbed SS in vivo. Klf2 mRNA and protein expression was diminished in the artery proximal to the fistula in proportion to increasing SS. Increased fistula diameter produces increased SS magnitude and frequency, consistent with disturbed SS in vivo. Disturbed SS is associated with decreased mRNA and protein expression of Klf2. Disturbed SS and reduced Klf2 expression near the fistula are potential therapeutic targets to improve AVF maturation. PMID:25780089

  6. Plasma graft of poly(ethylene glycol) methyl ether methacrylate (PEGMA) on RGP lens surface for reducing protein adsorption

    NASA Astrophysics Data System (ADS)

    Shiheng, Yin; Li, Ren; Yingjun, Wang

    2017-01-01

    Poly(ethylene glycol) methyl ether methacrylate (PEGMA) was grafted on fluorosilicone acrylate rigid gas permissible contact lens surface by means of argon plasma induced polymerization to improve surface hydrophilicity and reduce protein adsorption. The surface properties were characterized by contact angle measurement, x-ray photoelectron spectroscopy (XPS) and atomic force microscopy respectively. The surface protein adsorption was evaluated by lysozyme solution immersion and XPS analysis. The results indicated that a thin layer of PEGMA was successfully grafted. The surface hydrophilicity was bettered and surface free energy increased. The lysozyme adsorption on the lens surface was reduced greatly. The study was supported by National Natural Science Foundation of China (No. 51273072).

  7. Plasma graft of poly(ethylene glycol) methyl ether methacrylate (PEGMA) on RGP lens surface for reducing protein adsorption

    NASA Astrophysics Data System (ADS)

    Yin, Shiheng; Ren, Li; Wang, Yingjun

    2017-01-01

    Poly(ethylene glycol) methyl ether methacrylate (PEGMA) was grafted on fluorosilicone acrylate rigid gas permissible contact lens surface by means of argon plasma induced polymerization to improve surface hydrophilicity and reduce protein adsorption. The surface properties were characterized by contact angle measurement, x-ray photoelectron spectroscopy (XPS) and atomic force microscopy respectively. The surface protein adsorption was evaluated by lysozyme solution immersion and XPS analysis. The results indicated that a thin layer of PEGMA was successfully grafted. The surface hydrophilicity was bettered and surface free energy increased. The lysozyme adsorption on the lens surface was reduced greatly. The study was supported by National Natural Science Foundation of China (No. 51273072).

  8. Acute Heat Stress and Reduced Nutrient Intake Alter Intestinal Proteomic Profile and Gene Expression in Pigs

    PubMed Central

    Pearce, Sarah C.; Lonergan, Steven M.; Huff-Lonergan, Elisabeth; Baumgard, Lance H.; Gabler, Nicholas K.

    2015-01-01

    Heat stress and reduced feed intake negatively affect intestinal integrity and barrier function. Our objective was to compare ileum protein profiles of pigs subjected to 12 hours of HS, thermal neutral ad libitum feed intake, or pair-fed to heat stress feed intake under thermal neutral conditions (pair-fed thermal neutral). 2D-Differential In Gel Electrophoresis and gene expression were performed. Relative abundance of 281 and 138 spots differed due to heat stress, compared to thermal neutral and pair-fed thermal neutral pigs, respectively. However, only 20 proteins were different due to feed intake (thermal neutral versus pair-fed thermal neutral). Heat stress increased mRNA expression of heat shock proteins and protein abundance of heat shock proteins 27, 70, 90-α and β were also increased. Heat stress reduced ileum abundance of several metabolic enzymes, many of which are involved in the glycolytic or TCA pathways, indicating a change in metabolic priorities. Stress response enzymes peroxiredoxin-1 and peptidyl-prolyl cis-trans isomerase A were decreased in pair-fed thermal neutral and thermal neutral pigs compared to heat stress. Heat stress increased mRNA abundance markers of ileum hypoxia. Altogether, these data show that heat stress directly alters intestinal protein and mRNA profiles largely independent of reduced feed intake. These changes may be related to the reduced intestinal integrity associated with heat stress. PMID:26575181

  9. Surface modifications of photoanodes in dye sensitized solar cells: enhanced light harvesting and reduced recombination

    NASA Astrophysics Data System (ADS)

    Saxena, Vibha; Aswal, D. K.

    2015-06-01

    In a quest to harvest solar power, dye-sensitized solar cells (DSSCs) have potential for low-cost eco-friendly photovoltaic devices. The major processes which govern the efficiency of a DSSC are photoelectron generation, injection of photo-generated electrons to the conduction band (CB) of the mesoporous nanocrystalline semiconductor (nc-SC); transport of CB electrons through nc-SC and subsequent collection of CB electrons at the counter electrode (CE) through the external circuit; and dye regeneration by redox couple or hole transport layer (HTL). Most of these processes occur at various interfaces of the photoanode. In addition, recombination losses of photo-generated electrons with either dye or redox molecules take place at the interfaces. Therefore, one of the key requirements for high efficiency is to improve light harvesting of the photoanode and to reduce the recombination losses at various interfaces. In this direction, surface modification of the photoanode is the simplest method among the various other approaches available in the literature. In this review, we present a comprehensive discussion on surface modification of the photoanode, which has been adopted in the literature for not only enhancing light harvesting but also reducing recombination. Various approaches towards surface modification of the photoanode discussed are (i) fluorine-doped tin oxide (FTO)/nc-SC interface modified via a compact layer of semiconductor material which blocks exposed sites of FTO to electrolyte (or HTL), (ii) nc-SC/dye interface modification either through acid treatment resulting in enhanced dye loading due to a positively charged surface or by depositing insulating/semiconducting blocking layer on the nc-SC surface, which acts as a tunneling barrier for recombination, (iii) nc-SC/dye interface modified by employing co-adsorbents which helps in reducing the dye aggregation and thereby recombination, and (iv) dye/electrolyte (or dye/HTL) interface modification using

  10. Pharmacological concentrations of biotin reduce serum triglycerides and the expression of lipogenic genes.

    PubMed

    Larrieta, Elena; Velasco, Fidel; Vital, Paz; López-Aceves, Teresita; Lazo-de-la-Vega-Monroy, María Luisa; Rojas, Alberto; Fernandez-Mejia, Cristina

    2010-10-10

    Besides its role as a carboxylase prosthetic group, biotin regulates gene expression and has a wide repertoire of effects on systemic processes. Several studies have shown that pharmacological concentrations of biotin reduce hypertriglyceridemia. The molecular mechanisms by which pharmacological concentrations of biotin affect lipid metabolism are largely unknown. The present study analyzed the effects of pharmacological doses of biotin on triglyceridemia, insulin sensitivity and on mRNA expression of various lipogenic genes. Three-week-old male BALB/cAnN Hsd mice were fed a biotin-control or a biotin-supplemented diet (1.76 or 97.7mg of free biotin/kg diet, respectively) over a period of eight weeks. Serum triglyceride concentrations, insulin and glucose tolerance and mRNA abundance of various lipogenic genes were investigated. The biotin-supplemented group showed 35% less serum triglycerides than control mice. In the liver, we found a significant (P<0.05) reduction of mRNA levels of SREBP1-c, glucose transporter-2, phosphofructokinase-1, pyruvate kinase, acetyl-CoA carboxylase and fatty acid synthase, while glucose-6-phosphate dehydrogenase expression increased. No changes in glucokinase, stearoyl-CoA desaturase-1, FoxO1 or PPAR-gamma expression were observed. In adipose tissue, we found a decreased expression of SREBP1c, glucose-6-phosphate deshydrogenase, acetyl-CoA carboxylase, fatty acid synthase, stearoyl-CoA desaturase-1, phosphofructokinase-1 and PPAR-gamma, but no changes in FoxO1 expression. Moreover, the group fed a biotin-supplemented diet showed a significant decrease in adipose tissue weight. No differences in insulin sensitivity or serum insulin concentrations were observed between groups. Our results indicate that pharmacological concentrations of biotin decrease serum tryglyceride concentrations and lipogenic gene expression in liver and adipose tissues. Copyright 2010 Elsevier B.V. All rights reserved.

  11. Reduced Expression of IRF7 in Nasal Epithelial Cells from Smokers after Infection with Influenza

    PubMed Central

    Jaspers, Ilona; Horvath, Katherine M.; Zhang, Wenli; Brighton, Luisa E.; Carson, Johnny L.; Noah, Terry L.

    2010-01-01

    Smokers are more susceptible to respiratory viral infections, including influenza virus, but the mechanisms mediating this effect are unknown. To determine how epithelial cells contribute to the enhanced susceptibility seen in smokers, we established an in vitro model of differentiated nasal epithelial cells (NECs) from smokers, which showed enhanced mucin expression. The NECs from smokers responded to influenza infection with greater cytotoxicity, release of interleukin-6, and viral shedding than NECs from nonsmokers. Focusing on type I interferon (IFN) expression, we observed that influenza-infected NECs from smokers produced significantly less IFN-α than NECs from nonsmokers. Similarly, the expression of IRF7, a key transcription factor controlling the expression of IFN-α, was significantly decreased in influenza-infected and IFN-β–stimulated NECs from smokers. Furthermore, our data indicate that the DNA methylation of the IRF7 gene and expression of the DNA (cytosine-5-)-methyltransferase 1 was enhanced in NECs from smokers. To confirm these findings in vivo, we initiated a study in which smoking and nonsmoking healthy volunteers were inoculated nasally with the live-attenuated influenza virus (LAIV) vaccine, and nasal biopsies were obtained before and after the administration of LAIV. The LAIV-induced expression of IRF7 was lower in the nasal epithelium from smokers, supporting our in vitro observations. These data demonstrate that infection with influenza results in the reduced expression of transcription factor IRF7 in NECs from smokers, and that these effects may be mediated by an epigenetic modification of the IRF7 gene, thus providing a potential mechanism rendering smokers more susceptible to respiratory virus infections. PMID:19880818

  12. Inducing a Concurrent Motor Load Reduces Categorization Precision for Facial Expressions

    PubMed Central

    2015-01-01

    Motor theories of expression perception posit that observers simulate facial expressions within their own motor system, aiding perception and interpretation. Consistent with this view, reports have suggested that blocking facial mimicry induces expression labeling errors and alters patterns of ratings. Crucially, however, it is unclear whether changes in labeling and rating behavior reflect genuine perceptual phenomena (e.g., greater internal noise associated with expression perception or interpretation) or are products of response bias. In an effort to advance this literature, the present study introduces a new psychophysical paradigm for investigating motor contributions to expression perception that overcomes some of the limitations inherent in simple labeling and rating tasks. Observers were asked to judge whether smiles drawn from a morph continuum were sincere or insincere, in the presence or absence of a motor load induced by the concurrent production of vowel sounds. Having confirmed that smile sincerity judgments depend on cues from both eye and mouth regions (Experiment 1), we demonstrated that vowel production reduces the precision with which smiles are categorized (Experiment 2). In Experiment 3, we replicated this effect when observers were required to produce vowels, but not when they passively listened to the same vowel sounds. In Experiments 4 and 5, we found that gender categorizations, equated for difficulty, were unaffected by vowel production, irrespective of the presence of a smiling expression. These findings greatly advance our understanding of motor contributions to expression perception and represent a timely contribution in light of recent high-profile challenges to the existing evidence base. PMID:26618622

  13. Method and Apparatus for Reducing Noise from Near Ocean Surface Sources

    DTIC Science & Technology

    2001-10-01

    reducing the acoustic noise from near-surface 4 sources using an array processing technique that utilizes 5 Multiple Signal Classification ( MUSIC ...sources without 13 degrading the signal level and quality of the TOI. The present 14 invention utilizes a unique application of the MUSIC beamforming...specific algorithm that utilizes a 5 MUSIC technique and estimates the direction of arrival (DOA) of 6 the acoustic signal signals and generates output

  14. The MS Risk Allele of CD40 Is Associated with Reduced Cell-Membrane Bound Expression in Antigen Presenting Cells: Implications for Gene Function.

    PubMed

    Field, Judith; Shahijanian, Fernando; Schibeci, Stephen; Johnson, Laura; Gresle, Melissa; Laverick, Louise; Parnell, Grant; Stewart, Graeme; McKay, Fiona; Kilpatrick, Trevor; Butzkueven, Helmut; Booth, David

    2015-01-01

    Human genetic and animal studies have implicated the costimulatory molecule CD40 in the development of multiple sclerosis (MS). We investigated the cell specific gene and protein expression variation controlled by the CD40 genetic variant(s) associated with MS, i.e. the T-allele at rs1883832. Previously we had shown that the risk allele is expressed at a lower level in whole blood, especially in people with MS. Here, we have defined the immune cell subsets responsible for genotype and disease effects on CD40 expression at the mRNA and protein level. In cell subsets in which CD40 is most highly expressed, B lymphocytes and dendritic cells, the MS-associated risk variant is associated with reduced CD40 cell-surface protein expression. In monocytes and dendritic cells, the risk allele additionally reduces the ratio of expression of full-length versus truncated CD40 mRNA, the latter encoding secreted CD40. We additionally show that MS patients, regardless of genotype, express significantly lower levels of CD40 cell-surface protein compared to unaffected controls in B lymphocytes. Thus, both genotype-dependent and independent down-regulation of cell-surface CD40 is a feature of MS. Lower expression of a co-stimulator of T cell activation, CD40, is therefore associated with increased MS risk despite the same CD40 variant being associated with reduced risk of other inflammatory autoimmune diseases. Our results highlight the complexity and likely individuality of autoimmune pathogenesis, and could be consistent with antiviral and/or immunoregulatory functions of CD40 playing an important role in protection from MS.

  15. The MS Risk Allele of CD40 Is Associated with Reduced Cell-Membrane Bound Expression in Antigen Presenting Cells: Implications for Gene Function

    PubMed Central

    Field, Judith; Shahijanian, Fernando; Schibeci, Stephen; Johnson, Laura; Gresle, Melissa; Laverick, Louise; Parnell, Grant; Stewart, Graeme; McKay, Fiona; Kilpatrick, Trevor; Butzkueven, Helmut; Booth, David

    2015-01-01

    Human genetic and animal studies have implicated the costimulatory molecule CD40 in the development of multiple sclerosis (MS). We investigated the cell specific gene and protein expression variation controlled by the CD40 genetic variant(s) associated with MS, i.e. the T-allele at rs1883832. Previously we had shown that the risk allele is expressed at a lower level in whole blood, especially in people with MS. Here, we have defined the immune cell subsets responsible for genotype and disease effects on CD40 expression at the mRNA and protein level. In cell subsets in which CD40 is most highly expressed, B lymphocytes and dendritic cells, the MS-associated risk variant is associated with reduced CD40 cell-surface protein expression. In monocytes and dendritic cells, the risk allele additionally reduces the ratio of expression of full-length versus truncated CD40 mRNA, the latter encoding secreted CD40. We additionally show that MS patients, regardless of genotype, express significantly lower levels of CD40 cell-surface protein compared to unaffected controls in B lymphocytes. Thus, both genotype-dependent and independent down-regulation of cell-surface CD40 is a feature of MS. Lower expression of a co-stimulator of T cell activation, CD40, is therefore associated with increased MS risk despite the same CD40 variant being associated with reduced risk of other inflammatory autoimmune diseases. Our results highlight the complexity and likely individuality of autoimmune pathogenesis, and could be consistent with antiviral and/or immunoregulatory functions of CD40 playing an important role in protection from MS. PMID:26068105

  16. Pivotal Advance: Nonfunctional lung effectors exhibit decreased calcium mobilization associated with reduced expression of ORAI1

    PubMed Central

    Arimilli, Subhashini; Sharma, Sharad K.; Yammani, Rama; Reid, Sean D.; Parks, Griffith D.; Alexander-Miller, Martha A.

    2010-01-01

    CD8+ T cells play a critical role in the clearance of respiratory pathogens. Thus, it is surprising that functional inactivation of lung effectors has been observed in many models of viral infection. Currently, the molecular defect responsible for the shut-off of function in these cells is unknown. In the present study, we addressed this question using a model of respiratory infection with the paramyxovirus SV5. Nonfunctional cells were found to exhibit decreases in SOCE, resulting in reduced NFAT1 activation. Notably, function could be restored by the provision of increased levels of extracellular calcium. The reduced ability to mobilize calcium was associated with reduced expression of ORAI1, the CRAC channel subunit. These findings reveal a previously unknown mechanism for the negative regulation of function in effector T cells. PMID:20103768

  17. Reducing The Light Reflected by Silicon Surface Using ZnO/TS Antireflection Coating

    NASA Astrophysics Data System (ADS)

    Suhandi, Andi; Tayubi, Yuyu R.; Wibowo, Firmanul C.; Arifin, Pepen; Supriyatman

    2017-07-01

    Zinc Oxide (ZnO) thin films was coated on a texturized silicon (TS) surface using a spincoating technique. The TS layer was prepared by a wet etching method using 20 % KOH solution at temperature of 80°C for 5 minutes. To prepared precursor solution for ZnO layer, zinc acetate dehydrate, 2-methoxyethanol and monoethanolamine are used as a starting material, solvent and stabilizer, respectively. The XRD and SEM measurements confirmed that the thin films grown by spincoating technique have a single oriented crystal plane and homogenous surfaces. From photoluminescence measurement found that the optical band gap of grown films to be 3.44 eV. The optical reflectance of the grown films is characterized by UV-VIS spectrometry show that the presence of anti-reflection coating ZnO/TS is proven to reduce the reflection of solar radiation by silicon surface significantly.

  18. Spectroscopic study of surface enhanced Raman scattering of caffeine on borohydride-reduced silver colloids

    NASA Astrophysics Data System (ADS)

    Chen, Xiaomin; Gu, Huaimin; Shen, Gaoshan; Dong, Xiao; Kang, Jian

    2010-06-01

    The surface enhanced Raman scattering (SERS) of caffeine on borohydride-reduced silver colloids system under different aqueous solution environment has been studied in this paper. The relative intensity of SERS of caffeine significantly varies with different concentrations of sodium chloride and silver particles. However, at too high or too low concentration of sodium chloride and silver particle, the enhancement of SERS spectra is not evident. The SERS spectra of caffeine suggest that the contribution of the charge transfer mechanism to SERS may be dominant. The chloride ions can significantly enhance the efficiency of SERS, while the enhancement is selective, as the efficiency in charge transfer enhancement is higher than in electromagnetic enhancement. Therefore, it can be concluded that the active site of chloride ion locates on the bond between the caffeine and the silver surface. In addition, the SERS spectra of caffeine on borohydride-reduced and citrate-reduced silver colloids are different, which may be due to different states caffeine adsorbed on silver surface under different silver colloids.

  19. Impact of Reduced Rhodopsin Expression on the Structure of Rod Outer Segment Disc Membranes†

    PubMed Central

    Rakshit, Tatini; Park, Paul S.-H.

    2015-01-01

    Rhodopsin is the light receptor embedded in rod outer segment (ROS) disc membranes of photoreceptor cells that initiates vision via phototransduction. The relationship between rhodopsin expression and the formation of membrane structures in the ROS is unclear but important to better understand both normal function and pathological conditions. To determine the impact of reduced rhodopsin expression on the structure of ROS discs and the supramolecular organization of rhodopsin, ROS disc membrane samples from heterozygous rhodopsin knockout mice were examined by atomic force microscopy. Similar to rhodopsin in wild-type mice, rhodopsin formed nanodomains in ROS disc membranes of heterozygous knockout mice. The reduced rhodopsin expression in heterozygous knockout mice resulted in ROS disc membranes that were smaller compared to those in wild-type mice at all ages tested. Changes in ROS disc membrane properties were observed between 4 and 6 weeks of age in heterozygous knockout mice that were not present in age-matched wild-type mice. In 4 week-old mice, the number and density of rhodopsin in ROS disc membranes was lower than that in age-matched wild-type mice. In contrast, 6 and 8 week-old mice had more rhodopsin molecules present in disc membranes compared to 4 week-old mice, which resulted in rhodopsin densities similar to those found in age-matched wild-type mice. Thus, mechanisms appear to be present that maintain a constant density of rhodopsin within ROS disc membranes even when reducing the expression of the light receptor by about half. These adaptive mechanisms, however, only occur after 4 weeks of age. PMID:25881629

  20. Ab initio modeling of the bonding of benzotriazole corrosion inhibitor to reduced and oxidized copper surfaces.

    PubMed

    Kokalj, Anton

    2015-01-01

    The bonding of benzotriazole-an outstanding corrosion inhibitor for copper-on reduced and oxidized copper surfaces is discussed on the basis of density functional theory (DFT) calculations. Calculations reveal that benzotriazole is able to bond with oxide-free and oxidized copper surfaces and on both of them it bonds significantly stronger to coordinatively unsaturated Cu sites. This suggests that benzotriazole is able to passivate the reactive under-coordinated surface sites that are plausible microscopic sites for corrosion attack. Benzotriazole can adsorb in a variety of different forms, yet it forms a strong molecule-surface bond only in deprotonated form. The bonding is even stronger when the deprotonated form is incorporated into organometallic adcomplexes. This is consistent with existing experimental evidence that benzotriazole inhibits corrosion by forming protective organometallic complexes. It is further shown that adsorption of benzotriazole considerably reduces the metal work function, which is a consequence of a large permanent molecular dipole and a properly oriented adsorption structure. It is argued that such a pronounced effect on the work function might be relevant for corrosion inhibition, because it should diminish the anodic corrosion reaction, which is consistent with existing experimental evidence that benzotriazole, although a mixed type inhibitor, predominantly affects the anodic reaction.

  1. Reducing bacteria and macrophage density on nanophase hydroxyapatite coated onto titanium surfaces without releasing pharmaceutical agents

    NASA Astrophysics Data System (ADS)

    Bhardwaj, Garima; Yazici, Hilal; Webster, Thomas J.

    2015-04-01

    Reducing bacterial density on titanium implant surfaces has been a major concern because of the increasing number of nosocomial infections. Controlling the inflammatory response post implantation has also been an important issue for medical devices due to the detrimental effects of chronic inflammation on device performance. It has recently been demonstrated that manipulating medical device surface properties including chemistry, roughness and wettability can control both infection and inflammation. Here, we synthesized nanophase (that is, materials with one dimension in the nanoscale) hydroxyapatite coatings on titanium to reduce bacterial adhesion and inflammatory responses (as measured by macrophage functions) and compared such results to bare titanium and plasma sprayed hydroxyapatite titanium coated surfaces used clinically today. This approach is a pharmaceutical-free approach to inhibit infection and inflammation due to the detrimental side effects of any drug released in the body. Here, nanophase hydroxyapatite was synthesized in sizes ranging from 110-170 nm and was subsequently coated onto titanium samples using electrophoretic deposition. Results indicated that smaller nanoscale hydroxyapatite features on titanium surfaces alone decreased bacterial attachment in the presence of gram negative (P. aeruginosa), gram positive (S. aureus) and ampicillin resistant gram-negative (E. coli) bacteria as well as were able to control inflammatory responses; properties which should lead to their further investigation for improved medical applications.

  2. Reducing bacteria and macrophage density on nanophase hydroxyapatite coated onto titanium surfaces without releasing pharmaceutical agents.

    PubMed

    Bhardwaj, Garima; Yazici, Hilal; Webster, Thomas J

    2015-05-14

    Reducing bacterial density on titanium implant surfaces has been a major concern because of the increasing number of nosocomial infections. Controlling the inflammatory response post implantation has also been an important issue for medical devices due to the detrimental effects of chronic inflammation on device performance. It has recently been demonstrated that manipulating medical device surface properties including chemistry, roughness and wettability can control both infection and inflammation. Here, we synthesized nanophase (that is, materials with one dimension in the nanoscale) hydroxyapatite coatings on titanium to reduce bacterial adhesion and inflammatory responses (as measured by macrophage functions) and compared such results to bare titanium and plasma sprayed hydroxyapatite titanium coated surfaces used clinically today. This approach is a pharmaceutical-free approach to inhibit infection and inflammation due to the detrimental side effects of any drug released in the body. Here, nanophase hydroxyapatite was synthesized in sizes ranging from 110-170 nm and was subsequently coated onto titanium samples using electrophoretic deposition. Results indicated that smaller nanoscale hydroxyapatite features on titanium surfaces alone decreased bacterial attachment in the presence of gram negative (P. aeruginosa), gram positive (S. aureus) and ampicillin resistant gram-negative (E. coli) bacteria as well as were able to control inflammatory responses; properties which should lead to their further investigation for improved medical applications.

  3. Expression of a bacterial 3-dehydroshikimate dehydratase reduces lignin content and improves biomass saccharification efficiency

    DOE PAGES

    Eudes, Aymerick; Sathitsuksanoh, Noppadon; Baidoo, Edward E. K.; ...

    2015-01-13

    Lignin confers recalcitrance to plant biomass used as feedstocks in agro-processing industries or as source of renewable sugars for the production of bioproducts. The metabolic steps for the synthesis of lignin building blocks belong to the shikimate and phenylpropanoid pathways. Genetic engineering efforts to reduce lignin content typically employ gene knockout or gene silencing techniques to constitutively repress one of these metabolic pathways. Recently, new strategies have emerged offering better spatiotemporal control of lignin deposition, including the expression of enzymes that interfere with the normal process for cell wall lignification. In this study, we report that expression of a 3-dehydroshikimatemore » dehydratase (QsuB from Corynebacterium glutamicum) reduces lignin deposition in Arabidopsis cell walls. QsuB was targeted to the plastids to convert 3-dehydroshikimate – an intermediate of the shikimate pathway – into protocatechuate. Compared to wild-type plants, lines expressing QsuB contain higher amounts of protocatechuate, p-coumarate, p-coumaraldehyde and p-coumaryl alcohol, and lower amounts of coniferaldehyde, coniferyl alcohol, sinapaldehyde and sinapyl alcohol. 2D-NMR spectroscopy and pyrolysis-gas chromatography/mass spectrometry (pyro-GC/MS) reveal an increase of p-hydroxyphenyl units and a reduction of guaiacyl units in the lignin of QsuB lines. Size-exclusion chromatography indicates a lower degree of lignin polymerization in the transgenic lines. Therefore, our data show that the expression of QsuB primarily affects the lignin biosynthetic pathway. Finally, biomass from these lines exhibits more than a twofold improvement in saccharification efficiency. We conclude that the expression of QsuB in plants, in combination with specific promoters, is a promising gain-of-function strategy for spatiotemporal reduction of lignin in plant biomass.« less

  4. Expression of a bacterial 3-dehydroshikimate dehydratase reduces lignin content and improves biomass saccharification efficiency.

    PubMed

    Eudes, Aymerick; Sathitsuksanoh, Noppadon; Baidoo, Edward E K; George, Anthe; Liang, Yan; Yang, Fan; Singh, Seema; Keasling, Jay D; Simmons, Blake A; Loqué, Dominique

    2015-12-01

    Lignin confers recalcitrance to plant biomass used as feedstocks in agro-processing industries or as source of renewable sugars for the production of bioproducts. The metabolic steps for the synthesis of lignin building blocks belong to the shikimate and phenylpropanoid pathways. Genetic engineering efforts to reduce lignin content typically employ gene knockout or gene silencing techniques to constitutively repress one of these metabolic pathways. Recently, new strategies have emerged offering better spatiotemporal control of lignin deposition, including the expression of enzymes that interfere with the normal process for cell wall lignification. In this study, we report that expression of a 3-dehydroshikimate dehydratase (QsuB from Corynebacterium glutamicum) reduces lignin deposition in Arabidopsis cell walls. QsuB was targeted to the plastids to convert 3-dehydroshikimate - an intermediate of the shikimate pathway - into protocatechuate. Compared to wild-type plants, lines expressing QsuB contain higher amounts of protocatechuate, p-coumarate, p-coumaraldehyde and p-coumaryl alcohol, and lower amounts of coniferaldehyde, coniferyl alcohol, sinapaldehyde and sinapyl alcohol. 2D-NMR spectroscopy and pyrolysis-gas chromatography/mass spectrometry (pyro-GC/MS) reveal an increase of p-hydroxyphenyl units and a reduction of guaiacyl units in the lignin of QsuB lines. Size-exclusion chromatography indicates a lower degree of lignin polymerization in the transgenic lines. Therefore, our data show that the expression of QsuB primarily affects the lignin biosynthetic pathway. Finally, biomass from these lines exhibits more than a twofold improvement in saccharification efficiency. We conclude that the expression of QsuB in plants, in combination with specific promoters, is a promising gain-of-function strategy for spatiotemporal reduction of lignin in plant biomass.

  5. Thin microelectrodes reduce GFAP expression in the implant site in rodent somatosensory cortex

    NASA Astrophysics Data System (ADS)

    Stice, Paula; Gilletti, Aaron; Panitch, Alyssa; Muthuswamy, Jit

    2007-06-01

    The objective of this study was to test the hypothesis that neural implants with reduced cross-sectional areas will have less glial scarring associated with implantation injury in long-term experiments. In this study, we implanted nine adult rats with two different implants of 12 µm (n = 6), and 25 µm (n = 6) diameters (cross-sectional areas of 68 µm2, 232 µm2 respectively) and the expression of glial fibrilliary acidic protein (GFAP) was assessed after 2 weeks and 4 weeks of implantation. In order to facilitate implantation, the 12 µm diameter implants were coated with poly-glycolic acid (PGA), a biodegradable polymer that degraded within minutes of implantation. In n = 3 animals, 25 µm diameter implants also coated with PGA were implanted and assessed for GFAP expression at the end of 4 weeks of implantation. Statistical analysis of the GFAP expression around the different implants demonstrated that after 2 weeks of implantation there is no statistically significant difference in GFAP expression between the 12 µm and the 25 µm diameter implants. However, after 4 weeks of implantation the implant site of 12 µm diameter implants exhibited a statistically significant reduction in GFAP expression when compared to the implant sites of the 25 µm diameter implants (both with and without the PGA coating). We conclude that in neural implants that are tethered to the skull, implant cross-sectional areas of 68 µm2 and smaller could lead to a reduced glial scarring under chronic conditions. Future studies with longer implant durations can confirm if this observation remains consistent beyond 4 weeks.

  6. Irradiation of rat brain reduces P-glycoprotein expression and function.

    PubMed

    Bart, J; Nagengast, W B; Coppes, R P; Wegman, T D; van der Graaf, W T A; Groen, H J M; Vaalburg, W; de Vries, E G E; Hendrikse, N H

    2007-08-06

    The blood-brain barrier (BBB) hampers delivery of several drugs including chemotherapeutics to the brain. The drug efflux pump P-glycoprotein (P-gp), expressed on brain capillary endothelial cells, is part of the BBB. P-gp expression on capillary endothelium decreases 5 days after brain irradiation, which may reduce P-gp function and increase brain levels of P-gp substrates. To elucidate whether radiation therapy reduces P-gp expression and function in the brain, right hemispheres of rats were irradiated with single doses of 2-25 Gy followed by 10 mg kg(-1) of the P-gp substrate cyclosporine A (CsA) intravenously (i.v.), with once 15 Gy followed by CsA (10, 15 or 20 mg kg(-1)), or with fractionated irradiation (4 x 5 Gy) followed by CsA (10 mg kg(-1)) 5 days later. Additionally, four groups of three rats received 25 Gy once and were killed 10, 15, 20 or 25 days later. The brains were removed and P-gp detected immunohistochemically. P-gp function was assessed by [(11)C]carvedilol uptake using quantitative autoradiography. Irradiation increased [(11)C]carvedilol uptake dose-dependently, to a maximum of 20% above non irradiated hemisphere. CsA increased [(11)C]carvedilol uptake dose-dependently in both hemispheres, but more (P<0.001) in the irradiated hemisphere. Fractionated irradiation resulted in a lost P-gp expression 10 days after start irradiation, which coincided with increased [(11)C]carvedilol uptake. P-gp expression decreased between day 15 and 20 after single dose irradiation, and increased again thereafter. Rat brain irradiation results in a temporary decreased P-gp function.

