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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Journal of industrial microbiology and biotechnology 1 (1986), S. 1-8 
    ISSN: 1476-5535
    Keywords: Mycotoxins ; Aflatoxins ; Aflatoxicosis ; Syrian hamster ; Reversal ; Zinc
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Chronic exposure to aflatoxins (AFTs) below the LD50 can result in reduced weight gain, hepatocellular necrosis and bile duct cell proliferation. Here, we report whether dietary zinc (Zn2+) protects against both aflatoxicosis and precancer in male weanling hamsters fed either 14.6 mg/kg AFTs, 3000 mg/kg zinc carbonate, or both for 17 weeks. The AFTs (either alone or with Zn2+) reduced weight gains but not feed consumption. Whereas controls possessed 172.7±21.7 mg/100 ml plasma glucose, the AFTs and Zn2+ groups had 132.1±19.5 and 122.7 mg/100 ml, respectively. For plasma cholesterol, the AFTs plus Zn2+ group's was 26.5±4.3 compared to 32.3±3.0, 31.5±4.8 and 36.0±2.1 mg/100 ml for control, Zn2+ and AFTs groups, respectively. The latter exhibited bile duct cell hyperplasia, focal liver necrosis and hemorrhage but the AFTs plus Zn2+ group's livers had less damage. Meglahepatocytes indicated precancerous changes. These data suggest a trend toward Zn2+-induced reduction for AFTs-promoted liver damage.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1476-5535
    Keywords: Coriolus versicolor ; Wood-decay fungus ; Polyphenol oxidase ; Substrate specificity ; de novo Synthesis ; Partial purification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Coriolus versicolor, a white-rot Basidiomycete, secretes cellulolytic and ligninolytic enzymes as well as polyphenol oxidase (PPO). Whereas the former degrade wood polymers, the latter can convert diphenols to diquinones and oligomerize syringic acid, a lignin derivative. Certain phenolic compounds can serve as disease-resistance factors controlling the proliferation of wood-decay fungi within host tissues. BecauseC. vesicolor can be ‘batch-cultured’, overproduction and enhanced secretion of enzymes of biological and commercial interests are feasible. Reported here are the results of attempts to define the timed appearances of intracellular and extracellular PPO, to assess substrate specificity as well as distinguish synthesis versus activation of intracellular PPO and to partially purify extracellular PPO. These efforts were to provide data enabling cell-free synthesis of PPO, cloning of the gene(s) for the oxidase and the establishment of its subcellular route of secretion. Whereas two protein peaks (6 and 12 days in a 16 day time-course) were observed for dialyzed mycelial homogenates, the homogenates' PPO specific activity rose between 4 and 12 days and then declined. Total extracellular protein content climbed from 6 to 15 days for dialyzed growth medium and the medium's PPO specific activity rose at 4 days post-inoculation and except at 9 days increased linearly to 15 days. When aliquots of dialyzed 12 and 15 day media were added to PPO assay mixtures containing catechol and either syringic or gallic acids, statistically significant differences in PPO specific activity between phenolic substrates were noted. Supplementation of cultures with 1.91 μg cycloheximide ml growth medium−1 (control, growth medium only) together with 0.5 μCi [14C]-leucine revealed that cycloheximide inhibited PPO activity and suppressed [14C]-leucine incorporation into TCA-insoluble cytoplasmic protein. As for PPO partial purification, growth medium dialysis followed by 0–30% (NH4)2SO4 fractionation and subsequent 12 000×g dialyzate centrifugation yielded a 3.27-fold enhancement in PPO specific activity within the 12 000×g supernatant. Chromatography of the latter upon DEAE-Sephadex indicated that PPO exchanged with the DEAE counterion as it could be eluted with high ionic strength salt. These results suggest that: the occurrences of intracellular and extracellular PPO are time-dependent, intracellular PPO is de novo synthesized, the preferred substrate for extracellular PPO appears to be catechol and extracellular PPO can be partially purified by a combination of dialysis and ammonium sulfate fractionation as well as possibly DEAE chromatography and/or Sephadex G-150 gel filtration.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-0832
