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  • 1
    Electronic Resource
    Electronic Resource
    Woodbury, NY : American Institute of Physics (AIP)
    Applied Physics Letters 73 (1998), S. 954-956 
    ISSN: 1077-3118
    Source: AIP Digital Archive
    Topics: Physics
    Notes: We have investigated the dependence of leakage current and capacitance of Pt/Ba0.5Sr0.5TiO3/Pt capacitors on annealing temperature under high vacuum conditions. It is observed that leakage currents increase asymmetrically for negative and positive bias voltage with increasing annealing temperature. A model of leakage current and capacitance characteristics has been proposed on the assumption of generation of oxygen vacancies by annealing at the interfaces of the dielectric film adjacent to the Pt electrodes. The model predicts the oxygen vacancies of about 1020 cm−3. © 1998 American Institute of Physics.
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  • 2
    ISSN: 1432-203X
    Keywords: Key words ADP-glucose starch glycosyl transferase ; Amyloplast ; BY-2 ; Nicotiana tabacum ; Transcription/translation inhibitors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract When BY-2 cultured tobacco (Nicotiana tabacum L.) cells were transferred to auxin-depleted culture medium containing cytokinin (benzyladenine, 1 mg/l), the starch content per cell started increasing from 18 h of culture and amyloplasts had formed by 48 h. Pulse-treatment of the cells with actinomycin D and cycloheximide for the first 12 h (or longer) of culture significantly decreased the cellular starch content after 48 h, whereas the starch content did not decrease significantly when the cells were released from the inhibition within 6 h. This suggests that nuclear gene expression necessary for amyloplast formation begins 6–12 h after the transfer. Immunoblotting analysis of the accumulation of ADP-glucose starch glycosyl transferase (starch synthase) supported this inference.
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  • 3
    ISSN: 1432-2145
    Keywords: Key words Plastid ; Mitochondrion ; Biparental cytoplasmic inheritance ; Jasminum officinale ; Jasminum nudiflorum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Mature Jasminum officinale and J. nudiflorum pollen grains were stained with 4′,6-diamidino-2-phenylindole (DAPI) and examined by epifluorescence microscopy. The pollen grains were found to be trinucleate, and the sperm cells in both species contained a large number of epifluorescent spots that corresponded to cytoplasmic DNA aggregates (nucleoids). The nucleoids of J. nudiflorum were observed to be dimorphic under the epifluorescence microscope, indicating that the sperm cells might contain both plastid and mitochondrial DNA. The nucleoids of J. officinale presented a similar appearance when stained with DAPI, but electron microscopic examination of the sperm cells revealed that they contained both plastids and mitochondria. When analyzed by DNA immunogold electron microscopy, gold particles were detected on both plastids and mitochondria. These findings demonstrated the preservation of plastid and mitochondrial DNA in mature sperm cells and thus the potential for biparental cytoplasmic inheritance in J. officinale and J. nudiflorum.
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  • 4
    ISSN: 1615-6102
    Keywords: Golgi apparatus ; Cultured tobacco cell BY-2 ; DiOC6 ; Distribution ; Cell plate ; Caffeine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We developed a new method for distinguishing the Golgi apparatus from the other membranous organelles which contain DNA, such as mitochondria and chloroplasts, under a fluorescence microscope. Thin sections of cells embedded in Technovit 8100 resin were stained with both 3,3′-dihexyloxacarbocyanine iodide (DiOC6) and 4′,6-diamidino-3-phenylindole (DAPI), and those three membranous organelles were observed under an epifluorescence microscope. The Golgi apparatus, which do not contain DNA, were easily recognized when the two images stained with DiOC6 and DAPI were superimposed using an image processor. Using this method, we investigated the dynamics of cellular membranes and organelles during the mitotic cycle of synchronized cultured tobacco cells BY-2 (Nicotiana tabacum L. cv. Bright Yellow 2). The Golgi apparatus did not accumulate in the rim of the formating early cell plate at anaphase, while it accumulated near the maturing cell plate at telophase, and this accumulation seemed to be related to the maturation of cell plates. To confirm this hypothesis, synchronized BY-2 cells were treated with caffeine, which is known to inhibit the cell plate formation. Most of the cells treated with caffeine remained in a phase in which Golgi vesicles were accumulated at the equatorial plate, but the cell plate was only partially maturing. The Golgi apparatus accumulated only near the partially maturing cell plate, but not by the equatorial plate where the Golgi vesicles had accumulated.
