- Volume 112, Issue 2, 1979
Volume 112, Issue 2, 1979
- Biochemistry
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Characterization of Peptidoglycan from the Cyanelles of Cyanophora paradoxa
More LessSUMMARY: Peptidoglycan has been purified from the photosynthetic organelles (‘cyanelles’) of Cyanophora paradoxa. The results indicate the presence of N-acetylmuramic acid, N-acetylglucosamine, alanine, glutamic acid and diaminopimelic acid in the molar ratio of 1:1:1·6:1:1. The peptidoglycan may be present in a lipoprotein-peptidoglycan complex. The structure of the peptidoglycan appears to be identical to that of cyanobacteria and the other Gram-negative bacteria so far studied. Peptidoglycan was also partly purified from two species of cyanobacteria, Gloeothece 7109 and Gloeobacter violaceus. Since the genome of the cyanelle closely resembles that of chloroplasts, the results suggest that the cyanelle of Cyanophora paradoxa is an intermediate form between an endosymbiotic cyanobacterium and a chloroplast.
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The Control of the Synthesis of Isocitrate Lyase in a Thermophilic Bacillus
More LessSUMMARY: From a strain of Bacillus stear othermophilus, devoid of active pyruvate carboxylase, a mutant (NG-15) was selected that grew on acetate in the presence of glucose. This mutant differed from its parent organism in possessing high activities of isocitrate lyase when grown on all carbon sources tested except nutrient broth, in possessing unusually low activities of NADP+-dependent isocitrate dehydrogenase and in containing increased amounts of isocitrate. Revertants of mutant NG-15 which regained the ability to synthesize active pyruvate carboxylase also synthesized isocitrate lyase and isocitrate dehydrogenase to the same extent as the wild-type strain. These results suggest that the regulatory mechanism for the synthesis of isocitrate lyase in the thermophile may be different from that in mesophilic bacilli.
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- Development And Structure
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The Study of Homogeneous Populations of Caulobacter Stalked (Mother) Cells
U. Swoboda and C. S. DowSUMMARY: An experimental procedure is described which facilitates the study of homogeneous populations of Caulobacter stalked (mother) cells. These populations are derived from synchronized swarm cell cultures, the preparation of which is described. From this study it appears that cross-band formation in Caulobacter NCIB 9083 is not linked to the reproductive cell cycle.
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Ultrastructure of Cytophaga johnsonae and C. aquatilis by Freeze-etching
More LessSUMMARY: Cytophaga johnsonae and C. aquatilis (ATCC 29551), examined by frozen-surface replica and freeze-etch techniques, displayed undulated and particulated surfaces. They produced copious amounts of extracellular slime when cultured. Cytophaga aquatilis contained large, complex mesosomes in freeze-etched cells whereas C. johnsonae contained none. In non-cryoprotected freeze-etch preparations, C. aquatilis contained two major fracture planes whereas the C. johnsonae cell envelope fractured in three distinct planes. Linear fibres, such as the peripheral fibrils previously described in Flexibacter columnaris, were not observed in the outer membrane of freeze-etched C. aquatilis. Cytophaga johnsonae contained periplasmic linear fibrils; however, these appeared to be associated with the outer surface of the cytoplasmic membrane or an intermediate layer and not with peripheral fibrils.
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- Ecology
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Evidence of an Indigenous Microbiota (Yeast) in the Dry Valleys of Antarctica
More LessSUMMARY: Soil samples from sites in the Dry Valleys of South Victoria Land, Antarctica, similar to some sites previously considered abiotic, contained a variety of microbes. The fungi recovered included Tilletiopsis washingtonensis, Sporobolomyces holsaticus, Sp. roseus (all previously unreported from Antarctica), Cryptococcus laurentii var. laurentii and Cr. vishniacii, as well as some unremarkable filamentous fungi imperfecti. Cryptococcus vishniacii occurs only in these soils, has a distribution suggesting indigenous origin and shows appropriate psychrophilic character and energy requirements for life in this highly stressed environment. Populations of Cr. vishniacii cloned and grown at 4 °C are heterogeneous in temperature response, some cells being capable of growth at 20 °C or 23 °C with loss of the ability to assimilate succinate or citrate. Cryptococcus vishniacii and endolithic cyano-bacteria could constitute a minimal community in Dry Valley soils.
