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  • 1
    Electronic Resource
    Electronic Resource
    Restriction fragment length polymorphisms in Septoria tritici occur at a high frequency (1990)
    Springer
    Current genetics 17 (1990), S. 133-138 
    Details
    ISSN: 1432-0983
    Keywords: RFLPs ; Septoria tritici ; DNA fingerprinting ; Genetic variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A set of probes that detect restriction fragment length polymorphisms (RFLPs) in nuclear DNA has been developed for genetic studies of the phytopathogenic fungus Septoria tritici. Two plasmid libraries containing 0.5–1.3 or 1.3–2.4 kb fragments of S. tritici nuclear DNA were constructed. Seventeen random clones from each library were used as probes to screen for RFLP variation among a geographically-diverse group of six S. tritici isolates. Among the 196 probe-enzyme combinations tested, 145 detected RFLPs among the six isolates. The restriction enzymes EcoRV and PstI detected RFLPs most efficiently. Three probes detected deletions. A ribosomal DNA probe from yeast did not detect a significant amount of variation. These probes will be useful for studying genetic variation, population genetics, and genome organization of S. tritici.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00312858
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  • 2
    Electronic Resource
    Electronic Resource
    RFLPs in mitochondrial and nuclear DNA indicate low levels of genetic diversity in the oak wilt pathogen Ceratocystis fagacearum (1995)
    Springer
    Current genetics 27 (1995), S. 373-378 
    Details
    ISSN: 1432-0983
    Keywords: RFLPs ; Genetic diversity ; Population genetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genetic diversity in the oak wilt pathogen Ceratocystis fagacearum was assessed using restriction fragment length polymorphisms (RFLPs) of the mitochondrial DNA (mtDNA) and anonymous RFLP loci in the nuclear DNA (nuDNA). No genetic variation was detected in the mtDNA among 27 isolates sampled from a broad geographical area. Southern hybridization to 100 anonymous, random, nuDNA probes detected a low level of variation among nine of the isolates. Only 35 out of 437 probe-enzyme combinations detected RFLPs. Most of the RFLPs appeared to result from insertions and deletions of less than 200 bp. A composite multilocus haplotype based on hybridization to six anonymous probes could differentiate each of the nine isolates tested, suggesting that these probes may be useful for further studies of the population biology and epidemiology of this pathogen. Hypotheses are presented to account for the low level of genetic variation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00352107
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  • 3
    Electronic Resource
    Electronic Resource
    The role of selection on the genetic structure of pathogen populations: Evidence from field experiments with Mycosphaerella graminicola on wheat (1996)
    Springer
    Euphytica 92 (1996), S. 73-80 
    Details
    ISSN: 1573-5060
    Keywords: coevolution ; DNA fingerprints ; Mycosphaerella graminicola ; RFLPs ; Septoria tritici
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Coevolution refers to reciprocal genetic changes that occur in two or more ecologically interacting species. In agricultural ecosystems, we are especially concerned with the genetic response of pathogen populations to resistant cultivars produced by plant breeding programs. It would be useful to be able to predict whether disease resistance is likely to be durable or ephemeral before a cultivar is widely grown. Though it may not be possible to predict durability in advance, knowledge of the genetic structure of pathogen populations may prove useful for making predictions about the rate at which pathogens adapt to resistant varieties. Much has been learned about the genetic structure of populations of obligate fungal pathogens such as rusts and mildews, which have become paradigms for plant pathology. We have focused our effort on the population genetics of the less known, non-specialized, necrotrophic pathogens, such as the Septorias of small grains. Our approach has been to use DNA fingerprinting and RFLP analysis to conduct field experiments that elucidate how populations of fungal pathogens adapt in agroecosystems. Our results suggest that mating system may have a greater impact than natural selection on the genetic structure of populations of Mycosphaerella graminicola (anamorph Septoria tritici).