  7. T cell interleukin-15 surface expression in chimpanzees infected with human immunodeficiency virus.

    PubMed

    Rodriguez, Annette R; Hodara, Vida; Murthy, Kruthi; Morrow, LaShayla; Sanchez, Melissa; Bienvenu, Amy E; Murthy, Krishna K

    2014-01-01

    Interleukin-15 (IL-15) contributes to natural killer cell development and immune regulation. However, IL-15 and interferon-gamma (IFN-γ) production are significantly reduced during progression to AIDS. We have previously reported that HIV infected chimpanzees (Pan troglodytes) express CD3-CD8+ IFN-γ+ natural killer (NK) cells with an inverse correlation to plasma HIV viral load. To expand on our initial study, we examined a larger population of HIV infected chimpanzees (n=10). Whole blood flow cytometry analyses showed that recombinant gp120 (rgp120) or recombinant IL-15 induces specific CD3-CD8+ IFN-γ+ NK cells at higher levels than CD3+CD8+ IFN-γ+ T cells in HIV infected specimens. Interestingly, peripheral blood T cells exhibited 0.5-3% IL-15 surface Tcell/NKT cell phenotypes, and rIL-15 stimulation significantly (P<0.007) up-regulated CD4+CD25+ T cell expression. Importantly, these data demonstrate novel T cell interleukin-15 expression and indicate a plausible regulatory mechanism for this cell-type during viral infection.

  8. Reduced iNOS expression in adenoids from children with otitis media with effusion.

    PubMed

    Granath, Anna; Norrby-Teglund, Anna; Uddman, Rolf; Cardell, Lars-Olaf

    2010-12-01

    Nitric oxide (NO) is a key mediator in the local immune response of human airways. Inducible NO-synthases (iNOS), and endothelial NO-synthases (eNOS) are two enzymes known to regulate its production. The role of NO in middle ear disease is not fully known. Previous studies suggest that NO might have a dual role, both promoting and suppressing middle ear inflammation. The aim of the present study was to compare the eNOS and iNOS expression in adenoids obtained from children with otitis media with effusion (OME) with the expression seen in adenoids derived from children without middle ear disease. In addition, the expression of IL-1β and TNF-α were analyzed, because of their role in the iNOS-induction pathway. The iNOS and eNOS expression were analyzed with real-time PCR in 8 OME and 11 control adenoids. The corresponding proteins were demonstrated by immunohistochemical staining of adenoid tissue. A Luminex(®) assay was performed to analyze IL-1β and TNF-α in nasopharyngeal secretion in 10 OME and 8 controls, and immunohistochemistry was performed on adenoid tissue and imprints from the adenoid surface. Children with OME exhibited lower levels of iNOS than controls without middle ear disease. No such difference was seen for eNOS. The corresponding proteins were found mainly in conjunction with surface epithelium. No significant changes were seen among the cytokines tested. The present results indicate that local induction of iNOS in adenoids might be of importance for preventing development of OME. © 2010 John Wiley & Sons A/S.

  9. Drought response transcriptomes are altered in poplar with reduced tonoplast sucrose transporter expression

    PubMed Central

    Xue, Liang-Jiao; Frost, Christopher J.; Tsai, Chung-Jui; Harding, Scott A.

    2016-01-01

    Transgenic Populus tremula x alba (717-1B4) plants with reduced expression of a tonoplast sucrose efflux transporter, PtaSUT4, exhibit reduced shoot growth compared to wild type (WT) under sustained mild drought. The present study was undertaken to determine whether SUT4-RNAi directly or indirectly altered poplar predisposition and/or response to changes in soil water availability. While sucrose and hexose levels were constitutively elevated in shoot organs, expression responses to drought were most altered in the root tips of SUT4-RNAi plants. Prior to any drought treatment, constitutively elevated transcript levels of abscisic acid biosynthetic genes and bark/vegetative storage proteins suggested altered metabolism in root tips of RNAi plants. Stronger drought-stimulation of stress-inducible genes encoding late-embryogenesis-abundant proteins in transgenic roots was consistent with increased vulnerability to soil drying. Transcript evidence suggested an RNAi effect on intercellular water trafficking by aquaporins in stem xylem during soil drying and recovery. Co-expression network analysis predicted altered integration of abscisic acid sensing/signaling with ethylene and jasmonate sensing/signaling in RNAi compared to WT roots. The overall conclusion is that steepened shoot-root sugar gradient in RNAi plants increased sensitivity of root tips to decreasing soil water availability. PMID:27641356

  10. Intermittent compressive strain may reduce aggrecanase expression in cartilage: a study of chondrocytes in agarose gel.

    PubMed

    Mio, Kensuke; Saito, Seiji; Tomatsu, Taisuke; Toyama, Yoshiaki

    2005-04-01

    We tested the hypothesis that intermittent compressive strain reduces the catabolic actions of interleukin-1 beta on chondrocyte metabolism at the gene expression level. We investigated the effects of intermittent compressive strain on mRNA expression in bovine chondrocytes cultured in agarose gel supplemented with or without interleukin-1 beta. Fifteen percent compressive strain amplitude was applied to agarose-chondrocyte constructs at a frequency of 1 Hz. In the absence of interleukin-1 beta, the strain caused an increase in the mRNA levels of anabolic factors like aggrecan and Type II collagen, compared with the levels of anabolic factors in unstrained constructs. These results suggest that the strain may stimulate matrix production in normal cartilage. Interleukin-1 beta is a powerful catabolic agent; in unstrained agarose-chondrocyte constructs, interleukin-1 beta caused a decrease in the mRNA levels of anabolic factors. However, interleukin-1 beta also caused an increase in the mRNA of catabolic factors like aggrecanase-1, aggrecanase-2, and matrix metalloproteinase-3. In the presence of interleukin-1 beta, the strain reduced the mRNA levels of aggrecanase-1 and aggrecanase-2. These results suggest that intermittent compressive strain may protect cartilage by suppressing the expression of aggrecanase-1 and aggrecanase-2, which are thought to be the major matrix-degrading enzymes responsible for cleaving aggrecan.

  11. Semaphorin 3F expression is reduced in pregnancy complicated by preeclampsia. An observational clinical study.

    PubMed

    Stallone, Giovanni; Matteo, Maria; Netti, Giuseppe Stefano; Infante, Barbara; Di Lorenzo, Adelaide; Prattichizzo, Clelia; Carlucci, Stefania; Trezza, Federica; Gesualdo, Loreto; Greco, Pantaleo; Grandaliano, Giuseppe

    2017-01-01

    Preeclampsia is a systemic disorder, affecting 2-10% of pregnancies, characterized by a deregulated pro- and anti-angiogenic balance. Semaphorin 3F is an angiogenesis inhibitor. We aimed to investigate whether semaphorin 3F expression is modulated in preeclampsia. We performed two observational single center cohort studies between March 2013 and August 2014. In the first we enrolled 110 consecutive women, undergoing an elective cesarean section; in the second we included 150 consecutive women undergoing amniocentesis for routine clinical indications at 16-18 week of gestation. Semaphorin 3F concentration was evaluated in maternal peripheral blood, venous umbilical blood and amniotic fluid, along with its placenta protein expression at the time of delivery in the first study group and in the amniotic fluid at 16-18 weeks of gestation in the second study group. In the first study 19 patients presented at delivery with preeclampsia. Semaphorin 3F placenta tissue expression was significantly reduced in preeclampsia. In addition, semaphorin 3F level at delivery was significantly lower in serum, amniotic fluid and venous umbilical blood of preeclamptic patients compared with normal pregnant women. In the prospective cohort study 14 women developed preeclampsia. In this setting, semaphorin 3F amniotic level at 16-18 weeks of gestation was reduced in women who subsequently developed preeclampsia compared to women with a normal pregnancy. ROC curve analysis showed that semaphorin 3F amniotic levels could identify women at higher risk of preeclampsia. Semaphorin 3F might represent a predictive biomarker of preeclampsia.

  12. Reduced effects of thyroid hormone on gene expression and metamorphosis in a paedomorphic plethodontid salamander.

    PubMed

    Aran, Robert P; Steffen, Michael A; Martin, Samuel D; Lopez, Olivia I; Bonett, Ronald M

    2014-07-01

    It has been over a century since Gudernatsch (1912, Wilhelm Roux Arch Entwickl Mech Org 35:457-483) demonstrated that mammalian thyroid gland extracts can stimulate tadpole metamorphosis. Despite the tremendous developmental diversity of amphibians, mechanisms of metamorphosis have mostly been studied in a few model systems. This limits our understanding of the processes that influence the evolution of developmental aberrations. Here we isolated thyroid hormone receptors alpha (TRα) and beta (TRβ) from Oklahoma salamanders (Eurycea tynerensis), which exhibit permanently aquatic (paedomorphic) or biphasic (metamorphic) developmental modes in different populations. We found that TRα and TRβ were upregulated by thyroid hormone (T3 ) in tail tissues of larvae from metamorphic populations, but basal levels of TR expression and T3 responsiveness were reduced in larvae from paedomorphic populations. Likewise, we found that T3 treatment resulted in complete loss of larval epibranchials in larvae from metamorphic populations, but little to no epibranchial remodeling occurred in larvae from paedomorphic populations over the same duration. This is the first study to directly demonstrate reduced gene expression and metamorphic responses to T3 in a paedomorphic plethodontid compared to metamorphic conspecifics, and the first salamander system to show differential expression of thyroid hormone receptors associated with alternative developmental patterns.

  13. Reduced myelin basic protein and actin-related gene expression in visual cortex in schizophrenia.

    PubMed

    Matthews, Paul R; Eastwood, Sharon L; Harrison, Paul J

    2012-01-01

    Most brain gene expression studies of schizophrenia have been conducted in the frontal cortex or hippocampus. The extent to which alterations occur in other cortical regions is not well established. We investigated primary visual cortex (Brodmann area 17) from the Stanley Neuropathology Consortium collection of tissue from 60 subjects with schizophrenia, bipolar disorder, major depression, or controls. We first carried out a preliminary array screen of pooled RNA, and then used RT-PCR to quantify five mRNAs which the array identified as differentially expressed in schizophrenia (myelin basic protein [MBP], myelin-oligodendrocyte glycoprotein [MOG], β-actin [ACTB], thymosin β-10 [TB10], and superior cervical ganglion-10 [SCG10]). Reduced mRNA levels were confirmed by RT-PCR for MBP, ACTB and TB10. The MBP reduction was limited to transcripts containing exon 2. ACTB and TB10 mRNAs were also decreased in bipolar disorder. None of the transcripts were altered in subjects with major depression. Reduced MBP mRNA in schizophrenia replicates findings in other brain regions and is consistent with oligodendrocyte involvement in the disorder. The decreases in expression of ACTB, and the actin-binding protein gene TB10, suggest changes in cytoskeletal organisation. The findings confirm that the primary visual cortex shows molecular alterations in schizophrenia and extend the evidence for a widespread, rather than focal, cortical pathophysiology.

  14. Side-stream smoking reduces intestinal inflammation and increases expression of tight junction proteins

    PubMed Central

    Wang, Hui; Zhao, Jun-Xing; Hu, Nan; Ren, Jun; Du, Min; Zhu, Mei-Jun

    2012-01-01

    AIM: To investigate the effect of side-stream smoking on gut microflora composition, intestinal inflammation and expression of tight junction proteins. METHODS: C57BL/6 mice were exposed to side-stream cigarette smoking for one hour daily over eight weeks. Cecal contents were collected for microbial composition analysis. Large intestine was collected for immunoblotting and quantitative reverse transcriptase polymerase chain reaction analyses of the inflammatory pathway and tight junction proteins. RESULTS: Side-stream smoking induced significant changes in the gut microbiota with increased mouse intestinal bacteria, Clostridium but decreased Fermicutes (Lactoccoci and Ruminococcus), Enterobacteriaceae family and Segmented filamentous baceteria compared to the control mice. Meanwhile, side-stream smoking inhibited the nuclear factor-κB pathway with reduced phosphorylation of p65 and IκBα, accompanied with unchanged mRNA expression of tumor necrosis factor-α or interleukin-6. The contents of tight junction proteins, claudin3 and ZO2 were up-regulated in the large intestine of mice exposed side-stream smoking. In addition, side-stream smoking increased c-Jun N-terminal kinase and p38 MAPK kinase signaling, while inhibiting AMP-activated protein kinase in the large intestine. CONCLUSION: Side-stream smoking altered gut microflora composition and reduced the inflammatory response, which was associated with increased expression of tight junction proteins. PMID:22611310

  15. Drought response transcriptomes are altered in poplar with reduced tonoplast sucrose transporter expression.

    PubMed

    Xue, Liang-Jiao; Frost, Christopher J; Tsai, Chung-Jui; Harding, Scott A

    2016-09-19

    Transgenic Populus tremula x alba (717-1B4) plants with reduced expression of a tonoplast sucrose efflux transporter, PtaSUT4, exhibit reduced shoot growth compared to wild type (WT) under sustained mild drought. The present study was undertaken to determine whether SUT4-RNAi directly or indirectly altered poplar predisposition and/or response to changes in soil water availability. While sucrose and hexose levels were constitutively elevated in shoot organs, expression responses to drought were most altered in the root tips of SUT4-RNAi plants. Prior to any drought treatment, constitutively elevated transcript levels of abscisic acid biosynthetic genes and bark/vegetative storage proteins suggested altered metabolism in root tips of RNAi plants. Stronger drought-stimulation of stress-inducible genes encoding late-embryogenesis-abundant proteins in transgenic roots was consistent with increased vulnerability to soil drying. Transcript evidence suggested an RNAi effect on intercellular water trafficking by aquaporins in stem xylem during soil drying and recovery. Co-expression network analysis predicted altered integration of abscisic acid sensing/signaling with ethylene and jasmonate sensing/signaling in RNAi compared to WT roots. The overall conclusion is that steepened shoot-root sugar gradient in RNAi plants increased sensitivity of root tips to decreasing soil water availability.

  16. Vitamin D improves endothelial dysfunction and restores myeloid angiogenic cell function via reduced CXCL-10 expression in systemic lupus erythematosus.

    PubMed

    Reynolds, John A; Haque, Sahena; Williamson, Kate; Ray, David W; Alexander, M Yvonne; Bruce, Ian N

    2016-03-01

    Patients with systemic lupus erythematosus (SLE) have accelerated cardiovascular disease and dysfunctional endothelial repair mechanisms. Myeloid angiogenic cells (MACs), derived from circulating monocytes, augment vascular repair by paracrine secretion of pro-angiogenic factors. We observed that SLE MACs are dysfunctional and secrete pro-inflammatory cytokines. We also found that the vitamin D receptor was transiently expressed during MAC differentiation and that in vitro, calcitriol increased differentiation of monocytes into MACs in both SLE and in a model using the prototypic SLE cytokine, interferon-alpha. The active form of vitamin D (calcitriol) restored the SLE MAC phenotype towards that of healthy subjects with reduced IL-6 secretion, and normalised surface marker expression. Calcitriol also augmented the angiogenic capacity of MACs via the down-regulation of CXCL-10. In SLE patients treated with cholecalciferol for 12 weeks, the improvement in endothelial function correlated with increase in serum 25(OH)D concentrations independently of disease activity. We also show that MACs were able to positively modulate eNOS expression in human endothelial cells in vitro, an effect further enhanced by calcitriol treatment of SLE MACs. The results demonstrate that vitamin D can positively modify endothelial repair mechanisms and thus endothelial function in a population with significant cardiovascular risk.

  17. Vitamin D improves endothelial dysfunction and restores myeloid angiogenic cell function via reduced CXCL-10 expression in systemic lupus erythematosus

    PubMed Central

    Reynolds, John A.; Haque, Sahena; Williamson, Kate; Ray, David W.; Alexander, M. Yvonne; Bruce, Ian N.

    2016-01-01

    Patients with systemic lupus erythematosus (SLE) have accelerated cardiovascular disease and dysfunctional endothelial repair mechanisms. Myeloid angiogenic cells (MACs), derived from circulating monocytes, augment vascular repair by paracrine secretion of pro-angiogenic factors. We observed that SLE MACs are dysfunctional and secrete pro-inflammatory cytokines. We also found that the vitamin D receptor was transiently expressed during MAC differentiation and that in vitro, calcitriol increased differentiation of monocytes into MACs in both SLE and in a model using the prototypic SLE cytokine, interferon-alpha. The active form of vitamin D (calcitriol) restored the SLE MAC phenotype towards that of healthy subjects with reduced IL-6 secretion, and normalised surface marker expression. Calcitriol also augmented the angiogenic capacity of MACs via the down-regulation of CXCL-10. In SLE patients treated with cholecalciferol for 12 weeks, the improvement in endothelial function correlated with increase in serum 25(OH)D concentrations independently of disease activity. We also show that MACs were able to positively modulate eNOS expression in human endothelial cells in vitro, an effect further enhanced by calcitriol treatment of SLE MACs. The results demonstrate that vitamin D can positively modify endothelial repair mechanisms and thus endothelial function in a population with significant cardiovascular risk. PMID:26930567

  18. Reducing the surface roughness of dental acrylic resins by using an eggshell abrasive material.

    PubMed

    Onwubu, Stanley C; Vahed, Anisa; Singh, Shalini; Kanny, Krishnan M

    2017-02-01

    Excessive surface roughness of denture base resins adversely impacts oral health. The purpose of this in vitro study was to examine the abrasive potential of eggshell powder in reducing the surface roughness of denture base resins. Thirty poly(methyl methacrylate) specimens were fabricated and polished with eggshell powders of different particle sizes and with pumice. The average surface roughness (Ra) after polishing was measured with a profilometer. Scanning electron microscope and optical electron microscope techniques were used to assess the surface roughness morphology of the specimens. ANOVA was used to analyze the Ra values. The Tukey honest significant differences and Bonferroni tests were used to identify differences between the 2 abrasive materials (α=.05). Significant differences in the Ra values were observed between the fine and medium eggshell powder abrasives (P<.05). Similarly, significant differences were found between pumice and the fine eggshell powder abrasives (P<.001). No significant differences were found between pumice and the medium eggshell powder abrasive (P>.05). Specimens polished with pumice had the highest Ra values, whereas specimens polished with the fine eggshell powder abrasive had the lowest Ra values. By connecting the Ra values to the threshold limit value of 0.2 μm, eggshell powder abrasive finished denture acrylic resin surfaces better than pumice. Copyright © 2016 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.

  19. Morphology of gas cavities on patterned hydrophobic surfaces under reduced pressure

    NASA Astrophysics Data System (ADS)

    Xue, Yahui; Lv, Pengyu; Liu, Ying; Shi, Yipeng; Lin, Hao; Duan, Huiling

    2015-09-01

    Gas cavities trapped on structured hydrophobic surfaces play important roles in realizing functionalities such as superhydrophobicity, drag reduction, and surface cleaning. The morphology of the cavities exhibits strong dependence on system parameters which impact the performance of these surfaces. In this work, a complete theoretical analysis is presented to predict cavity morphological change under reduced liquid pressure, on a submerged hydrophobic surface patterned with cylindrical pores. Equilibrium solutions are derived for five different phases, namely, (I) pinned recession, (II) depinned recession, (III) Cassie-Baxter, (IV) expansion, and (V) coalescence; their stabilities are also analyzed. A phase map is developed outlining the different regimes with respect to the gas amount and liquid pressure. Importantly, phase (IV) exhibits a complex stability behavior that leads to two possible routes to coalescence, which lends two different mechanisms of cavitation. Accordingly, the threshold pressure for cavitation can be calculated. The theoretical model is supported by direct experimental measurements via confocal microscopy and demonstrates good quantitative accuracy. This work provides a predictive tool for the design of functional structured hydrophobic surfaces.

  20. MCT8 expression in human fetal cerebral cortex is reduced in severe intrauterine growth restriction.

    PubMed

    Chan, Shiao Y; Hancox, Laura A; Martín-Santos, Azucena; Loubière, Laurence S; Walter, Merlin N M; González, Ana-Maria; Cox, Phillip M; Logan, Ann; McCabe, Christopher J; Franklyn, Jayne A; Kilby, Mark D

    2014-02-01

    The importance of the thyroid hormone (TH) transporter, monocarboxylate transporter 8 (MCT8), to human neurodevelopment is highlighted by findings of severe global neurological impairment in subjects with MCT8 (SLC16A2) mutations. Intrauterine growth restriction (IUGR), usually due to uteroplacental failure, is associated with milder neurodevelopmental deficits, which have been partly attributed to dysregulated TH action in utero secondary to reduced circulating fetal TH concentrations and decreased cerebral thyroid hormone receptor expression. We postulate that altered MCT8 expression is implicated in this pathophysiology; therefore, in this study, we sought to quantify changes in cortical MCT8 expression with IUGR. First, MCT8 immunohistochemistry was performed on occipital and parietal cerebral cortex sections obtained from appropriately grown for gestational age (AGA) human fetuses between 19 weeks of gestation and term. Secondly, MCT8 immunostaining in the occipital cortex of stillborn IUGR human fetuses at 24-28 weeks of gestation was objectively compared with that in the occipital cortex of gestationally matched AGA fetuses. Fetuses demonstrated widespread MCT8 expression in neurons within the cortical plate and subplate, in the ventricular and subventricular zones, in the epithelium of the choroid plexus and ependyma, and in microvessel wall. When complicated by IUGR, fetuses showed a significant fivefold reduction in the percentage area of cortical plate immunostained for MCT8 compared with AGA fetuses (P<0.05), but there was no significant difference in the proportion of subplate microvessels immunostained. Cortical MCT8 expression was negatively correlated with the severity of IUGR indicated by the brain:liver weight ratios (r(2)=0.28; P<0.05) at post-mortem. Our results support the hypothesis that a reduction in MCT8 expression in the IUGR fetal brain could further compromise TH-dependent brain development.

  1. MCT8 expression in human fetal cerebral cortex is reduced in severe intrauterine growth restriction

    PubMed Central

    Chan, Shiao Y; Hancox, Laura A; Martín-Santos, Azucena; Loubière, Laurence S; Walter, Merlin N M; González, Ana-Maria; Cox, Phillip M; Logan, Ann; McCabe, Christopher J; Franklyn, Jayne A; Kilby, Mark D

    2014-01-01

    The importance of the thyroid hormone (TH) transporter, monocarboxylate transporter 8 (MCT8), to human neurodevelopment is highlighted by findings of severe global neurological impairment in subjects with MCT8 (SLC16A2) mutations. Intrauterine growth restriction (IUGR), usually due to uteroplacental failure, is associated with milder neurodevelopmental deficits, which have been partly attributed to dysregulated TH action in utero secondary to reduced circulating fetal TH concentrations and decreased cerebral thyroid hormone receptor expression. We postulate that altered MCT8 expression is implicated in this pathophysiology; therefore, in this study, we sought to quantify changes in cortical MCT8 expression with IUGR. First, MCT8 immunohistochemistry was performed on occipital and parietal cerebral cortex sections obtained from appropriately grown for gestational age (AGA) human fetuses between 19 weeks of gestation and term. Secondly, MCT8 immunostaining in the occipital cortex of stillborn IUGR human fetuses at 24–28 weeks of gestation was objectively compared with that in the occipital cortex of gestationally matched AGA fetuses. Fetuses demonstrated widespread MCT8 expression in neurons within the cortical plate and subplate, in the ventricular and subventricular zones, in the epithelium of the choroid plexus and ependyma, and in microvessel wall. When complicated by IUGR, fetuses showed a significant fivefold reduction in the percentage area of cortical plate immunostained for MCT8 compared with AGA fetuses (P<0.05), but there was no significant difference in the proportion of subplate microvessels immunostained. Cortical MCT8 expression was negatively correlated with the severity of IUGR indicated by the brain:liver weight ratios (r2=0.28; P<0.05) at post-mortem. Our results support the hypothesis that a reduction in MCT8 expression in the IUGR fetal brain could further compromise TH-dependent brain development. PMID:24204008

  2. Blood-Gene Expression Reveals Reduced Circadian Rhythmicity in Individuals Resistant to Sleep Deprivation

    PubMed Central

    Arnardottir, Erna S.; Nikonova, Elena V.; Shockley, Keith R.; Podtelezhnikov, Alexei A.; Anafi, Ron C.; Tanis, Keith Q.; Maislin, Greg; Stone, David J.; Renger, John J.; Winrow, Christopher J.; Pack, Allan I.

    2014-01-01

    Study Objectives: To address whether changes in gene expression in blood cells with sleep loss are different in individuals resistant and sensitive to sleep deprivation. Design: Blood draws every 4 h during a 3-day study: 24-h normal baseline, 38 h of continuous wakefulness and subsequent recovery sleep, for a total of 19 time-points per subject, with every 2-h psychomotor vigilance task (PVT) assessment when awake. Setting: Sleep laboratory. Participants: Fourteen subjects who were previously identified as behaviorally resistant (n = 7) or sensitive (n = 7) to sleep deprivation by PVT. Intervention: Thirty-eight hours of continuous wakefulness. Measurements and Results: We found 4,481 unique genes with a significant 24-h diurnal rhythm during a normal sleep-wake cycle in blood (false discovery rate [FDR] < 5%). Biological pathways were enriched for biosynthetic processes during sleep. After accounting for circadian effects, two genes (SREBF1 and CPT1A, both involved in lipid metabolism) exhibited small, but significant, linear changes in expression with the duration of sleep deprivation (FDR < 5%). The main change with sleep deprivation was a reduction in the amplitude of the diurnal rhythm of expression of normally cycling probe sets. This reduction was noticeably higher in behaviorally resistant subjects than sensitive subjects, at any given P value. Furthermore, blood cell type enrichment analysis showed that the expression pattern difference between sensitive and resistant subjects is mainly found in cells of myeloid origin, such as monocytes. Conclusion: Individual differences in behavioral effects of sleep deprivation are associated with differences in diurnal amplitude of gene expression for genes that show circadian rhythmicity. Citation: Arnardottir ES, Nikonova EV, Shockley KR, Podtelezhnikov AA, Anafi RC, Tanis KQ, Maislin G, Stone DJ, Renger JJ, Winrow CJ, Pack AI. Blood-gene expression reveals reduced circadian rhythmicity in individuals resistant to

  3. Reduced IL-37 Production Increases Spontaneous Chemokine Expressions in Colon Epithelial Cells.

    PubMed

    Günaltay, Sezin; Ghiboub, Mohammed; Hultgren, Olof; Hörnquist, Elisabeth Hultgren

    2017-05-01

    Microscopic colitis, comprising collagenous colitis and lymphocytic colitis, is a common cause of chronic diarrhea. Previously, we showed enhanced chemokine productions in microscopic colitis patients, indicating dysregulated immune cell chemotaxis in the immunopathogenesis. We also showed decreased mRNA of IL-37, mainly regarded as an anti-inflammatory cytokine, in the colonic mucosa of these patients, potentially an important factor for the chronicity of the colitis. Our aim in this study was to understand the possible role of IL-37 in chemokine production using a cell line model. A colon epithelial cell line, T84, was stimulated with the TLR5 ligand flagellin. IL-37 protein production was reduced 20% using the CRISPR/Cas9 system, and the changes in chemokine mRNA and protein expressions were compared to cells transfected with empty plasmid. The 20% reduction in IL-37 protein levels spontaneously increased CCL5, CXCL8, CXCL10, and CXCL11 mRNA and protein expressions. CCL2 mRNA and protein levels were enhanced upon TLR5 stimulation. CCL3, CCL20, and CX3CL1 mRNA expressions were increased either spontaneously or following TLR5 stimulation, whereas CCL4 and CCL22 mRNA expressions were significantly decreased. Even a minor decrease in the ability of colon epithelial cells to produce IL-37 results in altered chemokine expression, mainly an increase in the production of several chemokines. Our results indicate that a decreased IL-37 expression by colon epithelial cells may be an important factor for increasing the recruitment of immune cells and subsequently developing microscopic colitis.

  4. Reducing phosphorus loading of surface water using iron-coated sand.

    PubMed

    Groenenberg, Jan E; Chardon, Wim J; Koopmans, Gerwin F

    2013-01-01

    Phosphorus losses from agricultural soils is an important source of P in surface waters leading to surface water quality impairment. In addition to reducing P inputs, mitigation measures are needed to reduce P enrichment of surface waters. Because drainage of agricultural land by pipe drainage is an important pathway of P to surface waters, removing P from drainage water has a large potential to reduce P losses. In a field trial, we tested the performance of a pipe drain enveloped with Fe-coated sand, a side product of the drinking water industry with a high ability to bind P, to remove P from the drainage water. The results of this trial, encompassing more than one hydrological season, are very encouraging because the efficiency of this mitigation measure to remove P amounted to 94%. During the trial, the pipe drains were below the groundwater level for a prolonged time. Nevertheless, no reduction of Fe(III) in the Fe-coated sand occurred, which was most likely prevented by reduction of Mn oxides present in this material. The enveloped pipe drain was estimated to be able to lower the P concentration in the effluent to the desired water quality criterion for about 14 yr. Manganese oxides are expected to be depleted after 5 to 10 yr. The performance of the enveloped pipe drain, both in terms of its ability to remove P to a sufficiently low level and the stability of the Fe-coated sand under submerged conditions in the long term, needs prolonged experimental research. Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.

  5. Hypoxia Reduces the Pathogenicity of Pseudomonas aeruginosa by Decreasing the Expression of Multiple Virulence Factors.

    PubMed

    Schaible, Bettina; Rodriguez, Javier; Garcia, Amaya; von Kriegsheim, Alexander; McClean, Siobhán; Hickey, Caitríona; Keogh, Ciara E; Brown, Eric; Schaffer, Kirsten; Broquet, Alexis; Taylor, Cormac T

    2017-05-01

    Our understanding of how the course of opportunistic bacterial infection is influenced by the microenvironment is limited. We demonstrate that the pathogenicity of Pseudomonas aeruginosa strains derived from acute clinical infections is higher than that of strains derived from chronic infections, where tissues are hypoxic. Exposure to hypoxia attenuated the pathogenicity of strains from acute (but not chronic) infections, implicating a role for hypoxia in regulating bacterial virulence. Mass spectrometric analysis of the secretome of P. aeruginosa derived from an acute infection revealed hypoxia-induced repression of multiple virulence factors independent of altered bacterial growth. Pseudomonas aeruginosa lacking the Pseudomonas prolyl-hydroxylase domain-containing protein, which has been implicated in bacterial oxygen sensing, displays reduced virulence factor expression. Furthermore, pharmacological hydroxylase inhibition reduces virulence factor expression and pathogenicity in a murine model of pneumonia. We hypothesize that hypoxia reduces P. aeruginosa virulence at least in part through the regulation of bacterial hydroxylases. © The Author 2017. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.

  6. Lactobacilli Reduce Helicobacter pylori Attachment to Host Gastric Epithelial Cells by Inhibiting Adhesion Gene Expression

    PubMed Central

    de Klerk, Nele; Maudsdotter, Lisa; Gebreegziabher, Hanna; Saroj, Sunil D.; Eriksson, Beatrice; Eriksson, Olaspers Sara; Roos, Stefan; Lindén, Sara; Sjölinder, Hong

    2016-01-01

    The human gastrointestinal tract, including the harsh environment of the stomach, harbors a large variety of bacteria, of which Lactobacillus species are prominent members. The molecular mechanisms by which species of lactobacilli interfere with pathogen colonization are not fully characterized. In this study, we aimed to study the effect of lactobacillus strains upon the initial attachment of Helicobacter pylori to host cells. Here we report a novel mechanism by which lactobacilli inhibit adherence of the gastric pathogen H. pylori. In a screen with Lactobacillus isolates, we found that only a few could reduce adherence of H. pylori to gastric epithelial cells. Decreased attachment was not due to competition for space or to lactobacillus-mediated killing of the pathogen. Instead, we show that lactobacilli act on H. pylori directly by an effector molecule that is released into the medium. This effector molecule acts on H. pylori by inhibiting expression of the adhesin-encoding gene sabA. Finally, we verified that inhibitory lactobacilli reduced H. pylori colonization in an in vivo model. In conclusion, certain Lactobacillus strains affect pathogen adherence by inhibiting sabA expression and thereby reducing H. pylori binding capacity. PMID:26930708

  7. Lactobacilli Reduce Helicobacter pylori Attachment to Host Gastric Epithelial Cells by Inhibiting Adhesion Gene Expression.