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Selective Advantage of a Spirillum sp. in a Carbon-limited Environment. Accumulation of Poly-β-hydroxybutyric Acid and Its Role in Starvation
More LessSUMMARY: A freshwater Spirillum sp., which apparently belongs to a niche of low nutritional status (Matin & Veldkamp, 1978 ), accumulated poly-β-hydroxybutyric acid (PHB) during lactate-limited growth in continuous culture. The PHB content varied in a complex manner with the dilution rate (D), but was greatest at the lowest D value examined: about 18% (w/w) at D = 0.025 h−1. It is not known what mechanism accounted for PHB accumulation during carbon-limited growth. The resistance of cultures of Spirillum sp. to starvation after growth at various D values was compared with that of a Pseudomonas sp. which appears to belong to relatively richer environments (Matin & Veldkamp, 1978 ) and does not accumulate PHB. In Spirillum sp., resistance correlated directly with the PHB content of the culture subjected to starvation, whereas in Pseudomonas sp. it increased with RNA content. Further, after growth at D = 0·03 to 0·05 h−1, the Spirillum sp. was much more resistant to starvation than was the Pseudomonas sp. Since the microflora of oligotrophic environments are probably often subjected to starvation conditions, PHB accumulation by Spirillum sp. during growth in such environments may assist survival. PHB in Spirillum sp. was rapidly degraded during starvation but it had no sparing effect on RNA degradation. It is not known how PHB enhanced resistance to starvation.
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- Genetics And Molecular Biology
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Fusion of the lac Genes to the Proximal Promoters of the deo Operon of Escherichia coli K12
More LessSUMMARY: Fusions of the lac structural genes to the proximal promoters deoP and cytP of the deo operon have been isolated using a λ (lac, Mu') phage. This phage was inserted into a heat-inducible Mu prophage which had itself been inserted into the deoA gene. Selection was made for heat-resistant derivatives of this lysogen which maximally expressed the lac genes when an inducer of the deo operon was present; these were shown to have the lac genes fused to the deo operon - in some cases with a concomitant deletion of the deoC gene, whilst in others deoC was retained. The level of β-galactosidase activity in these strains was induced by growth in the presence of deoxyadenosine or cytidine, both of which lead to induction of the deo enzymes - deoxyadenosine through the deoOP system and cytidine through the cytOP system. The level of induction of β-galactosidase was of the same order as the level of induction of deoxyriboaldolase, the deoC gene product, in those strains retaining an intact deoC + gene. Plaque-forming λ phages carrying the deo-lac fusions were isolated by inducing the lysogens with mitomycin C.
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Transduction of Fimbriation Demonstrating Common Ancestry in FIRN Strains of Salmonella typhimurium
More LessSUMMARY: The production of fimbriate (Fim+) recombinants was observed in transductional crosses between different pairs of wild-type strains of different biotypes of Salmonella typhimurium. Fim+ recombinants were readily produced in transductions from Fim+ donor strains to Fim− recipient strains and, less frequently, between some pairs of Fim− strains, for example, between almost any strain of the FIRN biogroup (Fim− Inl− Rha− Bxyl−) and many strains of the non-FIRN Fim− biogroup. None of numerous crosses between different pairs of FIRN strains gave Fim+ recombinants, suggesting that the fim mutation was present at the same intragenic site in all FIRN strains. FIRN strains are thought to have descended from a single ancestral FIRN bacterium which originated by a series of mutations from a strain of the common biotype la (Fim+ Inl+ Rha+ Bxyl+). Two FIRN-like (Fim− Inl+ Rha− Bxyl−) strains that did not yield Fim+ recombinants in crosses with FIRN strains were probably wild-type Inl+ mutants from FIRN strains.
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Genetic Instability of Sporulation-associated Characters in a Bacillus subtilis Mutant: Analysis of the Segregation Pattern and Genetic Studies
More LessSUMMARY: A Bacillus subtilis mutant which formed dark-brown ‘medusa’ (M) colonies was obtained. It sporulated at a high frequency, overproduced extracellular protease during sporulation and possessed a high genetic instability with a complex segregation pattern. Segregation was maintained after repeated re-isolation of single M colonies. The major wild-type-like class of segregants (B) was stable, sporulated normally and produced normal amounts of protease. Occasionally segregants were obtained which produced extremely high amounts of protease, sporulated poorly, formed transparent colonies and were either highly unstable (TD) or stable (TDst). Rarely B(D) (stable, normal sporulation and protease overproduction) and W and T (both stable and asporogenous) segregants were produced. The M phenotype was transmitted as a single factor by transformation but not by transduction. The results of transduction experiments suggest the presence of two mutations, ScoC and ScoD. It is proposed that this new segregating system in B. subtilis may result from tandem duplication of part of the bacterial chromosome.
