41 results on '"Huflejt M"'
Search Results
2. Changes in the repertoire of natural antibodies caused by immunization with bacterial antigens
- Author
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Shilova, N. V., Navakouski, M. J., Huflejt, M., Kuehn, A., Grunow, R., Blixt, O., and Bovin, N. V.
- Published
- 2011
- Full Text
- View/download PDF
3. Glycoarrays for diagnosis and therapy of the disorders of the female reproductive system
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Shilova, N. V., primary, Bovin, N. V., additional, Nokel, A. Yu., additional, Ziganshina, M. M., additional, Khasbiullina, N. R., additional, Vuskovic, M., additional, and Huflejt, M. E., additional
- Published
- 2021
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4. 2-Aminopyridine—a label for bridging of oligosaccharides HPLC profiling and glycoarray printing
- Author
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Shilova, N. V., Galanina, O. E., Rubina, A. Yu., Butvilovskaya, V. I., Huflejt, M. E., Chambers, J., Roucoux, A., and Bovin, N. V.
- Published
- 2008
- Full Text
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5. Changes in membrane lipid composition during saline growth of the fresh water cyanobacterium Synechococcus 6311
- Author
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Huflejt, M. E, Tremolieres, A, Pineau, B, Lang, J. K, Hatheway, J, and Packer, L
- Subjects
Man/System Technology And Life Support - Abstract
Growth of Synechococcus 6311 in the presence of 0.5 molar NaCl is accompanied by significant changes in membrane lipid composition. Upon transfer of the cells from a low salt' (0.015 molar NaCl) to high salt' (0.5 molar NaCl) growth medium at different stages of growth, a rapid decrease in palmitoleic acid (C16:1 delta 9) content was accompanied by a concomitant increase in the amount of the two C18:1 acids (C18:1 delta 9, C18:1 delta 11), with the higher increase in oleic acid C18:1 delta 9 content. These changes began to occur within the first hour after the sudden elevation of NaCl and progressed for about 72 hours. The percentage of palmitic acid (C16:0) and stearic acid (C18:0) remained almost unchanged in the same conditions. High salt-dependent changes within ratios of polar lipid classes also occurred within the first 72 hours of growth. The amount of monogalactosyl diacylglycerol (bilayer-destabilizing lipid) decreased and that of the digalactosyl diacylglycerol (bilayer-stabilizing lipid) increased. Consequently, in the three day old cells, the ratio of monogalactosyl diacylglycerol to digalactosyl diacylglycerol in the membranes of high salt-grown cells was about half of that in the membranes of low salt-grown cells. The total content of anionic lipids (phosphatidylglycerol and sulfoquinovosyl diacylglycerol) was always higher in the isolated membranes and the whole cells from high salt-grown cultures compared to that in the cells and membranes from low salt-grown cultures. All the observed rearrangements in the lipid environment occurred in both thylakoid and cytoplasmic membranes. Similar lipid composition changes, however, to a much lesser extent, were also observed in the aging, low salt-grown cultures. The observed changes in membrane fatty acids and lipids composition correlate with the alterations in electron and ion transport activities, and it is concluded that the rearrangement of the membrane lipid environment is an essential part of the process by which cells control membrane function and stability.
- Published
- 1990
6. High-grade serous ovarian and peritoneal cancers display distinct genetic and post-translational signatures – a criterion to treat them differently?
- Author
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Jacob, F, primary, Anugraham, M, additional, Schötzau, A, additional, Everest-Dass, A, additional, Bovin, N, additional, Huflejt, M, additional, Fedier, A, additional, Hacker, N, additional, Fink, D, additional, Packer, N, additional, and Heinzelmann-Schwarz, V, additional
- Published
- 2016
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7. The glycosphingolipid P1 is an ovarian cancer-associated carbohydrate antigen involved in migration
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Jacob, F, primary, Anugraham, M, additional, Pochechueva, T, additional, Tse, B W C, additional, Alam, S, additional, Guertler, R, additional, Bovin, N V, additional, Fedier, A, additional, Hacker, N F, additional, Huflejt, M E, additional, Packer, N, additional, and Heinzelmann-Schwarz, V A, additional
- Published
- 2014
- Full Text
- View/download PDF
8. Changes in the repertoire of natural antibodies caused by immunization with bacterial antigens
- Author
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Shilova, N V, Navakouski, M J, Huflejt, M, Kuehn, A, Grunow, R, Blixt, Klas Ola, Bovin, N V, Shilova, N V, Navakouski, M J, Huflejt, M, Kuehn, A, Grunow, R, Blixt, Klas Ola, and Bovin, N V
- Abstract
The repertoire of natural anti-glycan antibodies in naïve chickens and in chickens immunized with bacteria Burkholderia mallei, Burkholderia pseudomallei, and Francisella tularensis as well as with peptides from an outer membrane protein of B. pseudomallei was studied. A relatively restricted pattern of natural antibodies (first of all IgY against bacterial cell wall peptidoglycan fragments, L-Rha, and core N-acetyllactosamine) shrank and, moreover, the level of detectable antibodies decreased as a result of immunization.
- Published
- 2011
9. Comparison of printed glycan array, suspension array and ELISA in the detection of human anti-glycan antibodies.
- Author
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Pochechueva, T, Jacob, F, Goldstein, D R, Huflejt, M E, Chinarev, A, Caduff, R, Fink, D, Hacker, N, Bovin, N V, Heinzelmann-Schwarz, V, Pochechueva, T, Jacob, F, Goldstein, D R, Huflejt, M E, Chinarev, A, Caduff, R, Fink, D, Hacker, N, Bovin, N V, and Heinzelmann-Schwarz, V
- Abstract
Anti-glycan antibodies represent a vast and yet insufficiently investigated subpopulation of naturally occurring and adaptive antibodies in humans. Recently, a variety of glycan-based microarrays emerged, allowing high-throughput profiling of a large repertoire of antibodies. As there are no direct approaches for comparison and evaluation of multi-glycan assays we compared three glycan-based immunoassays, namely printed glycan array (PGA), fluorescent microsphere-based suspension array (SA) and ELISA for their efficacy and selectivity in profiling anti-glycan antibodies in a cohort of 48 patients with and without ovarian cancer. The ABO blood group glycan antigens were selected as well recognized ligands for sensitivity and specificity assessments. As another ligand we selected P(1), a member of the P blood group system recently identified by PGA as a potential ovarian cancer biomarker. All three glyco-immunoassays reflected the known ABO blood groups with high performance. In contrast, anti-P(1) antibody binding profiles displayed much lower concordance. Whilst anti-P(1) antibody levels between benign controls and ovarian cancer patients were significantly discriminated using PGA (p = 0.004), we got only similar results using SA (p = 0.03) but not for ELISA. Our findings demonstrate that whilst assays were largely positively correlated, each presents unique characteristic features and should be validated by an independent patient cohort rather than another array technique. The variety between methods presumably reflects the differences in glycan presentation and the antigen/antibody ratio, assay conditions and detection technique. This indicates that the glycan-antibody interaction of interest has to guide the assay selection.