    PubMed

    de Klerk, Nele; Maudsdotter, Lisa; Gebreegziabher, Hanna; Saroj, Sunil D; Eriksson, Beatrice; Eriksson, Olaspers Sara; Roos, Stefan; Lindén, Sara; Sjölinder, Hong; Jonsson, Ann-Beth

    2016-05-01

    The human gastrointestinal tract, including the harsh environment of the stomach, harbors a large variety of bacteria, of which Lactobacillus species are prominent members. The molecular mechanisms by which species of lactobacilli interfere with pathogen colonization are not fully characterized. In this study, we aimed to study the effect of lactobacillus strains upon the initial attachment of Helicobacter pylori to host cells. Here we report a novel mechanism by which lactobacilli inhibit adherence of the gastric pathogen H. pylori In a screen with Lactobacillus isolates, we found that only a few could reduce adherence of H. pylori to gastric epithelial cells. Decreased attachment was not due to competition for space or to lactobacillus-mediated killing of the pathogen. Instead, we show that lactobacilli act on H. pylori directly by an effector molecule that is released into the medium. This effector molecule acts on H. pylori by inhibiting expression of the adhesin-encoding gene sabA Finally, we verified that inhibitory lactobacilli reduced H. pylori colonization in an in vivo model. In conclusion, certain Lactobacillus strains affect pathogen adherence by inhibiting sabA expression and thereby reducing H. pylori binding capacity.

  8. Nuclear protein import is reduced in cells expressing nuclear envelopathy-causing lamin A mutants

    SciTech Connect

    Busch, Albert; Kiel, Tilman; Heupel, Wolfgang-M.; Wehnert, Manfred; Huebner, Stefan

    2009-08-15

    Lamins, which form the nuclear lamina, not only constitute an important determinant of nuclear architecture, but additionally play essential roles in many nuclear functions. Mutations in A-type lamins cause a wide range of human genetic disorders (laminopathies). The importance of lamin A (LaA) in the spatial arrangement of nuclear pore complexes (NPCs) prompted us to study the role of LaA mutants in nuclear protein transport. Two mutants, causing prenatal skin disease restrictive dermopathy (RD) and the premature aging disease Hutchinson Gilford progeria syndrome, were used for expression in HeLa cells to investigate their impact on the subcellular localization of NPC-associated proteins and nuclear protein import. Furthermore, dynamics of the LaA mutants within the nuclear lamina were studied. We observed affected localization of NPC-associated proteins, diminished lamina dynamics for both LaA mutants and reduced nuclear import of representative cargo molecules. Intriguingly, both LaA mutants displayed similar effects on nuclear morphology and functions, despite their differences in disease severity. Reduced nuclear protein import was also seen in RD fibroblasts and impaired lamina dynamics for the nucleoporin Nup153. Our data thus represent the first study of a direct link between LaA mutant expression and reduced nuclear protein import.

  9. The Inflammatory Cytokines TWEAK and TNFα Reduce Renal Klotho Expression through NFκB

    PubMed Central

    Moreno, Juan A.; Izquierdo, Maria C.; Sanchez-Niño, Maria D.; Suárez-Alvarez, Beatriz; Lopez-Larrea, Carlos; Jakubowski, Aniela; Blanco, Julia; Ramirez, Rafael; Selgas, Rafael; Ruiz-Ortega, Marta; Egido, Jesus; Sanz, Ana B.

    2011-01-01

    Proinflammatory cytokines contribute to renal injury, but the downstream effectors within kidney cells are not well understood. One candidate effector is Klotho, a protein expressed by renal cells that has antiaging properties; Klotho-deficient mice have an accelerated aging-like phenotype, including vascular injury and renal injury. Whether proinflammatory cytokines, such as TNF and TNF-like weak inducer of apoptosis (TWEAK), modulate Klotho is unknown. In mice, exogenous administration of TWEAK decreased expression of Klotho in the kidney. In the setting of acute kidney injury induced by folic acid, the blockade or absence of TWEAK abrogated the injury-related decrease in renal and plasma Klotho levels. TWEAK, TNFα, and siRNA-mediated knockdown of IκBα all activated NFκB and reduced Klotho expression in the MCT tubular cell line. Furthermore, inhibition of NFκB with parthenolide prevented TWEAK- or TNFα-induced downregulation of Klotho. Inhibition of histone deacetylase reversed TWEAK-induced downregulation of Klotho, and chromatin immunoprecipitation showed that TWEAK promotes RelA binding to the Klotho promoter, inducing its deacetylation. In conclusion, inflammatory cytokines, such as TWEAK and TNFα, downregulate Klotho expression through an NFκB-dependent mechanism. These results may partially explain the relationship between inflammation and diseases characterized by accelerated aging of organs, including CKD. PMID:21719790

  10. Decreased adrenoceptor stimulation in heart failure rats reduces NGF expression by cardiac parasympathetic neurons.

    PubMed

    Hasan, Wohaib; Smith, Peter G

    2014-04-01

    Postganglionic cardiac parasympathetic and sympathetic nerves are physically proximate in atrial cardiac tissue allowing reciprocal inhibition of neurotransmitter release, depending on demands from central cardiovascular centers or reflex pathways. Parasympathetic cardiac ganglion (CG) neurons synthesize and release the sympathetic neurotrophin nerve growth factor (NGF), which may serve to maintain these close connections. In this study we investigated whether NGF synthesis by CG neurons is altered in heart failure, and whether norepinephrine from sympathetic neurons promotes NGF synthesis. NGF and proNGF immunoreactivity in CG neurons in heart failure rats following chronic coronary artery ligation was investigated. NGF immunoreactivity was decreased significantly in heart failure rats compared to sham-operated animals, whereas proNGF expression was unchanged. Changes in neurochemistry of CG neurons included attenuated expression of the cholinergic marker vesicular acetylcholine transporter, and increased expression of the neuropeptide vasoactive intestinal polypeptide. To further investigate norepinephrine's role in promoting NGF synthesis, we cultured CG neurons treated with adrenergic receptor (AR) agonists. An 82% increase in NGF mRNA levels was detected after 1h of isoproterenol (β-AR agonist) treatment, which increased an additional 22% at 24h. Antagonist treatment blocked isoproterenol-induced increases in NGF transcripts. In contrast, the α-AR agonist phenylephrine did not alter NGF mRNA expression. These results are consistent with β-AR mediated maintenance of NGF synthesis in CG neurons. In heart failure, a decrease in NGF synthesis by CG neurons may potentially contribute to reduced connections with adjacent sympathetic nerves.

  11. Preconditioning reduces tissue complement gene expression in the rabbit isolated heart.

    PubMed

    Tanhehco, E J; Yasojima, K; McGeer, P L; Washington, R A; Kilgore, K S; Homeister, J W; Lucchesi, B R

    1999-12-01

    Both preconditioning and inhibition of complement activation have been shown to ameliorate myocardial ischemia-reperfusion injury. The recent demonstration that myocardial tissue expresses complement components led us to investigate whether preconditioning affects complement expression in the isolated heart. Hearts from New Zealand White rabbits were exposed to either two rounds of 5 min global ischemia followed by 10 min reperfusion (ischemic preconditioning) or 10 microM of the ATP-dependent K+ (KATP) channel opener pinacidil for 30 min (chemical preconditioning) before induction of 30 min global ischemia followed by 60 min of reperfusion. Both ischemic and chemical preconditioning significantly (P < 0.05) reduced myocardial C1q, C1r, C3, C8, and C9 mRNA levels. Western blot and immunohistochemistry demonstrated a similar reduction in C3 and membrane attack complex protein expression. The K(ATP) channel blocker glyburide (10 microM) reversed the depression of C1q, C1r, C3, C8, and C9 mRNA expression observed in the pinacidil-treated hearts. The results suggest that reduction of local tissue complement production may be one means by which preconditioning protects the ischemic myocardium.

  12. Nandrolone reduces activation of Notch signaling in denervated muscle associated with increased Numb expression

    SciTech Connect

    Liu, Xin-Hua; Yao, Shen; Qiao, Rui-Fang; Levine, Alice C.; Kirschenbaum, Alexander; Pan, Jiangping; Wu, Yong; Qin, Weiping; Bauman, William A.; Cardozo, Christopher P.

    2011-10-14

    Highlights: {yields} Nerve transection increased Notch signaling in paralyzed muscle. {yields} Nandrolone prevented denervation-induced Notch signaling. {yields} Nandrolone induced the expression of an inhibitor of the Notch signaling, Numb. {yields} Reduction of denervation-induced Notch signaling by nandrolone is likely through upregulation of Numb. -- Abstract: Nandrolone, an anabolic steroid, slows denervation-atrophy in rat muscle. The molecular mechanisms responsible for this effect are not well understood. Androgens and anabolic steroids activate Notch signaling in animal models of aging and thereby mitigate sarcopenia. To explore the molecular mechanisms by which nandrolone prevents denervation-atrophy, we investigated the effects of nandrolone on Notch signaling in denervated rat gastrocnemius muscle. Denervation significantly increased Notch activity reflected by elevated levels of nuclear Notch intracellular domain (NICD) and expression of Hey1 (a Notch target gene). Activation was greatest at 7 and 35 days after denervation but remained present at 56 days after denervation. Activation of Notch in denervated muscle was prevented by nandrolone associated with upregulated expression of Numb mRNA and protein. These data demonstrate that denervation activates Notch signaling, and that nandrolone abrogates this response associated with increased expression of Numb, suggesting a potential mechanism by which nandrolone reduces denervation-atrophy.

  13. Expression of Arabidopsis Hexokinase in Citrus Guard Cells Controls Stomatal Aperture and Reduces Transpiration

    PubMed Central

    Lugassi, Nitsan; Kelly, Gilor; Fidel, Lena; Yaniv, Yossi; Attia, Ziv; Levi, Asher; Alchanatis, Victor; Moshelion, Menachem; Raveh, Eran; Carmi, Nir; Granot, David

    2015-01-01

    Hexokinase (HXK) is a sugar-phosphorylating enzyme involved in sugar-sensing. It has recently been shown that HXK in guard cells mediates stomatal closure and coordinates photosynthesis with transpiration in the annual species tomato and Arabidopsis. To examine the role of HXK in the control of the stomatal movement of perennial plants, we generated citrus plants that express Arabidopsis HXK1 (AtHXK1) under KST1, a guard cell-specific promoter. The expression of KST1 in the guard cells of citrus plants has been verified using GFP as a reporter gene. The expression of AtHXK1 in the guard cells of citrus reduced stomatal conductance and transpiration with no negative effect on the rate of photosynthesis, leading to increased water-use efficiency. The effects of light intensity and humidity on stomatal behavior were examined in rooted leaves of the citrus plants. The optimal intensity of photosynthetically active radiation and lower humidity enhanced stomatal closure of AtHXK1-expressing leaves, supporting the role of sugar in the regulation of citrus stomata. These results suggest that HXK coordinates photosynthesis and transpiration and stimulates stomatal closure not only in annual species, but also in perennial species. PMID:26734024

  14. Expression of Arabidopsis Hexokinase in Citrus Guard Cells Controls Stomatal Aperture and Reduces Transpiration.

    PubMed

    Lugassi, Nitsan; Kelly, Gilor; Fidel, Lena; Yaniv, Yossi; Attia, Ziv; Levi, Asher; Alchanatis, Victor; Moshelion, Menachem; Raveh, Eran; Carmi, Nir; Granot, David

    2015-01-01

    Hexokinase (HXK) is a sugar-phosphorylating enzyme involved in sugar-sensing. It has recently been shown that HXK in guard cells mediates stomatal closure and coordinates photosynthesis with transpiration in the annual species tomato and Arabidopsis. To examine the role of HXK in the control of the stomatal movement of perennial plants, we generated citrus plants that express Arabidopsis HXK1 (AtHXK1) under KST1, a guard cell-specific promoter. The expression of KST1 in the guard cells of citrus plants has been verified using GFP as a reporter gene. The expression of AtHXK1 in the guard cells of citrus reduced stomatal conductance and transpiration with no negative effect on the rate of photosynthesis, leading to increased water-use efficiency. The effects of light intensity and humidity on stomatal behavior were examined in rooted leaves of the citrus plants. The optimal intensity of photosynthetically active radiation and lower humidity enhanced stomatal closure of AtHXK1-expressing leaves, supporting the role of sugar in the regulation of citrus stomata. These results suggest that HXK coordinates photosynthesis and transpiration and stimulates stomatal closure not only in annual species, but also in perennial species.

  15. Acute digoxin loading reduces ABCA8A mRNA expression in the mouse liver.

    PubMed

    Wakaumi, Michi; Ishibashi, Kenichi; Ando, Hitoshi; Kasanuki, Hiroshi; Tsuruoka, Shuichi

    2005-12-01

    Human ABCA8, a new member of the ATP binding cassette (ABC) transporter family, transports certain lipophilic drugs, such as digoxin. To investigate the roles of this transporter, we cloned a mouse homologue of ABCA8, from a mouse heart cDNA library, named ABCA8a. The deduced mouse ABCA8a protein is 66% identical with that of human ABCA8 and possesses features common to the ABC superfamily. It was found that ABCA8a was mainly expressed in the liver and heart, similar to human ABCA8. We further evaluated the effect of acute digoxin (a substrate for ABCA8) intoxication on the mRNA expression of ABCA8 using northern blotting with a 3' non-coding region as a probe to avoid cross-hybridization with other ABCA genes. Following acute digoxin infusion, the mRNA expression of ABCA8 was significantly reduced in the liver 12-24 h after injection (14.7% of vehicle treatment), but not in the heart and kidney. Real-time quantitative polymerase chain reaction analysis confirmed the reduction in ABCA8a mRNA. Similar reductions in ABCA5, ABCA7, ABCA8b and ABCA9 mRNA were also observed. A comparable amount of digitoxin did not affect ABCA8a mRNA expression in the liver. The results suggest that ABCA8 may play a role in digoxin metabolism in the liver.

  16. Reduced folate carrier (RFC-1) gene expression in normal and psoriatic skin.

    PubMed

    Sprecher, E; Bergman, R; Sprecher, H; Maor, G; Reiter, I; Krivoy, N; Drori, S; Assaraf, Y G; Friedman-Birnbaum, R

    1998-12-01

    Methotrexate is widely used in the treatment of severe psoriasis. However, little is currently known about the mechanisms underlying its therapeutic activity in the skin. Methotrexate has been shown to be carried into cells through the reduced folate carrier (RFC-1). The recent cloning and characterization of the human gene encoding this transmembranal carrier enabled us to investigate RFC-1 gene expression in human skin. Biopsies were obtained from the skin of healthy and psoriatic volunteers. RNA extracted from these biopsies was analyzed by the reverse transcriptase-polymerase chain reaction technique. While RFC-1 gene expression was barely detectable in the uninvolved skin of psoriatic patients and in the skin of healthy volunteers, high levels of RFC-1 transcripts were found in biopsies obtained from psoriatic plaques. To further investigate this pattern of gene expression, we studied skin biopsies by in situ hybridization with a labeled antisense riboprobe specific for the RFC-1 gene. The RFC-1 gene was found to be weakly expressed in the epidermis, in biopsies obtained from the skin of healthy subjects as well as in those from the uninvolved skin of psoriatic patients. In contrast, in biopsies obtained from psoriatic plaques, high levels of RFC-1 gene transcripts were found mostly in the spinous layer of the epidermis. These results suggest the existence of a specific methotrexate carrier in the human epidermis, and may bear relevance to the cutaneous manifestations of methotrexate toxicity.

  17. Reduced Airway Surface pH Impairs Bacterial Killing in the Porcine Cystic Fibrosis Lung

    PubMed Central

    Pezzulo, Alejandro A.; Tang, Xiao Xiao; Hoegger, Mark J.; Abou Alaiwa, Mahmoud H.; Ramachandran, Shyam; Moninger, Thomas O.; Karp, Phillip H.; Wohlford-Lenane, Christine L.; Haagsman, Henk P.; van Eijk, Martin; Bánfi, Botond; Horswill, Alexander R.; Stoltz, David A.; McCray, Paul B.; Welsh, Michael J.; Zabner, Joseph

    2012-01-01

    Cystic fibrosis (CF) is a life-shortening disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene 1. Although bacterial lung infection and the resulting inflammation cause most of the morbidity and mortality, how loss of CFTR first disrupts airway host defense has remained uncertain 2–6. We asked what abnormalities impair eradication when a bacterium lands on the pristine surface of a newborn CF airway? To investigate these defects, we interrogated the viability of individual bacteria immobilized on solid grids and placed on the airway surface. As a model we studied CF pigs, which spontaneously develop hallmark features of CF lung disease 7,8. At birth, their lungs lack infection and inflammation, but have a reduced ability to eradicate bacteria 8. Here we show that in newborn wild-type pigs, the thin layer of airway surface liquid (ASL) rapidly killed bacteria in vivo, when removed from the lung, and in primary epithelial cultures. Lack of CFTR reduced bacterial killing. We found that ASL pH was more acidic in CF, and reducing pH inhibited the antimicrobial activity of ASL. Reducing ASL pH diminished bacterial killing in wild-type pigs, and increasing ASL pH rescued killing in CF pigs. These results directly link the initial host defense defect to loss of CFTR, an anion channel that facilitates HCO3− transport 9–13. Without CFTR, airway epithelial HCO3− secretion is defective, ASL pH falls and inhibits antimicrobial function, and thereby impairs killing of bacteria that enter the newborn lung. These findings suggest that increasing ASL pH might prevent the initial infection in patients with CF and that assaying bacterial killing could report on the benefit of therapeutic interventions. PMID:22763554

  18. Root surface conditioning with nicotine or cotinine reduces viability and density of fibroblasts in vitro.

    PubMed

    Martinez, Aurora Esmeralda Traverso; Silverio, Karina Gonzales; Fogo, Jose Carlos; Kirkwood, Keith Lough; Rossa, Carlos

    2005-09-01

    The purpose of study was to evaluate fibroblast attachment and cellular morphology on root surfaces chemically conditioned with nicotine or cotinine. A secondary objective was to determine if mechanical scaling and root planning of these chemically conditioned surfaces would alter cellular attachment. Root surface dentin specimens were prepared from uniradicular teeth of non-smoking patients. Specimens were randomly assigned to two experimental groups: no treatment (chemical conditioning only) and scaling and root planning after conditioning (SRPC). The concentrations of the tested substances were in the range of 0-1 mg/mL (nicotine) and 0-1 g/mL (cotinine). After a 24-h conditioning period, dentin slices were incubated with continuous lineage of fibroblastic cells from rat (McCoy cells) for another 24 h. Specimens were prepared for SEM analysis and microphotographs. The statistical analysis of the data indicated significant alteration of cellular morphology on fibroblasts that were grown on root surface exposed to nicotine concentrations greater than 1 ?g/mL. This effect of nicotine was not reduced by SRPC. On the other hand, in the SRPC group cellular density was greater. For cotinine-conditioned specimens, the greater concentrations also led to alteration on morphology, and these alterations were observed in the SRPC group as well. Cotinine did not induce significant changes on cellular density. The results indicated that fibroblasts are negatively influenced by nicotine present on the dentin substrate and also that scaling may reduce these effects. Cotinine treatment on root surfaces may alter cell morphology and density but these effects were less severe than that promoted by nicotine, and were not affected by scaling.

  19. Huoxue Rongluo Tablet reduces matrix metalloproteinase-9 expression in infarcted brain tissue.

    PubMed

    Zhou, Desheng; Li, Mei; Hu, Hua; Chen, Yao; Yang, Yang; Zhong, Jie; Liu, Lijuan

    2013-12-05

    Huoxue Rongluo Tablet was made of tall gastrodis tuber, dahurian angelica root, honeysuckle stem, grassleaf sweetflag rhizome, common flowering quince fruit, figwort root, red peony root and peach seed at a ratio of 3:2:6:2:3:3:3:3. Huoxue Rongluo Tablet is a well-established and common pre-scription for the treatment of cerebral infarction. In this study, a rat model of cerebral ischemia was established and the animals were intragastrically administered Huoxue Rongluo Tablet. This treat-ment reduced infarct volume, decreased matrix metalloproteinase-9 expression, and improved neurological function. Moreover, the effects of Huoxue Rongluo Tablet were better than those of buflomedil pyridoxal phosphate. These results indicate that Huoxue Rongluo Tablet is effective in treating cerebral infarction by regulating matrix metalloproteinase-9 protein expression.

  20. Huoxue Rongluo Tablet reduces matrix metalloproteinase-9 expression in infarcted brain tissue

    PubMed Central

    Zhou, Desheng; Li, Mei; Hu, Hua; Chen, Yao; Yang, Yang; Zhong, Jie; Liu, Lijuan

    2013-01-01

    Huoxue Rongluo Tablet was made of tall gastrodis tuber, dahurian angelica root, honeysuckle stem, grassleaf sweetflag rhizome, common flowering quince fruit, figwort root, red peony root and peach seed at a ratio of 3:2:6:2:3:3:3:3. Huoxue Rongluo Tablet is a well-established and common pre-scription for the treatment of cerebral infarction. In this study, a rat model of cerebral ischemia was established and the animals were intragastrically administered Huoxue Rongluo Tablet. This treat-ment reduced infarct volume, decreased matrix metalloproteinase-9 expression, and improved neurological function. Moreover, the effects of Huoxue Rongluo Tablet were better than those of buflomedil pyridoxal phosphate. These results indicate that Huoxue Rongluo Tablet is effective in treating cerebral infarction by regulating matrix metalloproteinase-9 protein expression. PMID:25206642

  1. Inducible nitric oxide synthase expression is reduced in cystic fibrosis murine and human airway epithelial cells.

    PubMed Central

    Kelley, T J; Drumm, M L

    1998-01-01

    It has been reported that exhaled nitric oxide levels are reduced in cystic fibrosis (CF) patients. We have examined the inducible isoform of nitric oxide synthase (iNOS) in the airways by immunostaining and found that iNOS is constitutively expressed in the airway epithelia of non-CF mouse and human tissues but essentially absent in the epithelium of CF airways. We explored potential consequences of lost iNOS expression and found that iNOS inhibition significantly increases mouse nasal trans-epithelial potential difference, and hindered the ability of excised mouse lungs to prevent growth of Pseudomonas aeruginosa. The absence of continuous nitric oxide production in epithelial cells of CF airways may play a role in two CF-associated characteristics: hyperabsorption of sodium and susceptibility to bacterial infections. PMID:9739054

  2. Surface expression of the hRSV nucleoprotein impairs immunological synapse formation with T cells

    PubMed Central

    Céspedes, Pablo F.; Bueno, Susan M.; Ramírez, Bruno A.; Gomez, Roberto S.; Riquelme, Sebastián A.; Palavecino, Christian E.; Mackern-Oberti, Juan Pablo; Mora, Jorge E.; Depoil, David; Sacristán, Catarina; Cammer, Michael; Creneguy, Alison; Nguyen, Tuan H.; Riedel, Claudia A.; Dustin, Michael L.; Kalergis, Alexis M.

    2014-01-01

    Human respiratory syncytial virus (hRSV) is the leading cause of bronchiolitis and pneumonia in young children worldwide. The recurrent hRSV outbreaks and reinfections are the cause of a significant public health burden and associate with an inefficient antiviral immunity, even after disease resolution. Although several mouse- and human cell-based studies have shown that hRSV infection prevents naïve T-cell activation by antigen-presenting cells, the mechanism underlying such inhibition remains unknown. Here, we show that the hRSV nucleoprotein (N) could be at least partially responsible for inhibiting T-cell activation during infection by this virus. Early after infection, the N protein was expressed on the surface of epithelial and dendritic cells, after interacting with trans-Golgi and lysosomal compartments. Further, experiments on supported lipid bilayers loaded with peptide-MHC (pMHC) complexes showed that surface-anchored N protein prevented immunological synapse assembly by naive CD4+ T cells and, to a lesser extent, by antigen-experienced T-cell blasts. Synapse assembly inhibition was in part due to reduced T-cell receptor (TCR) signaling and pMHC clustering at the T-cell−bilayer interface, suggesting that N protein interferes with pMHC−TCR interactions. Moreover, N protein colocalized with the TCR independently of pMHC, consistent with a possible interaction with TCR complex components. Based on these data, we conclude that hRSV N protein expression at the surface of infected cells inhibits T-cell activation. Our study defines this protein as a major virulence factor that contributes to impairing acquired immunity and enhances susceptibility to reinfection by hRSV. PMID:25056968

  3. Neurotransplantation of stem cells genetically modified to express human dopamine transporter reduces alcohol consumption

    PubMed Central

    2010-01-01

    Introduction Regulated neurotransmitter actions in the mammalian central nervous system determine brain function and control peripheral organs and behavior. Although drug-seeking behaviors, including alcohol consumption, depend on central neurotransmission, modification of neurotransmitter actions in specific brain nuclei remains challenging. Herein, we report a novel approach for neurotransmission modification in vivo by transplantation of stem cells engineered to take up the neurotransmitter dopamine (DA) efficiently through the action of the human dopamine transporter (hDAT). As a functional test in mice, we used voluntary alcohol consumption, which is known to release DA in nucleus accumbens (NAC), an event hypothesized to help maintain drug-seeking behavior. We reasoned that reducing extracellular DA levels, by engrafting into NAC DA-sequestering stem cells expressing hDAT, would alter alcohol intake. Methods We have generated a neural stem cell line stably expressing the hDAT. Uptake kinetics of DA were determined to select a clone for transplantation. These genetically modified stem cells (or cells transfected with a construct lacking the hDAT sequence) were transplanted bilaterally into the NAC of wild-type mice trained to consume 10% alcohol in a two-bottle free-choice test for alcohol consumption. Alcohol intake was then ascertained for 1 week after transplantation, and brain sections through the NAC were examined for surviving grafted cells. Results Modified stem cells expressed hDAT and uptaken DA selectively via hDAT. Mice accustomed to drinking 10% ethanol by free choice reduced their alcohol consumption after being transplanted with hDAT-expressing stem cells. By contrast, control stem cells lacked that effect. Histologic examination revealed surviving stem cells in the NAC of all engrafted brains. Conclusions Our findings represent proof of principle suggesting that genetically engineered stem cells can be useful for exploring the role of

  4. Reduced Adipogenic Gene Expression in Thigh Adipose Tissue Precedes Human Immunodeficiency Virus-Associated Lipoatrophy

    PubMed Central

    Kratz, Mario; Purnell, Jonathan Q.; Breen, Patricia A.; Thomas, Katherine K.; Utzschneider, Kristina M.; Carr, Darcy B.; Kahn, Steven E.; Hughes, James P.; Rutledge, Elizabeth A.; Van Yserloo, Brian; Yukawa, Michi; Weigle, David S.

    2008-01-01

    Context: The expression of adipogenic genes in sc adipose tissue has been reported to be lower among patients with HIV-associated lipoatrophy than HIV-uninfected controls. It is unclear whether this is a result or cause of lipoatrophy. Objective: The objective of the study was to investigate the temporal relationships among changes in adipogenic gene expression in sc adipose tissue and changes in body fat distribution and metabolic complications in HIV-infected subjects on antiretroviral therapy. Design: This was a prospective longitudinal study. Setting: The study was conducted at HIV clinics in Seattle, Washington. Participants: The study population included 31 HIV-infected and 12 control subjects. Interventions: Subjects were followed up for 12 months after they initiated or modified their existing antiretroviral regimen. Main Outcome Measures: Changes in body composition, plasma lipids, insulin sensitivity, and gene expression in sc abdominal and thigh adipose tissue. Results: Subjects who developed lipoatrophy (n = 10) had elevated fasting triglycerides [3.16 (sd 2.79) mmol/liter] and reduced insulin sensitivity as measured by frequently sampled iv glucose tolerance test [1.89 (sd 1.27) × 10−4 min−1/μU·ml] after 12 months, whereas those without lipoatrophy (n = 21) did not show any metabolic complications [triglycerides 1.32 (sd 0.58) mmol/liter, P = 0.01 vs. lipoatrophy; insulin sensitivity 3.52 (sd 1.91) × 10−4 min−1/μU·ml, P = 0.01 vs. lipoatrophy]. In subjects developing lipoatrophy, the expression of genes involved in adipocyte differentiation, lipid uptake, and local cortisol production in thigh adipose tissue was significantly reduced already at the 2-month visit, several months before any loss of extremity fat mass was evident. Conclusions: In HIV-infected subjects, lipoatrophy is associated with elevated fasting triglycerides and insulin resistance and might be caused by a direct or indirect effect of antiretroviral drugs on sc adipocyte

  5. Tomatidine inhibits invasion of human lung adenocarcinoma cell A549 by reducing matrix metalloproteinases expression.

    PubMed

    Yan, Kun-Huang; Lee, Liang-Ming; Yan, Shao-Han; Huang, Hsiang-Ching; Li, Chia-Chen; Lin, Hui-Ting; Chen, Pin-Shern

    2013-05-25

    Tomatidine is an aglycone of glycoalkaloid tomatine in tomato. Tomatidine is found to possess anti-inflammatory properties and may serve as a chemosensitizer in multidrug-resistant tumor cells. However, the effect of tomatidine on cancer cell metastasis remains unclear. This study examines the effect of tomatidine on the migration and invasion of human lung adenocarcinoma A549 cell in vitro. The data demonstrates that tomatidine does not effectively inhibit the viability of A549 cells. When treated with non-toxic doses of tomatidine, cell invasion is markedly suppressed by Boyden chamber invasion assay, while cell migration is not affected. Tomatidine reduces the mRNA level of matrix metalloproteinase-2 (MMP-2), MMP-9 and increases the expression of reversion-inducing cysteine-rich protein with kazal motifs (RECK), as well as tissue inhibitor of metalloproteinase-1 (TIMP-1). The immunoblotting assays indicate that tomatidine is very effective in suppressing the phosphorylation of Akt and extracellular signal regulating kinase (ERK). In addition, tomatidine significantly decreases the nuclear level of nuclear factor kappa B (NF-κB), which suggests that tomatidine inhibits NF-κB activity. Furthermore, the treatment of inhibitors specific for PI3K/Akt (LY294002), ERK (U0126), or NF-κB (pyrrolidine dithiocarbamate) to A549 cells reduced cell invasion and MMP-2/9 expression. The results suggest that tomatidine inhibits the invasion of A549 cells by reducing the expression of MMPs. It also inhibits ERK and Akt signaling pathways and NF-κB activity. These findings demonstrate a new therapeutic potential for tomatidine in anti-metastatic therapy.

  6. Green tea extracts reduce adipogenesis by decreasing expression of transcription factors C/EBPα and PPARγ

    PubMed Central

    Yang, Xiuling; Yin, Lei; Li, Tang; Chen, Zhihong

    2014-01-01

    Objectives: This study is to determine if green tea (Camellia sinensis) extracts (GTE) affects adipogenesis and further investigate the related molecular mechanisms. Methods: Patients with metabolic syndrome were recruited in this study. Of them, 70 patients received GTE and 64 received water to serve as the control group. The human serum adiponectin, visfatin, and leptin concentrations were determined by enzyme-linked immunosorbent assay. Adipogenesis of 3T3-L1 preadipocytes was induced with reagents and then the cells were treated with GTE. The lipids were stained with Oil Red O for analysis of adipogenesis of 3T3-L1 preadipocytes. The 3T3-L1 preadipocytes were treated with increasing concentrations (0.2-0.5%, w/v) of GTE for 2 days and the cell viability was determined by MTT assay. Reverse transcription real-time PCR and immunoblotting assays were performed to determine RNA and protein levels of relative molecules. Results: GTE increases the serum concentrations of adiponectin but decreases visfatin levels in patients received GTE. The leptin concentrations in serum were not significantly affected. The GTE reduces the adipogenesis-induced lipid accumulation in 3T3-L1 preadipocytes. GTE decreases the mRNA and protein expression of adipogenic transcription factors C/EBPα and PPARγ in 3T3-L1 cells. Expression levels of the adipocyte-specific genes encoding adipocyte protein 2, lipoprotein lipase, and glucose transporter 4 were also decreased by GTE. Furthermore, it was found that GTE reduces phosphorylation of Akt during adipocyte differentiation. Conclusions: GTE reduces adipogenesis by decreasing expression of transcription factors C/EBPα and PPARγ by reduction of phosphorylation of Akt during adipocyte differentiation. PMID:25663987

  7. Highly reflective roof surfaces reduce cooling energy use and peak demand

    SciTech Connect

    George, K.L.

