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  • 1
    Electronic Resource
    Electronic Resource
    Photosynthesis of isolated chloroplasts and protoplasts under osmotic stress (1981)
    Springer
    Planta 151 (1981), S. 375-380 
    Details
    ISSN: 1432-2048
    Keywords: Chloroplast volume ; Photosynthesis and chloroplast volume ; Spinacia ; Water stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract 1. Isolated intact spinach chloroplasts respond to changes of the sorbitol concentration of the suspending medium as near-perfect osmometers within a large range of osmotic potentials. Under isotonic conditions (π=9–10 bar), their average osmotic volume is 24 μm3 and the total volume 36 μm3. The osmotic volume can be increased to 63 μm3 by lowering the sorbitol concentration until a critical osmotic potential of π=4 bar is reached. Below that value chloroplasts rupture. Between 10 bar and 4 bar, volume changes are reversible. 2. Increasing the chloroplast volume above 24 μm3 causes inhibition of photosynthesis, with 50% inhibition occurring at an osmotic potential of π=5–6 bar. This corresponds to an osmotic volume of 45–55 μm3. Depending on the duration of hypotonic treatment, inhibition of photosynthesis is more or less reversible. 3. Between 4 and 10 bar, the chloroplast envelope exhibits a very low permeability for ferricyanide, many metabolites, and soluble stroma proteins. 4. Electron transport is not inhibited by swelling of chloroplasts. Also, the ATP/ADP-ratio remains unchanged. 5. The solute concentration in the chloroplasts appears to be optimal for photosynthesis at 10 bar. Increasing the chloroplast volume causes inhibition of photosynthesis by dilution effects.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00393294
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  • 2
    Electronic Resource
    Electronic Resource
    The role of monovalent cations for photosynthesis of isolated intact chloroplasts (1980)
    Springer
    Planta 149 (1980), S. 170-175 
    Details
    ISSN: 1432-2048
    Keywords: Cations and photosynthesis ; Chloroplast (low-salt effects) ; Light activation (photosynthesis enzymes) ; Photosynthesis ; Spinacia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The role of monovalent cations in the photosynthesis of isolated intact spinach chloroplasts was investigated. When intact chloroplasts were assayed in a medium containing only low concentrations of mono- and divalent cations (about 3 mval l-1), CO2-fixation was strongly inhibited although the intactness of chloroplasts remained unchanged. Addition of K+, Rb+, or Na+ (50–100 mM) fully restored photosynthesis. Both the degree of inhibition and restoration varied with the plant material and the storage time of the chloroplasts in “low-salt” medium. In most experiments the various monovalent cations showed a different effectiveness in restoring photosynthesis of low-salt chloroplasts (K+〉Rb+〉Na+). Of the divalent cations tested, Mg2+ also restored photosynthesis, but to a lesser extent than the monovalent cations. In contrast to CO2-fixation, reduction of 3-phosphoglycerate was not ihibited under low-salt conditions. In the dark, CO2-fixation of lysed chloroplasts supplied with ATP, NADPH, and 3-phosphoglycerate strictly required the presence of Mg2+ but was independent of monovalent cations. This finding excludes a direct inactivation of Calvin cycle enzymes as a possible basis for the inhibition of photosynthesis under low-salt conditions. Light-induced alkalization of the stroma and an increase in the concentration of freely exchangeable Mg2+ in the stroma, which can be observed in normal chloroplasts, did not occur under low-salt conditions but were strongly enhanced after addition of monovalent cations (50–100 mM) or Mg2+ (20–50 mM). The relevance of a light-triggered K+/H+ exchange at the chloroplast envelope is discussed with regard to the light-induced increase in the pH and the Mg2+ concentration in the stroma, which are thought to be obligatory for light activation of Calvincycle enzymes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00380879
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  • 3
    Electronic Resource
    Electronic Resource
    Photosynthesis under osmotic stress (1981)
    Springer
    Planta 153 (1981), S. 430-435 
    Details
    ISSN: 1432-2048