- Published
- 2011
10. The prognostic relevance of altered antiglycan antibody profiles in the sera of primary melanoma patients.
- Author
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Haimovic, A., primary, Ma, M. W., additional, Vuskovic, M. I., additional, Miller, T., additional, DiBenedetto, M., additional, Grossman, J., additional, Shapiro, R. L., additional, Pavlick, A. C., additional, Berman, R. S., additional, Pass, H. I., additional, Huflejt, M., additional, and Osman, I., additional
- Published
- 2011
- Full Text
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11. 2-Aminopyridine—a label for bridging of oligosaccharides HPLC profiling and glycoarray printing
- Author
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Shilova, N. V., primary, Galanina, O. E., additional, Rubina, A. Yu., additional, Butvilovskaya, V. I., additional, Huflejt, M. E., additional, Chambers, J., additional, Roucoux, A., additional, and Bovin, N. V., additional
- Published
- 2007
- Full Text
- View/download PDF
12. The glycosphingolipid P1 is an ovarian cancer-associated carbohydrate antigen involved in migration.
- Author
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Jacob, F, Anugraham, M, Pochechueva, T, Tse, B W C, Alam, S, Guertler, R, Bovin, N V, Fedier, A, Hacker, N F, Huflejt, M E, Packer, N, and Heinzelmann-Schwarz, V A
- Subjects
OVARIAN cancer patients ,WOMEN'S health ,GLYCOSPHINGOLIPIDS ,CARBOHYDRATES ,ANTIGENS ,BLOOD plasma ,CANCER relapse - Abstract
Background:The level of plasma-derived naturally circulating anti-glycan antibodies (AGA) to P
1 trisaccharide has previously been shown to significantly discriminate between ovarian cancer patients and healthy women. Here we aim to identify the Ig class that causes this discrimination, to identify on cancer cells the corresponding P1 antigen recognised by circulating anti-P1 antibodies and to shed light into the possible function of this glycosphingolipid.Methods:An independent Australian cohort was assessed for the presence of anti-P1 IgG and IgM class antibodies using suspension array. Monoclonal and human derived anti-glycan antibodies were verified using three independent glycan-based immunoassays and flow cytometry-based inhibition assay. The P1 antigen was detected by LC-MS/MS and flow cytometry. FACS-sorted cell lines were studied on the cellular migration by colorimetric assay and real-time measurement using xCELLigence system.Results:Here we show in a second independent cohort (n=155) that the discrimination of cancer patients is mediated by the IgM class of anti-P1 antibodies (P=0.0002). The presence of corresponding antigen P1 and structurally related epitopes in fresh tissue specimens and cultured cancer cells is demonstrated. We further link the antibody and antigen (P1 ) by showing that human naturally circulating and affinity-purified anti-P1 IgM isolated from patients ascites can bind to naturally expressed P1 on the cell surface of ovarian cancer cells. Cell-sorted IGROV1 was used to obtain two study subpopulations (P1 -high, 66.1%; and P1 -low, 33.3%) and observed that cells expressing high P1 -levels migrate significantly faster than those with low P1 -levels.Conclusions:This is the first report showing that P1 antigen, known to be expressed on erythrocytes only, is also present on ovarian cancer cells. This suggests that P1 is a novel tumour-associated carbohydrate antigen recognised by the immune system in patients and may have a role in cell migration. The clinical value of our data may be both diagnostic and prognostic; patients with low anti-P1 IgM antibodies present with a more aggressive phenotype and earlier relapse. [ABSTRACT FROM AUTHOR]- Published
- 2014
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13. Fas ligand- mediated killing by intestinal intraepithelial lymphocytes. Participation in intestinal graft-versus-host disease.
- Author
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Lin, T, primary, Brunner, T, additional, Tietz, B, additional, Madsen, J, additional, Bonfoco, E, additional, Reaves, M, additional, Huflejt, M, additional, and Green, D R, additional
- Published
- 1998
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14. Regulated Cl transport, K and Cl permeability, and exocytosis in T84 cells.
- Author
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Huflejt, M E, primary, Blum, R A, additional, Miller, S G, additional, Moore, H P, additional, and Machen, T E, additional
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- 1994
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15. L-29, a soluble lactose-binding lectin, is phosphorylated on serine 6 and serine 12 in vivo and by casein kinase I.
- Author
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Huflejt, M E, primary, Turck, C W, additional, Lindstedt, R, additional, Barondes, S H, additional, and Leffler, H, additional
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- 1993
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16. Primary structure of the soluble lactose binding lectin L-29 from rat and dog and interaction of its non-collagenous proline-, glycine-, tyrosine-rich sequence with bacterial and tissue collagenase.
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Herrmann, J, primary, Turck, C W, additional, Atchison, R E, additional, Huflejt, M E, additional, Poulter, L, additional, Gitt, M A, additional, Burlingame, A L, additional, Barondes, S H, additional, and Leffler, H, additional
- Published
- 1993
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17. ACUTE EFFECTS OF NEAR ULTRAVIOLET AND VISIBLE LIGHT ON THE CUTANEOUS ANTIOXIDANT DEFENSE SYSTEM.
- Author
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Fuchs, J., Huflejt, M. E., Rothfuss, L. M., Wilson, D. S., Carcamo, G., and Packer, L.
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- 1989
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18. Strikingly different localization of galectin-3 and galectin-4 in human colon adenocarcinoma T84 cells. Galectin-4 is localized at sites of cell adhesion.
- Author
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Huflejt, M E, Jordan, E T, Gitt, M A, Barondes, S H, and Leffler, H
- Abstract
Two beta-galactoside-binding proteins were found to be prominently expressed in the human colon adenocarcinoma T84 cell line. Cloning and sequencing of one, a 36-kDa protein, identified it as the human homolog of galectin-4, a protein containing two carbohydrate binding domains and previously found only in the epithelial cells of the rat and porcine alimentary tract. The other, a 29-kDa protein, is galectin-3, containing a single carbohydrate binding domain, previously found in a number of different cell types including human intestinal epithelium. Despite the marked similarities in the carbohydrate binding domains of these two galectins, their cellular distribution patterns are strikingly different and vary with cellular conditions. In confluent T84 cells, galectin-4 is mostly cytosolic and concentrated at the basal membrane, whereas galectin-3 tends to be concentrated in large granular inclusions mostly at the apical membrane. In subconfluent T84 cells, each galectin is distributed to specific domains of lamellipodia, with galectin-4 concentrated in the leading edge and galectin-3 more proximally. Such different localization of galectins-4 and -3 within T84 cells implies different targeting mechanisms, ligands, and functions. The localization of galectin-4 suggests a role in cell adhesion which is also supported by the ability of immobilized recombinant galectin-4 to stimulate adhesion of T84 cells.