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00022831
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  • 4
    Electronic Resource
    Electronic Resource
    Cytoskeletal architecture and motility in a giant freshwater amoeba, Reticulomyxa (1986)
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 6 (1986), S. 521-533 
    Details
    ISSN: 0886-1544
    Keywords: intracellular organelle transport ; microtubules ; microfilaments ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Reticulomyxa is a large, multinucleated freshwater protozoan with striking intracellular transport. Cyloplasmic streaming and saltatory movements of individual organelles (at rates of up to 25 μm/sec) are observed within the naked cell body and the extensive reticulate peripheral network of fine cytoplasmic strands. As demonstrated by video-enhanced light microscopy, individual organelles move only when associated with cytoskeletal linear elements. The linear elements are composed of mixed colinear bundles of microtubules and actin filaments, which form the backbone of the reticulopodial network. The constant branching, sprouting, and fusion of network stands suggest unique membrane properties and an unusually dynamic cytoskeleton. The electrophoretic mobility of Reticulomyxa tubulins and the lack of crossreactivity with several antibodies known to react with many plant and animal tubulins suggest that they may differ from other tubulins more widely than might be expected. Reticulomyxa's large size, the rapidity and pervasiveness of the two forms of transport, and the simple and ordered cytoskeleton make the organism well suited for future studies on the mechanisms of intracellular transport.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cm.970060511
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  • 5
    Electronic Resource
    Electronic Resource
    Differential effects of tamoxifen-like compounds on osteoclastic bone degradation, H+-ATPase activity, calmodulin-dependent cyclic nucleotide phosphodiesterase activity, and calmodulin binding (1997)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 66 (1997), S. 358-369 
    Details
    ISSN: 0730-2312
    Keywords: calmodulin antagonists ; calmodulin binding proteins ; osteoclast ; phosphodiesterase ; H+-ATPase ; trifluoperazine ; centchroman ; cischroman ; calcium ; bone resorption ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: We studied effects of calmodulin antagonists on osteoclastic activity and calmodulin-dependent HCl transport. The results were compared to effects on the calmodulin-dependent phosphodiesterase and antagonist-calmodulin binding affinity. Avian osteoclast degradation of labeled bone was inhibited ∼40% by trifluoperazine or tamoxifen with half-maximal effects at 1-3 μM. Four benzopyrans structurally resembling tamoxifen were compared: d-centchroman inhibited resorption 30%, with half-maximal effect at ∼100 nM, cischroman and CDRI 85/287 gave 15-20% inhibition, and l-centchroman was ineffective. No benzopyran inhibited cell attachment or protein synthesis below 10 μM. However, ATP-dependent membrane vesicle acridine transport showed that H+-ATPase activity was abolished by all compounds with 50% effects at 0.25-1 μM. All compounds also inhibited calmodulin-dependent cyclic nucleotide phosphodiesterase at micromolar calcium. Relative potency varied with assay type, but d- and l-centchroman, surprisingly, inhibited both H+-ATPase and phosphodiesterase activity at similar concentrations. However, d- and l-centchroman effects in either assay diverged at nanomolar calcium. Of benzopyrans tested, only the d-centchroman effects were calcium-dependent. Interaction of compounds with calmodulin at similar concentrations were confirmed by displacement of labeled calmodulin from immobilized trifluoperazine. Thus, the compounds tested all interact with calmodulin directly to varying degrees, and the observed osteoclast inhibition is consistent with calmodulin-mediated effects. However, calmodulin antagonist activity varies between specific reactions, and free calcium regulates specificity of some interactions. Effects on whole cells probably also reflect other properties, including transport into cells. J. Cell. Biochem. 66:358-369, 1997. © 1997 Wiley-Liss, Inc.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Tyrosine phosphorylation of phosphatase inhibitor 2 (1995)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 57 (1995), S. 415-422 
    Details
    ISSN: 0730-2312
    Keywords: insulin ; dephosporylation ; type 1 phosphatase ; tyrosine kinase ; polylysine ; insulin receptor ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Inhibitor 2 is a heat-stable protein that complexes with the catalytic subunit of type-1 protein phosphatase. The reversible phosphorylation of Thr 72 of the inhibitor in this complex has been shown to regulate phosphatase activity. Here we show that inhibitor 2 can also be phosphorylated on tyrosine residues. Inhibitor 2 was 32P-labeled by the insulin receptor kinase in vitro, in the presence of polylysine. Phosphorylation of inhibitor 2 was accompanied by decreased electrophoretic mobility. Dephosphorylation of inhibitor 2 by tyrosine phosphatase 1B, restored normal electrophoretic mobility. Phosphotyrosine in inhibitor 2 was detected by immunoblotting with antiphosphotyrosine antibodies and phosphoamino acid analysis. In addition, following tryptic digestion, one predominant phosphopeptide was recovered at the anode. The ability of inhibitor 2 to inhibit type-1 phosphatase activity was diminished with increasing phosphorylation up to a stoichiometry of 1 mole phosphate incorporated/mole of inhibitor 2, where inhibitory activity was completely lost. These data demonstrate that inhibitor 2 can be phosphorylated on tyrosine residues by the insulin receptor kinase, resulting in a molecule with decreased ability to inhibit type-1 phosphatase activity.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcb.240570307
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  • 7
    Electronic Resource
    Electronic Resource
    Improved preservation of ultrastructure in difficult-to-fix organisms by high pressure freezing and freeze substitution: I. Drosophila melanogaster and Strongylocentrotus purpuratus embryos (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 24 (1993), S. 465-473 
    Details
    ISSN: 1059-910X
    Keywords: High pressure freezing ; Freeze substitution ; Drosophila ; Sea urchin ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: In this study, we have applied the techniques of high pressure freezing and freeze substitution to embryonic cell types which are usually difficult to fix properly for electron microscopy. In both Drosophila and Strongylocentrotus purpuratus, we see improved preservation of both membrane systems and cytoskeleton when compared to published results on the same cells using conventional electron microscope (EM) fixation methods. Finally, we have seen that postembedding labelling of sections is possible even after light osmium fixation during freeze substitution. © 1993 Wiley-Liss, Inc.