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Genetic Instability of Sporulation-associated Characters in a Bacillus subtilis Mutant: Relationship between Sporulation, Segregation and the Synthesis of Extracellular Enzymes (Kinetic Studies)
More LessSUMMARY: In the genetically unstable, protease-overproducing ‘medusa’ (M) strains of Bacillus subtilis, segregation of stable, wild-type-like B cells occurred mainly during sporulation. After the end of the exponential growth phase, a small fraction of M cells sporulated quickly and formed M spores, while the majority of the cells, after a ‘critical period’, gave rise to B segregants which sporulated after a delay. Segregation occurred without cell division. Delayed sporulation, segregation and protease overproduction are related. Similar but more complex results were obtained with the highly unstable TD strains. Sporulation and the kinetics of protease overproduction were also followed in several stable segregants. Depending on the strain, either the rate of protease production or both the rate and time course were affected. The results are interpreted in terms of sequential activation and de-activation of sporulation genes. The production of the alkaline and the neutral proteases was, in general, under common genetic control. In some strains α-amylase was also overproduced.
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Effect of Bacteriophage C5 on Ultraviolet Light Survival in Pseudomonas aeruginosa
More LessSUMMARY: Bacteriophage C5 of Pseudomonas aeruginosa is able to reactivate ultraviolet (u.v.)-irradiated phage E79 in coinfection experiments and decrease the u.v.-sensitivity of a host-cell reactivation deficient mutant. These properties suggest that phage C5 has a gene(s) which is involved in the repair of u.v.-damaged DNA. The isolation of two u.v.-sensitive mutants of C5 supports this hypothesis.
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- Physiology And Growth
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The Effect of Transition Metal Ions on the Resistance of Bacterial Spores to Hydrogen Peroxide and to Heat
More LessSUMMARY: The presence of 10 μm-Cu2+ increased the lethal effect of hydrogen peroxide on spores of Clostridium bifermentans but not on those of Clostridium sporogenes PA 3679, Clostridium perfringens, Bacillus cereus or Bacillus subtilis var. niger. Cu2+ at 100 μm. also increased the lethal effect of heat on spores of C. bifermentans but not on those of B. subtilis var. niger. The rate and extent of Cu2+ uptake by spores of C. bifermentans and B. subtilis var. niger were similar, but examination of unstained sections of spores by electron microscopy suggested that Cu2+ is bound by the protoplasts of spores of C. bifermentans but not of B. subtilis var. niger.
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Trimethylamine Oxide: A Terminal Electron Acceptor in Anaerobic Respiration of Bacteria
More LessSUMMARY: Trimethylamine oxide (TMAO) stimulated both the anaerobic growth rate and the growth yield of Proteus NTHC 153. The molar growth yield from glucose and pyruvate in tryptone/ yeast extract medium doubled in the presence of TMAO, and the organism grew anaerobically on the non-fermentable substrates l-lactate and formate when TMAO was added to the medium. We conclude that TMAO stimulated growth by serving as a terminal electron acceptor in an oxidative phosphorylation process.
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Location of Binding Sites on Common Type 1 Fimbriae from Escherichia coli
More LessSUMMARY: Common type 1 fimbriae were isolated from Escherichia coli and their length distribution profile was determined before and after treatment with ultrasound. As fimbriae were shortened, so their haemagglutinating capacity decreased, but their ability to bind to erythrocytes did not decrease to the same extent. Isolated fimbriae did not agglutinate inside-out vesicles prepared from horse erythrocytes or liposomes, suggesting that the binding mechanism was not based on non-specific hydrophobic interactions. The results support a lateral rather than a terminal location for the fimbrial binding site responsible for haemagglutination.
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The Role of Manganese in Growth and Sporulation of Bacillus subtilis
More LessSUMMARY: Phosphoglycerate phosphomutase of Bacillus subtilis, Bacillus cereus and Bacillus megaterium required Mn2+ as cofactor, whereas the wheat germ and rabbit liver enzymes did not. In the absence of Mn2+, B. subtilis did not sporulate in normal sporulation media but it did sporulate if the proper ratio of glucose or glycerol and malate was used. Decoyinine, an inhibitor of guanosine monophosphate synthesis, induced sporulation in the presence of excess glucose and malate to the same extent with and without Mn2+. Apparently, phosphoglycerate phosphomutase is the only strictly Mn2+-requiring enzyme needed for optimal sporulation in normal sporulation media.