- Published
- 1997
19. EPR signals of redox active copper in EDTA washed membrane of the cyanobacterium Synechococcus 6311
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Fry, I.V., primary, Peschek, G.A., additional, Huflejt, M., additional, and Packer, L., additional
- Published
- 1985
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20. The role of respiration during adaptation of the freshwater cyanobacterium Synechococcus 6311 to salinity
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Fry, I.V., primary, Huflejt, M., additional, Erber, W.W.A., additional, Peschek, G.A., additional, and Packer, L., additional
- Published
- 1986
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21. EPR signals of redox active copper in EDTA washed membrane of the cyanobacterium [formula omitted] 6311
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Fry, I.V., Peschek, G.A., Huflejt, M., and Packer, L.
- Published
- 1985
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22. Migration of human mesenchymal stem cells stimulated with pulsed electric field and the dynamics of the cell surface glycosylation.
- Author
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Jezierska-Woźniak K, Lipiński S, Huflejt M, Grabarczyk Ł, Barczewska M, Habich A, and Maksymowicz W
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- Biomarkers, Cell Differentiation, Humans, Cell Movement, Glycosylation, Mesenchymal Stem Cells
- Abstract
Background: The analysis of the stem cells' glycome dynamics at different stages of differentiation and migration makes possible the exploration of the cell surface glycans as markers of the stem cell functional status, and, in the future, compatibility between transplanted cell and host environment., Objectives: The objective of our study was to develop novel techniques of investigating cell motility and to assess whether the electric field of the therapeutic spinal cord stimulation system used in vivo contributes to the migration of human mesenchymal stem cells (hMSCs) in vitro., Material and Methods: We have investigated the electrotaxis of bone marrow-derived MSCs using pulsed electric field (PEF) in the range of 16-80 mV/mm and the frequency of 130 Hz and 240 Hz. The PEF-related dynamics of the cell surface glycosylation was evaluated using 6 plant lectins recognizing individual glycans., Results: Pulsed electric field at physiological levels (10 mV/mm; 130 Hz) did not influence cellular motility in vitro, which may correspond to the maintenance of the transplanted cells at the lesion site in vivo. An increase of the PEF intensity and the frequency exceeding physiological levels resulted in an increase in the cellular migration rate in vitro. Pulsed electric field elevated above physiological intensity and frequency (40-80 mV/mm; 240 Hz), but not at physiological levels, resulted in changes of the cell surface glycosylation., Conclusions: We found the described approach convenient for investigations and for the in vitro modeling of the cellular systems intended for the regenerative cell transplantations in vivo. Probing cell surface glycomes may provide valuable biomarkers to assess the competence of transplanted cells.
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- 2018
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23. An Automated Confocal Micro-Extensometer Enables in Vivo Quantification of Mechanical Properties with Cellular Resolution.
- Author
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Robinson S, Huflejt M, Barbier de Reuille P, Braybrook SA, Schorderet M, Reinhardt D, and Kuhlemeier C
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- Automation, Biomechanical Phenomena, Cell Wall drug effects, Cell Wall physiology, Elasticity, Gibberellins pharmacology, Hypocotyl cytology, Hypocotyl drug effects, Hypocotyl growth & development, Hypocotyl radiation effects, Light, Models, Biological, Plant Cells drug effects, Stress, Physiological drug effects, Arabidopsis cytology, Microscopy, Confocal instrumentation, Plant Cells chemistry
- Abstract
How complex developmental-genetic networks are translated into organs with specific 3D shapes remains an open question. This question is particularly challenging because the elaboration of specific shapes is in essence a question of mechanics. In plants, this means how the genetic circuitry affects the cell wall. The mechanical properties of the wall and their spatial variation are the key factors controlling morphogenesis in plants. However, these properties are difficult to measure and investigating their relation to genetic regulation is particularly challenging. To measure spatial variation of mechanical properties, one must determine the deformation of a tissue in response to a known force with cellular resolution. Here, we present an automated confocal micro-extensometer (ACME), which greatly expands the scope of existing methods for measuring mechanical properties. Unlike classical extensometers, ACME is mounted on a confocal microscope and uses confocal images to compute the deformation of the tissue directly from biological markers, thus providing 3D cellular scale information and improved accuracy. Additionally, ACME is suitable for measuring the mechanical responses in live tissue. As a proof of concept, we demonstrate that the plant hormone gibberellic acid induces a spatial gradient in mechanical properties along the length of the Arabidopsis thaliana hypocotyl., (© 2017 American Society of Plant Biologists. All rights reserved.)
- Published
- 2017
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24. Measuring the mechanical properties of plant cells by combining micro-indentation with osmotic treatments.
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Weber A, Braybrook S, Huflejt M, Mosca G, Routier-Kierzkowska AL, and Smith RS
- Subjects
- Elasticity, Microscopy, Atomic Force, Models, Theoretical, Osmotic Pressure, Stress, Mechanical, Nicotiana growth & development, Cell Wall physiology, Plant Cells physiology, Nicotiana physiology
- Abstract
Growth in plants results from the interaction between genetic and signalling networks and the mechanical properties of cells and tissues. There has been a recent resurgence in research directed at understanding the mechanical aspects of growth, and their feedback on genetic regulation. This has been driven in part by the development of new micro-indentation techniques to measure the mechanical properties of plant cells in vivo. However, the interpretation of indentation experiments remains a challenge, since the force measures results from a combination of turgor pressure, cell wall stiffness, and cell and indenter geometry. In order to interpret the measurements, an accurate mechanical model of the experiment is required. Here, we used a plant cell system with a simple geometry, Nicotiana tabacum Bright Yellow-2 (BY-2) cells, to examine the sensitivity of micro-indentation to a variety of mechanical and experimental parameters. Using a finite-element mechanical model, we found that, for indentations of a few microns on turgid cells, the measurements were mostly sensitive to turgor pressure and the radius of the cell, and not to the exact indenter shape or elastic properties of the cell wall. By complementing indentation experiments with osmotic experiments to measure the elastic strain in turgid cells, we could fit the model to both turgor pressure and cell wall elasticity. This allowed us to interpret apparent stiffness values in terms of meaningful physical parameters that are relevant for morphogenesis., (© The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology.)