    Keywords: Chloroplast ; Photosynthesis (stress recovery) ; Protoplast ; Spinacia ; Water stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The reversibility of the inhibition of photosynthetic reactions by water stress was examined with four systems of increasing complexity—stromal enzymes, intact chloroplasts, mesophyll protoplasts, and leaf slices. The inhibition of soluble chloroplast enzymes by high solute concentrations was instantly relieved when solutes were properly diluted. In contrast, photosynthesis was not restored but actually more inhibited when isolated chloroplasts exposed to hypertonic stress were transferred to conditions optimal for photosynthesis of unstressed chloroplasts. Upon transfer, chloroplast volumes increased beyond the volumes of unstressed chloroplasts, and partial envelope rupture occurred. In protoplasts and leaf slices, considerable and rapid, but incomplete restoration of photosynthesis was observed during transfer from hypertonic to isotonic conditions. Chloroplast envelopes did not rupture in situ during water uptake. It is concluded that inhibition of photosynthesis by severe water stress is at the biochemical level brought about in part by reversible inhibition of chloroplast enzymes and in part by membrane damage which requires repair mechanisms for reversibility. Both soluble enzymes and membranes appear to be affected by the increased concentration of internal solutes, which is caused by dehydration.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00394981
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  • 4
    Electronic Resource
    Electronic Resource
    Inactivation of nitrate reductase involves NR-protein phosphorylation and subsequent ‘binding’ of an inhibitor protein (1995)
    Springer
    Planta 195 (1995), S. 514-518 
    Details
    ISSN: 1432-2048
    Keywords: Inhibitor protein ; Nitrate reductase ; Protein phosphorylation ; Protein kinase ; Spinacia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The function of two proteins (P67 and P100) required for the MgATP-dependent inactivation of nitrate reductase (NR) from spinach leaves (Spinacia oleracea L.) was studied. When NR was incubated with γ-[32P]ATP and P67, NR-protein was phosphorylated, but without a change in NR activity. Protein P100 by itself was neither able to phosphorylate nor to inactivate NR, and when added together with P67 it did not change the extent of NR phosphorylation. However, when NR was first phosphorylated with MgATP and P67, subsequent addition of P100 after removal of unreacted ATP caused an immediate NR inactivation. In presence of both P67 and P100 the time-course of ATP-dependent NR phosphorylation paralleled the time course of inactivation. The extent of NR phosphorylation and of NR inactivation (in the presence of P67 plus P100) was similarly affected by metabolites or high salt concentrations. Magnesium (Mg2+) played a dual role in the inactivation process: the phosphorylation of NR by P67 was strictly Mg2+-dependent. Further, phospho-NR (+P100) was inactive only in the presence of Mg2+, but active in the presence of excess EDTA. Dephospho-NR appeared to be Mg2+-insensitive. The observations suggest that phosphorylation of NR by P67 is obligatory, but not sufficient for inactivation. In addition to protein phosphorylation, inactivation requires “binding” of an inhibitor protein (P100) to phospho-NR.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00195708
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  • 5
    Electronic Resource
    Electronic Resource
    Acid-base-modulation of nitrate reductase in leaf tissues (1995)
    Springer
    Planta 196 (1995), S. 1-6 
    Details
    ISSN: 1432-2048
    Keywords: Acid-base loading ; Nitrate reductase ; pH regulation (intracellular) ; Protein phosphorylation ; Spinacia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of acid or base-loading of spinach (Spinacia oleracea L.) leaf discs on the activation status of nitrate reductase (NR) in the dark and in the light was investigated. Activity of NR (NRA), measured in crude extracts of leaf discs with removed lower epidermis, which had been floating on Mes-buffer [2-(N-morpholino)ethane sulfonic acid] pH 5.2 in the dark, was at a similar low level as in whole, darkened leaves. By addition of acetate or propionic acid, butyric acid or benzoic acid, NR was activated to or beyond the light level. The pH of crude tissue extracts was decreased by 0.5–1 pH units. Tissue acidification caused an inhibition of photosynthesis and of dark CO2 fixation. The acid-induced activation of NR in vivo was largely prevented by okadaic acid, an inhibitor of Type 1 and Type 2A protein phosphatases. This indicates that acid-induced activation was mediated by protein dephosphorylation. When, on the other hand, leaf discs were illuminated on Ches-buffer (2-[ N-cyclohexylamino]ethane sulfonic acid) pH 9 in the presence of bicarbonate (80 mM), their NR was as active as in intact leaves. Addition of ammonium chloride (up to 6 mM) caused a pH increase of the tissue extract up to 0.9 pH units. At the same time NR was inactivated to the dark level. Methionine sulfoximine did not prevent the ammonium effect. Photosynthesis and dark CO2 fixation were stimulated at pH 9 by ammonium chloride (1–2· mol· m −3) and were only slightly inhibited by up to 6 mol· m−3. The modulation of NR by acid-base treatment in vivo was fully reversible. The response of the NR system to acid or base treatment is consistent with a proposed role of nitrate reduction in the cellular pH-stat. The observation also indicates that cytosolic pH changes may be involved the signal chain triggering the modulation of NR.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00193210