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  • 5
    ISSN: 1615-6102
    Keywords: Giant mitochondria ; Mitochondrial nuclei ; Three-dimensional reconstruction ; Megasporogenesis ; Megagametogenesis ; Egg cell
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The preferential development of giant mitochondria and their nuclei (nucleoids) in the egg cells ofPelargonium zonale Ait. during megasporogenesis and megagametogenesis was examined by fluorescence microscopy, after Technovit embedding and 4′,6-diamidino-2-phenylindole (DAPI) staining, fluorimetry for DNA content, using a video-intensified microscope photon-counting system (VIMPICS), and by three-dimensional reconstruction of mitochondrial nuclei (mt-nuclei). Reproductive cells during the megaspore mother cell, meiosis, tetrad, and functioning megaspore stages contained many small mitochondria with characteristic, uniformly DAPI-stained mt-nuclei about 0.3 μm in diameter, containing a small amount of DNA (0.3 Mbp). During formation of the 2-, 4-, and 8-nucleate embryo sac, mt-nuclei did not markedly change in shape or DNA content. When the embryo sac formed and differentiation of each cell began, mitochondria and their nuclei in the egg cell took on a small ring or string-like shape. Accompanying the maturation of the embryo sac, they underwent progressive enlargement and gradually altered to long thick strings, or stacks of concentric or half concentric rings. By flower opening, they have developed to an extremely large size. One of these stacks of mt-nuclei was reconstructed in three dimensions; each ring in the stack was cup- or plate-shaped; 5 to 10 rings made up the stack, though each remained discontinuous from the others. From serial sections, we counted 44 mitochondria in one egg cell. Fluorometry using VIMPICS revealed that DNA amount within the stacked mitochondrion increased to 40 times that of the megaspore mother cell stage; a single stack of mitochondria contained 340–1700 Mbp DNA; which means that one egg cell contains at least 15000 Mbp mt-DNA, a value greater than the cell-nuclear DNA content.
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  • 6
    ISSN: 1615-6102
    Keywords: Brilliant sulfoflavin ; Cyanidioschyzon merolae ; Fenton reaction ; Fluorescence microscopy ; Hydrogen peroxide ; Microbody
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A novel procedure is described for fluorescence staining of microbodies, which can be applied quickly and easily. We developed this technique of microbody staining with the unicellular red algaCyanidioschyzon merolae. Cyanidioschyzon merolae only contains a single chloroplast, mitochondrion, and microbody per cell, and the mitotic cycle and the organelle division cycle are easily synchronized. Knowing that the concentration of H2O2 in the microbody is higher than it is in the cytosol and other cell components, we attempted to visualize the microbody by using fluorescence microscopy to detect H2O2. Brilliant sulfoflavin (BSF), used for detecting Fe2+ in analytical chemistry, fluoresces when it reacts with Fe2+ and H2C2. We were able to specifically stain microbodies with BSF, under acidic conditions (pH 3.0 or pH 2.5) with blue-light excitation. Using this procedure, we observed division of the microbody and the effect of aphidicolin on the microbody. We also discovered that microbody division is regulated by the cell nucleus and follows division of the cell nucleus.
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  • 7
    ISSN: 1615-6102
    Keywords: Mitochondrion-dividing ring ; Plastid-dividing ring ; Mitochondrial division ; Plastid division ; Microbody ; Cyanidioschyzon merolae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We present strong electron microscopic evidence that the mitochondrial-dividing ring (MD-ring) forms as a closed ring about 50 nm wide and 10 nm thick, at the contact point where the micro-body attaches to the mitochondrion. This ring forms in the cytoplasm around an equatorial plane perpendicular to the major axis of a mitochondrion. As the MD-ring increases in both width and thickness, the mitochondrion becomes dumbbell-shaped with a narrow interconnecting isthmus. Then, by successive contractions of the ring, the dumbbell-shaped mitochondrion separates to generate two daughter mitochondria. We also observed formation of an electron dense plastid-dividing ring (PD-ring) during plastid divisions. We noted too the behaviour of the MB in relation to the contraction of MD-rings and PD-rings.