- Published
- 2015
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25. Cross-platform comparison of glycan microarray formats.
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Wang L, Cummings RD, Smith DF, Huflejt M, Campbell CT, Gildersleeve JC, Gerlach JQ, Kilcoyne M, Joshi L, Serna S, Reichardt NC, Parera Pera N, Pieters RJ, Eng W, and Mahal LK
- Subjects
- Binding Sites, Carbohydrates biosynthesis, Carrier Proteins chemistry, Concanavalin A chemistry, Concanavalin A metabolism, Lectins chemistry, Lectins metabolism, Phytohemagglutinins chemistry, Phytohemagglutinins metabolism, Polysaccharides chemistry, Wheat Germ Agglutinins chemistry, Wheat Germ Agglutinins metabolism, Carrier Proteins metabolism, Microarray Analysis, Polysaccharides metabolism
- Abstract
Carbohydrates participate in almost every aspect of biology from protein sorting to modulating cell differentiation and cell-cell interactions. To date, the majority of data gathered on glycan expression has been obtained via analysis with either anti-glycan antibodies or lectins. A detailed understanding of the specificities of these reagents is critical to the analysis of carbohydrates in biological systems. Glycan microarrays are increasingly used to determine the binding specificity of glycan-binding proteins (GBPs). In this study, six different glycan microarray platforms with different modes of glycan presentation were compared using five well-known lectins; concanavalin A, Helix pomatia agglutinin, Maackia amurensis lectin I, Sambucus nigra agglutinin and wheat germ agglutinin. A new method (universal threshold) was developed to facilitate systematic comparisons across distinct array platforms. The strongest binders of each lectin were identified using the universal threshold across all platforms while identification of weaker binders was influenced by platform-specific factors including presentation of determinants, array composition and self-reported thresholding methods. This work compiles a rich dataset for comparative analysis of glycan array platforms and has important implications for the implementation of microarrays in the characterization of GBPs.
- Published
- 2014
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26. Plasma Anti-Glycan Antibody Profiles Associated with Nickel level in Urine.
- Author
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Vuskovic M, Barbuti AM, Goldsmith-Rooney E, Glassman L, Bovin N, Pass H, Tchou-Wong KM, Chen M, Yan B, Niu J, Qu Q, Costa M, and Huflejt M
- Abstract
Nickel (Ni) compounds are widely used in industrial and commercial products including household and cooking utensils, jewelry, dental appliances and implants. Occupational exposure to nickel is associated with an increased risk for lung and nasal cancers, is the most common cause of contact dermatitis and has an extensive effect on the immune system. The purpose of this study was two-fold: (i) to evaluate immune response to the occupational exposure to nickel measured by the presence of anti-glycan antibodies (AGA) using a new biomarker-discovery platform based on printed glycan arrays (PGA), and (ii) to evaluate and compile a sequence of bioinformatics and statistical methods which are specifically relevant to PGA-derived information and to identification of putative "Ni toxicity signature". The PGAs are similar to DNA microarrays, but contain deposits of various carbohydrates (glycans) instead of spotted DNAs. The study uses data derived from a set of 89 plasma specimens and their corresponding demographic information. The study population includes three subgroups: subjects directly exposed to Nickel that work in a refinery, subjects environmentally exposed to Nickel that live in a city where the refinery is located and subjects that live in a remote location. The paper describes the following sequence of nine data processing and analysis steps: (1) Analysis of inter-array reproducibility based on benchmark sera; (2) Analysis of intra-array reproducibility; (3) Screening of data - rejecting glycans which result in low intra-class correlation coefficient (ICC), high coefficient of variation and low fluorescent intensity; (4) Analysis of inter-slide bias and choice of data normalization technique; (5) Determination of discriminatory subsamples based on multiple bootstrap tests; (6) Determination of the optimal signature size (cardinality of selected feature set) based on multiple cross-validation tests; (7) Identification of the top discriminatory glycans and their individual performance based on nonparametric univariate feature selection; (8) Determination of multivariate performance of combined glycans; (9) Establishing the statistical significance of multivariate performance of combined glycan signature. The above analysis steps have delivered the following results: inter-array reproducibility ρ =0.920 ± 0.030; intra-array reproducibility ρ =0.929 ± 0.025; 249 out of 380 glycans passed the screening at ICC>80%, glycans in selected signature have ICC ≥ 88.7%; optimal signature size (after quantile normalization)=3; individual significance for the signature glycans p =0.00015 to 0.00164, individual AUC values 0.870 to 0.815; observed combined performance for three glycans AUC =0.966, p =0.005, CI= [0.757, 0947]; specifity =94.4%, sensitivity =88.9%; predictive (cross-validated) AUC value 0.836.
- Published
- 2013
- Full Text
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27. Fibulin-3 as a blood and effusion biomarker for pleural mesothelioma.
- Author
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Pass HI, Levin SM, Harbut MR, Melamed J, Chiriboga L, Donington J, Huflejt M, Carbone M, Chia D, Goodglick L, Goodman GE, Thornquist MD, Liu G, de Perrot M, Tsao MS, and Goparaju C
- Subjects
- Aged, Biomarkers blood, Case-Control Studies, Diagnosis, Differential, Female, Humans, Kaplan-Meier Estimate, Male, Mesothelioma blood, Middle Aged, Pleural Effusion blood, Pleural Effusion diagnosis, Pleural Effusion, Malignant blood, Pleural Effusion, Malignant diagnosis, Pleural Neoplasms blood, ROC Curve, Sensitivity and Specificity, Asbestos adverse effects, Extracellular Matrix Proteins blood, Mesothelioma diagnosis, Occupational Exposure, Pleural Neoplasms diagnosis
- Abstract
Background: New biomarkers are needed to detect pleural mesothelioma at an earlier stage and to individualize treatment strategies. We investigated whether fibulin-3 in plasma and pleural effusions could meet sensitivity and specificity criteria for a robust biomarker., Methods: We measured fibulin-3 levels in plasma (from 92 patients with mesothelioma, 136 asbestos-exposed persons without cancer, 93 patients with effusions not due to mesothelioma, and 43 healthy controls), effusions (from 74 patients with mesothelioma, 39 with benign effusions, and 54 with malignant effusions not due to mesothelioma), or both. A blinded validation was subsequently performed. Tumor tissue was examined for fibulin-3 by immunohistochemical analysis, and levels of fibulin-3 in plasma and effusions were measured with an enzyme-linked immunosorbent assay., Results: Plasma fibulin-3 levels did not vary according to age, sex, duration of asbestos exposure, or degree of radiographic changes and were significantly higher in patients with pleural mesothelioma (105±7 ng per milliliter in the Detroit cohort and 113±8 ng per milliliter in the New York cohort) than in asbestos-exposed persons without mesothelioma (14±1 ng per milliliter and 24±1 ng per milliliter, respectively; P<0.001). Effusion fibulin-3 levels were significantly higher in patients with pleural mesothelioma (694±37 ng per milliliter in the Detroit cohort and 636±92 ng per milliliter in the New York cohort) than in patients with effusions not due to mesothelioma (212±25 and 151±23 ng per milliliter, respectively; P<0.001). Fibulin-3 preferentially stained tumor cells in 26 of 26 samples. In an overall comparison of patients with and those without mesothelioma, the receiver-operating-characteristic curve for plasma fibulin-3 levels had a sensitivity of 96.7% and a specificity of 95.5% at a cutoff value of 52.8 ng of fibulin-3 per milliliter. In a comparison of patients with early-stage mesothelioma with asbestos-exposed persons, the sensitivity was 100% and the specificity was 94.1% at a cutoff value of 46.0 ng of fibulin-3 per milliliter. Blinded validation revealed an area under the curve of 0.87 for plasma specimens from 96 asbestos-exposed persons as compared with 48 patients with mesothelioma., Conclusions: Plasma fibulin-3 levels can distinguish healthy persons with exposure to asbestos from patients with mesothelioma. In conjunction with effusion fibulin-3 levels, plasma fibulin-3 levels can further differentiate mesothelioma effusions from other malignant and benign effusions. (Funded by the Early Detection Research Network, National Institutes of Health, and others.).