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  • 6
    Electronic Resource
    Electronic Resource
    Photosynthesis under osmotic stress (1981)
    Springer
    Planta 153 (1981), S. 423-429 
    Details
    ISSN: 1432-2048
    Keywords: Chloroplast envelope (permeability) ; Chloroplast stroma enzymes ; Spinacia ; Water stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract 1. Increasing the sorbitol concentration in a suspension of intact chloroplasts induced a fast, transient and not very specific efflux of metabolites from chloroplasts to the medium. Stroma proteins were retained by the chloroplasts. 2. Within the first 30 s following hypertonic stress, the chloroplast volume decreased according to the Boyle-Mariotte relation. A subsequent and transient increase suggested some influx of external solute. 3. Dark reactions of intact chloroplasts such as starch degradation and formation of labelled 3-phosphoglycerate from dihydroxyacetone phosphate or ribose-5-phosphate and 14CO2 were inhibited at low water potentials. After chloroplast rupture, the activity of stromal enzymes was decreased by high solute concentrations. Ribulose bisphosphate carboxylase exhibited a decrease of Vmax, while KmCO 2 remained unaltered. With sorbitol, sucrose, glycerol or glycinebetaine, 50% inhibition of enzymes was observed at osmotic potentials between 40 and 50 bar, with ethyleneglycol at about 70 bar. With salts such as KCl, 50% inhibition was found at 15 to 20 bar. 4. A comparison between inhibition of photosynthesis in intact chloroplasts and inhibition of enzymes in stroma extracts by solutes supports the notion that inhibition of photosynthesis at high osmotic potentials is mainly a solute effect. Another factor contributing to inhibition of photosynthesis in isolated chloroplasts is the loss of intermediates and cofactors which occurs during rapid osmotic dehydration.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00394980
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  • 7
    Electronic Resource
    Electronic Resource
    Modulation of nitrate reductase in vivo and in vitro: Effects of phosphoprotein phosphatase inhibitors, free Mg2+ and 5′-AMP (1994)
    Springer
    Planta 193 (1994), S. 358-364 
    Details
    ISSN: 1432-2048
    Keywords: AMP ; Cation (divalent) ; Nitrate reductase ; Pisum ; Protein phosphatase ; Spinacia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrate reductase in spinach (Spinacia oleracea L.) leaves was rapidly inactivated in the dark and reactivated by light, whereas in pea (Pisum sativum L.), roots, hyperoxic conditions caused inactivation, and anoxia caused reactivation. Reactivation in vivo, both in leaves and roots, was prohibited by high concentrations (10–30 μM) of the serine/threonine-protein phosphatase inhibitors okadaic acid or calyculin, consistent with the notion that protein dephosphorylation catalyzed by type-1 or type-2A phosphatases was the mechanism for the reactivation of NADH-nitrate reductase (NR). Following inactivation of leaf NR in vivo, spontaneous reactivation in vitro (in desalted extracts) was slow, but was drastically accelerated by removal of Mg2+ with excess ethylenediaminetetraacetic acid (EDTA), or by desalting in a buffer devoid of Mg2+. Subsequent addition of either Mg2+, Mn2+ or Ca2+ inhibited the activation of NR in vitro. Reactivation of NR (at pH 7.5) in vitro in the presence of Mg2+ was also accelerated by millimolar concentrations of AMP or other nucleoside monophosphates. The EDTA-mediated reactivation in desalted crude extracts was completely prevented by protein-phosphatase inhibitors whereas the AMP-mediated reaction was largely unaffected by these toxins. The Mg2+-response profile of the AMP-accelerated reactivation suggested that okadaic acid, calyculin and microcystin-LR were rather ineffective inhibitors in the presence of divalent cations. However, with partially purified enzyme preparations (5–15% polyethyleneglycol fraction) the AMPmediated reactivation was also inhibited (65–80%) by microcystin-LR. Thus, the dephosphorylation (activation) of NR in vitro is inhibited by divalent cations, and protein phosphatases of the PP1 or PP2A type are involved in both the EDTA and AMP-stimulated reactions. Evidence was also obtained that divalent cations may regulate NR-protein phosphatase activity in vivo. When spinach leaf slices were incubated in Mg2+ -and Ca2+-free buffer solutions in the dark, extracted