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Aflatoxins (AFTs) are hepatocarcinogens, mutagens, teratogens and toxins. Isolates of AFTs-producing strains of Aspergillus flavus can grow upon both autoclaved soybeans and to a lesser extent field-grown beans. Besides inhibiting the elongation of excised, in vitro cultured soybean roots, the AFTs can impair the roots' ability to remove [14C]-leucine from a culture medium and to incorporate the amino acid into acidinsoluble, cytoplasmic protein. In this connection, exogenous AFTs once taken-up and compartmentalized by the excised roots can reduce the acid-insoluble protein content of what appears to be a non-enriched plasmalemma fraction. Here, we report a combined biochemical and microscopical attempt to determine whether the protein reduction occurs throughout the excised, in vitro-cultured root and whether the plasmalemma proteins which are affected by AFTs can be both solubilized and characterized. Histochemistry of Carnoy-fixed roots revealed a reduction in Ninhydrin-Schiff-stainability throughout the AFTs-treated root. Transmission electron microscopy of 80 000 × g pellets derived from homogenates of both non-treated (NT) and treated (T) roots provided further evidence to our previously reported marker enzyme analyses for the occurrence of the plasmalemma in the 80 000 × g pellet. Treatment of the latter with the Laemmli SDS procedure released 〉85% of the protein associated with the pellets obtained from homogenates of either NT or T roots. Gel permeation chromatography on Biogel-100 of released proteins from 80 000 × g pellets of both NT and T roots yielded both void volume and retarded peaks. Both the amplitude and the quantities of protein recovered within the void volume peak were less within the void volume of the T than the NT root. Polyacrylamide tube gel electrophoresis of G-100 void volume peaks of chromatographed, SDS-released proteins from the pellets revealed quantitative but not qualitative differences in Coomassie Blue-gel staining patterns between T and NT roots. These results suggest that the SDS methods which we employed could solubilize the 80 000 × g pellet proteins but that combined gel permeation chromatography and tube gel electrophoresis were not sensitive enough to reveal whether AFTs could alter the types of proteins associated with the 80 000 × g pellet (purported plasmalemma).
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  • 4
    ISSN: 1573-0832
    Keywords: Aflatoxin ; soya-bean root ; leucine ; mycotoxin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The present work reports a portion of our continuing effort to determine the mechanism(s) whereby aflatoxins cause toxic responses in in vitro cultured plant tissues. Few investigations have dealt with the mode of action of aflatoxin B1 (AFB1) in excised plant tissues. Here is detailed AFB1 influence on growth, uptake and incorporation of 14C-leucine by excised, incubated soya-bean roots. Pure AFB1 was added to culture medium prior to autoclaving. One gram fresh weight portions of roots from three-day old soya-bean seedlings were excised and incubated for 4, 8, 12 and 24 hours. Growth was assayed by following changes in root dry weight. Aflatoxin B1 inhibited root dry weight at both 20 and 30 μg/ml. Uptake of 14C-leucine was checked by following its depletion from the medium. Reduced 14C-leucine uptake by roots exposed to 20 μg/ml AFB1 suggests that the toxin may alter the plasmalemma. A possible role for AFB1 in modification of membrane-associated amino acid transport mechanisms is discussed. Incorporation of 14C-leucine into trichloroacetic acid-precipitable cytoplasm was assayed. Inhibition of this incorporation at 20 μg/ml AFB1 was most apparent at 12 hours. Thus, AFB1 may also impair the ability of excised soya-bean roots to carry out protein synthesis.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1573-0832