    Additional Material: 13 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070240603
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  • 8
    Electronic Resource
    Electronic Resource
    Binding of cytochalasin B to platelets (1989)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 39 (1989), S. 65-73 
    Details
    ISSN: 0730-2312
    Keywords: cytochalasin B ; platelets ; cytochalasin binding ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The binding of cytochalasin B (CB) to human platelets and to isolated platelet cytosol and membranes has been analyzed with [3H]CB. High- and low affinity classes of saturable binding sites were associated with intact platelets. Binding at very low concentrations of CB (i.e., high-affinity binding) was partially prevented by 100 mM D-galatose or D-glucose and to a much lesser extent by L-glucose. Binding to platelet cytosol also involved two classes of sites with affinities and capacities similar to those observed with the whole cells. None of this binding, however, was affected by 100 mM D-galactose. Saturable binding to platelet membranes occurred at sites with a uniform binding affinity. Approximately 52% of this binding was prevented by 1 M D-galactose and another 15% by cytochalasin E (CE). We hypothesize that binding in the cytosol is to monomeric (low-affinity) and polymerized (high-affinity) actin, whereas membrane binding (high-affinity only) occurs primarily at sites involved with galactose transport.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcb.240390108
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  • 9
    Electronic Resource
    Electronic Resource
    Immature megakaryocytes in the mouse: In vitro relationship to megakaryocyte progenitor cells and mature megakaryocytes (1982)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 112 (1982), S. 339-344 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: An assay describing conditions for the maturation of single immature megakaryocytes in vitro is reported. Enriched populations of small, relatively immature megakaryocytes have been found to develop into single, mature megakaryocytes by 60 hours in semisolid agar cultures. Continued incubation of these cells did not lead to the formation of colonies within 5-7 days. Maturation was indicated by increasing cell size and cytoplasmic and acetylcholinesterase content. Factors stimulating the development of immature megakaryocytes were found in preparations of human embryonic kidney cell-conditioned media (a source of in vivo Thrombopoietic Stimulatory Factor), peritoneal exudate cell-conditioned medium, lung-conditioned medium, or bone marrow cellular sources of activity (adherent cells or cells that sediment at 5-6 mm hr-1). Immature megakaryocytes cultured serum free responded to sources of an auxiliary megakaryocyte potentiating activity by developing into single, large megakaryocytes but did not respond to a megakaryocyte colony-stimulating factor devoid of detectable potentiator activity present in WEHl-3-conditioned medium. In contrast, serum-free proliferation of the megakaryocyte progenitor cell required both megakaryocyte colony-stimulating factor and the auxiliary potentiator activity. In the presence of megakaryocyte colony-stimulating factor alone, progenitor cells did not form colonies of easily detectable megakaryocytes. However, groups of cells comprised entirely of small acetylcholinesterase containing immature megakaryocytes were observed, thus establishing that megakaryocyte colony development passes through a stage of immature cells prior to detectable megakaryocyte development and that some acetylcholinesterase-containing cells can undergo cellular division.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041120305
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  • 10
    Electronic Resource
    Electronic Resource
    Membrane potentials of BHK (baby hamster kidney) cell line: Ionic and metabolic determinants (1972)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 80 (1972), S. 347-358 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The transmembrane potential of cells from a continuous cell line (BHK-21) has been investigated by a combination of electrophysiological and flame photometric techniques. The ratio of sodium permeability to potassium permeability (PNa/PK) determined from membrane potentials recorded at varying external potassium concentrations was 0.082; from membrane potential measurements and the intracellular sodium and potassium concentrations of cells in 6.8 mM K+ media the value was 0.075. The PNa/PK ratio was not temperature dependent. Dinitrophenol (1 mM) did not significantly alter the membrane potential of cells incubated for one hour with the inhibitor. However, iodoacetate (1 mM) and sodium fluoride (30 mM) caused a significant depolarization during a one-hour incubation. Measurements of sodium and potassium concentrations during incubation at 4°C showed a decrease in internal potassium and an increase in internal sodium accompanied by a decreased membrane potential. Ion concentrations and membrane potentials were measured in cells recovering at 37°C following 24 hours at 4°C. Membrane potentials in excess of EK during the first ten minutes of recovery may indicate electrogenic pumping.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1040800305
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