NR was inactive. After addition of the Ca2+ /Mg2+-ionophore A 23187 plus EDTA to the leaf slices, NR was activated in the dark. It was again inactivated upon addition of divalent cations (Mg2+ or Ca2+). It is tentatively suggested that Mg2+ fulfills several roles in the regulatory system of NR: it is required for active NR-protein kinase, it inactivates the protein phosphatase and is, at the same time, necessary to keep phospho-NR in the inactive state. The EDTA- and AMP-mediated reactivation of NR in vitro had different pH optima, suggesting that two different protein phosphatases may be involved. At pH 6.5, the activation of NR was relatively slow and the addition or removal of Mg2+ had no effect. However, 5′-AMP was a potent activator of the reaction with an apparent K m of 0.5 mM. There was also considerable specificity for 5′AMP relative to 3′- or 2′-AMP or other nucleoside monophoposphates. We conclude that, depending upon conditions, the signals triggering NR modulation in vivo could be either metabolic (e.g. 5′-AMP) or physical (e.g. cytosolic [Mg2+]) in nature.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00201813
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  • 8
    Electronic Resource
    Electronic Resource
    Increased nitrate reductase activity in leaf tissue after application of the fungicide Kresoxim-methyl (1999)
    Springer
    Planta 207 (1999), S. 442-448 
    Details
    ISSN: 1432-2048
    Keywords: Key words: Nitrate reductase ; Respiration inhibitors ; Spinacia ; Strobilurin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Spinach (Spinacia oleracea L.) leaf discs floating on buffer solution were treated with Kresoxim-methyl (KROM), an inhibitor of respiratory electron transport. In the leaf tissue, actual and maximal nitrate reductase (NR) activities, nitrite content and ATP levels were determined. In darkened leaf discs incubated without KROM (control) actual NR activity decreased to 20% after 6 h in the dark. Treatment with 10 μg ml−1 (corresponding to 32 μM) KROM totally prevented inactivation of NR in the dark and also diminished NR-protein degradation during prolonged darkness. Due to restricted nitrite reduction in darkened leaf tissues, nitrite accumulated in KROM-treated discs. Inhibition of respiration decreased ATP and increased AMP levels in KROM-treated discs. In illuminated leaf discs, NR was highly activated to 65%. Nevertheless, KROM-treatment caused an additional activation of NR (activation state 76%) in the light. Possible side-effects of KROM on nitrite reduction and photosynthesis were also checked in the leaf-disc system. Neither nitrite reduction nor photosynthesis were altered in KROM-treated discs. The extent of KROM-induced activation of NR was dependent on the applied concentration and on the pH of the external medium. The highest activation of NR was achieved at an external pH of 4.8, confirming previous results (Kaiser and Brendle-Behnisch, 1995, Planta 196: 1–6) that cytosolic acidification might play an important role in the modulation of NR activity.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004250050503
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  • 9
    Electronic Resource
    Electronic Resource
    Das Klarzellchondrosarkom (2000)
    Springer
    Der Pathologe 21 (2000), S. 449-455 
    Details
    ISSN: 1432-1963
    Keywords: Schlüsselwörter Knochentumoren ; Klarzellchondrosarkom ; Histologie ; Differenzialdiagnose ; Keywords Bone tumors ; Clear-cell chondrosarcoma ; Histology ; Differential diagnosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Abstract Clear-cell chondrosarcoma is a rare, low-malignant bone tumor. The lesion most commonly occurs in adults, generally in the 3rd or 4th decade. Clear-cell chondrosarcoma has a predilection for the epiphyses of long tubular bones, particularly the femoral head. Radiologically, it is a sharply defined radiolucent lesion. Histologically, it is characterized by large tumor cells with distinct boundaries and a clear cytoplasm. In addition to areas of conventional chondrosarcoma, there are partially mineralized trabecular osteoid formations. On the basis of 16 cases of clear-cell chondrosarcoma, we present the observations of the Hamburg Bone Tumor Register. The cases were examined according to age distribution, location, and radiological and pathomorphological criteria. The proximal femur was the most frequent location; rare manifestations in the rib, os ilium, and distal phalanx of the hand were also present. In 50% of the cases high-malignant osteosarcoma or conventional chondrosarcoma was considered. In addition to the epiphysis of the proximal femur, clear-cell chondrosarcoma can also involve other, rather unusual sites in the skeleton. Knowledge of the histomorphology of this tumor is therefore of crucial importance for the diagnosis.