    Keywords: Aflatoxin ; Germination ; Elongation ; Metabolism ; Pollen ; Soybeans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Aflatoxin producing strains of Aspergillus grow on soybeans thereby contaminating the latter through secretion of the toxin. Investigations dealing with either soybean seed germination or intact seedling growth responses to aflatoxin (B1) are lacking. Similarly, a possible interaction of aflatoxins with phosphate in the germination and elongation of both soybeans and pollen as well as roots of the former and tubes of the latter has not been examined. Imbibition of Glycine max, cv. ‘Essex’ seeds for 18 hours in solutions containing 0.38, 2.90, 5.80 or 11.60 μg/ml (AFB1) yielded% germination inhibitions of 5, 20, 40 and 80, respectively. By 36 hours these were 6, 4, 13 and 19 % for the same toxin concentration series. At 140 hours attached root elongation was inhibited 26, 35 and 50 % for 2.90, 5.80 and 11.60 μg/ml AFB1. No effect was noted at 0.38 μg/ml AFB1. Incubation of excised roots in medium containing 3.0 mM KH2PO4 stimulated their elongation 3.2 fold. Addition of 33.28 μg/ml mixed aflatoxins together with KH2PO4 resulted in only a 1.5 fold stimulation. When KH2PO4 was added to a culture medium lacking AFB1, Lilium longiflorum, cv. ‘Ace’ pollen germination was enhanced 50%. Withholding KH2PO2 but supplying AFB1 did not markedly affect germination. However, supplementing the medium with KH2PO4 while simultaneously adding AFB1 did not inhibit germination at 5 and 10 μg/ml but caused 27.3 and 45.1 % declines at 25 and 30 μg/ml. In the absence of KH2PO4 AFB1 stimulated pollen tube elongation 7.5, 14.3, 16.5 and 13.2 % at 5, 10, 15 and 20 μg/ml but 30 μg/ml inhibited it 11.1%. In contrast, tube elongation was suppressed at all AFB1 concentrations (maximum 36.1% at 30 μg/ml) tested upon KH2PO4 addition. Results derived from germinating pollen in medium supplemented with KH2PO4 or NaH2PO4 indicate that the phosphate anion does not preferentially promote aflatoxin-induced inhibition of tube elongation.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1573-0832
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Isolates of aflatoxin-producing strains of Aspergillus grow on autoclaved and field-grown (lesser extent) Glycine max beans. Both mixed and aflatoxin B1 inhibit G. max, cv. ‘Essex’ bean germination and elongation of either attached or excised cultured roots. Because B1 impairs the latter roots' ability to intracellularize [14C]-leucine, it may alter plasmalemma structure and/or function. To determine whether incubation of excised roots for 18 hours in toxin-containing medium could affect cellular membrane chemical content, organelles were isolated by differential centrifugation (1 000, 40 000, and 80 000 xg) of homogenates and characterized chemically. Statistically significant differences between treated and untreated roots in acid insoluble protein but not either sterol or lipid phosphorus levels were observed for both 40,000 and 80,000 xg pellets. Protein and sterol recoveries were 81 (treated) and 84 (untreated) % for the former and 77 (treated) and 79 (untreated) % for the latter. Lipid phosphorus recoveries were 87.3 (treated) and 136 (untreated) % with and 96 (treated) and 83 (untreated) without membrane stabilization. Protein:sterol:lipid phosphorus were 35.7∶4.5∶1 (1 000 xg), 18.9∶3.6∶1 (40000 xg), 26.3∶4.6∶1 (80 000 xg) and 1,010∶29∶1 (80 000 xg supernatant) for untreated and 36.9∶3.3∶1 (1,000 xg), 23.1∶3.8∶1 (40 000 xg), 36.2∶4.8∶1 (80 000 xg) and 1,053∶21.7∶1 (80 000 xg supernatant) for treated roots. Significant differences in RNA content between treated and untreated roots were found for both 1 000 and 40 000 xg pellets but not for the 80 000 xg pellet and its supernatant. Whereas a significant increase in the 1 000 xg pellet occurred upon treatment, a decrease was noted for the 40 000 xg pellet but not for the 80 000 xg pellet and its supernatant. Similar pH 6 (plasmalemma marker enzyme) and 9 (mitochondrial marker enzyme) K+-stimulated ATPase activities were demonstrated for 40 000 and 80 000 xg pellets. The 1 000 xg pellet contained greater than 50% of the NADH-cytochrome c-reductase activity (endoplasmic reticulum marker enzyme) recovered from fractions examined for this activity which was absent from the 40 000 xg pellet. Both the 80 000 xg pellet and its supernatant possessed equivalent reductase activities. Inosine diphosphatase activity (dictyosome marker enzyme) was not present in 1 000 xg pellets obtained from either treated or untreated roots but was in both 40 000 and 80 000 xg pellets. Based on these results, a tentative assignment of organelles to each fraction (xg force) is reported.