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  • 8
    ISSN: 1615-6102
    Keywords: Cell cycle ; Chloroplast cycle ; Chloroplast fission ; Chloroplast nuclei ; Fluorodeoxyuridine ; Scenedesmus quadricauda
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary FdUrd (5-fluorodeoxyuridine), a specific inhibitor of thymidylate synthase, was used to study the relationship between reproductive processes in chloroplast and nucleocytoplasmic compartments of the chlorococcal algaScenedesmus quadricauda. The courses of DNA replication and nuclear division in both the compartments were followed in populations synchronised by the alternation of light and dark periods. DAPI-staining of DNA-containing structures was used for their visualisation and quantification. In contrast with cellular reproductive events, those in chloroplasts were not substantially affected by the presence of FdUrd (25 μg/ml). It was shown that FdUrd specifically blocked nucDNA replication but not ptDNA replication. Thus, cells which had attained commitment to ptDNA replication, fission of pt-nuclei and chloroplast kinesis triggered and terminated these processes while the corresponding cellular processes were blocked. The courses of reproductive processes in chloroplasts were also substantially unaffected in cells grown in the presence of FdUrd for the whole cell cycle. This provided evidence that attainment of commitment to and termination of the entire sequence of reproductive events, including chloroplast fission, were controlled by different mechanisms than the reproductive processes in the nucleocytoplasmic compartment.
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  • 9
    ISSN: 1615-6102
    Keywords: Mitochondria ; Physarum polycephalum ; Polymerase chain reaction ; Optical trap ; Microdissection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We attempted to amplify a specific region of mitochondrial DNA (mtDNA) using the polymerase chain reaction (PCR) from fewer than ten mitochondria isolated individually by microdissection or use of an optical tweezer. We selected preliminarily isolated mitochondria fromPhysarum polycephalum as the model materials and tried to amplify the mtDNA region corresponding to the specific mitochondrial plasmid of this true slime mould. For separation of a few mitochondria from the mitochondrial population, we initially used a destruction method in which excluded mitochondria were disrupted by a UV laser. However, mtDNA was still amplified, although weakly, from mitochondria that had been destroyed by the UV laser. Therefore, we used an optical tweezer to trap individual mitochondria and separate them from the others. The required number of mitochondria were separated from the mitochondrial suspension through a narrow canal of isolation buffer and used directly for PCR amplification. The results showed that the mtDNA could be amplified from at least 9 mitochondria trapped by the optical tweezer.
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  • 10
    ISSN: 1615-6102
    Keywords: Actin ; Contractile ring ; Cyanidium caldarium RK-1 ; Cytokinesis ; Immunoelectron microscopy ; Indirect immunofluorescence microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In the primitive red algaCyanidium caldarium RK-1, cytokinesis is controlled by a simple contractile ring, as in animal cells. To clarify the mechanism of formation of the contractile ring, we isolated actin genes and performed an immunocytological study.C. caldarium RK-1 has two actin genes encoding proteins with the same sequence of 377 amino acids. The primary structure indicated that the actin molecules ofC. caldarium RK-1 are typical, despite the fact that the organism is considered to be phylogenetically primitive. We prepared antiserum against aC. caldarium RK-1 actin fusion protein and indirect immunofluorescence staining was performed. In interphase cells, many actin dots were observed in the cytoplasm but none at the future cleavage plane. Prior to cytokinesis, some of these dots appeared and became aligned along the equatorial plane. At the same time, a thin “immature” contractile ring was observed to appear to be formed by connection of the aligned actin dots. This immature contractile ring thickened to nearly its maximum size by the time cytokinesis began. The formation of the contractile ring seemed to be a result of de novo assembly of actin monomers, rather than a result of the accumulation and bundling of pre-existing actin filaments. During the constriction of the contractile ring, no actin dots were observed in the cytoplasm. These observations suggest that actin dots are responsible for the formation of the contractile ring, but are not necessary for its disintegration. Furthermore, immunogold localization specific for actin revealed at electron microscopy level that fine filaments running just beneath the cleavage furrow are, in fact, actin filaments.
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