    Notes: Zusammenfassung Das Klarzellchondrosarkom ist ein seltener niedrigmaligner Knochentumor. Betroffen sind v. a. Patienten in der 3. und 4. Lebensdekade. Bevorzugte Lokalisation sind die Epiphysen langer Röhrenknochen, insbesondere der Femurkopf. Radiologisch imponiert es meist als eine epiphysär gelegene, scharf begrenzte lytische Läsion. Histologisch charakteristisch sind die großen Tumorzellen mit deutlichen Zellgrenzen und einem hellen Zytoplasma. Daneben finden sich auch Areale mit dem histologischen Bild eines konventionellen Chondrosarkoms sowie unterschiedlich breite, z. T. mineralisierte trabekuläre Osteoidformationen. Anhand von 16 Fällen mit Klarzellchondrosarkom werden die Erfahrungen und Beobachtungen des Hamburger Knochentumor-Registers dargestellt. Die Fälle wurden nach Altersverteilung, Lokalisation sowie radiologischen und pathomorphologischen Kriterien untersucht. Das Hauptmanifestationsalter lag innerhalb der 5.–7. Lebensdekade. Das proximale Femur stellte den Hauptlokalisationsort dar. Daneben fanden sich auch eher seltene Manifestationen im Bereich der Rippe, Os ilium und Grundphalanx der Hand. In 50% der Fälle wurde auswärtig zunächst differenzialdiagnostisch ein hochmalignes Osteosarkom oder ein Chondrosarkom diskutiert. Da das Klarzellchondrosarkom außer in der Epiphyse des proximalen Femurs auch an anderen, eher ungewöhnlichen Stellen des Skeletts auftreten kann, ist die Kenntnis der Histomorphologie dieses Tumors von entscheidender Bedeutung für die Diagnosefindung.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002920000417
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  • 10
    Electronic Resource
    Electronic Resource
    Das periostale Osteosarkom Histologische Charakteristika, Präparationstechnik, Wachstumsmuster und Differentialdiagnose* (1996)
    Springer
    Der Pathologe 17 (1996), S. 86-91 
    Details
    ISSN: 1432-1963
    Keywords: Schlüsselwörter Periostales Osteosarkom ; Histologie ; Präparation ; Grading ; Key words Periosteal osteosarcoma ; Histology ; Preparation ; Grading
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Periosteal osteosarcoma is a distinct bone tumor entity with characteristic morphological features within the group of juxtacortical osteosarcoma. Periosteal osteosarcoma is predominantly located in the long tubular bones, especially in the tibia and femur and is situated on the outer circumference of the tumor-bearing bone (saucerization phenomenon). In contrast to parosteal osteosarcoma, periosteal osteosarcoma is less differentiated and is believed to have a worse prognosis. In this work the histological features are described with predominantly chondroblastic differentiation of 14 cases with periosteal osteosarcoma. A horizontal preparation technique of periosteal osteosarcoma specimens allows comparison with computed tomography and is the optimal method to detect an invasion of the medullary cavity. Further studies are necessary to clarify if neoadjuvant chemotherapy could improve the prognosis of certain patients.
    Notes: Zusammenfassung Das periostale Osteosarkom stellt eine eigene Entität dar, so daß der Tumor von der Gruppe der juxtakortikalen Osteosarkome eindeutig abgegrenzt werden sollte. Der Tumor tritt überwiegend an den langen Röhrenknochen, speziell Tibia und Femur auf. Er ist durch ein breitbasiges Aufsitzen an der äußeren Circumferenz des tumortragenden Skelettabschnittes charakterisiert (Saucieren-Muster). Im Gegensatz zum parossalen Osteosarkom ist das periostale Osteosarkom niedriger differenziert und hat offenbar eine schlechtere Prognose. Die histologischen Kriterien mit einer überwiegend chondroiden Differenzierung werden anhand von 14 Fällen eigener Beobachtung beschrieben. Der horizontalen Präparation von Tumorresektaten kommt eine besondere Bedeutung im Vergleich zu den bildgebenden Verfahren und zur Erfassung von Markraumeinbrüchen zu. Es gilt in zukünftigen Studien zu klären, bei welchen Patienten mit einer neoadjuvanten Chemotherapie die Prognose verbessert werden kann.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002920050140
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