- Published
- 2012
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28. Repertoire of human natural anti-glycan immunoglobulins. Do we have auto-antibodies?
- Author
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Bovin N, Obukhova P, Shilova N, Rapoport E, Popova I, Navakouski M, Unverzagt C, Vuskovic M, and Huflejt M
- Subjects
- Autoantibodies blood, Carbohydrate Sequence, Cohort Studies, Epitope Mapping, Glycomics methods, Humans, Immunoglobulins blood, Models, Biological, Molecular Sequence Data, Polysaccharides blood, Protein Array Analysis, Protein Binding, Autoantibodies analysis, Immunoglobulins analysis, Polysaccharides immunology
- Abstract
Background: Profiling of donor's antibodies using glycan arrays demonstrated presence of antibodies capable of binding to >100 mammalian glycans or their fragments. For example, relatively high binding to Galα1-4Galβ1-4GlcNAc (P(1)), Galα1-4Galβ1-4Glc (P(k)), Galβ1-3GlcNAc (Le(c)), 4-O-SuGalβ1-4GlcNAc, and GalNAcα1-3GalNAc (Fs) was found in all tested individuals. Affinity isolation using hapten-specific chromatography in combination with epitope mapping revealed their glycotopes. Notably, a significant part of the antibodies was capable of recognizing a fragment of larger glycans, for example, -Galβ1-4Glc of glycolipids, or Fucα1-3GlcNAc motif of Le(X)/Le(Y) antigens. Their epitope specificity did not vary between different healthy individuals. Nominally, all the mentioned immunoglobulins could be classified as auto-antibodies., Methods: In this work we re-evaluated results published earlier and analyzed new data to address the question why autologous antibodies found in healthy individuals do not cause severe auto-immune reactions., Results: In all cases the presumably "auto" antibodies were found to bind short fragments "subtracted" from larger glycans whereas recognition of the same fragment in the context of the whole natural chain was completely abolished. Thus, in spite of numerous formally positive signals observed on the printed glycan array, we are yet unable to identify in blood serum of healthy individuals true auto-antibodies capable of binding carbohydrate chains in their naturally occurring form., General Significance: The identified natural anti-glycan antibodies were found to be specific, high-titer and population conservative immunoglobulins - all of this suggesting as yet unknown biological role(s) of the studied proteins. This article is part of a Special Issue entitled Glycoproteomics., (Copyright © 2012 Elsevier B.V. All rights reserved.)
- Published
- 2012
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29. Detection of integrin-linked kinase in the serum of patients with malignant pleural mesothelioma.
- Author
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Watzka SB, Posch F, Pass HI, Huflejt M, Bernhard D, Hannigan GE, and Müller MR
- Subjects
- Aged, Biomarkers blood, Cell Transformation, Neoplastic, Enzyme-Linked Immunosorbent Assay, Female, Humans, Male, Mesothelioma diagnosis, Mesothelioma enzymology, Middle Aged, Pleural Neoplasms diagnosis, Pleural Neoplasms enzymology, Prognosis, Sensitivity and Specificity, Mesothelioma blood, Pleural Neoplasms blood, Protein Serine-Threonine Kinases blood
- Abstract
Objective: Integrin-linked kinase, which is relevant to neoplastic transformation, is highly expressed in malignant pleural mesothelioma. Recently, detection of integrin-linked kinase in serum of patients with ovarian cancer has been reported. This study asks whether integrin-linked kinase can also be detected in serum of patients with malignant pleural mesothelioma and whether serum level has diagnostic or prognostic relevance for that disease., Methods: A sandwich enzyme-linked immunosorbent assay was designed to detect integrin-linked kinase and applied to serum samples from 46 patients with malignant pleural mesothelioma, 98 patients with other malignant chest disease, and 23 patients with benign chest disease. Integrin-linked kinase serum concentration and clinical data were correlated statistically., Results: Median serum integrin-linked kinase concentration was significantly higher in malignant pleural mesothelioma (8.89 ng/mL) than in other malignant chest disease (0.66 ng/mL) or benign chest disease (0.78 ng/mL, P < .001). There was no relevant correlation of serum integrin-linked kinase with cell lysis parameters (R(2) < 0.1). Serum integrin-linked kinase concentration greater than 2.48 ng/mL had diagnostic sensitivity of 80%, specificity of 95%, positive predictive value of 85.7%, negative predictive value of 92.7%, and overall accuracy of 91% for distinction between malignant pleural mesothelioma and other diseases. Serum integrin-linked kinase concentration in malignant pleural mesothelioma was independent of histologic subtype or asbestos exposure. There was no statistically significant impact of serum integrin-linked kinase concentration on prognosis., Conclusions: Integrin-linked kinase can be detected in serum of patients with malignant pleural mesothelioma and may be a diagnostic marker for the disease., (Copyright © 2011 The American Association for Thoracic Surgery. Published by Mosby, Inc. All rights reserved.)
- Published
- 2011
- Full Text
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30. Reliable motion artifact detection for ECG monitoring systems with dry electrodes.