    1993-12-31

    Light-colored building surfaces are a time-honored method of staying cool, familiar to anyone who has seen the white-washed buildings in the Mediterranean, the Caribbean, and other warm climes. This heat rejecting strategy has been largely ignored in many places since the advent of mechanical air conditioning. However, recent monitoring experiments suggest that utilities could reduce summer cooling loads significantly by encouraging wide-scale application of light-colored, highly reflective roof surfaces on homes and small commercial buildings. Recent field tests in California and Florida show that application of light-colored coatings to poorly insulated roof systems can reduce cooling energy use and peak demand by more than one-third. Installation of highly reflective, or high ``albedo,`` roofs can be a cost-effective and low-risk utility DSM measure if done during new construction or when buildings are scheduled for reroofing. Most commercial building roofing materials are available in white versions or are easily coated. The residential market is more problematic, however, since asphalt shingles, which constitute more than half of the US residential roofing market, are not highly reflective. More research and market development is warranted to help realize the potential of highly reflective roofs. This includes additional product testing, labeling of highly reflective roofing materials, development of nonwhite reflective surfaces, additional performance monitoring, and implementation of utility pilot programs.

  8. Wear-reducing Surface Functionalization of Implant Materials Using Ultrashort Laser Pulses

    NASA Astrophysics Data System (ADS)

    Oldorf, P.; Peters, R.; Reichel, S.; Schulz, A.-P.; Wendlandt, R.

    The aim of the project called "EndoLas" is the development of a reproducible and reliable method for a functionalization of articulating surfaces on hip joint endoprostheses due to a reduction of abrasion and wear by the generation of micro structures using ultrashort laser pulses. On the one hand, the microstructures shall ensure the capture of abraded particles, which cause third-body wear and thereby increase aseptic loosening. On the other hand, the structures shall improve or maintain the tribologically important lubricating film. Thereby, the cavities serve as a reservoir for the body's own synovial fluid. The dry friction, which promotes abrasion and is a part of the mixed friction in the joint, shall therefore be reduced. In experimental setups it was shown, that the abrasive wear can be reduced significantly due to micro-structuring the articulating implant surfaces. To shape the fine and deterministic cavities on the surfaces, an ultra-short pulsed laser, which is integrated in a high-precision, 5-axes micro-machining system, was used. The laser system, based on an Yb:YAG thin-disk regenerative amplifier, has an average output power of 50 W at the fundamental wavelength of 1030 nm, a maximum repetition rate of 400 kHz and a pulse duration of 6 ps. Due to this, a maximum pulse energy of 125 μJ is achievable. Furthermore external second and third harmonic generation enables the usage of wavelengths in the green and violet spectral range.

  9. Mitigation strategies to reduce pesticide inputs into ground- and surface water and their effectiveness; a review.

    PubMed

    Reichenberger, Stefan; Bach, Martin; Skitschak, Adrian; Frede, Hans-Georg

    2007-10-01

    In this paper, the current knowledge on mitigation strategies to reduce pesticide inputs into surface water and groundwater, and their effectiveness when applied in practice is reviewed. Apart from their effectiveness in reducing pesticide inputs into ground- and surface water, the mitigation measures identified in the literature are evaluated with respect to their practicability. Those measures considered both effective and feasible are recommended for implementing at the farm and catchment scale. Finally, recommendations for modelling are provided using the identified reduction efficiencies. Roughly 180 publications directly dealing with or being somehow related to mitigation of pesticide inputs into water bodies were examined. The effectiveness of grassed buffer strips located at the lower edges of fields has been demonstrated. However, this effectiveness is very variable, and the variability cannot be explained by strip width alone. Riparian buffer strips are most probably much less effective than edge-of-field buffer strips in reducing pesticide runoff and erosion inputs into surface waters. Constructed wetlands are promising tools for mitigating pesticide inputs via runoff/erosion and drift into surface waters, but their effectiveness still has to be demonstrated for weakly and moderately sorbing compounds. Subsurface drains are an effective mitigation measure for pesticide runoff losses from slowly permeable soils with frequent waterlogging. For the pathways drainage and leaching, the only feasible mitigation measures are application rate reduction, product substitution and shift of the application date. There are many possible effective measures of spray drift reduction. While sufficient knowledge exists for suggesting default values for the efficiency of single drift mitigation measures, little information exists on the effect of the drift reduction efficiency of combinations of measures. More research on possible interactions between different drift

  10. 1,25-Dihydroxyvitamin D(3) Inhibits Podocyte uPAR Expression and Reduces Proteinuria

    PubMed Central

    Liu, Shuangxin; Xie, Shaoting; Yang, Yun; Ma, Juan; Deng, Yujun; Wang, Wenjian; Xu, Lixia; Li, Ruizhao; Zhang, Li; Yu, Chunping; Shi, Wei

    2013-01-01

    Background Accumulating studies have demonstrated that 1,25-Dihydroxyvitamin D(3) (1,25(OH)2D3) reduces proteinuria and protects podocytes from injury. Recently, urokinase receptor (uPAR) and its soluble form have been shown to cause podocyte injury and focal segmental glomerulosclerosis (FSGS). Here, our findings showed that 1,25(OH)2D3 did inhibit podocyte uPAR expression and attenuate proteinuria and podocyte injury. Methodology/Principal Findings In this study, the antiproteinuric effect of 1,25(OH)2D3 was examined in the lipopolysaccharide mice model of transient proteinuria (LPS mice) and in the 5/6 nephrectomy rat FSGS model(NTX rats). uPAR protein expression were tested by flow cytometry, immune cytochemistry and western blot analysis, and uPAR mRNA expression by real-time quantitative PCR in cultured podocytes and kidney glomeruli isolated from mice and rats. Podocyte motility was observed by transwell migration assay and wound healing assay. Podocyte foot processes effacement was identified by transmission electron microscopy. We found that 1,25(OH)2D3 inhibited podocyte uPAR mRNA and protein synthesis in LPS-treated podocytes, LPS mice and NTX rats, along with 1,25(OH)2D3 reducing proteinuria in NTX rats and LPS mice.1,25(OH)2D3 reduced glomerulosclerosis in NTX rats and alleviated podocyte foot processes effacement in LPS mice. Transwell migration assay and wound healing assay showed that LPS-induced podocyte motility, irrespective of random or directed motility, were substantially reduced by 1,25(OH)2D3. Conclusions/Significance Our results demonstrated that 1,25(OH)2D3 inhibited podocyte uPAR expression in vitro and in vivo, which may be an unanticipated off target effect of 1,25(OH)2D3 and explain its antiproteinuric effect in the 5/6 nephrectomy rat FSGS model and the LPS mouse model of transient proteinuria. PMID:23741418

  11. Preparation of Reduced Iron Powders from Mill Scale with Microwave Heating: Optimization Using Response Surface Methodology

    NASA Astrophysics Data System (ADS)

    Ye, Qianxu; Zhu, Hongbo; Peng, Jinhui; Srinivasa Kannan, C.; Chen, Jian; Dai, Linqing; Liu, Peng

    2013-12-01

    Preparation of the reduced iron powder has been attempted with mill scale as the iron-bearing material and with wood charcoal as the reducing agent through microwave heating. The response surface methodology (RSM) is used to optimize the process conditions, with wood charcoal, process temperature, and holding time being the three process parameters. The regressed model equation eliminating the insignificant parameters through an analysis of variance (ANOVA) was used to optimize the process conditions. The optimum process parameters for the preparation of reduced iron powders have been identified to be the wood charcoal of 13.8 pct, a process temperature of 1391 K (1118 °C), and a holding time of 43 minutes. The optimum conditions resulted in reduced iron powders with a total iron content of 98.60 pct and a metallization ratio of 98.71 pct. X-ray fluorescence (XRF) was used to estimate the elemental contents of the reduced iron powder, which meets the specification of the HY100.23 first-class iron powder standard. Additionally X-ray diffraction (XRD), energy-dispersive spectroscopy (EDS), and scanning electron microscopy (SEM) analysis were performed and the results are compiled.

  12. Apoptosis is associated with reduced expression of complement regulatory molecules, adhesion molecules and other receptors on polymorphonuclear leucocytes: functional relevance and role in inflammation.

    PubMed Central

    Jones, J; Morgan, B P

    1995-01-01

    Human polymorphonuclear leucocytes (PMN) express proteins that protect them from damage by homologous complement. Protection may be particularly important when these cells migrate to inflammatory sites where complement activation is taking place. Resolution of inflammation involves removal of these PMN. The major mechanism of removal is likely to involve PMN apoptosis followed by recognition and engulfment by macrophages. However, little attention has been paid to the possible relevance of apoptosis to PMN susceptibility to immune effectors. Here we describe a reduction in cell surface expression of two complement regulatory proteins, CD59, an inhibitor of the membrane attack complex and CD55 (decay accelerating factor), an inhibitor of the C3/C5 convertase, on a subpopulation of PMN aged in culture. Loss of these proteins, both attached to the membrane by glycosyl phosphatidylinositol (GPI) anchors, correlated closely with the appearance of apoptotic morphology. We also observed a marked reduction in expression of the GPI-anchored molecule CD16 on apoptotic PMN. Reduced expression of membrane proteins was not confined to those anchored through GPI--several transmembrane molecules including CD11a CD11b and CD18 were also reduced on apoptotic PMN, whilst other were little changed (CD35, CD46). The precipitous fall in CD16 surface expression on PMN was not specific for apoptosis--in vitro incubation of PMN with lipopolysaccharide-inhibited apoptosis but caused a reduction in CD16 expression to 'apoptotic' levels. Images Figure 2 PMID:8567034

  13. MicroRNA-22 Can Reduce Parathymosin Expression in Transdifferentiated Hepatocytes

    PubMed Central

    Chen, Chun-Ming; Chu, Tien-Hua; Shih, Chiaho

    2012-01-01

    Pancreatic acinar cells AR42J-B13 can transdifferentiate into hepatocyte-like cells permissive for efficient hepatitis B virus (HBV) replication. Here, we profiled miRNAs differentially expressed in AR42J-B13 cells before and after transdifferentiation to hepatocytes, using chip-based microarray. Significant increase of miRNA expression, including miR-21, miR-22, and miR-122a, was confirmed by stem-loop real-time PCR and Northern blot analyses. In contrast, miR-93, miR-130b, and a number of other miRNAs, were significantly reduced after transdifferentiation. To investigate the potential significance of miR-22 in hepatocytes, we generated cell lines stably expressing miR-22. By 2D-DIGE, LC-MS/MS, and Western blot analyses, we identified several potential target genes of miR-22, including parathymosin. In transdifferentiated hepatocytes, miR-22 can inhibit both mRNA and protein expression of parathymosin, probably through a direct and an indirect mechanism. We tested two computer predicted miR-22 target sites at the 3′ UTR of parathymosin, by the 3′ UTR reporter gene assay. Treatment with anti-miR-22 resulted in significant elevation of the reporter activity. In addition, we observed an in vivo inverse correlation between miR-22 and parathymosin mRNA in their tissue distribution in a rat model. The phenomenon that miR-22 can reduce parathymosin protein was also observed in human hepatoma cell lines Huh7 and HepG2. So far, we detected no major effect on several transdifferentiation markers when AR42J-B13 cells were transfected with miR-22, or anti-miR-22, or a parathymosin expression vector, with or without dexamethasone treatment. Therefore, miR-22 appears to be neither necessary nor sufficient for transdifferentiation. We discussed the possibility that altered expression of some other microRNAs could induce cell cycle arrest leading to transdifferentiation. PMID:22493679

  14. Successful Surface Treatments for Reducing Instabilities in Advanced Nickel-base Superalloys for Turbine Blades

    NASA Technical Reports Server (NTRS)

    Locci, Ivan E.; MacKay, Rebecca A.; Garg, Anita; Ritzert, Frank J.

    2004-01-01

    An optimized carburization treatment has been developed to mitigate instabilities that form in the microstructures of advanced turbine airfoil materials. Current turbine airfoils consist of a single crystal superalloy base that provides the mechanical performance of the airfoil, a thermal barrier coating (TBC) that reduces the temperature of the base superalloy, and a bondcoat between the superalloy and the TBC, that improves the oxidation and corrosion resistance of the base superalloy and the spallation resistance of the TBC. Advanced nickel-base superalloys containing high levels of refractory metals have been observed to develop an instability called secondary reaction zone (SRZ), which can form beneath diffusion aluminide bondcoats. This instability between the superalloy and the bondcoat has the potential of reducing the mechanical properties of thin-wall turbine airfoils. Controlled gas carburization treatments combined with a prior stress relief heat treatment and adequate surface preparation have been utilized effectively to minimize the formation of SRZ. These additional processing steps are employed before the aluminide bondcoat is deposited and are believed to change the local chemistry and local stresses of the surface of the superalloy. This paper presents the detailed processing steps used to reduce SRZ between platinum aluminide bondcoats and advanced single crystal superalloys.

  15. Response Surface Energy Modeling of an Electric Vehicle over a Reduced Composite Drive Cycle

    SciTech Connect

    Jehlik, Forrest; LaClair, Tim J.

    2014-04-01

    Response surface methodology (RSM) techniques were applied to develop a predictive model of electric vehicle (EV) energy consumption over the Environmental Protection Agency's (EPA) standardized drive cycles. The model is based on measurements from a synthetic composite drive cycle. The synthetic drive cycle is a minimized statistical composite of the standardized urban (UDDS), highway (HWFET), and US06 cycles. The composite synthetic drive cycle is 20 minutes in length thereby reducing testing time of the three standard EPA cycles by over 55%. Vehicle speed and acceleration were used as model inputs for a third order least squared regression model predicting vehicle battery power output as a function of the drive cycle. The approach reduced three cycles and 46 minutes of drive time to a single test of 20 minutes. Application of response surface modeling to the synthetic drive cycle is shown to predict energy consumption of the three EPA cycles within 2.6% of the actual measured values. Additionally, the response model may be used to predict energy consumption of any cycle within the speed/acceleration envelope of the synthetic cycle. This technique results in reducing test time, which additionally provides a model that may be used to expand the analysis and understanding of the vehicle under consideration.

  16. Shared requirement for dynein function and intact microtubule cytoskeleton for normal surface expression of cardiac potassium channels.

    PubMed

    Loewen, Matthew E; Wang, Zhuren; Eldstrom, Jodene; Dehghani Zadeh, Alireza; Khurana, Anu; Steele, David F; Fedida, David

    2009-01-01

    Potassium channels at the cardiomyocyte surface must eventually be internalized and degraded, and changes in cardiac potassium channel expression are known to occur during myocardial disease. It is not known which trafficking pathways are involved in the control of cardiac potassium channel surface expression, and it is not clear whether all cardiac potassium channels follow a common pathway or many pathways. In the present study we have surveyed the role of retrograde microtubule-dependent transport in modulating the surface expression of several cardiac potassium channels in ventricular myocytes and heterologous cells. The disruption of microtubule transport in rat ventricular myocytes with nocodazole resulted in significant changes in potassium currents. A-type currents were enhanced 1.6-fold at +90 mV, rising from control densities of 20.9 +/- 2.8 to 34.0 +/- 5.4 pA/pF in the nocodazole-treated cells, whereas inward rectifier currents were reduced by one-third, perhaps due to a higher nocodazole sensitivity of Kir channel forward trafficking. These changes in potassium currents were associated with a significant decrease in action potential duration. When expressed in heterologous human embryonic kidney (HEK-293) cells, surface expression of Kv4.2, known to substantially underlie A-type currents in rat myocytes, was increased by nocodazole, by the dynein inhibitor erythro-9-(2-hydroxy-3-nonyl) adenine hydrochloride, and by p50 overexpression, which specifically interferes with dynein motor function. Peak current density was 360 +/- 61.0 pA/pF in control cells and 658 +/- 94.5 pA/pF in cells overexpressing p50. The expression levels of Kv2.1, Kv3.1, human ether-a-go-go-related gene, and Kir2.1 were similarly increased by p50 overexpression in this system. Thus the regulation of potassium channel expression involves a common dynein-dependent process operating similarly on the various channels.

  17. Reduced expression of GNA11 and silencing of MCT1 in human breast cancers.

    PubMed

    Asada, Kiyoshi; Miyamoto, Kazuaki; Fukutomi, Takashi; Tsuda, Hitoshi; Yagi, Yukiko; Wakazono, Kuniko; Oishi, Shoko; Fukui, Hiroshi; Sugimura, Takashi; Ushijima, Toshikazu

    2003-01-01

    Alteration in the methylation status of a gene is often associated with its altered expression. Based on a genome scanning technique for differences in CpG methylations, methylation-sensitive representational difference analysis, DNA fragments hypermethylated in a human breast cancer were isolated. A DNA fragment was isolated from intron 1 of guanine-nucleotide-binding protein alpha-11 (GNA11). mRNA expression of GNA11 was shown to be decreased in 10 of 16 breast cancers by RT-PCR analysis, and the immunoreactivity of the GNA11 product, Galpha11 subunit of heterotrimeric G-protein, was observed to be reduced in 14 of the 16 cancers by immunohistochemistry. Methylation of a CpG island (CGI) in the 5' region of GNA11 or that of intron 1 did not show a clear correlation with its decreased expression. Another DNA fragment was isolated from a CGI in the 5' upstream region of monocarboxylate transporter 1 (MCT1), and was methylated in 4 of 20 breast cancers. The CGI was also methylated in a human breast cancer cell line, MDA-MB-231, and quantitative RT-PCR showed that its expression was almost lost in the cell line. By treatment of the cells with a demethylating agent, 5-aza-2'-deoxycytidine, the methylation was removed and the expression was restored. GNA11 is involved in signalling of gonadotropin-releasing hormone receptor, which negatively regulates cell growth. MCT1 is involved in cellular transportation of butyrate, which induces cellular differentiation. Downregulation of these two genes was suggested to be involved in human breast cancers. Copyright 2003 S. Karger AG, Basel

  18. Adiponectin expression in subcutaneous adipose tissue is reduced in women with cellulite.

    PubMed

    Emanuele, Enzo; Minoretti, Piercarlo; Altabas, Karmela; Gaeta, Elio; Altabas, Velimir

    2011-04-01

    Cellulite, which appears as orange peel-type or cottage cheese-like dimpling of the skin on the thighs and buttocks, is a complex, multifactorial, cosmetic disorder of the subcutaneous fat layer and the overlying superficial skin. Adiponectin is an adipocyte-derived hormone mainly produced by subcutaneous fat that shows important protective anti-inflammatory and vasodilatory effects. We hypothesized that adiponectin expressed in the subcutaneous adipose tissue (SAT) might play a role in the pathogenesis of cellulite. We reasoned that a reduction in the expression of adiponectin - a humoral vasodilator - in the SAT of cellulite areas might contribute to the altered microcirculation frequently found in these regions. A total of 15 lean (body mass index [BMI] < 25 kg/m(2) ) women with cellulite and 15 age- and BMI-matched women without cellulite participated in this study. Real-time reverse transcription polymerase chain reaction (RT-PCR) was used to assess adiponectin gene expression. Plasma adiponectin levels were measured using a commercial enzyme immunoassay kit. Adiponectin mRNA expression in the SAT of the gluteal region was significantly lower in areas with cellulite compared with those without (12.6 ± 3.1 AU versus 16.6 ± 4.1 AU; P=0.006). However, plasma adiponectin levels did not differ between women with (20.3 ± 7.3 μg/ml) and without (19.3 ± 6.1 μg/ml) cellulite (P=0.69). Adiponectin expression is significantly reduced in the SAT in areas affected by cellulite. Our findings provide novel insights into the nature of cellulite and may give clues to the treatment of this cosmetic issue. © 2011 The International Society of Dermatology.

  19. Over-expression of TRESK K(+) channels reduces the excitability of trigeminal ganglion nociceptors.

    PubMed

    Guo, Zhaohua; Cao, Yu-Qing

    2014-01-01

    TWIK-related spinal cord K(+) (TRESK) channel is abundantly expressed in trigeminal ganglion (TG) and dorsal root ganglion neurons and is one of the major background K(+) channels in primary afferent neurons. Mutations in TRESK channels are associated with familial and sporadic migraine. In rats, both chronic nerve injury and inflammation alter the expression level of TRESK mRNA. Functional studies indicate that reduction of endogenous TRESK channel activity results in hyper-excitation of primary afferent neurons, suggesting that TRESK is a potential target for the development of new analgesics. However, whether and how enhancing TRESK channel activity would decrease the excitability of primary afferent neurons has not been directly tested. Here, we over-expressed TRESK subunits in cultured mouse TG neurons by lipofectamine-mediated transfection and investigated how this altered the membrane properties and the excitability of the small-diameter TG population. To account for the heterogeneity of neurons, we further divided small TG neurons into two groups, based on their ability to bind to fluorescently-labeled isolectin B (IB4). The transfected TG neurons showed a 2-fold increase in the level of TRESK proteins. This was accompanied by a significant increase in the fraction of lamotrigine-sensitive persistent K(+) currents as well as the size of total background K(+) currents. Consequently, both IB4-positive and IB4-negative TG neurons over-expressing TRESK subunits exhibited a lower input resistance and a 2-fold increase in the current threshold for action potential initiation. IB4-negative TG neurons over-expressing TRESK subunits also showed a significant reduction of the spike frequency in response to supra-threshold stimuli. Importantly, an increase in TRESK channel activity effectively inhibited capsaicin-evoked spikes in TG neurons. Taken together, our results suggest that potent and specific TRESK channel openers likely would reduce the excitability of

  20. Hepatic AQP9 expression in male rats is reduced in response to PPARα agonist treatment.

    PubMed

    Lebeck, Janne; Cheema, Muhammad Umar; Skowronski, Mariusz T; Nielsen, Søren; Praetorius, Jeppe

    2015-02-01

    The peroxisome proliferator receptor α (PPARα) is a key regulator of the hepatic response to fasting with effects on both lipid and carbohydrate metabolism. A role in hepatic glycerol metabolism has also been found; however, the results are somewhat contradictive. Aquaporin 9 (AQP9) is a pore-forming transmembrane protein that facilitates hepatic uptake of glycerol. Its expression is inversely regulated by insulin in male rodents, with increased expression during fasting. Previous results indicate that PPARα plays a crucial role in the induction of AQP9 mRNA during fasting. In the present study, we use PPARα agonists to explore the effect of PPARα activation on hepatic AQP9 expression and on the abundance of enzymes involved in glycerol metabolism using both in vivo and in vitro systems. In male rats with free access to food, treatment with the PPARα agonist WY 14643 (3 mg·kg(-1)·day(-1)) caused a 50% reduction in hepatic AQP9 abundance with the effect being restricted to AQP9 expressed in periportal hepatocytes. The pharmacological activation of PPARα had no effect on the abundance of GlyK, whereas it caused an increased expression of hepatic GPD1, GPAT1, and L-FABP protein. In WIF-B9 and HepG2 hepatocytes, both WY 14643 and another PPARα agonist GW 7647 reduced the abundance of AQP9 protein. In conclusion, pharmacological PPARα activation results in a marked reduction in the abundance of AQP9 in periportal hepatocytes. Together with the effect on the enzymatic apparatus for glycerol metabolism, our results suggest that PPARα activation in the fed state directs glycerol into glycerolipid synthesis rather than into de novo synthesis of glucose. Copyright © 2015 the American Physiological Society.

  1. Mutations in the main cytoplasmic loop of the GABA(A) receptor α4 and δ subunits have opposite effects on surface expression.

    PubMed

    Bracamontes, John R; Li, Ping; Akk, Gustav; Steinbach, Joe Henry

    2014-07-01

    We examined the role of putative trafficking sequences in two GABA(A) receptor subunits: α4 and δ. These subunits assemble with a β subunit to form a subtype of GABA(A) receptor involved in generating the "tonic" outward current. Both α4 and δ subunits contain dibasic retention motifs in homologous positions. When basic residues are mutated to alanine in the α4 subunit, surface expression of epitope-tagged δ subunits is increased. When basic residues in homologous regions of the δ subunit are mutated, however, surface expression is reduced. We focused on the mutants that had the maximal effects to increase (in α4) or reduce (in δ) surface expression. The total expression of δ subunits is significantly decreased by the δ mutation, suggesting an effect on subunit maturation. We also examined surface expression of the β2 subunit. Expression of the mutated α4 subunit resulted in increased surface expression of β2 compared with wild-type α4, indicating enhanced forward trafficking. In contrast, mutated δ resulted in decreased surface expression of β2 compared with wild-type δ and to α4 and β2 in the absence of any δ. This observation suggests that the mutated δ incorporates into multimeric receptors and reduces the overall forward trafficking of receptors. These observations indicate that the roles of trafficking motifs are complex, even when located in homologous positions in related subunits. The physiologic properties of receptors containing mutated subunits were not significantly affected, indicating that the mutations in the α4 subunit will be useful to enhance surface expression.

  2. Tumor targeting RGD conjugated bio-reducible polymer for VEGF siRNA expressing plasmid delivery.

    PubMed

    Kim, Hyun Ah; Nam, Kihoon; Kim, Sung Wan

    2014-08-01

    Targeted delivery of therapeutic genes to the tumor site is critical for successful and safe cancer gene therapy. The arginine grafted bio-reducible poly (cystamine bisacrylamide-diaminohexane, CBA-DAH) polymer (ABP) conjugated poly (amido amine) (PAMAM), PAM-ABP (PA) was designed previously as an efficient gene delivery carrier. To achieve high efficacy in cancer selective delivery, we developed the tumor targeting bio-reducible polymer, PA-PEG1k-RGD, by conjugating cyclic RGDfC (RGD) peptides, which bind αvβ3/5 integrins, to the PAM-ABP using polyethylene glycol (PEG, 1 kDa) as a spacer. Physical characterization showed nanocomplex formation with bio-reducible properties between PA-PEG1k-RGD and plasmid DNA (pDNA). In transfection assays, PA-PEG1k-RGD showed significantly higher transfection efficiency in comparison with PAM-ABP or PA-PEG1k-RAD in αvβ3/5 positive MCF7 breast cancer and PANC-1 pancreatic cancer cells. The targeting ability of PA-PEG1k-RGD was further established using a competition assay. To confirm the therapeutic effect, the VEGF siRNA expressing plasmid was constructed and then delivered into cancer cells using PA-PEG1k-RGD. PA-PEG1k-RGD showed 20-59% higher cellular uptake rate into MCF7 and PANC-1 than that of non-targeted polymers. In addition, MCF7 and PANC-1 cancer cells transfected with PA-PEG1k-RGD/pshVEGF complexes had significantly decreased VEGF gene expression (51-71%) and cancer cell viability (35-43%) compared with control. These results demonstrate that a tumor targeting bio-reducible polymer with an anti-angiogenic therapeutic gene could be used for efficient and safe cancer gene therapy. Copyright © 2014 Elsevier Ltd. All rights reserved.

  3. Tumor targeting RGD conjugated bio-reducible polymer for VEGF siRNA expressing plasmid delivery

    PubMed Central

    Kim, Hyun Ah; Nam, Kihoon; Kim, Sung Wan

    2014-01-01

    Targeted delivery of therapeutic genes to the tumor site is critical for successful and safe cancer gene therapy. The arginine grafted bio-reducible poly (cystamine bisacrylamide-diaminohexane, CBA-DAH) polymer (ABP) conjugated poly (amido amine) (PAMAM), PAM-ABP (PA) was designed previously as an efficient gene delivery carrier. To achieve high efficacy in cancer selective delivery, we developed the tumor targeting bio-reducible polymer, PA-PEG1k-RGD, by conjugating cyclic RGDfC (RGD) peptides, which bind αvβ3/5 integrins, to the PAM-ABP using polyethylene glycol (PEG,1kDa) as a spacer. Physical characterization showed nanocomplex formation with bio-reducible properties between PA-PEG1k-RGD and plasmid DNA (pDNA). In transfection assays, PA-PEG1k-RGD showed significantly higher transfection efficiency in comparison with PAM-ABP or PA-PEG1k-RGD in αvβ3/5 positive MCF7 breast cancer and PANC-1 pancreatic cancer cells. The targeting ability of PA-PEG1k-RGD was further established using a competition assay. To confirm the therapeutic effect, the VEGF siRNA expressing plasmid was constructed and then delivered into cancer cells using PA-PEG1k-RGD. PA-PEG1k-RGD showed 20-59% higher cellular uptake rate into MCF7 and PANC-1 than that of non-targeted polymers. In addition, MCF7 and PANC-1 cancer cells transfected with PA-PEG1k-RGD/pshVEGF complexes had significantly decreased VEGF gene expression (51-71%) and cancer cell viability (35-43%) compared with control. These results demonstrate that a tumor targeting bio-reducible polymer with an anti-angiogenic therapeutic gene could be used for efficient and safe cancer gene therapy. PMID:24894645

  4. Surface expression of subglacial meltwater movement, Bering Glacier, Alaska

    SciTech Connect

    Cadwell, D.H. ); Fleisher, P.J. . Dept. of Earth Sciences); Bailey, P.K. )

    1993-03-01

    Longitudinal topographic profiles (1988--1992) across the thermokarst terminus of the Grindle Hills Ice-tongue and interlobate moraine of the Bering Piedmont Glacier document annual changes in crevasse patterns and fluctuations in surface elevation related to subglacial water movement. A semi-continuous record of aerial photos (1978--1990), plus field observations (1988--1992), reveal the progressive enlargement of two lateral collapse basin on both sides of the thermokarst, connected by a transverse collapse trough. Seasonally generated meltwater at depth rises within the glacier, fills the basins and other depressions and lifts the thermokarst terminus of the ice-tongue a few meters by buoyancy and hydrostatic pressure. The resulting surface tension creates a chaotic crevasse pattern unrelated to normal glacier movement. The crevasses open (2 m wide, 8--10 m deep) in response to increased water accumulation at depth and close during subsidence as the ice-tongue settles following evacuation of subglacier water. A network of open conduits (>10 m diameter), exposed by surface ablation, provides evidence for the scale of englacial passageways beneath the thermokarst and represents a form of subglacial ablation that leads to removal of support and collapse in stagnant glacier masses.

  5. Effectiveness of unfertilized buffer strips for reducing nitrogen loads from agricultural lowland to surface waters.

    PubMed

    Noij, Ignatius G A M; Heinen, Marius; Heesmans, Hanneke I M; Thissen, Jac T N M; Groenendijk, Piet

    2012-01-01

    Unfertilized buffer strips (BS) are widely accepted to reduce nitrogen (N) loads from agricultural land to surface water. However, the relative reduction of N load or concentration (BS effectiveness, BSE), varies with management and local conditions, especially hydrogeology. We present novel experimental evidence on BSE for 5-m-wide grass BS on intensively drained and managed plain agricultural lowland with varying hydrogeology. We selected characteristic sites for five major hydrogeological classes of the Netherlands and installed paired 5-m-wide unfertilized grass (BS) and reference (REF) treatments along the ditch. The REF was managed like the adjacent field, and BS was only harvested. Treatments were equipped with reservoirs in the ditch to collect and measure discharge and flow proportional N concentration for 3 or 4 yr. In addition, N concentration in upper groundwater was measured. We found a statistically significant BSE of 10% on the peat site. At the other sites, BSE for N was low and statistically insignificant. Low BSE was explained by denitrification between adjacent field and ditch, as well as by the site-specific hydrologic factors including low proportion of shallow groundwater flow, downward seepage, low residence time in the BS, and surface runoff away from the ditch. We emphasize that a REF treatment is needed to evaluate BSE in agriculture and recommend reservoirs if drainage patterns are unknown. Introduction of a 5-m-wide BS is ineffective for mitigating N loads from lowland agriculture to surface waters. We expect more from BS specifically designed to abate surface runoff.