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  • 7
    ISSN: 1573-0832
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The present paper reports our attempts to determine whether the inclusion of 0.0014 mM Zn++ within a hydroponic culture medium affects the ability of 12-day-old Zea mays, cv. ‘SS-522’ to take-up [3H]-aflatoxin B1. Data from the corollary experiment, i.e., whether inclusion of aflatoxin affects the ability of Zea mays, cvs. ‘Truckers White’, ‘X-Sweet’ and ‘Merit’ to take-up 65ZnCl2 are presented also. This report is a preliminary to one regarding an in-progress analysis of whether pollutant levels of Zn++ affect aflatoxin uptake and distribution. In the absence of irrigating seedlings, which were grown in Perlite containing 65ZnCl2, with a solution containing mixed aflatoxins, the stem contained the greatest amount of label with root plus seed the next highest and the leaf the least for each of the cvs. In contrast, when the seedlings were irrigated with a solution containing mixed aflatoxins, the root plus seed contained either an amount nearly identical to (cv. ‘Truckers White’) or in excess of that within the stem (cvs. ‘X-Sweet’ and ‘Merit’). Calculation of the percentages of aflatoxin-induced diminutions in leaf, stem and root label suggested that the aflatoxins interfered with the translocation of 65ZnCl2 from the root to the stem and leaf, at least for cvs ‘X-Sweet’ and ‘Merit’. When 0.0014mM Zn++ as ZnSO4 was added to an incubation medium in which 12-day-old seedlings were suspended and plant growth assessed over 72 hours, a 15% increase (significant at 0.05 level) in seedling height over that of Zn++-deficient plants was observed. No differences in [3H]-aflatoxin B1 uptake were noted between those seedlings which were grown in either Zn++-containing or lacking media. Less than one % of the[3H]-aflatoxin B1 which was taken-up was recovered within chloroform extracts of the seedlings. The distributions of radioactivity from [3H]-aflatoxin B1 for leaf, stem, seed and root were 0, 57, 26 and 19% and 0, 26, 58 and 18% for Zn++-containing and -lacking media, ‘respectively’.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Mycopathologia 81 (1983), S. 83-94 
    ISSN: 1573-0832
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1573-0832
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Maximum inhibition of Glycine max, cv. ‘Essex’ seed germination occurred at 10 μg/ml following 72 hr imbibition in constant light. Seeds imbided 108 hr in constant darkness at this concentration showed a 20% rise in germination over that of the control. Imbibition of G. max, cv. ‘Williams’ seeds in either light or dark for 96 hr did not suppress germination. Imbibition of ‘Essex’ seeds in either light or dark at 2.5 through 10 μg/ml stimulated root elongation except for 10 μg/ml at 96 hr (light). Maximum inhibition of ‘Williams’ root elongation under constant light was at 48 and 72 hr with 10 μg/ml. Statistically significant differences in cotyledon, leaf and stem lengths between non-treated (NT) and treated (T) seedlings were not found except for ‘Williams’ stem length at 2.5 μ/ ml. Root elongation was stimulated 1.2- and 1.1-folds, respectively, at 5.0 (‘Essex’) and 2.5 (‘Williams’) μg/ml. Toxin at 2.5 through 10.0 μg/ml did not markedly alter either cotyledon or leaf widths with the exception of ‘Williams” leaf width at 2.5 μg/ml. Medium supplementation with 2.5 through 10.0 μg/ml resulted in cotyledon, leaf and root weight enhancements for ‘Essex’ seedlings. Stem weight was not markedly affected. An 18% rise in ‘Williams’ cotyledon weight above that of the control was seen at 2.5 μg/ml. ‘Williams’ leaf weights were increased 1.75- and 1.25-folds, respectively, at 2.5 and 10.0 μg/ml. Aflatoxin B1, at 2.5 μg/ml promoted ‘Williams’ stem and root elongation 1.20- and 1.09-folds, respectively. Most of the radioactivity from 65Zn-ZnCl2 recovered within organs was found within ‘Essex’ roots for both T and NT seedlings. A higher amount of radioactivity was recovered within roots at each toxin concentration than was without toxin. However, this was not statistically significant. Significant differences in the distribution of radioactivity within roots between NT and T ‘Williams’ seedlings were not observed. Generally, AFB1 failed to affect significantly these two varieties of soybeans based on the tests relating to germination, growth and radiolabel uptake.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    BioEssays 4 (1986), S. 27-30 
    ISSN: 0265-9247
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: ‘Yellow Rain’, an alleged biological warfare agent thought to be utilized in parts of both South East Asia and Afghanistan, may be composed in part of the mycotoxins, trichothecenes. However, more recent analyses suggest that the ‘Rain’ was mainly honey bee excreta. The history of the controversy together with the biological effects, chemistry as well as the fungi producing these mycotoxins and agricultural commodities affected by trichothecenes are reviewed.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
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