- Author
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Ottenbacher J, Kirst M, Jatobá L, Huflejt M, Grossmann U, and Stork W
- Subjects
- Acceleration, Algorithms, Biomedical Engineering, Electric Impedance, Electrodes, Humans, Motion, ROC Curve, Sensitivity and Specificity, Signal Processing, Computer-Assisted, Electrocardiography instrumentation, Electrocardiography statistics & numerical data
- Abstract
Reliable signals are the basic prerequisite for most mobile ECG monitoring applications. Especially when signals are analyzed automatically, capable motion artifact detection algorithms are of great importance. This article presents different artifact detection algorithms for ECG systems with dry electrodes. The algorithms are based on the measurement of additional parameters that are correlated with the artifacts. We describe a mobile measurement system and the procedure used for the evaluation of these algorithms. The algorithms are assessed based upon their effect on QRS detection. The best algorithm improved sensitivity (Se) from 98.7% to 99.8% and positive predictive value (+P) from 98.3% to 99.9%, while 15% of the signal was marked as artifact. This corresponds to a decrease in false positive and false negative detected beats by 89.9%. Different metrics to evaluate the performance of an artifact detection algorithm are presented.
- Published
- 2008
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31. Fluorescence polarization as an analytical tool to evaluate galectin-ligand interactions.
- Author
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Sörme P, Kahl-Knutsson B, Huflejt M, Nilsson UJ, and Leffler H
- Subjects
- Animals, Binding Sites, Carbohydrate Metabolism, Carbohydrates pharmacology, Fluorescein, Fluorescent Dyes chemistry, Galectin 1 antagonists & inhibitors, Galectin 1 chemistry, Galectin 1 metabolism, Galectin 3 antagonists & inhibitors, Galectin 3 chemistry, Galectin 3 metabolism, Galectin 4 antagonists & inhibitors, Galectin 4 chemistry, Galectin 4 metabolism, Galectins antagonists & inhibitors, Galectins metabolism, Humans, Hydrogen Bonding, Ligands, Protein Structure, Tertiary, Rats, Carbohydrates chemistry, Fluorescence Polarization, Fluorescent Dyes chemical synthesis, Galectins chemistry
- Abstract
Galectins are a family of beta-galactose binding lectins associated with functions such as immunological and malignant events. To study the binding affinity of galectins for natural and artificial saccharides and glycoconjugates we have developed an assay using fluorescence polarization. A collection of fluorescein-conjugated saccharides was synthesized and used as probes with galectins-1 and -3 and the two carbohydrate recognition domains of galectin-4. Direct binding of a fixed probe amount with different amounts of each galectin defined specificity and selectivity and permitted selection of the optimal probe for inhibition studies. Then fixed amounts of galectin and selected probe were used to screen the inhibitory potency of a library of nonfluorescent compounds. As the assay is in solution and does not require separation of free and bound probe, it is simple and rapid and can easily be applied to different unlabeled galectins. As all interaction components are known, K(d) values for galectin-inhibitor interaction can be directly calculated without approximation other than the assumption of a simple one-site competition.
- Published
- 2004
- Full Text
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32. Regulation of mucosal immune responses by recombinant interleukin 10 produced by intestinal epithelial cells in mice.
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De Winter H, Elewaut D, Turovskaya O, Huflejt M, Shimeld C, Hagenbaugh A, Binder S, Takahashi I, Kronenberg M, and Cheroutre H
- Subjects
- Animals, Antibody Formation drug effects, Colitis prevention & control, Cytokines biosynthesis, Gene Targeting, Immunoglobulin A biosynthesis, Interleukin-10 genetics, Intestinal Mucosa cytology, Intestinal Mucosa metabolism, Intestine, Small cytology, Lymphocyte Count, Lymphocytes cytology, Lymphocytes metabolism, Mice, Mice, Inbred Strains, Mice, Knockout genetics, Mice, Transgenic genetics, Rats, Recombinant Proteins pharmacology, T-Lymphocytes, Helper-Inducer metabolism, Tissue Distribution, Transforming Growth Factor beta metabolism, Transforming Growth Factor beta1, Interleukin-10 pharmacology, Intestinal Mucosa immunology
- Abstract
Background & Aims: Interleukin (IL)-10 is a cytokine with anti-inflammatory properties. The aim of this study was to explore the effect of a site-specific delivery of IL-10 on intestinal immune responses., Methods: Transgenic mice were created in which IL-10 is expressed by the intestinal epithelial cells., Results: Transgenic mice showed a marked increase in the number of intraepithelial lymphocytes in the small intestine. Mucosal lymphocytes of transgenic animals produced fewer T helper type 1 cytokines than wild-type lymphocytes. By contrast, the production of transforming growth factor beta was increased. Moreover, the epithelial layer in transgenic mice was significantly enriched for CD4(+)CD25(+) T cells. Furthermore, transgenic mice had increased numbers of immunoglobulin A-producing B cells in the small intestine. These effects were local because splenic lymphocytes were not affected. Studies in models of inflammatory bowel disease showed that transgenic IL-10 was able to attenuate the acute colitis induced by dextran sodium sulfate administration or by adoptive transfer of CD4(+)CD45RB(high) splenocytes, with a modest effect on the chronic intestinal inflammation arising spontaneously in IL-10(-/-) mice., Conclusions: These observations provide evidence for an in vivo lymphoepithelial cross talk, by which cytokines locally produced by epithelial cells can regulate immune responses in the intestine without systemic modifications.
- Published
- 2002
- Full Text
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33. The role of Thomsen-Friedenreich antigen in adhesion of human breast and prostate cancer cells to the endothelium.
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Glinsky VV, Glinsky GV, Rittenhouse-Olson K, Huflejt ME, Glinskii OV, Deutscher SL, and Quinn TP
- Subjects
- Amino Acid Sequence, Antigens, Differentiation metabolism, Antigens, Tumor-Associated, Carbohydrate biosynthesis, Bone Marrow blood supply, Breast Neoplasms immunology, Cell Adhesion physiology, Endothelium, Vascular metabolism, Epitopes immunology, Female, Galectin 3, Humans, Male, Microscopy, Confocal, Molecular Mimicry, Molecular Sequence Data, Peptide Fragments immunology, Prostatic Neoplasms immunology, Antigens, Neoplasm physiology, Antigens, Tumor-Associated, Carbohydrate physiology, Breast Neoplasms pathology, Endothelium, Vascular cytology, Prostatic Neoplasms pathology
- Abstract
Interactions of metastatic cancer cells with vasculatory endothelium are critical during early stages of cancer metastasis. Understanding the molecular underpinnings of these interactions is essential for the development of new efficacious cancer therapies. Here we demonstrate that cancer-associated carbohydrate T antigen plays a leading role in docking breast and prostate cancer cells onto endothelium by specifically interacting with endothelium-expressed beta-galactoside-binding protein, galectin-3. Importantly, T antigen-bearing glycoproteins are also capable of mobilizing galectin-3 to the surface of endothelial cells, thus priming them for harboring metastatic cancer cells. The T antigen-mediated, tumor-endothelial cell interactions could be efficiently disrupted using synthetic compounds either mimicking or masking this carbohydrate structure. High efficiency of T antigen-mimicking and T antigen-masking inhibitors of tumor cell adhesion warrants their further development into antiadhesive cancer therapeutics.