  6. Reducing dissolved inorganic nitrogen in surface runoff water from sugarcane production systems.

    PubMed

    Webster, A J; Bartley, R; Armour, J D; Brodie, J E; Thorburn, P J

    2012-01-01

    Nitrogen (N) lost from farms, especially as the highly bioavailable dissolved inorganic form, may be damaging Australia's Great Barrier Reef (GBR). As sugarcane is the dominant cropping system in GBR catchments, its N management practises are coming under increasing scrutiny. This study measured dissolved inorganic N lost in surface runoff water and sugarcane productivity over 3 years. The experiment compared the conventional fertiliser N application rate to sugarcane (average 180kg N/ha/year) and a rate based on replacing N exported in the previous crop (average 94kg N/ha/year). Dissolved inorganic N losses in surface water were 72%, 48% and 66% lower in the three monitored years in the reduced N fertiliser treatment. There was no significant difference in sugarcane yield between the two fertiliser N treatments, nor any treatment difference in soil mineral N - both of these results are indicators of the sustainability of the lower fertiliser N applications. Copyright © 2012 Elsevier Ltd. All rights reserved.

  7. Probing Surface Oxidation of Reduced Uranium Dioxide Thin Film Using Synchrotron Radiation

    SciTech Connect

    Senanayake,S.; Waterhouse, G.; Chan, A.; Madey , T.; Mullins, D.; Idriss, H.

    2007-01-01

    The work presents a study of the formation of defects created by argon ion sputtering of UO{sub 2} thin film surfaces using high-resolution X-ray photoelectron spectroscopy with photon energy of 625 eV. Stable-defected surfaces of UO{sub 2-x} were monitored by the uranium core level lines found at the low-binding energy side of the U{sup +4} features. The lower end of these states gave binding energies at 377.1 and 387.7 eV attributed to U4f{sub 7/2} and U4f{sub 5/2} core levels peaks, respectively, for metallic uranium (U{sup {circ}}). In between both states (U{sup {circ}} and U{sup +4}), the clear presence of other electronic states is observed. These lines are attributed to intermediate, metastable states of uranium ions between U{sup {circ}} and U{sup +4}. Temperature-programmed X-ray photoelectron spectroscopy is performed to observe the oxidation process of these reduced states with increasing temperature. The increase in the U{sup +4} intensity is not correlated with the decrease of U ions signal with lower oxidation states indicating that the limitation is the photoelectron escape depth. In other words, sputtering has created reduced clusters or ridges on the surface of average dimensions larger than the escape depth of the 4f photoelectron with kinetic energy of 225 eV. The activation energy for surface oxidation to U{sup +4} is {approx}20 kJ mol{sup -1}. This value is close to other values reported for vacancy-type diffusion mechanism in the fluorite structure.

  8. Reducing intrinsic loss in superconducting resonators by surface treatment and deep etching of silicon substrates

    SciTech Connect

    Bruno, A.; Lange, G. de; Asaad, S.; Enden, K. L. van der; Langford, N. K.; DiCarlo, L.

    2015-05-04

    We present microwave-frequency NbTiN resonators on silicon, systematically achieving internal quality factors above 1 M in the quantum regime. We use two techniques to reduce losses associated with two-level systems: an additional substrate surface treatment prior to NbTiN deposition to optimize the metal-substrate interface and deep reactive-ion etching of the substrate to displace the substrate-vacuum interfaces away from high electric fields. The temperature and power dependence of resonator behavior indicate that two-level systems still contribute significantly to energy dissipation, suggesting that more interface optimization could further improve performance.

  9. Gene expression by the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough grown on an iron electrode under cathodic protection conditions.

    PubMed

    Caffrey, Sean M; Park, Hyung Soo; Been, Jenny; Gordon, Paul; Sensen, Christoph W; Voordouw, Gerrit

    2008-04-01

    The genome sequence of the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough was reanalyzed to design unique 70-mer oligonucleotide probes against 2,824 probable protein-coding regions. These included three genes not previously annotated, including one that encodes a c-type cytochrome. Using microarrays printed with these 70-mer probes, we analyzed the gene expression profile of wild-type D. vulgaris grown on cathodic hydrogen, generated at an iron electrode surface with an imposed negative potential of -1.1 V (cathodic protection conditions). The gene expression profile of cells grown on cathodic hydrogen was compared to that of cells grown with gaseous hydrogen bubbling through the culture. Relative to the latter, the electrode-grown cells overexpressed two hydrogenases, the hyn-1 genes for [NiFe] hydrogenase 1 and the hyd genes, encoding [Fe] hydrogenase. The hmc genes for the high-molecular-weight cytochrome complex, which allows electron flow from the hydrogenases across the cytoplasmic membrane, were also overexpressed. In contrast, cells grown on gaseous hydrogen overexpressed the hys genes for [NiFeSe] hydrogenase. Cells growing on the electrode also overexpressed genes encoding proteins which promote biofilm formation. Although the gene expression profiles for these two modes of growth were distinct, they were more closely related to each other than to that for cells grown in a lactate- and sulfate-containing medium. Electrochemically measured corrosion rates were lower for iron electrodes covered with hyn-1, hyd, and hmc mutant biofilms than for wild-type biofilms. This confirms the importance, suggested by the gene expression studies, of the corresponding gene products in D. vulgaris-mediated iron corrosion.

  10. Surface grafting of reduced graphene oxide using nanocrystalline cellulose via click reaction

    NASA Astrophysics Data System (ADS)

    Kabiri, Roya; Namazi, Hassan

    2014-07-01

    Reduced graphene oxide (RGO) sheet was functionalized with nanocrystalline cellulose (NCC) via click coupling between azide-functionalized graphene oxide (GO-N3) and terminal propargyl-functionalized nanocrystalline cellulose (PG-NCC). First, the reactive azide groups were introduced on the surface of GO with azidation of 2-chloroethyl isocyanate-treated graphene oxide (GO-Cl). Then, the resulted compounds were reacted with PG-NCC utilizing copper-catalyzed azide-alkyne cycloaddition. During the click reaction, GO was simultaneously reduced to graphene. The coupling was confirmed by Fourier transform infrared, Raman, DEPT135, and 13C NMR spectroscopy, and the complete exfoliation of graphene in the NCC matrix was confirmed with X-ray diffraction measurement. The degree of functionalization from the gradual mass loss of RGO-NCC suggests that around 23 mass % has been functionalized covalently. The size of both NCC and GO was found to be in nanometric range, which decreased after click reaction.

  11. Global Pressure and Temperature Surface Measurements on a NACA 0012 Airfoil in Oscillatory Compressible Flow at Low Reduced Frequencies

    DTIC Science & Technology

    2012-01-01

    Global Pressure and Temperature Surface Measurements on a NACA 0012 Airfoil in Oscillatory Compressible Flow at Low Reduced Frequencies...SUBTITLE Global Pressure And Temperature Surface Measurements On A NACA 0012 Airfoil In Oscillatory Compressible Flow At Low Reduced Frequencies 5a...31 Section 4.2: Lifetime- vs . Intensity-Based PSP Techniques

  12. Daily disinfection of high-touch surfaces in isolation rooms to reduce contamination of healthcare workers' hands.

    PubMed

    Kundrapu, Sirisha; Sunkesula, Venkata; Jury, Lucy A; Sitzlar, Brett M; Donskey, Curtis J

    2012-10-01

    In a randomized nonblinded trial, we demonstrated that daily disinfection of high-touch surfaces in rooms of patients with Clostridium difficile infection and methicillin-resistant Staphylococcus aureus colonization reduced acquisition of the pathogens on hands after contacting high-touch surfaces and reduced contamination of hands of healthcare workers caring for the patients.

  13. Deafness and permanently reduced potassium channel gene expression and function in hypothyroid Pit1dw mutants.

    PubMed

    Mustapha, Mirna; Fang, Qing; Gong, Tzy-Wen; Dolan, David F; Raphael, Yehoash; Camper, Sally A; Duncan, R Keith

    2009-01-28

    The absence of thyroid hormone (TH) during late gestation and early infancy can cause irreparable deafness in both humans and rodents. A variety of rodent models have been used in an effort to identify the underlying molecular mechanism. Here, we characterize a mouse model of secondary hypothyroidism, pituitary transcription factor 1 (Pit1(dw)), which has profound, congenital deafness that is rescued by oral TH replacement. These mutants have tectorial membrane abnormalities, including a prominent Hensen's stripe, elevated beta-tectorin composition, and disrupted striated-sheet matrix. They lack distortion product otoacoustic emissions and cochlear microphonic responses, and exhibit reduced endocochlear potentials, suggesting defects in outer hair cell function and potassium recycling. Auditory system and hair cell physiology, histology, and anatomy studies reveal novel defects of hormone deficiency related to deafness: (1) permanently impaired expression of KCNJ10 in the stria vascularis of Pit1(dw) mice, which likely contributes to the reduced endocochlear potential, (2) significant outer hair cell loss in the mutants, which may result from cellular stress induced by the lower KCNQ4 expression and current levels in Pit1(dw) mutant outer hair cells, and (3) sensory and strial cell deterioration, which may have implications for thyroid hormone dysregulation in age-related hearing impairment. In summary, we suggest that these defects in outer hair cell and strial cell function are important contributors to the hearing impairment in Pit1(dw) mice.

  14. Phenanthrene exposure induces cardiac hypertrophy via reducing miR-133a expression by DNA methylation

    PubMed Central

    Huang, Lixing; Xi, Zhihui; Wang, Chonggang; Zhang, Youyu; Yang, Zhibing; Zhang, Shiqi; Chen, Yixin; Zuo, Zhenghong

    2016-01-01

    Growing evidence indicates that there is an emerging link between environmental pollution and cardiac hypertrophy, while the mechanism is unclear. The objective of this study was to examine whether phenanthrene (Phe) could cause cardiac hypertrophy, and elucidate the molecular mechanisms involved. We found that: 1) Phe exposure increased the heart weight and cardiomyocyte size of rats; 2) Phe exposure led to enlarged cell size, and increased protein synthesis in H9C2 cells; 3) Phe exposure induced important markers of cardiac hypertrophy, such as atrial natriuretic peptide, B-type natriuretic peptide, and c-Myc in H9C2 cells and rat hearts; 4) Phe exposure perturbed miR-133a, CdC42 and RhoA, which were key regulators of cardiac hypertrophy, in H9C2 cells and rat hearts; 5) Phe exposure induced DNA methyltransferases (DNMTs) in H9C2 cells and rat hearts; 6) Phe exposure led to methylation of CpG sites within the miR-133a locus and reduced miR-133a expression in H9C2 cells; 7) DNMT inhibition and miR-133a overexpression could both alleviate the enlargement of cell size and perturbation of CdC42 and RhoA caused by Phe exposure. These results indicated that Phe could induce cardiomyocyte hypertrophy in the rat and H9C2 cells. The mechanism might involve reducing miR-133a expression by DNA methylation. PMID:26830171

  15. Reduced RANKL expression impedes osteoclast activation and tooth eruption in alendronate-treated rats.

    PubMed

    Bradaschia-Correa, Vivian; Moreira, Mariana M; Arana-Chavez, Victor E

    2013-07-01

    The creation of the eruption pathway requires the resorption of the occlusal alveolar bone by osteoclasts and signaling events between bone and dental follicle are necessary. The aim of the present study has been to evaluate the effect of alendronate on osteoclastogenesis and the expression of the regulator proteins of osteoclast activation, namely RANK, RANKL and OPG, in the bone that covers the first molar germ. Newborn Wistar rats were treated daily with 2.5 mg/kg alendronate for 4, 8, 14, 21 and 28 days, whereas controls received sterile saline solution. At the time points cited, maxillae were fixed, decalcified and processed for light and electron microscopic analysis. TRAP histochemistry was performed on semi-serial sections and the osteoclasts in the occlusal half of the bony crypt surface were counted. TUNEL analysis was carried out on paraffin sections. The occlusal bone that covers the upper first molar was removed in additional 4- and 8-day-old alendronate-treated and control rats in which the expression of RANK, RANKL and OPG was analyzed by SDS-polyacrylamide gel electrophoresis and Western blotting. TRAP-positive osteoclasts were more numerous in the alendronate group at all time points, despite their unactivated phenotype and the presence of apoptotic cells. RANKL expression in the alendronate specimens was inhibited at all time points, unlike in controls. Our findings indicate that the expression of RANKL in the occlusal portion of the bony crypt is unrelated to osteoclast recruitment and differentiation but is crucial to their activation during the creation of the eruption pathway.

  16. Differential expression of surface antigens on activated endothelium.

    PubMed

    Favaloro, E J

    1993-12-01

    Endothelial cells lining the vasculature participate in a variety of physiological processes. Following cell activation, functional changes are accompanied by changes in the surface structure (or phenotype) of these cells. Studies to date have tended to concentrate on selective changes induced with one or two surface molecules. The following study uses a different approach, having assessed potential changes to the endothelial cell surface using a large number (> 120) of previously untested monoclonal antibodies, and the cytokines TNF-alpha and gamma-IFN, as well as the proteolytic enzyme thrombin. Antibody representatives from all cluster of differentiation groups CD1 through to CD54 were assessed in these studies, which used human umbilical vein endothelial cells. In line with previous observations, antibodies within CD9, CD13, CD26, CD29, CD31, CD34, CD44, CD46, CD47, CD49, CD51 and CD54 gave significant and consistent reactivity using non-stimulated ('quiescent') endothelium. Using parallel cells differentially stimulated with TNF-alpha, gamma-IFN or thrombin, antibodies within CD1 through to CD15, CDw17 to CD19, CD21 to CD23, CD26, CD27, CD29, CD30, CD33 to CD35, CD37, CD38, CD40, CD43 to CD46, CD48, CD51 to CD53 failed to provide any consistent alteration to reactivity patterns compared to non-stimulated cells. There did, however, appear to be some activation induced changes using antibodies within the other CD groups (i.e. CD16, CD20, CD24, CD25, CD28, CD31, CD32, CD36, CD39, CD41, CD42, CD47, CD49, CD50 and CD54) which ranged from minor to significant in scope and magnitude.

  17. Influences of reduced expression of maternal bone morphogenetic protein 2 on mouse embryonic development.

    PubMed

    Singh, A P; Castranio, T; Scott, G; Guo, D; Harris, M A; Ray, M; Harris, S E; Mishina, Y

    2008-01-01

    Bone morphogenetic protein 2 (BMP2) was originally found by its osteoinductive ability, and recent genetic analyses have revealed that it plays critical roles during early embryogenesis, cardiogenesis, decidualization as well as skeletogenesis. In the course of evaluation of the conditional allele for Bmp2, we found that the presence of a neo cassette, a selection marker needed for gene targeting events in embryonic stem cells, in the 3' untranslated region of exon 3 of Bmp2, reduced the expression levels of Bmp2 both in embryonic and maternal mouse tissues. Some of the embryos that were genotyped as transheterozygous for the floxed allele with the neo cassette over the conventional null allele (fn/-) showed a lethal phenotype including defects in cephalic neural tube closure and ventral abdominal wall closure. The number of embryos exhibiting these abnormalities was increased when, due to different genotypes, expression levels of Bmp2 in maternal tissues were lower. These results suggest that the expression levels of Bmp2 in both embryonic and maternal tissues influence the normal neural tube closure and body wall closure with different thresholds.

  18. A 3 base pair deletion in TBX1 leads to reduced protein expression and transcriptional activity

    PubMed Central

    Xu, Yuejuan; Fang, Shaohai; Zhang, Erge; Pu, Tian; Cao, Ruixue; Fu, Qihua; Li, Fen; Chen, Sun; Sun, Kun; Xu, Rang

    2017-01-01

    Transcription factor TBX1 plays a pivotal role in heart development and has been implicated in 22q11.2 deletion syndrome. The structure of this protein has been elucidated, and several mutations have been identified that disrupt TBX1 localization, DNA/protein-binding, or mRNA expression. This study reports a mutation in the TBX1 gene that leads to significantly reduced expression of the mutant protein. A total of 773 conotruncal heart defect patients and 516 unrelated healthy control individuals were enrolled, none of which harbored a 22q11.2 deletion or duplication. We identified a mutation, c.303-305delGAA, located in the third exon of TBX1 that does not disrupt TBX1 mRNA expression or DNA binding activity, but results in decreased TBX1 protein levels and transcriptional activity. Through protein degradation studies we demonstrated that TBX1 is degraded primarily in proteasomes. Although the c.303-305delGAA mutation leads to low levels of the mutant protein, we found that increased protein degradation was not the cause, and we hypothesize that an alternate mechanism, such as translational inhibition, may be the cause. PMID:28272434

  19. Reduced expression of glycolate oxidase leads to enhanced disease resistance in rice.

    PubMed

    Chern, Mawsheng; Bai, Wei; Chen, Xuewei; Canlas, Patrick E; Ronald, Pamela C

    2013-01-01

    Glycolate oxidase (GLO) is a key enzyme in photorespiration, catalyzing the oxidation of glycolate to glyoxylate. Arabidopsis GLO is required for nonhost defense responses to Pseudomonas syringae and for tobacco Pto/AvrPto-mediated defense responses. We previously described identification of rice GLO1 that interacts with a glutaredoxin protein, which in turn interacts with TGA transcription factors. TGA transcription factors are well known to participate in NPR1/NH1-mediated defense signaling, which is crucial to systemic acquired resistance in plants. Here we demonstrate that reduction of rice GLO1 expression leads to enhanced resistance to Xanthomonas oryzae pv oryzae (Xoo). Constitutive silencing of GLO1 leads to programmed cell death, resulting in a lesion-mimic phenotype and lethality or reduced plant growth and development, consistent with previous reports. Inducible silencing of GLO1, employing a dexamethasone-GVG (Gal4 DNA binding domain-VP16 activation domain-glucocorticoid receptor fusion) inducible system, alleviates these detrimental effects. Silencing of GLO1 results in enhanced resistance to Xoo, increased expression of defense regulators NH1, NH3, and WRKY45, and activation of PR1 expression.

  20. Does expressive writing reduce stress and improve health for family caregivers of older adults?

    PubMed

    Mackenzie, Corey S; Wiprzycka, Ursula J; Hasher, Lynn; Goldstein, David

    2007-06-01

    We examined whether written emotional disclosure reduces stress and improves health outcomes for family caregivers of physically frail and cognitively impaired older adults, as it has been shown to do for certain student and clinical populations. Primary caregivers of older adults attending a day program were randomly assigned to expressive-writing (n = 14), time-management (n = 13), or history-writing (n = 13) conditions. Participants wrote for 20 minutes on four occasions over a 2-week period, and they completed self-report measures of caregiver burden and health prior to the intervention, immediately afterward, and at 1-month follow-up. Contrary to expectations, expressive-writing and history-writing participants performed similarly across outcomes. Only caregiver participants in the time-management condition experienced significant mental and physical health improvements after writing. The results of this study add to a growing body of research demonstrating equivocal effects of expressive writing with clinical samples, and they suggest the potential benefit of written time management for stressed caregivers.

  1. TGF-beta and metalloproteinases differentially suppress NKG2D ligand surface expression on malignant glioma cells.

    PubMed

    Eisele, Günter; Wischhusen, Jörg; Mittelbronn, Michel; Meyermann, Richard; Waldhauer, Inja; Steinle, Alexander; Weller, Michael; Friese, Manuel A

    2006-09-01

    NKG2D ligands (NKG2DL) are expressed by infected and transformed cells. They transmit danger signals to NKG2D-expressing immune cells, leading to lysis of NKG2DL-expressing cells. We here report that the NKG2DL MHC class I-chain-related molecules A and B (MICA/B) and UL16-binding proteins (ULBP) 1-3 are expressed in human brain tumours in vivo, while expression levels are low or undetectable in normal brain. MICA and ULBP2 expression decrease with increasing WHO grade of malignancy, while MICB and ULBP1 are expressed independently of tumour grade. We further delineate two independent mechanisms that can explain these expression patterns: (i) transforming growth factor-beta (TGF-beta) is upregulated during malignant progression and selectively downregulates MICA, ULBP2 and ULBP4 expression, while MICB, ULBP1 and ULBP3 are unaffected. (ii) Cleavage of MICA and ULBP2 is reduced by inhibition of metalloproteinases (MP), whereas no changes in the expression levels of other NKG2DL were detected. Consequently, NKG2DL-dependent NK cell-mediated lysis is enhanced by depletion of TGF-beta or inhibition of MP. Thus, escape from NKG2D-mediated immune surveillance of malignant gliomas in vivo may be promoted by the inhibition of MICA and ULBP2 expression via an autocrine TGF-beta loop and by MP-dependent shedding from the cell surface. Loss of MICA and ULBP2, in contrast to other NKG2DL, may be particularly important in glioma immune escape, and differential regulation of human NKG2DL expression is part of the immunosuppressive properties of human malignant glioma cells.

  2. Surface-Displayed IL-10 by Recombinant Lactobacillus plantarum Reduces Th1 Responses of RAW264.7 Cells Stimulated with Poly(I:C) or LPS.

    PubMed

    Cai, Ruopeng; Jiang, Yanlong; Yang, Wei; Yang, Wentao; Shi, Shaohua; Shi, Chunwei; Hu, Jingtao; Gu, Wei; Ye, Liping; Zhou, Fangyu; Gong, Qinglong; Han, Wenyu; Yang, Guilian; Wang, Chunfeng

    2016-02-01

    Recently, poly-γ-glutamic acid synthetase A (pgsA) has been applied to display exogenous proteins on the surface of Lactobacillus casei or Lactococcus lactis, which results in a surfacedisplayed component of bacteria. However, the ability of carrying genes encoded by plasmids and the expression efficiency of recombinant bacteria can be somewhat affected by the longer gene length of pgsA (1,143 bp); therefore, a truncated gene, pgsA, was generated based on the characteristics of pgsA by computational analysis. Using murine IL-10 as an exogenous gene, recombinant Lactobacillus plantarum was constructed and the capacity of the surface-displayed protein and functional differences between exogenous proteins expressed by these strains were evaluated. Surface expression of IL-10 on both recombinant bacteria with anchorins and the higher expression levels in L. plantarum-pgsA'-IL-10 were confirmed by western blot assay. Most importantly, up-regulation of IL-1β, IL-6, TNF-α, IFN-γ, and the nuclear transcription factor NF-κB p65 in RAW264.7 cells after stimulation with Poly(I:C) or LPS was exacerbated after co-culture with L. plantarum-pgsA. By contrast, IL-10 expressed by these recombinant strains could reduce these factors, and the expression of these factors was associated with recombinant strains that expressed anchorin (especially in L. plantarum-pgsA'-IL-10) and was significantly lower compared with the anchorin-free strains. These findings indicated that exogenous proteins could be successfully displayed on the surface of L. plantarum by pgsA or pgsA', and the expression of recombinant bacteria with pgsA' was superior compared with bacteria with pgsA.

  3. Effectiveness of the compactness of clay and/or surface fines as a means of reducing acid mine drainage

    SciTech Connect

    Aspden, N.L.; Aharrah, E.C.

    1982-12-01

    The question of compaction of soil to reduce infiltration and subsequent acid-mine drainage is examined. Plots were constructed to direct surface flow into flumes. The flumes drained through weirs equipped with flow meters. Percolate was collected from wells and tension lysimeters. Runoff, percolate, and vegetation was analyzed. The results indicate the best treatment to be a lush vegetative cover. The least disturbance of spoil surface reduces erosion and sedimentation and also reduces the introduction of unweathered toxic materials to the surface layer. Compaction of surface fines and/or impervious clay is not the answer to mine reclamation.

  4. Optimization of Fat-Reduced Puff Pastry Using Response Surface Methodology

    PubMed Central

    Silow, Christoph; Zannini, Emanuele; Axel, Claudia; Belz, Markus C. E.; Arendt, Elke K.

    2017-01-01

    Puff pastry is a high-fat bakery product with fat playing a key role, both during the production process and in the final pastry. In this study, response surface methodology (RSM) was successfully used to evaluate puff pastry quality for the development of a fat-reduced version. The technological parameters modified included the level of roll-in fat, the number of fat layers (50–200) and the final thickness (1.0–3.5 mm) of the laminated dough. Quality characteristics of puff pastry were measured using the Texture Analyzer with an attached Extended Craft Knife (ECK) and Multiple Puncture Probe (MPP), the VolScan and the C-Cell imaging system. The number of fat layers and final dough thickness, in combination with the amount of roll-in fat, had a significant impact on the internal and external structural quality parameters. With technological changes alone, a fat-reduced (≥30%) puff pastry was developed. The qualities of fat-reduced puff pastries were comparable to conventional full-fat (33 wt %) products. A sensory acceptance test revealed no significant differences in taste of fatness or ‘liking of mouthfeel’. Additionally, the fat-reduced puff pastry resulted in a significant (p < 0.05) positive correlation to ‘liking of flavor’ and overall acceptance by the assessors. PMID:28231095

  5. Optimization of Fat-Reduced Puff Pastry Using Response Surface Methodology.

    PubMed

    Silow, Christoph; Zannini, Emanuele; Axel, Claudia; Belz, Markus C E; Arendt, Elke K

    2017-02-22

    Puff pastry is a high-fat bakery product with fat playing a key role, both during the production process and in the final pastry. In this study, response surface methodology (RSM) was successfully used to evaluate puff pastry quality for the development of a fat-reduced version. The technological parameters modified included the level of roll-in fat, the number of fat layers (50-200) and the final thickness (1.0-3.5 mm) of the laminated dough. Quality characteristics of puff pastry were measured using the Texture Analyzer with an attached Extended Craft Knife (ECK) and Multiple Puncture Probe (MPP), the VolScan and the C-Cell imaging system. The number of fat layers and final dough thickness, in combination with the amount of roll-in fat, had a significant impact on the internal and external structural quality parameters. With technological changes alone, a fat-reduced (≥30%) puff pastry was developed. The qualities of fat-reduced puff pastries were comparable to conventional full-fat (33 wt %) products. A sensory acceptance test revealed no significant differences in taste of fatness or 'liking of mouthfeel'. Additionally, the fat-reduced puff pastry resulted in a significant (p < 0.05) positive correlation to 'liking of flavor' and overall acceptance by the assessors.

  6. Chemical Inhibition of Kynureninase Reduces Pseudomonas aeruginosa Quorum Sensing and Virulence Factor Expression.

    PubMed

    Kasper, Stephen H; Bonocora, Richard P; Wade, Joseph T; Musah, Rabi Ann; Cady, Nathaniel C

    2016-04-15

    The opportunistic pathogen Pseudomonas aeruginosa utilizes multiple quorum sensing (QS) pathways to coordinate an arsenal of virulence factors. We previously identified several cysteine-based compounds inspired by natural products from the plant Petiveria alliacea which are capable of antagonizing multiple QS circuits as well as reducing P. aeruginosa biofilm formation. To understand the global effects of such compounds on virulence factor production and elucidate their mechanism of action, RNA-seq transcriptomic analysis was performed on P. aeruginosa PAO1 exposed to S-phenyl-l-cysteine sulfoxide, the most potent inhibitor from the prior study. Exposure to this inhibitor down-regulated expression of several QS-regulated virulence operons (e.g., phenazine biosynthesis, type VI secretion systems). Interestingly, many genes that were differentially regulated pertain to the related metabolic pathways that yield precursors of pyochelin, tricarboxylic acid cycle intermediates, phenazines, and Pseudomonas quinolone signal (PQS). Activation of the MexT-regulon was also indicated, including the multidrug efflux pump encoded by mexEF-oprN, which has previously been shown to inhibit QS and pathogenicity. Deeper investigation of the metabolites involved in these systems revealed that S-phenyl-l-cysteine sulfoxide has structural similarity to kynurenine, a precursor of anthranilate, which is critical for P. aeruginosa virulence. By supplementing exogenous anthranilate, the QS-inhibitory effect was reversed. Finally, it was shown that S-phenyl-l-cysteine sulfoxide competitively inhibits P. aeruginosa kynureninase (KynU) activity in vitro and reduces PQS production in vivo. The kynurenine pathway has been implicated in P. aeruginosa QS and virulence factor expression; however, this is the first study to show that targeted inhibition of KynU affects P. aeruginosa gene expression and QS, suggesting a potential antivirulence strategy.

  7. Early hypermethylation of hepatic Igfbp2 results in its reduced expression preceding fatty liver in mice

    PubMed Central

    Kammel, Anne; Saussenthaler, Sophie; Jähnert, Markus; Jonas, Wenke; Stirm, Laura; Hoeflich, Andreas; Staiger, Harald; Fritsche, Andreas; Häring, Hans-Ulrich; Joost, Hans-Georg; Schürmann, Annette; Schwenk, Robert W.

    2016-01-01

    Obesity and ectopic fat disposition are risk factors for metabolic disease. Recent data indicate that IGFBP2 expression in liver is epigenetically inhibited during hepatic steatosis. The aim of this study was to investigate if epigenetic de-regulation of hepatic Igfbp2 occurs already early in life and is associated with increased risk for diet-induced obesity (DIO) during adolescence. Male C57BL/6J mice received a high-fat diet. After 3 weeks on this diet (age of 6 weeks), DIO-susceptible (responder, Resp) and DIO-resistant (non-responder, nResp) mice were identified by early weight gain. At the age of 6 weeks, Resp mice exhibited elevated blood glucose (p < 0.05), plasma insulin (p < 0.01), HOMA-IR and leptin/adiponectin ratio, whereas liver triglycerides were identical but significantly increased (p < 0.01) in Resp mice at 20 weeks of age. Igfbp2 expression was reduced in young Resp compared with nResp mice (p < 0.01), an effect that correlated with elevated DNA methylation of intronic CpG2605 (p < 0.01). The epigenetic inhibition of Igfbp2 was stable over time and preceded DIO and hepatosteatosis in adult mice. In vitro studies demonstrated that selective methylation of CpG2605 significantly reduced reporter activity by ∼85%, indicating that Igfbp2 expression is modulated by methylation. In human whole blood cells, methylation of IGFBP2 at the homologous CpG site was increased in obese men with impaired glucose tolerance. In conclusion, our data show that increased methylation of hepatic Igfbp2 during infancy predicts the development of fatty liver later in life and is linked to deterioration of glucose metabolism. PMID:27126637

  8. An Anacardiaceae preparation reduces the expression of inflammation-related genes in murine macrophages.

    PubMed

    Leiro, J; García, D; Arranz, J A; Delgado, R; Sanmartín, M L; Orallo, F

    2004-08-01

    This study investigated the effects of an aqueous extract of the stem bark of Mangifera indica L. (Anacardiaceae; Vimang), which contains a defined mixture of components including polyphenols (principally mangiferin, MA), triterpenes, phytosteroids, fatty acids and microelements, on expression of inflammation mediators in inflammatory murine macrophages after stimulation in vitro with lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma). In vitro treatment with Vimang at 4 microg/ml reduced levels of NOS-2 mRNA and NOS-2, while treatment at 40 microg/ml also reduced levels of COX-2 mRNA, COX-2, and prostaglandin E2 (PGE2). Results suggested that MA is involved in these effects. In vitro treatment with Vimang at 40 microg/ml also inhibited mRNA levels of the proinflammatory cytokines interleukin 1beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha) and colony-stimulating factor (GM-CSF), but did not affect mRNA levels of IL-6 or tumor growth factor-beta (TGF-beta). Extracellular release of TNF-alpha by inflammatory macrophages was inhibited by in vitro treatment with Vimang at the same concentrations that showed inhibition of TNF-alpha mRNA levels. The inhibition of TNF-alpha production appears to be at least partially attributable to MA. Vimang at 4 microg/ml decreased mRNA levels of nuclear factor-kappaB (NF-kappaB) but did not affect expression of the NF-kappaB inhibitor (IkappaB). These data indicate that the potent anti-inflammatory effects of Vimang are due to selective modulation of the expression of inflammation-related genes, leading to attenuation of macrophage activation.