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Thermosensory Behaviour in Paramecium tetraurelia: a Quantitative Assay and Some Factors that Influence Thermal Avoidance
More LessSUMMARY: The free-swimming ciliate Paramecium avoids warm regions. We have developed a quantitative assay of thermal avoidance, based on the interference between thermal avoidance and the normal tendency to swim upward (negative geotaxis). Paramecium tetraurelia swimming in a Tris/Ca2+ solution avoided a region warmer than about 40°C. Several factors influenced the strength of the response, including the concentrations of monovalent cations and Ca2+, the temperature of the test region, and the temperature at which the animals had been cultured. At 1 mm-Ca2+, increasing Na+ concentration enhanced avoidance of a 40°C region, and at a constant Na+ concentration (0·5 mm), avoidance improved with decreased Ca2+. When the ratio [Na+]/[Ca2+]1/2 was held constant, varying Na+ and Ca2+ concentrations did not affect thermal avoidance. Other monovalent cations (Cs+, Li+, Rb+) and hydroxylamine also enhanced thermal avoidance, and K+ was somewhat less effective than these. The strength of the avoidance response increased with increasing test temperature in the range of 37 to 42°C for cells grown at 28°C. Cells grown at 15°C had a lower threshold for thermal avoidance, and those grown at 35°C showed no avoidance at 40°C and only poor avoidance at higher temperatures. Cells cultured at 15 or 35°C and then shifted to 28°C acquired the thermal behaviour typical of cells cultured at 28°C. Behavioural mutants with defective Ca2+ channels (Pawns) are incapable of reversing their swimming direction and showed little or no thermal avoidance. We suggest that thermal avoidance is triggered by thermotropic phase transitions in the lipids of the excitable membrane of Paramecium tetraurelia.
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Nutrient Requirement for the Degradation of Cellulose to Methane by a Mixed Population of Anaerobes
More LessSUMMARY: A synthetic medium containing inorganic sources of N, P, Fe, S, Ca2+ and Mg2+, trace minerals and essential vitamins was developed for anaerobic degradation of cellulose to CH4. A mixed culture obtained from sewage sludge and grown in batch culture in this medium degraded about 4 g cellulose l−1 week−1 and produced about 710 ml total gas per g cellulose degraded. The gas produced contained between 51 and 56% CH4, the remainder being CO2. For maximum degradation of cellulose to CH4 and CO2, the requirement for essential salts other than HCO3 −, Fe and S2- was comparable to that of other anaerobes. NH4 + or Na2CO3 at 20 to 24 mm and total Fe at 0·4 to 0·6 mm gave optimum conversion of cellulose to CH4, while the inclusion of additional S2- in the presence of 1·75 mm-SO4 2- was not required.
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- Short Communication
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Glutamine Synthetase and Glutamate Synthase Activities During Growth and Sporulation in Bacillus subtilis
More LessSUMMARY: Glutamine synthetase in Bacillus subtilis 168 was repressed to a greater extent by l-glutamine or l-arginine than by ammonia when each was used as sole nitrogen source. It was derepressed when either l-glutamate or nitrate was used as nitrogen source. Glutamate synthase was repressed by l-glutamate or l-arginine and, to a lesser extent, by l-glutamine but was derepressed during growth with ammonia or nitrate. Glutamine synthetase activity was unaltered during the onset of sporulation. Glutamate synthase activity, however, underwent a small and apparently transient increase in bacteria induced to sporulate by nitrogen limitation.
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Influence of Oxygen Tension on Nitrate Reduction by a Klebsiella sp. Growing in Chemostat Culture
More LessSUMMARY: At dissolved oxygen tensions of 15 mmHg (2 kPa) and below, nitrate-limited continuous cultures of Klebsiella K312 synthesized nitrate reductase (NR) and nitrite reductase (NiR) and excreted ammonia. Under anaerobic conditions over 60% of the nitrate-nitrogen utilized was excreted as ammonia. In contrast, carbon-limited cultures excreted nitrite at dissolved oxygen tensions of 15 mmHg or below and synthesized NR but not NiR. Ammonia repressed neither NR nor NiR synthesis. These observations indicate that below a critical oxygen tension of 15 mmHg Klebsiella K312 utilizes oxygen and nitrate as electron acceptors. This oxygen tension correlates well with the critical oxygen tension observed for a change from oxidative to fermentative metabolism in cultures of Klebsiella aerogenes. The product of dissimilatory nitrate reduction is ammonia in nitrate-limited cultures but principally nitrite in carbon-limited (nitrate excess) cultures.