- Published
- 2001
34. Glycoprotein 90K/MAC-2BP interacts with galectin-1 and mediates galectin-1-induced cell aggregation.
- Author
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Tinari N, Kuwabara I, Huflejt ME, Shen PF, Iacobelli S, and Liu FT
- Subjects
- 3T3 Cells, Animals, Antigens, Differentiation metabolism, Cell Aggregation, Galectin 1, Galectin 3, Humans, Melanoma pathology, Mice, Molecular Weight, Tumor Cells, Cultured, Glycoproteins metabolism, Hemagglutinins metabolism
- Abstract
The glycoprotein 90K was originally described as a tumor-secreted antigen and subsequently found to have immunostimulatory activity as well as other possible functions. This protein interacts with an endogenous lectin, galectin-3, and may play a role in tumor metastasis through this interaction. Because 90K is heavily glycosylated, it may also interact with other members of the galectin family, which would contribute to the multifunctionality of 90K. To test this possibility, we studied the recognition of 90K by galectin-1, which, like galectin-3, has been associated with neoplastic transformation. In a solid-phase binding assay, human recombinant galectin-1 bound immobilized human recombinant 90K in a fashion that was inhibitable by lactose. Galectins 1 and 3 appeared to bind to separate sites on 90K because they did not affect the binding of each other. The dissociation constant of galectin-1 to 90K was on the order of 10(-7) M. Galectin-1 also induced aggregation of a human melanoma cell line, A375, in a carbohydrate-dependent manner, and this appeared to be mediated, at least in part, by 90K expressed on A375 cells, since it was inhibitable by a specific anti-90K monoclonal antibody. We conclude that 90K interacts with both galectin-1 and galectin-3 and both interactions contribute to the formation of multicell aggregates. Because both of these galectins as well as 90K are often over-expressed in neoplasm, these interactions may occur in the setting of various carcinomas and contribute to their progression and metastasis., (Copyright 2001 Wiley-Liss, Inc.)
- Published
- 2001
- Full Text
- View/download PDF
35. Effects of Thomsen-Friedenreich antigen-specific peptide P-30 on beta-galactoside-mediated homotypic aggregation and adhesion to the endothelium of MDA-MB-435 human breast carcinoma cells.
- Author
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Glinsky VV, Huflejt ME, Glinsky GV, Deutscher SL, and Quinn TP
- Subjects
- Amino Acid Sequence, Cell Aggregation drug effects, Endothelium drug effects, Endothelium metabolism, Female, Galectins, Hemagglutinins metabolism, Humans, Molecular Sequence Data, Neoplasm Metastasis prevention & control, Tumor Cells, Cultured, Antigens, Neoplasm metabolism, Antigens, Tumor-Associated, Carbohydrate metabolism, Breast Neoplasms pathology, Cell Adhesion drug effects, Galactosides metabolism, Peptides pharmacology
- Abstract
Both the ability of malignant cells to form multicellular aggregates via homotypic or heterotypic aggregation and their adhesion to the endothelium are important if not critical during early stages of cancer metastasis. The tumor-associated carbohydrate Thomsen-Friedenreich antigen (T antigen) and beta-galactoside binding lectins (galectins) have been implicated in tumor cell adhesion and tissue invasion. In this study, we demonstrate the involvement of T antigen in both homotypic aggregation of MDA-MB-435 human breast carcinoma cells and their adhesion to the endothelium. The T antigen-specific peptide P-30 (HGRFILPWWYAFSPS) selected from a bacteriophage display library was able to inhibit spontaneous homotypic aggregation of MDA-MB-435 cells up to 74% in a dose-dependent manner. Because T antigen has beta-galactose as a terminal sugar, the expression profile of beta-galactoside-binding lectins (galectins) in MDA-MB-435 cells was studied. Our data indicated the abundant expression of [35S]methionine/cysteine-labeled galectin-1 and galectin-3 in this cell line, which suggested possible interactions between galectins and T antigen. As revealed by laser confocal microscopy, both galectin-1 and galectin-3 also participate in the adhesion of the MDA-MB-435 cells to the endothelium. We observed the clustering of galectin-3 on endothelial cells at the sites of the contact with tumor cells, consistent with its possible interaction with T antigen on cancer cells The galectin-1 signal, however, strongly accumulated at the sites of cell-cell contacts predominantly on tumor cells. The T antigen-specific P-30 significantly (50%) inhibited this adhesion, which indicated that T antigen participates in the adhesion of MDA-MB-435 breast cancer cells to the endothelium. The ability of synthetic P-30 to inhibit both the spontaneous homotypic aggregation of MDA-MB-435 cells and their adhesion to the endothelium (>70 and 50%, respectively) suggests its potential functional significance for antiadhesive therapy of cancer metastasis.
- Published
- 2000
36. Membrane lipid composition, fluidity, and surface charge changes in response to growth of the fresh water cyanobacterium Synechococcus 6311 under high salinity.
- Author
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Khomutov G, Fry IV, Huflejt ME, and Packer L
- Subjects
- Acclimatization, Cell Fractionation, Cyanobacteria growth & development, Fresh Water, Kinetics, Organelles ultrastructure, Saline Solution, Hypertonic, Thermodynamics, Cell Membrane physiology, Cyanobacteria physiology, Membrane Fluidity, Membrane Lipids analysis
- Abstract
The effect of adaptation to saline growth of a fresh water cyanobacterium Synechococcus 6311 on components of the cytoplasmic membranes and thylakoids was investigated. Significant changes in membrane surface charge, lipid, fatty acid, and carotenoid composition were observed upon transfer of the cells from a low salt (0.015 M NaCl) to a high salt (0.50 M NaCl) growth medium. Very similar changes in the polar lipid classes and fatty acid composition were observed in both membranes, but changes in fluidity and surface charge and a significant shift in the protein to lipid ratio were only apparent in the cytoplasmic membranes. The fluidity and surface charge data correlate well with functional studies and we can attribute the cytoplasmic membrane as the major site of interaction and adaptation to the saline environment.