  9. Semaphorin 3F expression is reduced in pregnancy complicated by preeclampsia. An observational clinical study

    PubMed Central

    Stallone, Giovanni; Matteo, Maria; Netti, Giuseppe Stefano; Infante, Barbara; Di Lorenzo, Adelaide; Prattichizzo, Clelia; Carlucci, Stefania; Trezza, Federica; Gesualdo, Loreto; Greco, Pantaleo

    2017-01-01

    Background and objective Preeclampsia is a systemic disorder, affecting 2–10% of pregnancies, characterized by a deregulated pro- and anti-angiogenic balance. Semaphorin 3F is an angiogenesis inhibitor. We aimed to investigate whether semaphorin 3F expression is modulated in preeclampsia. Design, setting, participants, and measurements We performed two observational single center cohort studies between March 2013 and August 2014. In the first we enrolled 110 consecutive women, undergoing an elective cesarean section; in the second we included 150 consecutive women undergoing amniocentesis for routine clinical indications at 16–18 week of gestation. Semaphorin 3F concentration was evaluated in maternal peripheral blood, venous umbilical blood and amniotic fluid, along with its placenta protein expression at the time of delivery in the first study group and in the amniotic fluid at 16–18 weeks of gestation in the second study group. Results In the first study 19 patients presented at delivery with preeclampsia. Semaphorin 3F placenta tissue expression was significantly reduced in preeclampsia. In addition, semaphorin 3F level at delivery was significantly lower in serum, amniotic fluid and venous umbilical blood of preeclamptic patients compared with normal pregnant women. In the prospective cohort study 14 women developed preeclampsia. In this setting, semaphorin 3F amniotic level at 16–18 weeks of gestation was reduced in women who subsequently developed preeclampsia compared to women with a normal pregnancy. ROC curve analysis showed that semaphorin 3F amniotic levels could identify women at higher risk of preeclampsia. Conclusions Semaphorin 3F might represent a predictive biomarker of preeclampsia. PMID:28350837

  10. Surface functionalities of gold nanoparticles impact embryonic gene expression responses

    PubMed Central

    Truong, Lisa; Tilton, Susan C.; Zaikova, Tatiana; Richman, Erik; Waters, Katrina M.; Hutchison, James E.; Tanguay, Robert L.

    2012-01-01

    Incorporation of gold nanoparticles (AuNPs) into consumer products is increasing; however, there is a gap in available toxicological data to determine the safety of AuNPs. In this study, we utilised the embryonic zebrafish to investigate how surface functionalisation and charge influence molecular responses. Precisely engineered AuNPs with 1.5 nm cores were synthesised and functionalized with three ligands: 2-mercaptoethanesulfonic acid (MES), N,N,N-trimethylammoniumethanethiol (TMAT), or 2-(2-(2-mercaptoethoxy)ethoxy)ethanol. Developmental assessments revealed differential biological responses when embryos were exposed to the functionalised AuNPs at the same concentration. Using inductively coupled plasma–mass spectrometry, AuNP uptake was confirmed in exposed embryos. Following exposure to MES- and TMAT-AuNPs from 6 to 24 or 6 to 48 h post fertilisation, pathways involved in inflammation and immune response were perturbed. Additionally, transport mechanisms were misregulated after exposure to TMAT and MES-AuNPs, demonstrating that surface functionalisation influences many molecular pathways. PMID:22263968

  11. Atomic and electronic structure of unreduced and reduced CeO2 surfaces: a first-principles study.

    PubMed

    Yang, Zongxian; Woo, Tom K; Baudin, Micael; Hermansson, Kersti

    2004-04-22

    The atomic and electronic structure of (111), (110), and (100) surfaces of ceria (CeO2) were studied using density-functional theory within the generalized gradient approximation. Both stoichiometric surfaces and surfaces with oxygen vacancies (unreduced and reduced surfaces, respectively) have been examined. It is found that the (111) surface is the most stable among the considered surfaces, followed by (110) and (100) surfaces, in agreement with experimental observations and previous theoretical results. Different features of relaxation are found for the three surfaces. While the (111) surface undergoes very small relaxation, considerably larger relaxations are found for the (110) and (100) surfaces. The formation of an oxygen vacancy is closely related to the surface structure and occurs more easily for the (110) surface than for (111). The preferred vacancy location is in the surface layer for CeO2(110) and in the subsurface layer (the second O-atomic layer) for CeO2(111). For both surfaces, the O vacancy forms more readily than in the bulk. An interesting oscillatory behavior is found for the vacancy formation energy in the upper three layers of CeO2(111). Analysis of the reduced surfaces suggests that the additional charge resulting from the formation of the oxygen vacancies is localized in the first three layers of the surface. Furthermore, they are not only trapped in the 4f states of cerium. (c) 2004 American Institute of Physics

  12. Enhancing surface functionality of reduced graphene oxide biosensors by oxygen plasma treatment for Alzheimer's disease diagnosis.

    PubMed

    Chae, Myung-Sic; Kim, Jinsik; Jeong, Dahye; Kim, YoungSoo; Roh, Jee Hoon; Lee, Sung Min; Heo, Youhee; Kang, Ji Yoon; Lee, Jeong Hoon; Yoon, Dae Sung; Kim, Tae Geun; Chang, Suk Tai; Hwang, Kyo Seon

    2017-06-15

    We performed oxygen plasma treatment on reduced graphene oxide (rGO) to improve its surface reactivity with respect to biomolecular interactions. Oxygen-plasma-treated rGO surfaces were employed as reactive interfaces for the detection of amyloid-beta (Aβ) peptides, the pathological hallmarks of Alzheimer's disease (AD), as the target analytes. By measuring the changes in electrical characteristics and confirmation through topographic analysis, the oxygen-plasma-treated rGO sensors had enhanced surface functionality for better antibody immobilization and sensing performance, with a 3.33-fold steeper slope for the electrical responses versus analyte concentration curve (logarithmic scale) compared to the untreated. The elicited biomolecular reactivity of the rGO surfaces with the oxygen plasma treatment remained at 46-51% of the initial value even after aging for 6h in ambient conditions. This phenomenon was also confirmed by pretreating the rGO surfaces with a blocking agent and subsequently subjecting them to antibody immobilization. Finally, the feasibility of the oxygen-plasma-treated rGO sensors as a diagnostic tool was evaluated with clinical samples of neural-derived exosomal Aβ peptides extracted from apparent AD patients and normal controls (NC). In contrast to the untreated sensors (p=0.0460), the oxygen-plasma-treated rGO sensors showed a significant p-value in the identification of clinical samples of AD and NC subjects (p<0.001). These results suggest that oxygen plasma treatment improves sensor performance without complicated fabrication procedures and should aid in the development of novel diagnostic tools based on carbon nanomaterials.

  13. Reduced Connexin 43 expression is associated with tumor malignant behaviors and biochemical recurrence-free survival of prostate cancer

    PubMed Central

    Xu, Ning; Chen, Hui-Jun; Chen, Shao-Hao; Xue, Xue-Yi; Chen, Hong; Zheng, Qing-Shui; Wei, Yong; Li, Xiao-Dong; Huang, Jin-Bei; Cai, Hai; Sun, Xiong-Lin

    2016-01-01

    Connexin 43, a gap junction protein, coordinates cell-to-cell communication and adhesion. Altered Connexin 43 expression associated with cancer development and progression. In this study, we assessed Connexin 43 expression for association with clinicopathological features and biochemical recurrence of prostate cancer after radical prostatectomy. Pathological specimens were collected from 243 patients who underwent radical prostatectomy and from 60 benign prostatic hyperplasia (BPH) patients to construct tissue microarrays and immunohistochemical analysis of Connexin 43 expression. Kaplan-Meier curves and multivariable Cox proportion hazard model were performed to associate Connexin 43 expression with postoperative biochemical recurrence-free survival (BFS). Connexin 43 expression was significantly reduced or lost in tumor tissues compared to that of BPHs (39.1% vs. 96.7%, P<0.001). Reduced Connexin 43 expression was associated with high levels of preoperative PSA, high Gleason score, advanced pT stage, positive surgical margin, extracapsular extension, and seminal vesicle invasion (P < 0.05, for all). Kaplan–Meier curves showed that reduced Connexin 43 expression was associated with shortened postoperative BFS (P < 0.001). Multivariate analysis showed that reduced Connexin 43 expression, high Gleason score and advanced pT stage were independent predictors for BFS of patients (P < 0.05). Connexin 43 expression was significantly reduced or lost in prostate cancer tissues, which was associated with advanced clinicopathological features and poor BFS of patients after radical prostatectomy. PMID:27623212

  14. Tissue factor expression on the surface of monocytes from a patient with hereditary angioedema.

    PubMed

    Iwamoto, Kazumasa; Morioke, Satoshi; Yanase, Yuhki; Uchida, Kazue; Hide, Michihiro

    2014-10-01

    Hereditary angioedema (HAE) presents as severe angioedema, which is mostly due to the C1 inhibitor (C1-INH) gene mutations. Environmental factors, minor trauma and oral contraceptives have been reported to induce angioedema attack, but the trigger may often be uncertain. Activated factor XII controlled by C1-INH facilitates bradykinin generation and also regulates coagulation cascade, but the relationship between edema formation and coagulation is still unclear. We have described a 35-year-old female patient with HAE, presenting with frequent angioedema attacks in the absence of an apparent triggering factor. She showed higher levels of FDP and D-dimer during angioedema than those in remission. In addition, tissue factor (TF), an initiator of the extrinsic coagulation cascade, was expressed on the surface of monocytes. It was significantly higher than that of monocytes from healthy controls and tends to further increase during attacks. The expression of TF on monocytes may play a role in the induction of angioedema attacks in HAE by activating the coagulation pathway in association with reduced functions of C1-INH.

  15. SYNTAXIN 1A REGULATES DOPAMINE TRANSPORTER ACTIVITY, PHOSPHORYLATION AND SURFACE EXPRESSION

    PubMed Central

    Cervinski, Mark A.; Foster, James D.; Vaughan, Roxanne A.

    2010-01-01

    We investigated the functional relationship between the SNARE protein syntaxin 1A (syn 1A) and the dopamine transporter (DAT) by treating rat striatal tissue with Botulinum Neurotoxin C (BoNT/C) and co-transfecting syn 1A with DAT in non-neuronal cells, followed by analysis of DAT activity, phosphorylation, and regulation. Treatment of striatal slices with BoNT/C resulted in elevated dopamine (DA) transport Vmax and reduced DAT phosphorylation, while heterologous co-expression of syn 1A led to reduction in DAT surface expression and transport Vmax. Syn 1A was present in DAT immunoprecipitation complexes, supporting a direct or indirect interaction between the proteins. Phorbol ester regulation of DA transport activity was retained in BoNT/C-treated synaptosomes and syn 1A transfected cells, demonstrating that PKC and syn 1A effects occur through independent processes. These findings reveal a novel mechanism for regulation of DAT activity and phosphorylation, and suggest the potential for syn 1A to impact DA neurotransmission through effects on reuptake. PMID:20643191

  16. Two Patients with Dry Eye Disease Followed Up Using an Expression Assay of Ocular Surface Mucin

    PubMed Central

    Machida, Yumiko; Shoji, Jun; Harada, Natsuko; Inada, Noriko

    2016-01-01

    Purpose We report 2 patients with dry eye disease followed up using the expression levels of ocular surface mucin. Case Reports Patient 1: a 57-year-old woman with Sjögren's syndrome-associated dry eyes experienced severe dryness and foreign body sensation in both her eyes, and instilled sodium hyaluronate ophthalmic solution 0.3% about 10–15 times daily. We measured the expression levels of MUC5AC mRNA (MUC5AC) and MUC16 mRNA (MUC16) by using real-time reversed transcription polymerase chain reaction for the specimens of modified impression cytology. Expression levels of MUC5AC and MUC16 on her ocular surface were very low. Subjective symptoms and expression levels of ocular surface mucin improved after combined treatment of rebamipide (4 times daily) and fluorometholone (once daily) ophthalmic suspension. Patient 2: a 62-year-old man with chronic graft-versus-host disease-associated dry eye experienced severe foreign body sensation and developed superficial punctate keratopathy with mucous thread and filamentary keratitis. Expression level of MUC5AC was very high at baseline. Subjective symptoms and expression levels of ocular surface mucin improved by combined treatment of rebamipide (4 times daily) and fluorometholone (once daily) ophthalmic suspension. Conclusion Clinical test for MUC gene expression on the ocular surface was found to be useful in the follow-up of dry eye treatment. PMID:27194990

  17. Reduced neurofilament expression in cutaneous nerve fibers of patients with CMT2E.

    PubMed

    Pisciotta, Chiara; Bai, Yunhong; Brennan, Kathryn M; Wu, Xingyao; Grider, Tiffany; Feely, Shawna; Wang, Suola; Moore, Steven; Siskind, Carly; Gonzalez, Michael; Zuchner, Stephan; Shy, Michael E

    2015-07-21

    To investigate the effects of NEFL Glu396Lys mutation on the expression and assembly of neurofilaments (NFs) in cutaneous nerve fibers of patients with Charcot-Marie-Tooth disease type 2E (CMT2E). A large family with CMT2E underwent clinical, electrophysiologic, and skin biopsy studies. Biopsies were processed by indirect immunofluorescence (IF), electron microscopy (EM), and Western blot analysis. The clinical features demonstrated intrafamilial phenotypic variability, and the electrophysiologic findings revealed nerve conductions that were either slow or in the intermediate range. All patients had reduced or absent compound muscular action potential amplitudes. Skin biopsies showed axons labeled with the axonal markers protein gene product 9.5 and α-tubulin, but not with NFs. The results of Western blot analysis were consistent with those of IF, showing reduced or absent NFs and normal expression of α-tubulin. EM revealed clusters of regenerated fibers, in absence of myelin sheath abnormalities. Both IF and EM failed to show NF aggregates in dermal axons. The morphometric analysis showed a smaller axonal caliber in patients than in controls. The study of the nodal/paranodal architecture demonstrated that sodium channels and Caspr were correctly localized in patients with CMT2E. Decrease in NF abundance may be a pathologic marker of CMT2E. The lack of NF aggregates, consistent with prior studies, suggests that they occur proximally leading to subsequent alterations in the axonal cytoskeleton. The small axonal caliber, along with the normal molecular architecture of nodes and paranodes, explain the reduced velocities detected in patients with CMT2E. Our results also demonstrate that skin biopsy can provide evidence of pathologic and pathogenic abnormalities in patients with CMT2E. © 2015 American Academy of Neurology.

  18. Reduced neurofilament expression in cutaneous nerve fibers of patients with CMT2E

    PubMed Central

    Bai, Yunhong; Brennan, Kathryn M.; Wu, Xingyao; Grider, Tiffany; Feely, Shawna; Wang, Suola; Moore, Steven; Siskind, Carly; Gonzalez, Michael; Zuchner, Stephan; Shy, Michael E.

    2015-01-01

    Objective: To investigate the effects of NEFL Glu396Lys mutation on the expression and assembly of neurofilaments (NFs) in cutaneous nerve fibers of patients with Charcot-Marie-Tooth disease type 2E (CMT2E). Methods: A large family with CMT2E underwent clinical, electrophysiologic, and skin biopsy studies. Biopsies were processed by indirect immunofluorescence (IF), electron microscopy (EM), and Western blot analysis. Results: The clinical features demonstrated intrafamilial phenotypic variability, and the electrophysiologic findings revealed nerve conductions that were either slow or in the intermediate range. All patients had reduced or absent compound muscular action potential amplitudes. Skin biopsies showed axons labeled with the axonal markers protein gene product 9.5 and α-tubulin, but not with NFs. The results of Western blot analysis were consistent with those of IF, showing reduced or absent NFs and normal expression of α-tubulin. EM revealed clusters of regenerated fibers, in absence of myelin sheath abnormalities. Both IF and EM failed to show NF aggregates in dermal axons. The morphometric analysis showed a smaller axonal caliber in patients than in controls. The study of the nodal/paranodal architecture demonstrated that sodium channels and Caspr were correctly localized in patients with CMT2E. Conclusions: Decrease in NF abundance may be a pathologic marker of CMT2E. The lack of NF aggregates, consistent with prior studies, suggests that they occur proximally leading to subsequent alterations in the axonal cytoskeleton. The small axonal caliber, along with the normal molecular architecture of nodes and paranodes, explain the reduced velocities detected in patients with CMT2E. Our results also demonstrate that skin biopsy can provide evidence of pathologic and pathogenic abnormalities in patients with CMT2E. PMID:26109717

  19. Carboxymethylated chitin reduces MMP-1 expression in rabbit ACLT osteoarthritic cartilage

    PubMed Central

    Hongbin, W; Jingyuan, D; Linyun, C; Yuming, D

    2004-01-01

    Objective: To examine the in vivo effects of carboxymethylated chitin (CMC), intra-articularly administered, on cartilage degradation and the level and distribution of cartilage matrix metalloproteinase-1 (MMP-1). Methods: Osteoarthritis (OA) was induced in 20 rabbits by unilateral anterior cruciate ligament transection (ACLT). The experimental group, comprising 10 rabbits randomly selected, was given an intra-articular injection of 0.3 ml of 2% CMC solution at 1, 3, and 5 weeks after ACLT. A further 10 rabbits that received an intra-articular injection of 0.3 ml normal saline at the same time served as controls. All knees were harvested at 6 weeks after surgery. Cartilage degradation of femoral condyles was evaluated at two levels: macroscopic and light microscopic. Tissue level and distribution of MMP-1 was documented by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry. Results: Cartilage degradation in the control group was significantly more severe than that in the experimental group both on the macroscopic grading scale and on Mankin's grading scale. In RT-PCR the amount of MMP-1 was significantly reduced by the treatment of CMC. Immunohistochemical study showed that in the experimental group MMP-1 was predominantly expressed in the superficial and upper intermediate layers of cartilage, and the amount of MMP-1 in the experimental group was also lower than that in control group. Conclusion: CMC significantly reduces the severity of cartilage degradation and reduces the expression of MMP-1 in cartilage, both at the mRNA and the protein level, and thus may be a potential drug for the treatment of OA. PMID:15020329

  20. Corticosteroids Mediate Heart Failure-Induced Depression through Reduced σ1-Receptor Expression

    PubMed Central

    Bhuiyan, Md. Shenuarin; Hasegawa, Hideyuki; Kanai, Hiroshi; Zhang, Chen; Han, Feng; Fukunaga, Kohji

    2016-01-01

    Cardiovascular diseases are risk factors for depression in humans. We recently proposed that σ1 receptor (σ1R) stimulation rescued cardiac hypertrophy and heart failure induced by transverse aortic constriction (TAC) in mice. Importantly, σ1R stimulation reportedly ameliorates depression-like behaviors in rodents. Thus, we hypothesized that impaired σ1R activity in brain triggers depression-like behaviors in animals with cardiovascular disease. Indeed, here we found that cardiac hypertrophy and heart failure induced by TAC were associated with depression-like behaviors concomitant with downregulation of σ1R expression in brain 6 weeks after surgery. σ1R levels significantly decreased in astrocytes in both the hippocampal CA1 region and dentate gyrus. Oral administration of the specific σ1R agonist SA4503 (0.3–1.0mg/kg) significantly improved TAC-induced depression-like behaviors concomitant with rescued astrocytic σ1R expression in CA1 and the dentate gyrus. Plasma corticosterone levels significantly increased 6 weeks after TAC, and chronic treatment of mice with corticosterone for 3 weeks elicited depression-like behaviors concomitant with reduced astrocytic σ1R expression in hippocampus. Furthermore, the glucocorticoid receptor antagonist mifepristone antagonized depressive-like behaviors and ameliorated decreased hippocampal σ1R expression in TAC mice. We conclude that elevated corticosterone levels trigger hippocampal σ1R downregulation and that σ1R stimulation with SA4503 is an attractive therapy to improve not only cardiac dysfunction but depression-like behaviors associated with heart failure. PMID:27741227

  1. Hexavalent chromium reduces larval growth and alters gene expression in mummichog (Fundulus heteroclitus).

    PubMed

    Roling, Jonathan A; Bain, Lisa J; Gardea-Torresdey, Jorge; Bader, Julia; Baldwin, William S

    2006-10-01

    Hexavalent chromium [Cr(VI)] is a common bioavailable metal ion that causes oxidative stress, DNA adducts, and perturbs gene expression. Changes in gene expression are useful biomarkers of toxicant exposure that provide information about an organism's health, adaptability, and toxicant-specific effects. Therefore, we developed a cDNA array for the estuarine sentinel species mummichog (Fundulus heteroclitus). Mummichog larvae were exposed to concentrations ranging from 0 to 24 mg/L (462 microM) of Cr(VI) for 30 d, and growth was measured to determine the no-observable-effect concentration (1.5 mg/L) and the lowest-observable-effect concentration (3 mg/L). Body burdens from Cr(VI)-exposed fish showed a dose-dependent increase and were inversely correlated to body weight. Mummichog larvae exposed to Cr(VI) differentially expressed 16 genes in a dose-dependent manner, including GLUT-2, L-FABP, ATPase synthase 8, type II keratin, TBT-binding protein, and complement component C3-2. Many of these genes are involved in energy metabolism or growth, which is consistent with the reduced growth observed. In subsequent experiments, adults were exposed to Cr(VI) for 7 d at 0, 1.5, or 3 mg/L, because adult mummichog are used in monitoring Superfund sites. Hexavalent chromium altered the expression of 10 genes in adult liver, including HGFA, H-FABP, and complement component C3-2. Many of these genes also are involved in energy metabolism. The mummichog arrays provide a potential mechanism for the effects of Cr(VI) on growth. We anticipate using these arrays and the data they provide to monitor effects at polluted sites, to assess the bioavailability of chromium at these sites, and to investigate the efficacy of remediation in chromium-polluted estuaries.

  2. Hypertonic saline reduces lipopolysaccharide-induced mouse brain edema through inhibiting aquaporin 4 expression

    PubMed Central

    2012-01-01

    Introduction Three percent sodium chloride (NaCl) treatment has been shown to reduce brain edema and inhibited brain aquaporin 4 (AQP4) expression in bacterial meningitis induced by Escherichia coli. Lipopolysaccharide (LPS) is the main pathogenic component of E. coli. We aimed to explore the effect of 3% NaCl in mouse brain edema induced by LPS, as well as to elucidate the potential mechanisms of action. Methods Three percent NaCl was used to treat cerebral edema induced by LPS in mice in vivo. Brain water content, IL-1β, TNFα, immunoglobulin G (IgG), AQP4 mRNA and protein were measured in brain tissues. IL-1β, 3% NaCl and calphostin C (a specific inhibitor of protein kinase C) were used to treat the primary astrocytes in vitro. AQP4 mRNA and protein were measured in astrocytes. Differences in various groups were determined by one-way analysis of variance. Results Three percent NaCl attenuated the increase of brain water content, IL-1β, TNFα, IgG, AQP4 mRNA and protein in brain tissues induced by LPS. Three percent NaCl inhibited the increase of AQP4 mRNA and protein in astrocytes induced by IL-1β in vitro. Calphostin C blocked the decrease of AQP4 mRNA and protein in astrocytes induced by 3% NaCl in vitro. Conclusions Osmotherapy with 3% NaCl ameliorated LPS-induced cerebral edema in vivo. In addition to its osmotic force, 3% NaCl exerted anti-edema effects possibly through down-regulating the expression of proinflammatory cytokines (IL-1β and TNFα) and inhibiting the expression of AQP4 induced by proinflammatory cytokines. Three percent NaCl attenuated the expression of AQP4 through activation of protein kinase C in astrocytes. PMID:23036239

  3. Sulfone derivatives reduce growth, adhesion and aspartic protease SAP2 gene expression.

    PubMed

    Bondaryk, Małgorzata; Ochal, Zbigniew; Staniszewska, Monika

    2014-09-01

    Fungal virulence factors represent a strategy for the design of new compounds with effective activities against Candida spp. Dichloromethyl-4-chloro-3-nitrophenylsulfone (named Compound 1) and chlorodibromomethyl-4-hydrazino-3-nitrophenylsulfone (Compound 2) versus Candida albicans virulence factors (SAP2 expression and adhesion to Caco-2 cell line) were investigated. Candida albicans SC5314 and its mutants: Δsap9, Δsap10, Δsap9/10 were used. MICs of the Compounds (concentrated at 0.0313-16 µg/ml) were determined using M27-A3. Percentage of cell inhibition was assessed spectrophotometrically (OD405) after 48 h at 35 °C. The SAP2 expression was analyzed with the use of RT-PCR; relative quantification was normalized against ACT1 in cells grown in YEPD and on Caco-2. Adherence assay of C. albicans to Caco-2 was performed in a 24-well-plate. Compound 1 showed higher activity (% = 100 at 4 µg/ml) than Compound 2 (MIC90 = 16 µg/ml). Dichloromethyl at the para position of the phenyl ring exerted anti-Candidal potential. Under Compound 1, SAP2 was down-regulated in all the strains (P ≤ 0.05). Conversely, SAP2 was over-expressed in Δsap9-10 (untreated cells) compared with the wild-type. The Compounds significantly affected adherence to epithelium (P ≤ 0.05). The tested sulfones interfered with the adhesion of C. albicans cells to the epithelial tissues without affecting their viability after 90-min of incubation. The Compounds' mode of action was attributed to the reduced adhesiveness and the lower SAP2 expression. Saps9-10 play a role in C. albicans adhesion and they can be involved in the sulfone resistance mechanisms.

  4. Neural stem cells genetically-modified to express neprilysin reduce pathology in Alzheimer transgenic models.

    PubMed

    Blurton-Jones, Mathew; Spencer, Brian; Michael, Sara; Castello, Nicholas A; Agazaryan, Andranik A; Davis, Joy L; Müller, Franz-Josef; Loring, Jeanne F; Masliah, Eliezer; LaFerla, Frank M

    2014-04-16

    Short-term neural stem cell (NSC) transplantation improves cognition in Alzheimer's disease (AD) transgenic mice by enhancing endogenous synaptic connectivity. However, this approach has no effect on the underlying beta-amyloid (Aβ) and neurofibrillary tangle pathology. Long term efficacy of cell based approaches may therefore require combinatorial approaches. To begin to examine this question we genetically-modified NSCs to stably express and secrete the Aβ-degrading enzyme, neprilysin (sNEP). Next, we studied the effects of sNEP expression in vitro by quantifying Aβ-degrading activity, NSC multipotency markers, and Aβ-induced toxicity. To determine whether sNEP-expressing NSCs can also modulate AD-pathogenesis in vivo, control-modified and sNEP-NSCs were transplanted unilaterally into the hippocampus of two independent and well characterized transgenic models of AD: 3xTg-AD and Thy1-APP mice. After three months, stem cell engraftment, neprilysin expression, and AD pathology were examined. Our findings reveal that stem cell-mediated delivery of NEP provides marked and significant reductions in Aβ pathology and increases synaptic density in both 3xTg-AD and Thy1-APP transgenic mice. Remarkably, Aβ plaque loads are reduced not only in the hippocampus and subiculum adjacent to engrafted NSCs, but also within the amygdala and medial septum, areas that receive afferent projections from the engrafted region. Taken together, our data suggest that genetically-modified NSCs could provide a powerful combinatorial approach to not only enhance synaptic plasticity but to also target and modify underlying Alzheimer's disease pathology.

  5. Characterization of mutant strains of Candida albicans deficient in expression of a surface determinant.

    PubMed Central

    Chaffin, W L; Collins, B; Marx, J N; Cole, G T; Morrow, K J

    1993-01-01

    Monoclonal antibody (MAb) 17E4 reacts with a surface carbohydrate determinant and agglutinates cells of Candida albicans. Using this MAb, we have isolated N-methyl-N'-nitro-N-nitrosoguanidine-induced nonagglutinating mutants. Eleven of these were characterized for the presence and expression of the surface antigen recognized by MAb 17E4 by immunoblot analysis of whole cells and by fluorescence flow cytometry. Soluble cell wall extracts from five mutant strains were negative by immunoblot analysis. The reactivities of the strains with several other MAbs and commercial antisera (Candida Check; Iatron Laboratories, Tokyo, Japan) which also recognize carbohydrate determinants were examined by immunoblot analysis of whole cells. Mutant strains showed no or reduced expression of the MAb 17E4 antigen and could be placed into at least two distinct phenotypic classes. Recognition by the other MAbs tested showed a similar pattern, while recognition by the commercial antisera was unchanged in the mutant strains. 1H and 13C nuclear magnetic resonance spectral analysis of mannan prepared from the wild type and nonexpressing mutant-strain 4A showed that the spectra from the mutant strain were simpler than those of the wild type. Most of the beta-1,2 linkages and all of the C-1 phosphate linkages were absent in the 4A mannan spectra, which suggested that the mutant mannan lacked the phosphate-bound beta-1,2-linked mannooligosaccharides. The effect of the surface defect on the ability of mutant strain 4A to adhere and to invade host tissue was examined in two murine models. In ex vivo binding assays, strain 4A showed reduced binding to the marginal zone and increased binding to the white pulp of splenic tissue, decreased binding to kidney tissue, and no change in binding to liver tissue compared with the wild type. In vivo, no difference was observed in translocation of the wild type or strain 4A to liver following immuno-compromising treatment of infant mice which had been

  6. Surface functionalization of titanium substrates with cecropin B to improve their cytocompatibility and reduce inflammation responses.

    PubMed

    Xu, Dawei; Yang, Weihu; Hu, Yan; Luo, Zhong; Li, Jinghua; Hou, Yanhua; Liu, Yun; Cai, Kaiyong

    2013-10-01

    Bacteria-related inflammation is a common postoperative complication in orthopedic implantation. In this study, cecropin B (CecB), a cationic peptide, was immobilized onto the surfaces of titanium substrates to improve their cytocompatibility and reduce inflammation responses. Polydopamine film was coated onto the surfaces of titanium substrates as an intermediate layer for the further immobilization of the CecB, which was confirmed by scanning electron microscopy (SEM), atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS) and water contact angle measurement, respectively. Osteoblasts grown onto the CecB-immobilized titanium substrates displayed significantly higher (p<0.01) cell viability than that of native titanium substrates (controls). Gram-positive bacteria - Bacillus subtilis, Staphylococcus aureus and Gram-negative bacteria - Escherichia coli, Pseudomonas aeruginosa were employed for antibacterial characterization. Media-borne assay and anti-biofilm formation showed that CecB-immobilized titanium substrates inhibited the adhesion and growth of bacteria. Macrophages cultured onto CecB-immobilized titanium substrates demonstrated statistically lower (p<0.01) levels of tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) than those of the control groups. The results indicated that the immobilization of CecB onto titanium substrates was responsible for improved cytocompatibility and reduced inflammation responses. The approach presented here has great potential in the development of antibacterial titanium-based implants in clinical applications. Copyright © 2013 Elsevier B.V. All rights reserved.

  7. Ion implantation of Ti + C to reduce friction and wear on steel surfaces

    SciTech Connect

    Follstaedt, D.M.; Pope, L.E.

    1987-01-01

    Within the last decade, ion implantation has been investigated for use in improving the surface-related mechanical properties of metal alloys. The treatment has improved the performance of device components and is now being used commercially in selected applications. A notable accomplishment is the reduction of wear by nitrogen implantation of many steel components used in mild wear applications. Another treatment receiving intensive investigation is the implantation of Ti + C into steels. This procedure not only reduces wear of steels, but has two important benefits beyond those obtained with nitrogen implantation: (a) friction is also reduced and (b) the reductions extend to hard bearing steels. Benefits have been obtained for all ferrous materials examined with unlubricated pin-on-disk testing in air, which range from soft austenitic (Type 304) to hard martensitic (Types 52100 and 440C) steels. The reductions in friction and wear obtained with this treatment are due to the amorphous surface layer. Examination with TEM of wear tracks from tests exhibiting the reductions show that not only is implanted titanium still present, but the amorphous layer is intact across the track.

  8. Mars Express scientists find a different Mars underneath the surface

    NASA Astrophysics Data System (ADS)

    2006-12-01

    Observations by MARSIS, the first subsurface sounding radar used to explore a planet, strongly suggest that ancient impact craters lie buried beneath the smooth, low plains of Mars' northern hemisphere. The technique uses echoes of radio waves that have penetrated below the surface. MARSIS found evidence that these buried impact craters - ranging from about 130 to 470 kilometres in diameter - are present under much of the northern lowlands. The findings appear in the 14 December 2006 issue of the journal Nature. With MARSIS "it's almost like having X-ray vision," said Thomas R. Watters of the National Air and Space Museum's Center for Earth and Planetary Studies, Washington, and lead author of the results. "Besides finding previously unknown impact basins, we've also confirmed that some subtle, roughly circular, topographic depressions in the lowlands are related to impact features." Studies of how Mars evolved help in understanding early Earth. Some signs of the forces at work a few thousand million years ago are harder to detect on Earth because many of them have been obliterated by tectonic activity and erosion. The new findings bring planetary scientists closer to understanding one of the most enduring mysteries about the geological evolution and history of Mars. In contrast to Earth, Mars shows a striking difference between its northern and southern hemispheres. Almost the entire southern hemisphere has rough, heavily cratered highlands, while most of the northern hemisphere is smoother and lower in elevation. Since the impacts that cause craters can happen anywhere on a planet, the areas with fewer craters are generally interpreted as younger surfaces where geological processes have erased the impact scars. The surface of Mars' northern plains is young and smooth, covered by vast amounts of volcanic lava and sediment. However, the new MARSIS data indicate that the underlying crust is extremely old. “The number of buried impact craters larger than 200

  9. Rapamycin Inhibits Proliferation of Hemangioma Endothelial Cells by Reducing HIF-1-Dependent Expression of VEGF

    PubMed Central

    Medici, Damian; Olsen, Bjorn R.