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Reduction of Perturbations in Leucine Incorporation in Synchronous Cultures of Schizosaccharomyces pombe Made by Elutriation
More LessSUMMARY: Good synchronous cultures of the fission yeast Schizosaccharomyces pombe can be made by selecting small cells from the Beckman JE-6 elutriator rotor. More important, the rate of leucine incorporation in controls shows far less perturbation than after selection from tube gradients. Elutriation may therefore be the best available method for reducing perturbations in selection synchronized cultures.
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The Components of Mycoplasma salivarium and its Growth Medium that are Responsible for Film Formation
More LessSUMMARY: Studies on film production by mycoplasmas revealed that film was produced by completely disintegrated mycoplasma cells on Noble agar in the presence of horse serum. Film production was due to an enzymic reaction between mycoplasma lipase, possibly phospho-lipase A, and phospholipid in serum.
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Characterization of Ca2+-ATPase Activity in Streptomyces griseus
More LessSUMMARY: Ca2+-ATPase activity has been characterized in Streptomyces griseus. The enzyme has a pH optimum of 8·5 at 37°C. Its Ca2+ requirement can be substituted by Cd2+, Zn2+ and Mn2+. Mg2+ inhibits the enzyme non-competitively.
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Isolation of Atypical Lipopolysaccharides from Purified Cell Walls of Pseudomonas cepacia
More LessSUMMARY: Wall fragments were prepared from two strains of Pseudomonas cepacia and from P. aeruginosa, and their contents of readily extractable lipid, pronase-digestible protein and lipo-polysaccharide were measured. Lipopolysaccharide extracted from P. cepacia, although biologically active, contained no detectable 2-keto-3-deoxyoctonic acid, but contained phosphate, rhamnose, glucose, heptose and hexosamine in concentrations comparable to those found in P. aeruginosa.
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Description of Strains of Peptostreptococcus anaerobius Isolated from Subcutaneous Abscesses in Cats
More LessSUMMARY: Strains of Peptostreptococcus, Streptococcus and of a Gram-positive coccus, which was initially isolated as an anaerobe but grew subsequently as a facultative organism, were isolated from subcutaneous abscesses in cats. The cat strains of Peptostreptococcus gave metabolic fermentation products in combinations described for P. anaerobius. The Streptococcus strains conformed to the group S. intermedius. The facultative organism described had the metabolic products of P. anaerobius but the distinctly different biochemical characteristics of S. intermedius and fits neither of the genera strictly.
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An Examination of Rhizobium leguminosarum for the Production of Extracellular and Periplasmic Proteins
More LessSUMMARY: Rhizobium leguminosarum WU163 and WU235 did not release extracellular proteins into the environment but did synthesize periplasmic proteins, including alkaline phosphatase (EC 3.1.3.1), cyclic phosphodiesterase (EC 3.1.4.d) and inorganic pyrophosphatase (EC 3.6.1.1). Fourteen periplasmic proteins, recognized by polyacrylamide gel electrophoresis, were released by lysozyme/EDTA treatment. Four of these proteins, including the alkaline phosphatase, were repressed by phosphate.
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Factors Influencing Filament Length of Methanospirillum hungatii
More LessSUMMARY: Cultivation of Methanospirillum hungatii GP1 at 35°C in low PO4 3- (3·0 mm) medium resulted in growth as motile curved rods (6 to 8 × 0·5 μm). However, predominantly long filaments averaging 70 to 300 μm were observed in the presence of high concentrations of PO4 3- (43·7 mm) or Na+ (225 mm) or after incubation at 25°C in low PO4 3- medium. These conditions also resulted in decreased growth rates. Tests with cell-free extracts of long and short form filaments indicated that long filament formation is probably related to a deficiency in heat-labile factor(s) required for cell separation.
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- Taxonomy
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Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis of Cell-surface Proteins as an Aid to the Identification of the Bacteroides fragilis Group
More LessSUMMARY: The outer-membrane complexes from 40 strains of Bacteroides, representing eight of the species included in the Bacteroides fragilis group, were released by EDTA treatment. The component polypeptides were examined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis on slab gels. Within a species (biotype) the patterns produced indicated marked similarities in the structures of the surface proteins among the strains examined. The patterns produced by strains belonging to different species, however, showed fewer similarities. An unknown organism could therefore be identified to species level using this technique and a few selected biochemical tests.
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