- Published
- 1990
- Full Text
- View/download PDF
37. Dermatologic antioxidant therapy may be warranted to prevent ultraviolet induced skin damage.
- Author
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Fuchs J, Huflejt M, Rothfuss L, Wilson D, Carcamo G, and Packer L
- Subjects
- Animals, Antioxidants analysis, Chromatography, High Pressure Liquid, Female, Mice, Mice, Hairless, Skin chemistry, Skin Aging radiation effects, Ubiquinone therapeutic use, Vitamin E therapeutic use, Antioxidants therapeutic use, Skin Aging drug effects, Skin Diseases prevention & control, Ultraviolet Rays adverse effects
- Published
- 1990
- Full Text
- View/download PDF
38. Hydroperoxide metabolism in cyanobacteria.
- Author
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Tel-Or E, Huflejt ME, and Packer L
- Subjects
- Ascorbate Peroxidases, Ascorbic Acid metabolism, Catalase metabolism, Cyanobacteria enzymology, Glutathione metabolism, Oxidation-Reduction, Peroxidases metabolism, Substrate Specificity, Cyanobacteria metabolism, Hydrogen Peroxide metabolism
- Abstract
The enzymes involved in antioxidative activity and the cellular content of the antioxidants glutathione and ascorbate in the cyanobacteria Nostoc muscorum 7119 and Synechococcus 6311 have been examined for their roles in hydroperoxide removal. High activities of ascorbate peroxidase and catalase were found in vegetative cells of both species and in the heterocysts of N. muscorum. The affinity of ascorbate peroxidase for H2O2 was 15- to 25-fold higher than that of catalase. Increased activity of ascorbate peroxidase was observed in N. muscorum when H2O2 production was enhanced by photorespiration. Catalase activity was decreased in dilute cultures whereas ascorbate peroxidase activity increased. Ascorbate peroxidase activity also increased when the CO2 concentration was reduced. Ascorbate peroxidase appears to be a key enzyme in a cascade of reactions regenerating antioxidants. Dehydroascorbate reductase was found to regenerate ascorbate, and glutathione reductase recycled glutathione. In vegetative cells glutathione was present in high amounts (2-4 mM) whereas the ascorbate content was almost 100-fold lower (20-100 microM). Glutathione peroxidase was not detected in either cyanobacterium. It is concluded from the high activity of ascorbate peroxidase activity and the levels of antioxidants found that this enzyme can effectively remove low concentrations of peroxides. Catalase may remove H2O2 produced under photooxidative conditions where the peroxide concentration is higher.
- Published
- 1986
- Full Text
- View/download PDF
39. Intracellular pH of halobacteria can be determined by the fluorescent dye 2', 7'-bis(carboxyethyl)-5(6)-carboxyfluorescein.
- Author
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Tsujimoto K, Semadeni M, Huflejt M, and Packer L
- Subjects
- Calibration, Cytosol metabolism, Hydrogen-Ion Concentration, Indicators and Reagents, Monensin, Sodium Chloride, Spectrometry, Fluorescence, Body Fluids metabolism, Fluoresceins, Halobacterium metabolism, Intracellular Fluid metabolism
- Abstract
Determination of the internal pH of halobacterial cells grown in 4M salt solution has proven to be a difficult problem. We now report the steady state cytosolic pH of Halobacterium halobium S-9 to be 7.2. Intracellular pH was determined after the cells were loaded with the membrane permeable precursor of the pH sensitive dye 2', 7'-bis-(carboxyethyl)-5(6)-carboxyfluorescein-acetyxymethyl ester - (BCECF/AM). In order to minimize light-scattering in the measurement of the fluorescence, a thin cuvette was newly devised. This method should be suitable for studies of the cytosolic pH in other bacteria.
- Published
- 1988
- Full Text
- View/download PDF
40. The role of glutathione and ascorbate in hydroperoxide removal in cyanobacteria.
- Author
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Tel-Or E, Huflejt M, and Packer L
- Subjects
- Ascorbate Peroxidases, Benzene Derivatives metabolism, Glutathione Peroxidase metabolism, Oxidation-Reduction, Oxidoreductases metabolism, Peroxidases metabolism, Photosynthesis, Selenium metabolism, Ascorbic Acid physiology, Cyanobacteria metabolism, Glutathione physiology, Peroxides metabolism
- Abstract
The antioxidative potential of cyanobacteria to scavenge hydroperoxides formed as by-products of photosynthetic activity was investigated in Nostoc muscorum 7119 and Synechococcus 6311. These cells contained a high concentration of glutathione, 2-5 mM, and a low concentration of ascorbate, 20-100 uM. No glutathione peroxidase was detected while the activity of ascorbate peroxidase was high, reacting with hydrogen peroxide, t-butyl hydroperoxide, and cumene hydroperoxide. Dehydroascorbate reductase was active in recycling ascorbate and glutathione reductase regenerated glutathione from glutathione disulphide. The activity of these antioxidative enzymes in the cyanobacteria was sufficient to remove between 60-230 nmoles H2O2 .mg protein-1 min-1. It is suggested that in cyanobacteria an effective reaction sequence for removal of hydroperoxides involves ascorbate peroxidase and recycling of glutathione and ascorbate.
- Published
- 1985
- Full Text
- View/download PDF
41. Impairment of enzymic and nonenzymic antioxidants in skin by UVB irradiation.
- Author
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Fuchs J, Huflejt ME, Rothfuss LM, Wilson DS, Carcamo G, and Packer L
- Subjects
- Animals, Mice, Mice, Hairless, Skin analysis, Ultraviolet Rays, Antioxidants analysis, Skin radiation effects
- Abstract
Antioxidants may play a significant role in ameliorating or preventing photobiologic damage in skin that could lead to cutaneous disorders such as cancer and premature aging. The objective of this study was to assess the acute cutaneous enzymic and nonenzymic antioxidant response to a single exposure of large fluence (300 mJ/cm2) ultraviolet radiation (greater than 280 nm) in hairless mice. This treatment caused an immediate and statistically significant inhibition of glutathione reductase and catalase activity. Glutathione peroxidase and superoxide dismutase were not affected. Glutathione levels decreased and, conversely glutathione disulfide concentrations increased. A slight depletion of the total glutathione was observed, while the content of total ascorbic acid did not change. The lipophilic antioxidants alpha-tocopherol, ubiquinol 9 and ubiquinone 9 also decreased significantly, and the concentration of malondialdehyde remained constant. The free radical scavenging activity of epidermis, as assessed by reduction of the stable, cationic nitroxide radical [2,2,6,6-tetramethyl-1-piperidinoxy-4-(2',4',6'-trimethyl) methylpyridinium perchlorate] was considerably inhibited. The study indicates that immediately after exposure to a large fluence of ultraviolet radiation the enzymic and nonenzymic antioxidant capacity of skin decreases significantly.
- Published
- 1989
- Full Text
- View/download PDF
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