    2012-01-01

    Hemangiomas are tumors formed by hyper-proliferation of vascular endothelial cells. This is caused by elevated vascular endothelial growth factor (VEGF) signaling through VEGF receptor 2 (VEGFR2). Here we show that elevated VEGF levels produced by hemangioma endothelial cells are reduced by the mTOR inhibitor rapamycin. mTOR activates p70S6K, which controls translation of mRNA to generate proteins such as hypoxia inducible factor-1 (HIF-1). VEGF is a known HIF-1 target gene, and our data show that VEGF levels in hemangioma endothelial cells are reduced by HIF-1α siRNA. Over-expression of HIF-1α increases VEGF levels and endothelial cell proliferation. Furthermore, both rapamycin and HIF-1α siRNA reduce proliferation of hemangioma endothelial cells. These data suggest that mTOR and HIF-1 contribute to hemangioma endothelial cell proliferation by stimulating an autocrine loop of VEGF signaling. Furthermore, mTOR and HIF-1 may be therapeutic targets for the treatment of hemangiomas. PMID:22900063

  10. BMP15 Mutations Associated With Primary Ovarian Insufficiency Reduce Expression, Activity, or Synergy With GDF9.

    PubMed

    Patiño, Liliana C; Walton, Kelly L; Mueller, Thomas D; Johnson, Katharine E; Stocker, William; Richani, Dulama; Agapiou, David; Gilchrist, Robert B; Laissue, Paul; Harrison, Craig A

    2017-03-01

    Bone morphogenetic protein (BMP)15 is an oocyte-specific growth factor, which, together with growth differentiation factor (GDF) 9, regulates folliculogenesis and ovulation rate. Multiple mutations in BMP15 have been identified in women with primary ovarian insufficiency (POI), supporting a pathogenic role; however, the underlying biological mechanism of many of these mutants remains unresolved. To determine how mutations associated with ovarian dysfunction alter the biological activity of human BMP15. The effects of 10 mutations in BMP15 on protein production, activation of granulosa cells, and synergy with GDF9 were assessed. Sequencing of 35 patients with POI identified both an unrecognized BMP15 variant (c.986G>A, R329H) and a variant (c.581T>C, F194S) previously associated with the condition. Assessing expression and activity of these and 8 other BMP15 mutants identified: (1) multiple variants, including L148P, F194S, and Y235C, with reduced mature protein production; (2) three variants (R138H, A180T, and R329H) with ∼fourfold lower activity than wild-type BMP15; and (3) 3 variants (R68W, F194S, and N196K) with a significantly reduced ability to synergize with GDF9. Mutations in BMP15 associated with POI reduce mature protein production, activity, or synergy with GDF9. The latter effect is perhaps most interesting given that interactions with GDF9 most likely underlie the physiology of BMP15 in the human ovary.

  11. Subinhibitory concentrations of punicalagin reduces expression of virulence-related exoproteins by Staphylococcus aureus.

    PubMed

    Mun, Su-Hyun; Kong, Ryong; Seo, Yun-Soo; Zhou, Tian; Kang, Ok-Hwa; Shin, Dong-Won; Kwon, Dong-Yeul

    2016-11-01

    Staphylococcus aureus produces a number of virulence factors. The major virulence factors exhibited by S aureus include various antigens, enzymes, cytotoxins and exotoxins (e.g. hemolysins, enterotoxins and toxic shock syndrome toxin). In this report, we show the influence of punicalagin on the secretion of exoprotein from S aureus by western blotting, tumor necrosis factor (TNF) release assay and quantitative RT-PCR. When added to S aureus cultures at an OD600 of 0.9, graded subinhibitory concentrations of punicalagin reduced the production of α-toxin, SEA and SEB in methicillin-resistant Staphylococcus aureus in a dose-dependent manner. Consistently, punicalagin reduced TNF-inducing activity by S aureus culture supernatants. Here, the transcriptional level of agr (accessory gene regulator) in S aureus was inhibited by punicalagin, suggesting that the reduced transcription may affect the secretion of exotoxins. These findings suggest that the expression of α-toxin and enterotoxins in S aureus is sensitive to the action of punicalagin, which may be an advantageous candidate in the treatment of toxigenic staphylococcal disease.

  12. Reduced Cationic Nanoparticle Cytotoxicity Based on Serum Masking of Surface Potential

    PubMed Central

    McConnell, Kellie I.; Shamsudeen, Sabeel; Meraz, Ismail M.; Mahadevan, Thiruvillamalai S.; Ziemys, Arturas; Rees, Paul; Summers, Huw D.; Serda, Rita E.

    2016-01-01

    Functionalization of nanoparticles with cationic moieties, such as polyethyleneimine (PEI), enhances binding to the cell membrane; however, it also disrupts the integrity of the cell’s plasma and vesicular membranes, leading to cell death. Primary fibroblasts were found to display high surface affinity for cationic iron oxide nanoparticles and greater sensitivity than their immortalized counterparts. Treatment of cells with cationic nanoparticles in the presence of incremental increases in serum led to a corresponding linear decrease in cell death. The surface potential of the nanoparticles also decreased linearly as serum increased and this was strongly and inversely correlated with cell death. While low doses of nanoparticles were rendered non-toxic in 25% serum, large doses overcame the toxic threshold. Serum did not reduce nanoparticle association with primary fibroblasts, indicating that the decrease in nanoparticle cytotoxicity was based on serum masking of the PEI surface, rather than decreased exposure. Primary endothelial cells were likewise more sensitive to the cytotoxic effects of cationic nanoparticles than their immortalized counterparts, and this held true for cellular responses to cationic microparticles despite the much lower toxicity of microparticles compared to nanoparticles. PMID:27301181

  13. The effect of dentine surface preparation and reduced application time of adhesive on bonding strength.

    PubMed

    Saikaew, Pipop; Chowdhury, A F M Almas; Fukuyama, Mai; Kakuda, Shinichi; Carvalho, Ricardo M; Sano, Hidehiko

    2016-04-01

    This study evaluated the effects of surface preparation and the application time of adhesives on the resin-dentine bond strengths with universal adhesives. Sixty molars were cut to exposed mid-coronal dentine and divided into 12 groups (n=5) based on three factors; (1) adhesive: G-Premio Bond (GP, GC Corp., Tokyo, Japan), Clearfil Universal Bond (CU, Kuraray Noritake Dental Inc., Okayama, Japan) and Scotchbond Universal Adhesive (SB, 3M ESPE, St. Paul, MN, USA); (2) smear layer preparation: SiC paper ground dentine or bur-cut dentine; (3) application time: shortened time or as manufacturer's instruction. Fifteen resin-dentine sticks per group were processed for microtensile bond strength test (μTBS) according to non-trimming technique (1mm(2)) after storage in distilled water (37 °C) for 24h. Data were analyzed by three-way ANOVA and Dunnett T3 tests (α=0.05). Fractured surfaces were observed under scanning electron microscope (SEM). Another 12 teeth were prepared and cut into slices for SEM examination of bonded interfaces. μTBS were higher when bonded to SiC-ground dentine according to manufacturer's instruction. Bonding to bur-cut dentine resulted in significantly lower μTBS (p<0.000). Shortening the application time resulted in significantly lower bond strength for CU on SiC and GP on bur-cut dentine. SEM of fractured surfaces revealed areas with a large amount of porosities at the adhesive resin interface. This was more pronounced when adhesives were bonded with a reduced application time and on bur cut dentine. The performance of universal adhesives can be compromised on bur cut dentine and when applied with a reduced application time. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Reduced thrombogenicity of nitinol stents--in vitro evaluation of different surface modifications and coatings.

    PubMed

    Tepe, Gunnar; Schmehl, Joerg; Wendel, Hans P; Schaffner, Sivio; Heller, Stephan; Gianotti, Marc; Claussen, Claus D; Duda, Stephan H

    2006-02-01

    The material and the surface patterns of intravascular stents play a pivotal role in activating platelets and triggering adherence of inflammatory cells that consecutively leads to renarrowing caused by neointimal hyperplasia. To improve these features, besides mechanical and chemical modifications, ways of masking the stent by covering have been developed. In addition, polymer-coated stents are used as vehicle for local drug delivery. But as substances used for this application are described to possess an inflammatory potential, this aspect has to be evaluated. In the present study we compared different approaches to surface alterations applied to a nitinol stent design. Besides commonly used techniques like passivation and electropolishing, we evaluated coatings with heparin, aluminium and a polyurethane polymer regarding their thrombogenic and inflammatory characteristics. By weaving thin elastomer fibres a graft was generated. The previously described Chandler loop was used to simulate arterial flow conditions ex vivo using rotating PVC tubings filled with human blood. All stents received 120 min of blood contact. To determine thrombocyte activation and inflammatory reaction, the platelet count and levels of beta-TG, TAT and PMN-elastase were assessed. Scanning electron microscopy was used to visualize the reactions. Mechanical polishing and passivation did not improve the stent surface characteristics while sandblasting, electropolishing and aluminium covering decreased activation of the coagulation cascade. In terms of thrombogenicity, the heparin coating had no beneficial effect. The lowest thrombogenic potential was found in the Polyurethane-coated stent group. All stents showed similar levels of polymorph nuclear granulocyte elastase except for the membrane design. While mechanical and chemical modifications are able to reduce thrombogenicity, coating with this particular polyurethane polymer seems to be superior to these approaches regarding the

  15. Slotted surface coil with reduced g-factor for SENSE imaging.

    PubMed

    Ocegueda, K; Rodriguez, A O

    2006-01-01

    A new coil design inspired on the slot-and-hole magnetron tube is proposed for SENSE imaging. To investigate its g-factor behaviour: an SNR formula was derived using the quasi-static approach, and combined with the ultimate g-factor formula to compute the ultimate-factor-g-vs-depth plots. A g-factor expression was derived for the circular coil using the same approach for comparison purposes. SNR-vs-depth profiles of an 4-slot coil showed an important improvement over the circular coil. The 4-slot coil g-factor can be up to 58.32% lower than that of a single circular-shaped coil. This improvement makes the slotted surface coil a good choice for SENSE imaging.

  16. VARIABLE BOUND-SITE CHARGING CONTRIBUTIONS TO SURFACE COMPLEXATION MASS ACTION EXPRESSIONS

    EPA Science Inventory

    One and two pK models of surface complexation reactions between reactive surface sites (>SOH) and the proton (H+) use mass action expressions of the form: Ka={[>SOHn-1z-1]g>SOH(0-1)aH+EXP(-xeY/kT)}/{[>SOHnz]g>SOH(n)} where Ka=the acidity constant, [ ]=reactive species concentrati...

  17. VARIABLE BOUND-SITE CHARGING CONTRIBUTIONS TO SURFACE COMPLEXATION MASS ACTION EXPRESSIONS

    EPA Science Inventory

    One and two pK models of surface complexation reactions between reactive surface sites (>SOH) and the proton (H+) use mass action expressions of the form: Ka={[>SOHn-1z-1]g>SOH(0-1)aH+EXP(-xeY/kT)}/{[>SOHnz]g>SOH(n)} where Ka=the acidity constant, [ ]=reactive species concentrati...

  18. Biological variation among african trypanosomes: I. Clonal expression of virulence is not linked to the variant surface glycoprotein or the variant surface glycoprotein gene telomeric expression site.

    PubMed

    Inverso, Jill A; Uphoff, Timothy S; Johnson, Scott C; Paulnock, Donna M; Mansfield, John M

    2010-05-01

    The potential association of variant surface glycoprotein (VSG) gene expression with clonal expression of virulence in African trypanosomes was addressed. Two populations of clonally related trypanosomes, which differ dramatically in virulence for the infected host, but display the same apparent VSG surface coat phenotype, were characterized with respect to the VSG genes expressed as well as the chromosome telomeric expression sites (ES) utilized for VSG gene transcription. The VSG gene sequences expressed by clones LouTat 1 and LouTat 1A of Trypanosoma brucei rhodesiense were identical, and gene expression in both clones occurred precisely by the same gene conversion events (duplication and transposition), which generated an expression-linked copy (ELC) of the VSG gene. The ELC was present on the same genomic restriction fragments in both populations and resided in the telomere of a 330-kb chromosome; a single basic copy of the LouTat 1/1A VSG gene, present in all variants of the LouTat 1 serodeme, was located at an internal site of a 1.5-Mb chromosome. Restriction endonuclease mapping of the ES telomere revealed that the VSG ELC of clones LouTat 1 and 1A resides in the same site. Therefore, these findings provide evidence that the VSG gene ES and, potentially, any cotranscribed ES-associated genes do not play a role in the clonal regulation of virulence because trypanosome clones LouTat 1 and 1A, which differ markedly in their virulence properties, both express identical VSG genes from the same chromosome telomeric ES.

  19. Reduced Cortical Cannabinoid 1 Receptor Messenger RNA and Protein Expression in Schizophrenia

    PubMed Central

    Eggan, Stephen M.; Hashimoto, Takanori; Lewis, David A.

    2010-01-01

    Context Cannabis use is associated with both impaired cognitive functions, including working memory, and an increased risk of schizophrenia. Schizophrenia is characterized by impairments in working memory that are associated with reduced γ-aminobutyric acid (GABA) neurotransmission in the dorsolateral prefrontal cortex. The cannabinoid 1 receptor (CB1R) is highly expressed in the dorsolateral prefrontal cortex, is contained in the axon terminals of a subpopulation of perisomatic-targeting GABA neurons, and, when activated, suppresses the release of GABA. Objective To determine the potential relationship between CB1R signaling and altered GABA neurotransmission in schizophrenia by evaluating CB1R messenger RNA (mRNA) and protein expression in the dorsolateral pre-frontal cortex. Design In situ hybridization and immunocytochemistry techniques were used to examine the cortical levels of CB1R mRNA and protein, respectively. Setting Brain specimens were obtained from autopsies conducted at the Allegheny County Medical Examiner’s Office, Pittsburgh, Pennsylvania. Participants Postmortem brain specimens from 23 pairs of subjects with schizophrenia and age-, sex-, and postmortem interval–matched comparison subjects, as well as brain specimens from 18 macaque monkeys with long-term exposure to haloperidol, olanzapine, or placebo. Main Outcome Measures Optical density measures of CB1R mRNA expression and protein levels and correlations with previously reported glutamic acid decarboxylase 67 and cholecystokinin mRNA measures. Results Levels of CB1R mRNA were significantly lower by 14.8% in the subjects with schizophrenia. Similarly, CB1R protein levels, assessed by radioimmunocytochemistry and standard immunocytochemistry, were significantly decreased by 11.6% and 13.9%, respectively. Group differences in CB1R mRNA levels were significantly correlated with those in glutamic acid decarboxylase 67 and cholecystokinin mRNA levels. Expression of CB1R mRNA was not changed in

  20. Prolonged clopidogrel application reduces tissue factor expression after percutaneous coronary intervention in the porcine model.

    PubMed

    Ayral, Yunus; Rauch, Ursula; Goldin-Lang, Petra; Eisenreich, Andreas; Pepke, Wojciech; Deiner, Carolin; Schwimmbeck, Peter L; Schultheiss, Heinz-Peter; Pels, Klaus

    2011-01-01

    Late thrombotic events are important complications associated with intracoronary brachytherapy (ICBT) using ionizing radiation (IR) or with antiproliferative treatment modalities such as drug-eluting stents (DES). The mechanism mediating these thrombotic events is not well understood. This study assessed the effect of prolonged clopidogrel treatment on tissue factor (TF) expression in coronary arteries and on the circulating TF level after percutaneous transluminal coronary angioplasty /ICBT in a porcine coronary model. Pigs were treated with aspirin plus a 300 mg loading dose of clopidogrel one day before percutaneous coronary intervention (PCI), followed by a daily dose of clopidogrel and aspirin. During PCI one of the two balloon-injured arteries was treated by brachytherapy. Animals were sacrificed at different time points. The pigs, which were sacrificed 3 months post-PCI, were divided into two groups (Group I: clopidogrel for 3 months; Group II: clopidogrel for 1 month). Plasma TF was measured by enzyme-linked immunosorbent assay in blood samples taken from all pigs before and immediately after intervention and before sacrifice. Morphometric analysis was performed on digitalized images employing the software LUCIA G for TF staining. Vascular TF expression levels were assessed by quantitative real-time polymerase chain reaction. Prolonged clopidogrel application significantly reduced coronary TF at the protein (Group I vs. II, 8.975 ± 3.947% vs. 26.44 ± 5.375%, P = .007) and mRNA level [Group I vs. II, (0.3501 ± 0.0519) × 10(-3) vs. (0.7073 ± 0.0436) × 10(-3), P<.0005]. Circulating TF protein tended to be lower after 3 months than after 1 month clopidogrel treatment post-PCI (Group I vs. Group II, 488.3 ± 35.37 pg/ml vs. 572.3 ± 39.9 pg/ml, P = .130). Prolonged clopidogrel treatment reduced coronary TF expression and tended to reduce the blood TF level post-PCI, thus possibly modulating the risk of late thrombosis. Copyright © 2011 Elsevier Inc. All

  1. Surface Expression of Mantle Shortening at an Active Continental Margin

    NASA Astrophysics Data System (ADS)

    Bourguignon, S.; Stern, T.; Savage, M.

    2008-12-01

    We use teleseismic P-wave advances to show the mantle lithosphere has thickened beneath the central South Island of New Zealand. About 100 km of shortening has been absorbed within the South Island in the last 10 my or so. A 3-D ray tracing model is implemented to model teleseismic rays from a variety of azimuths. Our best fitting shape for the thickened mantle is for a sub-vertical body directly beneath the crustal root of the Southern Alps that has a strike sub-parallel to them. Dimensions of the body are 125 km thick and ~ 100 km, the centre around 110 km deep and an average Vp contrast with the regular mantle of ~0.6 km/s. We interpret the higher Vp within the thickened mantle as being due to colder temperatures compared to regular asthenosphere. Colder mantle will also be more dense than the asthenosphere, and hence the thickened mantle will act like a load pulling down on the overlying crust. Indirect evidence for such a loading is the observation that the crustal root is about 17 km thick, yet the mean elevation of the alps is only about 1600 m - an elevation about half that predicted by Airy isostasy. A further measure of the imbalance and suppression of surface topography comes from a pervasive negative isostatic gravity anomaly of about -30 mgals. In central and western North Island of New Zealand a similar amount (70 -100 km) of shortening occurred from late Oligocene to late Miocene times. Evidence for shortening comes from a Miocene fold and thrust belt with associated foreland basin. Subsidence curves from the basin provide an estimate of total shortening. At 5 Ma compression ceased and the region underwent a regional (400 km in extent) and rapid (few my) rock uplift of 2.5 km (or surface uplift of about 1100 m). Widespread volcanism followed thereafter. Today there is evidence that the mantle lithosphere has either gone from, or been highly attenuated beneath, much of the central and western North Island. In contrast to central South Island

  2. Cytomegalovirus m154 Hinders CD48 Cell-Surface Expression and Promotes Viral Escape from Host Natural Killer Cell Control

    PubMed Central

    Farré, Domènec; Tomic, Adriana; Borst, Eva Maria; Messerle, Martin; Jonjic, Stipan; Engel, Pablo; Angulo, Ana

    2014-01-01

    Receptors of the signalling lymphocyte-activation molecules (SLAM) family are involved in the functional regulation of a variety of immune cells upon engagement through homotypic or heterotypic interactions amongst them. Here we show that murine cytomegalovirus (MCMV) dampens the surface expression of several SLAM receptors during the course of the infection of macrophages. By screening a panel of MCMV deletion mutants, we identified m154 as an immunoevasin that effectively reduces the cell-surface expression of the SLAM family member CD48, a high-affinity ligand for natural killer (NK) and cytotoxic T cell receptor CD244. m154 is a mucin-like protein, expressed with early kinetics, which can be found at the cell surface of the infected cell. During infection, m154 leads to proteolytic degradation of CD48. This viral protein interferes with the NK cell cytotoxicity triggered by MCMV-infected macrophages. In addition, we demonstrate that an MCMV mutant virus lacking m154 expression results in an attenuated phenotype in vivo, which can be substantially restored after NK cell depletion in mice. This is the first description of a viral gene capable of downregulating CD48. Our novel findings define m154 as an important player in MCMV innate immune regulation. PMID:24626474

  3. Targeting Prostate Cancer Stemlike Cells Through Cell Surface-Expressed GRP78

    DTIC Science & Technology

    2015-10-01

    the cell surface GRP78-expressing subpopulation of cells supports nuclear Akt/GSK-3/ Snail -1 signaling. These findings are important because they are...original tasks outlined in the approved statement of work. 15. SUBJECT TERMS prostate cancer, cell surface GRP78, cancer stem cell, Snail -1 16. SECURITY...associated with cell surface GRP78 (Akt/GSK-3/ Snail -1) were upregulated in GRP78(+) relative to GRP78(-) prostate cancer cells. Our results in this

  4. Microbial methanogenesis in the sulfate-reducing zone of surface sediments traversing the Peruvian margin

    NASA Astrophysics Data System (ADS)

    Maltby, J.; Sommer, S.; Dale, A. W.; Treude, T.

    2015-09-01

    We studied the concurrence of methanogenesis and sulfate reduction in surface sediments (0-25 cm below sea floor, cmbsf) at six stations (70, 145, 253, 407, 770 and 1024 m) along the Peruvian margin (12° S). This oceanographic region is characterized by high carbon export to the seafloor, creating an extensive oxygen minimum zone (OMZ) on the shelf, both factors that could favor surface methanogenesis. Sediments sampled along the depth transect traversed areas of anoxic and oxic conditions in the bottom-near water. Net methane production (batch incubations) and sulfate reduction (35S-sulfate radiotracer incubation) were determined in the upper 0-25 cmbsf of multicorer cores from all stations, while deep hydrogenotrophic methanogenesis (> 30 cmbsf, 14C-bicarbonate radiotracer incubation) was determined in two gravity cores at selected sites (78 and 407 m). Furthermore, stimulation (methanol addition) and inhibition (molybdate addition) experiments were carried out to investigate the relationship between sulfate reduction and methanogenesis. Highest rates of methanogenesis and sulfate reduction in the surface sediments, integrated over 0-25 cmbsf, were observed on the shelf (70-253 m, 0.06-0.1 and 0.5-4.7 mmol m-2 d-1, respectively), while lowest rates were discovered at the deepest site (1024 m, 0.03 and 0.2 mmol m-2 d-1, respectively). The addition of methanol resulted in significantly higher surface methanogenesis activity, suggesting that the process was mostly based on non-competitive substrates, i.e., substrates not used by sulfate reducers. In the deeper sediment horizons, where competition was probably relieved due to the decline of sulfate, the usage of competitive substrates was confirmed by the detection of hydrogenotrophic activity in the sulfate-depleted zone at the shallow shelf station (70 m). Surface methanogenesis appeared to be correlated to the availability of labile organic matter (C / N ratio) and organic carbon degradation (DIC production

  5. Microbial methanogenesis in the sulfate-reducing zone of surface sediments traversing the Peruvian margin

    NASA Astrophysics Data System (ADS)

    Maltby, J.; Sommer, S.; Dale, A. W.; Treude, T.

    2016-01-01

    We studied the concurrence of methanogenesis and sulfate reduction in surface sediments (0-25 cm below sea floor) at six stations (70, 145, 253, 407, 990 and 1024 m) along the Peruvian margin (12° S). This oceanographic region is characterized by high carbon export to the seafloor creating an extensive oxygen minimum zone (OMZ) on the shelf, both factors that could favor surface methanogenesis. Sediments sampled along the depth transect traversed areas of anoxic and oxic conditions in the bottom-near water. Net methane production (batch incubations) and sulfate reduction (35S-sulfate radiotracer incubation) were determined in the upper 0-25 cm b.s.f. of multiple cores from all stations, while deep hydrogenotrophic methanogenesis (> 30 cm b.s.f., 14C-bicarbonate radiotracer incubation) was determined in two gravity cores at selected sites (78 and 407 m). Furthermore, stimulation (methanol addition) and inhibition (molybdate addition) experiments were carried out to investigate the relationship between sulfate reduction and methanogenesis.

    Highest rates of methanogenesis and sulfate reduction in the surface sediments, integrated over 0-25 cm b.s.f., were observed on the shelf (70-253 m, 0.06-0.1 and 0.5-4.7 mmol m-2 d-1, respectively), while lowest rates were discovered at the deepest site (1024 m, 0.03 and 0.2 mmol m-2 d-1, respectively). The addition of methanol resulted in significantly higher surface methanogenesis activity, suggesting that the process was mostly based on non-competitive substrates - i.e., substrates not used by sulfate reducers. In the deeper sediment horizons, where competition was probably relieved due to the decrease of sulfate, the usage of competitive substrates was confirmed by the detection of hydrogenotrophic activity in the sulfate-depleted zone at the shallow shelf station (70 m).

    Surface methanogenesis appeared to be correlated to the availability of labile organic matter (C / N ratio) and organic carbon

  6. Synthetic Strategies for Semiconductor Nanocrystals Expressing Localized Surface Plasmon Resonance.

    PubMed

    Niezgoda, J Scott; Rosenthal, Sandra J

    2016-03-03

    The field of semiconductor plasmonics has grown rapidly since its outset, only roughly six years ago, and now includes many crystalline substances ranging from GeTe to wide-bandgap transition-metal oxides. One byproduct of this proliferation is the sea of differing synthetic methods to realize localized surface plasmon resonances (LSPRs) based on the studied material. Strategies vary widely from material to material, but all have the common goal of introducing extremely high carrier densities to the semiconductor system. This doping results in tunable, size-quantized, and on/off-switchable LSPR modes, which are a complete departure from traditional metal-nanoparticle-based plasmon resonances. This Minireview will provide an overview of the current state of nanocrystal and quantum-dot plasmonics and the physical basis thereof, however its main purpose is to summarize the methods for realizing LSPRs in the various syntheses and systems that have been reported to date. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Do reducing regimens of fluorometholone for paediatric ocular surface disease cause glaucoma?

    PubMed

    Mataftsi, Asimina; Narang, Aman; Moore, Will; Nischal, Ken K

    2011-11-01

    Although fluorometholone (FML) is considered a steroid of minimal ocular penetration, reports in children have shown dose-dependent intraocular pressure (IOP) rise. The authors aimed to assess whether reducing regimens of FML for paediatric ocular surface disease have sustained clinically significant ocular hypertensive effects. Retrospective case-note review. Glaucoma was defined as an IOP of ≥ 21 mm Hg on at least two occasions or, in young children, moderate/firm digital IOP with one of the following: myopic shift, increased cup:disc ratio or corneal oedema. Exclusion criteria were other concurrent steroids or pre-existing optic nerve disease. 107 cases were included. The median age was 6 years (range 3 months to 17 years). The commonest indication for FML was blepharo-kerato-conjunctivitis. The maximal frequency prescribed was four times a day, gradually reduced to once weekly in cases of long-term treatment. The mean total number of eye-drop applications was 228 over a mean time span of 9 months. Post-FML IOP was formally documented in 51/107 casenotes (median age 6.85 years, range 4 months to 16 years) and it was <19 mm Hg in all cases. 56 cases did not allow IOP measurement (median age 5.9 years, range 3 months to 17 years), but none met the glaucoma definition. In this cohort, reducing regimens of FML proved to be a safe anti-inflammatory treatment in terms of avoiding steroid-induced glaucoma.

  8. Modeling effectiveness of agricultural BMPs to reduce sediment load and organophosphate pesticides in surface runoff.

    PubMed

    Zhang, Xuyang; Zhang, Minghua

    2011-04-15

    Quantifying effectiveness of agricultural BMPs at the watershed scale is a challenging issue, requiring robust algorithms to simulate not only the agricultural production system but also pollutant transport and fate. This research addresses the challenge to simulate performances of BMPs in reducing organophosphates (OPs) runoff at the watershed scale. The SWAT model is calibrated and validated following a sensitivity analysis combining Latin Hypercube sampling and One-factor-At-a-Time simulation. The calibrated model is then applied in the Orestimba Creek Watershed to simulate BMPs including buffer strips, sediment ponds, vegetated ditches, use reduction, and their combinations. BMP simulation suggested that sediment ponds trap 54-85% of sediment from field runoff, but less than 10% of dissolved diazinon and chlorpyrifos. Use reduction can reduce pesticide load in a close-to-linear fashion. Effectiveness of vegetated ditches and buffers depends on their physical dimension and vegetation cover. Combining individual BMPs provides enhanced mitigation effects. The combination of vegetated ditches, buffer strips and use reduction decreases diazinon and chlorpyrifos load by over 94%. This study has suggested that the SWAT model reasonably predicts BMP effectiveness at the watershed scale. Results will assist decision making in implementing BMPs to reduce pesticide loads in surface runoff. Published by Elsevier B.V.

  9. Analysis of copper surface features obtained using TEA CO2 laser at reduced air pressure

    NASA Astrophysics Data System (ADS)

    Momcilovic, M.; Trtica, M.; Ciganovic, J.; Savovic, J.; Stasic, J.; Kuzmanovic, M.

    2013-04-01

    Interaction of a transversely excited atmospheric (TEA) CO2 laser with rough copper surface, at reduced air pressure, was studied. Optical pulse duration of the laser employed was ∼2 μs, with the initial spike FWHM of ∼100 ns. Laser energy density of ∼32 J/cm2 (intensity ∼108 W/cm2) was above the plasma ignition threshold. Morphological features of the copper can be summarized as follows: (i) superficial damages, which take crater-shaped form at a higher number of accumulated laser pulses, (ii) development of melt pools with visible bubbles inside the damage region, (iii) formation of solid droplets at near periphery, and (iv) presence of “halo” effect at the irradiated surface. The laser induced surface changes were influenced by the target plasma formation. The formation of plasma influenced the laser-target interaction in two opposite ways: trough absorption of laser energy by the plasma, i.e. trough the effect of plasma shielding, and trough energy transfer from the plasma to the sample. Optical emission spectra were compared for laser induced plasma originated by a single and by cumulative laser pulses. It was found that plasma dimensions and emission intensities have a strong correlation with the number of accumulated laser pulses. Enhancement of both atomic and ionic copper lines was registered when laser induced plasma originated from a single pulse. Chemical analysis of the surface showed a tendency of copper content increase and oxygen content reduction when going from non-irradiated region to the central irradiated region. In the central damage zone, nearly pure copper was present which can be advantageous for some applications due to considerably lower contamination.

  10. Nifurtimox reduces N-Myc expression and aerobic glycolysis in neuroblastoma

    PubMed Central

    Cabanillas Stanchi, Karin Melanie; Bruchelt, Gernot; Handgretinger, Rupert; Holzer, Ursula

    2015-01-01

    Neuroblastoma is one of the most common solid tumors in childhood and usually accompanied with poor prognosis and rapid tumor progression when diagnosed with amplification of the proto-oncogene N-Myc. The amplification of N-Myc has major influence on the maintenance of aerobic glycolysis, also known as the Warburg effect. This specific switch in the conversion of pyruvate to lactate instead of the conversion of pyruvate to acetyl-coenzyme A even in the presence of oxygen has important benefits for the tumor, e.g. increased production of enzymes and enzyme substrates that are involved in tumor progression, angiogenesis and inhibition of apoptosis. The antiprotozoal drug nifurtimox, which is generally used for the treatment of infections with the parasitic protozoan Trypanosoma cruzi, has been reported to have cytotoxic properties in the therapy of neuroblastoma. However, its action of mechanism has not been described in detail yet. The presented in vitro study on the neuroblastoma cell lines LA-N-1, IMR-32, LS and SK-N-SH shows an increased production of oxidative stress, a reduced lactate dehydrogenase enzyme activity and reduced lactate production after nifurtimox treatment. Furthermore, nifurtimox leads to reduced mRNA and protein levels of the proto-oncogene protein N-Myc. Thus, the current work gives new insights into the effect of nifurtimox on tumor metabolism revealing a shifted glucose metabolism from production of lactate to oxidative phosphorylation and a reduced expression of the major molecular prognostic factor in neuroblastoma N-Myc, presenting nifurtimox as a possible adjuvant therapeutic agent against (high risk) neuroblastoma. PMID:26177922

  11. Over-Expression of TRESK K+ Channels Reduces the Excitability of Trigeminal Ganglion Nociceptors

    PubMed Central

    Guo, Zhaohua; Cao, Yu-Qing

    2014-01-01

    TWIK-related spinal cord K+ (TRESK) channel is abundantly expressed in trigeminal ganglion (TG) and dorsal root ganglion neurons and is one of the major background K+ channels in primary afferent neurons. Mutations in TRESK channels are associated with familial and sporadic migraine. In rats, both chronic nerve injury and inflammation alter the expression level of TRESK mRNA. Functional studies indicate that reduction of endogenous TRESK channel activity results in hyper-excitation of primary afferent neurons, suggesting that TRESK is a potential target for the development of new analgesics. However, whether and how enhancing TRESK channel activity would decrease the excitability of primary afferent neurons has not been directly tested. Here, we over-expressed TRESK subunits in cultured mouse TG neurons by lipofectamine-mediated transfection and investigated how this altered the membrane properties and the excitability of the small-diameter TG population. To account for the heterogeneity of neurons, we further divided small TG neurons into two groups, based on their ability to bind to fluorescently-labeled isolectin B (IB4). The transfected TG neurons showed a 2-fold increase in the level of TRESK proteins. This was accompanied by a significant increase in the fraction of lamotrigine-sensitive persistent K+ currents as well as the size of total background K+ currents. Consequently, both IB4-positive and IB4-negative TG neurons over-expressing TRESK subunits exhibited a lower input resistance and a 2-fold increase in the current threshold for action potential initiation. IB4-negative TG neurons over-expressing TRESK subunits also showed a significant reduction of the spike frequency in response to supra-threshold stimuli. Importantly, an increase in TRESK channel activity effectively inhibited capsaicin-evoked spikes in TG neurons. Taken together, our results suggest that potent and specific TRESK channel openers likely would reduce the excitability of primary

  12. Reduced inflammatory activity of RAW 264.7 macrophages on titania nanotube modified Ti surface.

    PubMed

    Neacsu, Patricia; Mazare, Anca; Cimpean, Anisoara; Park, Jung; Costache, Marieta; Schmuki, Patrik; Demetrescu, Ioana

    2014-10-01

    Macrophages play a pivotal role in the hosts response to biomaterials being considered as an essential cell type during both optimal tissue-implant integration and pathologic process of implant failure. Hence, understanding of their cellular activity on biomaterials is important for improving evaluation and design of biomaterials for biomedical applications. In the present study, we have comparatively investigated the interactions of titania nanotubes (78 nm diameter) and commercial pure Ti with RAW 264.7 macrophages in both standard and pro-inflammatory (stimulation with lipopolysaccharide, LPS) culture conditions. In vitro tests showed that TiO2 nanotubes exhibited significantly decreased inflammatory activity of macrophages with respect to cytokine and chemokine gene expression/protein secretion, induction of foreign body giant cells (FBGCs) and nitric oxide (NO) release thereby mitigating the inflammatory response induced by LPS as compared to flat Ti surface. Therefore, our results suggest a novel role of TiO2 nanotubes in modulating macrophage response in biomaterial-associated bacterial infections. Overall, the current study provides new insight into how TiO2 nanotubes can be involved in macrophage activation and supports the great promise of such surface modifications for biomedical applications. Copyright © 2014 Elsevier Ltd. All rights reserved.

  13. Targeting Prostate Cancer Stemlike Cells through Cell Surface Expressed GRP78

    DTIC Science & Technology

    2016-12-01

    Award Number: W81XWH-12-1-0478 TITLE: Targeting Prostate Cancer Stemlike Cells through Cell Surface-Expressed GRP78 PRINCIPAL INVESTIGATOR...Targeting Prostate Cancer Stemlike Cells through Cell Surface-Expressed GRP78 5a. CONTRACT NUMBER 5b. GRANT NUMBER W81XWH-12-1-0478 6. AUTHOR(S) Dr...NOTES 14. ABSTRACT This study investigated a function for cell surface GRP78 in regulating prostate cancer stem-like cells . In year 1, we showed that

  14. Polarized Axonal Surface Expression of Neuronal KCNQ Potassium Channels Is Regulated by Calmodulin Interaction with KCNQ2 Subunit

    PubMed Central

    Lee, Kwan Young; Kim, Edward H.; Issema, Rodal S.; Chung, Hee Jung

    2014-01-01

    KCNQ potassium channels composed of KCNQ2 and KCNQ3 subunits give rise to the M-current, a slow-activating and non-inactivating voltage-dependent potassium current that limits repetitive firing of action potentials. KCNQ channels are enriched at the surface of axons and axonal initial segments, the sites for action potential generation and modulation. Their enrichment at the axonal surface is impaired by mutations in KCNQ2 carboxy-terminal tail that cause benign familial neonatal convulsion and myokymia, suggesting that their correct surface distribution and density at the axon is crucial for control of neuronal excitability. However, the molecular mechanisms responsible for regulating enrichment of KCNQ channels at the neuronal axon remain elusive. Here, we show that enrichment of KCNQ channels at the axonal surface of dissociated rat hippocampal cultured neurons is regulated by ubiquitous calcium sensor calmodulin. Using immunocytochemistry and the cluster of differentiation 4 (CD4) membrane protein as a trafficking reporter, we demonstrate that fusion of KCNQ2 carboxy-terminal tail is sufficient to target CD4 protein to the axonal surface whereas inhibition of calmodulin binding to KCNQ2 abolishes axonal surface expression of CD4 fusion proteins by retaining them in the endoplasmic reticulum. Disruption of calmodulin binding to KCNQ2 also impairs enrichment of heteromeric KCNQ2/KCNQ3 channels at the axonal surface by blocking their trafficking from the endoplasmic reticulum to the axon. Consistently, hippocampal neuronal excitability is dampened by transient expression of wild-type KCNQ2 but not mutant KCNQ2 deficient in calmodulin binding. Furthermore, coexpression of mutant calmodulin, which can interact with KCNQ2/KCNQ3 channels but not calcium, reduces but does not abolish their enrichment at the axonal surface, suggesting that apo calmodulin but not calcium-bound calmodulin is necessary for their preferential targeting to the axonal surface. These findings

  15. In vivo detection of membrane protein expression using surface plasmon enhanced fluorescence spectroscopy (SPFS).

    PubMed

    Krupka, Simone S; Wiltschi, Birgit; Reuning, Ute; Hölscher, Kerstin; Hara, Masahiko; Sinner, Eva-Kathrin

    2006-08-15

    Surface plasmon enhanced fluorescence spectroscopy (SPFS) was applied for the detection of expression and functional incorporation of integral membrane proteins into plasma membranes of living cells in real time. A vesicular stomatitis virus (VSV) tagged mutant of photoreceptor bovine rhodopsin was generated for high level expression with the semliki forest virus (SFV) system. Adherent baby hamster kidney (BHK-21) cells were cultivated on fibronectin-coated gold surfaces and infected with genetically engineered virus driving the expression of rhodopsin. Using premixed fluorescently (Alexa Fluor 647) labeled anti-mouse secondary antibody and monoclonal anti-VSV primary antibody, expression of rhodopsin in BHK-21 cells was monitored by SPFS. Fluorescence enhancement by surface plasmons occurs exclusively in the close vicinity of the gold surface. Thus, only the Alexa Fluor 647 labeled antibodies binding to the VSV-tag at rhodopsin molecules exposed on the cell surface experienced fluorescence enhancement, whereas, unbound antibody molecules in the bulk solution were negligibly excited. With this novel technique, we successfully recorded an increase of fluorescence with proceeding rhodopsin expression. Thus, we were able to observe the incorporation of heterologously expressed rhodopsin in the plasma membrane of living cells in real time using a relatively simple and rapid method. We confirmed our results by comparison with conventional wide field fluorescence microscopy.

  16. Insulin increases surface expression of TRPC6 channels in podocytes: role of NADPH oxidases and reactive oxygen species

    PubMed Central

    Kim, Eun Young; Anderson, Marc

    2012-01-01

    Insulin receptors in podocytes are essential for normal kidney function. Here, we show that insulin evokes a rapid increase in the surface expression of canonical transient receptor potential-6 channel (TRPC6) channels in cultured podocytes, but caused a decrease in surface expression of TRPC5. These effects are accompanied by a marked increase in outwardly rectifying cationic currents that can be blocked by 10 μM SKF96365 or 100 μM La3+. Application of oleoyl-2-acetyl-sn-glycerol (OAG) also increased SKF96365- and La3+-sensitive cationic currents in podocytes. Importantly, current responses to a combination of OAG and insulin were the same amplitude as those evoked by either agent applied alone. This occlusion effect suggests that OAG and insulin are targeting the same population of channels. In addition, shRNA knockdown of TRPC6 markedly reduced cationic currents stimulated by insulin. The effects of insulin on TRPC6 were mimicked by treating podocytes with H2O2. Insulin treatment rapidly increased the generation of H2O2 in podocytes, and it increased the surface expression of the NADPH oxidase NOX4 in cultured podocytes. Basal and insulin-stimulated surface expression of TRPC6 were reduced by pretreatment with diphenylene iodonium, an inhibitor of NADPH oxidases and other flavin-dependent enzymes, by siRNA knockdown of NOX4, and by manganese (III) tetrakis (4-benzoic acid) porphyrin chloride, a membrane-permeable mimetic of superoxide dismutase and catalase. These observations suggest that insulin increases generation of ROS in part through activation of NADPH oxidases, and that this step contributes to modulation of podocyte TRPC6 channels. PMID:22031853

  17. Flunitrazepam rapidly reduces GABAA receptor subunit protein expression via a protein kinase C-dependent mechanism

    PubMed Central

    Johnston, Jonathan D; Price, Sally A; Bristow, David R

    1998-01-01

    Acute flunitrazepam (1 μM) exposure for 1 h reduced GABAA receptor α1 (22±4%, mean±s.e.mean) and β2/3 (21±4%) subunit protein levels in cultured rat cerebellar granule cells. This rapid decrease in subunit proteins was completely prevented by bisindolymaleimide 1 (1 μM), an inhibitor of protein kinase C, but not by N-[2-((p-bromocinnamyl)amino)ethyl]-5-isoquinolinesulfonamide (H-89, 4.8 μM), an inhibitor of protein kinases A and G. These results suggest the existence of a benzodiazepine-induced mechanism to rapidly alter GABAA receptor protein expression, that appears to be dependent on protein kinase C activity. PMID:9723942

  18. Room-temperature, atmospheric plasma needle reduces adenovirus gene expression in HEK 293A host cells

    NASA Astrophysics Data System (ADS)

    Xiong, Z.; Lu, X.; Cao, Y.; Ning, Q.; Ostrikov, K.; Lu, Y.; Zhou, X.; Liu, J.

    2011-12-01

    Room-temperature, atmospheric-pressure plasma needle treatment is used to effectively minimize the adenovirus (AdV) infectivity as quantified by the dramatic reduction of its gene expression in HEK 293A primary human embryonic kidney cells studied by green fluorescent protein imaging. The AdV titer is reduced by two orders of magnitude within only 8 min of the plasma exposure. This effect is due to longer lifetimes and higher interaction efficacy of the plasma-generated reactive species in confined space exposed to the plasma rather than thermal effects commonly utilized in pathogen inactivation. This generic approach is promising for the next-generation anti-viral treatments and imunotherapies.

  19. In vitro culture of mouse embryos reduces differential gene expression between inner cell mass and trophectoderm.

    PubMed

    Giritharan, G; Delle Piane, L; Donjacour, A; Esteban, F J; Horcajadas, J A; Maltepe, E; Rinaudo, P

    2012-03-01

    Differences in gene expression and imprinting have been reported, comparing in vivo versus in vitro generated preimplantation embryos. Furthermore, mouse studies have shown that placenta development is altered following in vitro culture. However, the molecular mechanisms underlying these findings are unknown. We therefore isolated trophectoderm (TE) and inner cell mass (ICM) cells from in vivo and in vitro fertilization (IVF) embryos and evaluated their transcriptome using microarrays. We found that the transcriptomes of in vitro produced ICM and TE cells showed remarkably few differences compared to ICM and TE cells of in vivo generated embryos. In vitro fertilization embryos showed a reduced number of TE cells compared to in vivo embryos. In addition, TE of IVF embryos showed significant downregulation of solute transporter genes and of genes involved in placenta formation (Eomesodermin, Socs3) or implantation (Hbegf). In summary, IVF and embryo culture significantly affects the transcriptome of ICM and TE cells.

  20. In Vitro Culture of Mouse Embryos Reduces Differential Gene Expression Between Inner Cell Mass and Trophectoderm

    PubMed Central

    Giritharan, G.; Piane, L. Delle; Donjacour, A.; Esteban, F. J.; Horcajadas, J. A.; Maltepe, E.; Rinaudo, P.

    2012-01-01

    Differences in gene expression and imprinting have been reported, comparing in vivo versus in vitro generated preimplantation embryos. Furthermore, mouse studies have shown that placenta development is altered following in vitro culture. However, the molecular mechanisms underlying these findings are unknown. We therefore isolated trophectoderm (TE) and inner cell mass (ICM) cells from in vivo and in vitro fertilization (IVF) embryos and evaluated their transcriptome using microarrays. We found that the transcriptomes of in vitro produced ICM and TE cells showed remarkably few differences compared to ICM and TE cells of in vivo generated embryos. In vitro fertilization embryos showed a reduced number of TE cells compared to in vivo embryos. In addition, TE of IVF embryos showed significant downregulation of solute transporter genes and of genes involved in placenta formation (Eomesodermin, Socs3) or implantation (Hbegf). In summary, IVF and embryo culture significantly affects the transcriptome of ICM and TE cells. PMID:22383776

  1. Altered cell surface expression of human MC1R variant receptor alleles associated with red hair and skin cancer risk.

    PubMed

    Beaumont, Kimberley A; Newton, Richard A; Smit, Darren J; Leonard, J Helen; Stow, Jennifer L; Sturm, Richard A

    2005-08-01

    The human melanocortin-1 receptor gene (MC1R) encodes a G-protein coupled receptor that is primarily expressed on melanocytes, where it plays a key role in pigmentation regulation. Variant alleles are associated with red hair colour and fair skin, known as the RHC phenotype, as well as skin cancer risk. The R151C, R160W and D294H alleles, designated 'R', are strongly associated with the RHC phenotype and have been proposed to result in loss of function receptors due to impaired G-protein coupling. We recently provided evidence that the R151C and R160W variants can efficiently couple to G-proteins in response to alpha-melanocyte stimulating hormone. The possibility that altered cellular localization of the R151C and R160W variant receptors could underlie their association with RHC was therefore considered. Using immunofluorescence and ligand binding studies, we found that melanocytic cells exogenously or endogenously expressing MC1R show strong surface localization of the wild-type and D294H alleles but markedly reduced cell surface expression of the R151C and R160W receptors. In additional exogenous expression studies, the R variant D84E and the rare I155T variant, also demonstrated a significant reduction in plasma membrane receptor numbers. The V60L, V92M and R163Q weakly associated RHC alleles, designated 'r', were expressed with normal or intermediate cell surface receptor levels. These results indicate that reduced receptor coupling activity may not be the only contributing factor to the genetic association between the MC1R variants and the RHC phenotype, with MC1R polymorphisms now linked to a change in receptor localization.

  2. VIP and CRF reduce ADAMTS expression and function in osteoarthritis synovial fibroblasts.

    PubMed

    Pérez-García, Selene; Carrión, Mar; Gutiérrez-Cañas, Irene; González-Álvaro, Isidoro; Gomariz, Rosa P; Juarranz, Yasmina

    2016-04-01

    ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) family is known to play an important role in the pathogenesis of osteoarthritis (OA), working on aggrecan degradation or altering the integrity of extracellular matrix (ECM). Thus, the main purpose of our study was to define the role of vasoactive intestinal peptide (VIP) and corticotrophin-releasing factor (CRF), as immunoregulatory neuropeptides, on ADAMTS production in synovial fibroblasts (SF) from OA patients and healthy donors (HD). OA- and HD-SF were stimulated with pro-inflammatory mediators and treated with VIP or CRF. Both neuropeptides decreased ADAMTS-4, -5, -7 and -12 expressions, aggrecanase activity, glycosaminoglycans (GAG), and cartilage oligomeric matrix protein (COMP) degradation after stimulation with fibronectin fragments (Fn-fs) in OA-SF. After stimulation with interleukin-1β, VIP reduced ADAMTS-4 and -5, and both neuropeptides decreased ADAMTS-7 production and COMP degradation. Moreover, VIP and CRF reduced Runx2 and β-catenin activation in OA-SF. Our data suggest that the role of VIP and CRF on ADAMTS expression and cartilage degradation could be related to the OA pathology since scarce effects were produced in HD-SF. In addition, their effects might be greater when a degradation loop has been established, given that they were higher after stimulation with Fn-fs. Our results point to novel OA therapies based on the use of neuropeptides, since VIP and CRF are able to stop the first critical step, the loss of cartilage aggrecan and the ECM destabilization during joint degradation.

  3. Hyperglycemia Reduces Functional Expression of Astrocytic Kir4.1 Channels and Glial Glutamate Uptake

    PubMed Central

    Rivera-Aponte, David E.; Méndez-González, Miguel P.; Rivera-Pagán, Aixa F.; Kucheryavykh, Yuriy V.; Kucheryavykh, Lilia Y.; Skatchkov, Serguei N.; Eaton, Misty J.

    2015-01-01

    Diabetics are at risk for a number of serious health complications including an increased incidence of epilepsy and poorer recovery after ischemic stroke. Astrocytes play a critical role in protecting neurons by maintaining extracellular homeostasis and preventing neurotoxicity through glutamate uptake and potassium buffering. These functions are aided by the presence of potassium channels, such as Kir4.1 inwardly rectifying potassium channels, in the membranes of astrocytic glial cells. The purpose of the present study was to determine if hyperglycemia alters Kir4.1 potassium channel expression and homeostatic functions of astrocytes. We used q-PCR, Western blot, patch-clamp electrophysiology studying voltage and potassium step responses and a colorimetric glutamate clearance assay to assess Kir4.1 channel levels and homeostatic functions of astrocytes grown in normal and high glucose conditions. We found that astrocytes grown in high glucose (25 mM) had an approximately 50% reduction in Kir4.1 mRNA and protein expression as compared with those grown in normal glucose (5 mM). These reductions occurred within 4 to 7 days of exposure to hyperglycemia, whereas reversal occurred between 7 to 14 days after return to normal glucose. The decrease in functional Kir channels in the astrocytic membrane was confirmed using barium to block Kir channels. In the presence of 100 μm barium, the currents recorded from astrocytes in response to voltage steps were reduced by 45%. Furthermore, inward currents induced by stepping extracellular [K+]o from 3 to 10 mM (reflecting potassium uptake) were 50% reduced in astrocytes grown in high glucose. In addition, glutamate clearance by astrocytes grown in high glucose was significantly impaired. Taken together, our results suggest that down-regulation of astrocytic Kir4.1 channels by elevated glucose may contribute to the underlying pathophysiology of diabetes-induced CNS disorders and contribute to the poor prognosis after stroke. PMID

  4. Disrupting Protein Expression with Peptide Nucleic Acids Reduces Infection by Obligate Intracellular Rickettsia

    PubMed Central

    Pelc, Rebecca S.; McClure, Jennifer C.; Kaur, Simran J.; Sears, Khandra T.; Rahman, M. Sayeedur; Ceraul, Shane M.

    2015-01-01

    Peptide Nucleic Acids (PNAs) are single-stranded synthetic nucleic acids with a pseudopeptide backbone in lieu of the phosphodiester linked sugar and phosphate found in traditional oligos. PNA designed complementary to the bacterial Shine-Dalgarno or start codon regions of mRNA disrupts translation resulting in the transient reduction in protein expression. This study examines the use of PNA technology to interrupt protein expression in obligate intracellular Rickettsia sp. Their historically intractable genetic system limits characterization of protein function. We designed PNA targeting mRNA for rOmpB from Rickettsia typhi and rickA from Rickettsia montanensis, ubiquitous factors important for infection. Using an in vitro translation system and competitive binding assays, we determined that our PNAs bind target regions. Electroporation of R. typhi and R. montanensis with PNA specific to rOmpB and rickA, respectively, reduced the bacteria’s ability to infect host cells. These studies open the possibility of using PNA to suppress protein synthesis in obligate intracellular bacteria. PMID:25781160

  5. Perinatal exposure to alcohol reduces the expression of complexins I and II.

    PubMed

    Zink, Mathias; Araç, Gülsemin; Frank, Stefanie Th; Gass, Peter; Gebicke-Härter, Peter J; Spanagel, Rainer

    2009-01-01

    Perinatal exposure to alcohol (PEA) induces general developmental and specific neuropsychiatric disturbances. Ethanol affects amino acid neurotransmission and synaptic plasticity. We were interested in the transcriptional effects of ethanol on the expression of complexins I and II, two synaptic vesicle proteins (SVP) with relevance for cognition and memory. We exposed pregnant Wistar inbred rats (N=4) and their pups until postnatal day 8 (P8) in vapor chambers and performed in situ-hybridizations regarding complexins I and II at P8 as well as neurobehavioral testing in adult animals of the same litters. At P8, serum ethanol levels of 281+/-58 mg/dl were achieved. PEA animals presented a pronounced retardation of postnatal growth. Significantly lower expression levels of complexin I was observed in CA1, together with trends of reductions in other hippocampal and cortical regions. Complexin II was found reduced in anterior cingulate, prefrontal and fronto-parietal cortex. Adult rats of exposed litters showed worse performance in hippocampus-dependent learning (Morris water maze). The observed suppression of complexins I and II reveals disturbed synaptic plasticity and corresponds with long lasting, ethanol-induced deficits of learning and memory. Further investigations should focus on other synaptic vesicle protein genes in order to unravel the molecular basis of ethanol-induced neurocognitive disabilities.

  6. Reduced proliferation and osteocalcin expression in osteoblasts of male idiopathic osteoporosis.

    PubMed

    Ruiz-Gaspà, Sílvia; Blanch-Rubió, Josep; Ciria-Recasens, Manuel; Monfort, Jordi; Tío, Laura; Garcia-Giralt, Natàlia; Nogués, Xavier; Monllau, Joan C; Carbonell-Abelló, Jordi; Pérez-Edo, Lluis

    2010-03-01

    Osteoporosis is characterized by low bone mineral density (BMD), resulting in increasing susceptibility to bone fractures. In men, it has been related to some diseases and toxic habits, but in some instances the cause of the primary--or idiopathic--osteoporosis is not apparent. In a previous study, our group compared histomorphometric measurements in cortical and cancellous bones from male idiopathic osteoporosis (MIO) patients to those of control subjects and found reduced bone formation without major differences in bone resorption. To confirm these results, this study analyzed the etiology of this pathology, examining the osteoblast behavior in vitro. We compared two parameters of osteoblast activity in MIO patients and controls: osteoblastic proliferation and gene expression of COL1A1 and osteocalcin, in basal conditions and with vitamin D(3) added. All these experiments were performed from a first-passage osteoblastic culture, obtained from osteoblasts that had migrated from the transiliac explants to the plate. The results suggested that the MIO osteoblast has a slower proliferation rate and decreased expression of genes related to matrix formation, probably due to a lesser or slower response to some stimulus. We concluded that, contrary to female osteoporosis, in which loss of BMD is predominantly due to increased resorption, low BMD in MIO seems to be due to an osteoblastic defect.

  7. Reduced expression of C5a receptors on neutrophils from cord blood

    PubMed Central

    Nybo, M.; Sorensen, O.; Leslie, R; Wang, P.

    1998-01-01

    AIM—To describe further functional deficiencies of neonatal neutrophils by measuring the expression of C5a receptors.
METHODS—C5a uptake was measured using flow cytometry with fluorescein isothiocynate labelled recombinant C5a. The response of neutrophils to stimulation with C5a and fMLP was tested by measuring migration and exocytosis of myeloperoxidase and lactoferrin.
RESULTS—C5a mean fluorescence on neutrophils from neonates was significantly lower (22.4 (SD 3.5)) than in adult controls (31.5 (3.1)). Neutrophils from neonates migrated poorly towards both C5a and fMLP compared with those from adult controls. Exocytosis of myeloperoxidase, but not lactoferrin from neonatal neutrophils stimulated with C5a, was significantly lower than in adult controls. fMLP stimulation, on the other hand, resulted in significantly higher exocytosis in neonates.
CONCLUSION—The lower expression of C5a receptors on neutrophils from neonates could be related to reduced C5a mediated exocytosis of myeloperoxidase.

 PMID:9577284

  8. HPV 5 and 8 E6 Expression Reduces ATM Protein Levels and Attenuates LINE-1 Retrotransposition

    PubMed Central

    Wallace, Nicholas A.; Gasior, Stephen L.; Faber, Zachary J.; Howie, Heather L.; Deininger, Prescott L.; Galloway, Denise A.

    2013-01-01

    The expression of the E6 protein from certain members of the HPV genus β (β HPV 5 and 8 E6) can disrupt p53 signaling by diminishing the steady state levels of two p53 modifying enzymes, ATR and p300. Here, we show that β-HPV 5 and 8 E6 are also capable of reducing the steady state levels of another p53 modifying enzyme, ATM, and as a result restrict LINE-1 retrotransposition. Furthermore, we show that the reduction of both ATM and LINE-1 retrotransposition is dependent upon the ability of β-HPV 8 E6 to bind and degrade p300. We use inhibitors and dominant negative mutants to confirm that ATM is needed for efficient LINE-1 retrotransposition. Furthermore, neither sensitivity to LINE-1 expression nor LINE-1 induced DSB formation is altered in an ATM deficient background. Together, these data illustrate the broad impact some β-HPVs have on DNA damage signaling by promoting p300 degradation. PMID:23706308

  9. Reduced expression of galectin-1 and galectin-9 by leucocytes in asthma patients.

    PubMed

    Sanchez-Cuellar, S; de la Fuente, H; Cruz-Adalia, A; Lamana, A; Cibrian, D; Giron, R M; Vara, A; Sanchez-Madrid, F; Ancochea, J

    2012-12-01

    Accumulating evidence shows that galectins play roles in the initiation and resolution phases of inflammatory responses by promoting anti- or proinflammatory effects. This study investigated the presence of three members of the galectin family (galectin-1, -3 and -9) in induced sputum samples of asthma patients, as well as their possible implication in the immunopathogenesis of human asthma. Levels of interleukin (IL)-5, IL-13, and galectins were determined in leucocytes isolated from induced sputum samples by reverse transcription-polymerase chain reaction (RT-PCR) immunofluorescence and flow cytometry. High levels of IL-5 and IL-13 mRNA were detected in sputum cells from asthma patients. In parallel, immunoregulatory proteins galectin-1 and galectin-9 showed a reduced expression on macrophages from sputum samples compared with cells from healthy donors. In-vitro immunoassays showed that galectin-1 and galectin-9, but not galectin-3, are able to induce the production of IL-10 by peripheral blood mononuclear cells from healthy donors. These findings indicate that macrophages from sputum samples of asthma patients express low levels of galectin-1 and galectin-9, favouring the exacerbated immune response observed in this disease.

  10. Expression of pro-inflammatory interleukin-8 is reduced by ayurvedic decoctions.

    PubMed

    Guerrini, Alessandra; Mancini, Irene; Maietti, Silvia; Rossi, Damiano; Poli, Ferruccio; Sacchetti, Gianni; Gambari, Roberto; Borgatti, Monica

    2014-08-01

    Eleven decoctions, obtained from indian plants widely used in ayurvedic medicine, have been investigated as a possible source of molecules exhibiting biological activity on the interaction between DNA and NF-kB, a transcription factor involved in the expression of proinflammatory genes. Cystic fibrosis (CF) cell line stimulated by TNF-α has been used as inflammatory cellular model to determinate interleukin-8 (IL-8), one of the most relevant pro-inflammatory mediator in CF regulated by the NF-kB. The chemical characterization of these 11 decoctions by spectrophotometric analysis and NMR fingerprinting highlighted that sugars and polyphenols seemed to be the main compounds. Our results demonstrated that Azadirachta indica, Terminalia bellerica, Terminalia chebula, Hemidesmus indicus, Emblica officinalis and Swertia chirata are the most active decoctions in inhibiting NF-kB/DNA interactions by EMSA assay and in reducing pro-inflammatory IL- 8 expression in CF cells at IC50 concentrations by Real-Time and Bio-plex analyses. Finally, we observed the increase of all inhibitory activities with the rise of total polyphenols, procyanidins and flavonoids, except for the levels of IL-8 mRNA accumulation, that were as high as flavonoid content grown up by the statistical multivariate analyses. In conclusion, these six decoctions might be interesting to explore new anti-inflammatory treatments for diseases, such as CF.

  11. Axin expression reduces staurosporine-induced mitochondria-mediated cell death in HeLa cells.

    PubMed

    Shin, Jee-Hye; Kim, Hyun-wook; Rhyu, Im Joo; Song, Ki-Joon; Kee, Sun-Ho

    2012-10-01

    Cytoplasmic axin expression frequently produces punctuate structures in cells, but the nature of axin puncta has not been fully elucidated. In an effort to analyze cytoplasmic axin puncta, we established HeLa cells expressing axin in a doxycycline-inducible manner (HeLa-Axin). We observed that axin accumulated in an aggregate-like pattern in perinuclear areas and appeared to be associated with mitochondria, Golgi apparatus, and endoplasmic reticulum (ER), but not lysosomes. Further biochemical analysis suggested that some part of the cytoplasmic axin pool was associated with mitochondria. In addition, mitochondrial proteins [i.e., cytochrome oxidase IV (CoxIV) and cytochrome c] were slightly higher in HeLa-Axin cells than in HeLa-EV cells, suggesting altered mitochondrial degradation. HeLa-Axin cells were then treated with staurosporine (STS) to determine if the mitochondria-induced apoptosis pathway was altered. Compared to STS-treated control cells (HeLa-EV), HeLa-Axin cells had less STS-induced cytotoxicity and reduced caspase-3 activation and PARP cleavage. Given that mitochondria outer membrane potential was unchanged, HeLa-Axin cells might be relatively resistant to STS-mediated mitochondrial damage. Mitochondria associated with axin aggregates were resistant to detergent-mediated permeabilization. These results suggest that axin forms aggregate-like structures in association with mitochondria, which render mitochondria resistant to STS-induced membrane damage and cytotoxicity.

  12. Aerobic expression of Vitreoscilla hemoglobin efficiently reduces overflow metabolism in Escherichia coli.

    PubMed

    Pablos, Tania E; Sigala, Juan Carlos; Le Borgne, Sylvie; Lara, Alvaro R

    2014-06-01

    Overflow metabolism is a prevalent problem for aerobic cultivations of Escherichia coli. Although several process and molecular approaches have been applied to prevent overflow metabolism, these approaches often result in reductions in growth rate, biomass yield or accumulation of other byproducts. In this report, we present an alternative approach based on increasing the efficiency of aerobic metabolism by the expression of the Vitreoscilla stercoraria hemoglobin (VHb) to avoid overflow metabolism. VHb is expected to increase the consumption of NADH in the respiratory chain, leading to increased activity of the tricarboxylic acid (TCA) cycle. This would result in a faster consumption of acetyl Co-A and a decrease in acetate production. When this strategy was tested in E. coli strains, acetate production decreased