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[
Biol Chem,
2003]
S-Adenosylmethionine decarboxylase (AdoMetDC) is a key enzyme in polyamine biosynthesis. In many eukaryotes its activity is stimulated specifically by putrescine. The AdoMetDC of the filarial parasite Onchocerca volvulus, however, is not only stimulated by putrescine but also by the naturally occuring polyamines spermidine and spermine. Several diamines, acetylated polyamines and polyamine analogues were used to analyse what molecular prerequisites are needed to stimulate nematode AdoMetDC activity. In the absence of an activator, the O. volvulus enzyme exhibits an extremely low specific activity. This fact, together with the unspecificity of activator binding, was thought to be useful for a new strategy to inhibit nematode AdoMetDC activity. Therefore, different polyamine analogues were tested as competitive inhibitors towards the stimulatory effect putrescine has on the O. volvulus and, in comparison, on the Caenorhabditis elegans and human AdoMetDC. Bis(aralkyl)- and bis(alkyl)-substituted polyamine analogues with a 3-7-3 backbone were found to inhibit AdoMetDC activities, however, probably without interfering with the putrescine stimulation. The best inhibitor, BW-1, was about 10-fold more effective against O. volvulus AdoMetDC than against the human enzyme. Unexpectedly, BW-1 was determined to be a competitive inhibitor with respect to AdoMet, having a Ki value of 310 microM for the putrescine-stimulated human AdoMetDC. Furthermore, we show for the O. volvulus and the human enzyme that the degree of inhibition by BW-1 depends on the actual putrescine concentration.
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[
Biol Chem,
2003]
S-Adenosylmethionine decarboxylase (AdoMetDC) is a key enzyme of the polyamine synthetic pathway providing decarboxylated S-adenosylmethionine for the formation of spermidine and spermine, respectively. The catalytic activity of the AdoMetDC from the free living nematode Caenorhabditis elegans highly depends on the presence of an activator molecule. Putrescine, a well-known stimulator of mammalian AdoMetDC activity, enhances the catalytic activity of the nematode enzyme 350-fold. Putrescine stimulation is discussed as a regulatory mechanism to relate putrescine abundance with the synthesis of spermidine and spermine. In contrast to any other known AdoMetDC, spermidine and spermine also represent significant activators of the nematode enzyme. However, the biological significance of the observed stimulation by these higher polyamines is unclear. Although C. elegans AdoMetDC exhibits a low specificity toward activator molecules, the amino acid residues that were shown to be involved in putrescine binding of the human enzyme are conserved in the nematode enzyme. Exchanging these residues by sitedirected mutagenesis indicates that at least three residues, Thr(192), Glu(194) and Glu(274), most likely contribute to activator binding in the C. elegans AdoMetDC. Interestingly, the mutant Glu(194)Gln exhibits a 100-fold enhanced basal activity in the absence of any stimulator, suggesting that this mutant protein mimics the conformational change usually induced by activator molecules. Furthermore, site directed mutagenesis revealed that at least Glu(33), Ser(83), Arg(91) and Lys(95) are involved in posttranslational processing of C. elegans
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[
J Helminthol,
2011]
Ethanolic and aqueous extracts of selected medicinal plants from Cameroon and Ghana were assessed for their in vitro anthelmintic activity by using the bovine filarial parasite Onchocerca ochengi and the free living nematode Caenorhabditis elegans, a model organism for research on nematode parasites. Worms were incubated in the presence of different concentrations of extracts and inhibitory effects were monitored at different time points. Among the extracts used in this study, ethanolic extracts of Anogeissus leiocarpus, Khaya senegalensis, Euphorbia hirta and aqueous extracts from Annona senegalensis and Parquetina nigrescens affected the growth and survival of C. elegans and O. ochengi significantly. The mortality was concentration dependent with an LC50 ranging between 0.38 and 4.00mg/ml for C. elegans (after 72h) and between 0.08 and 0.55mg/ml for O. ochengi after a 24h incubation time. Preliminary phytochemical screenings on these extracts revealed the presence of flavonoids, alkaloids, saponins, carbohydrates and tannins in the extracts. Accordingly, application of A. leiocarpus, K. senegalensis, E. hirta and A. senegalensis extracts could provide alternatives in the control of helminthic infections.
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[
Vet Parasitol,
2012]
The ethanolic extract of Anogeissus leiocarpus was assessed for the in vitro anthelmintic activity by using the cattle parasite nematode Onchocerca ochengi as well as levamisole-, ivermectin- and albendazole-resistant mutant strains of the free-living nematode Caenorhabditis elegans, a model organism for research on nematode parasites. Worms were incubated in the presence of different concentrations of the plant extract and effects on survival were monitored after each 12 h to 96 h. The A. leiocarpus extract affected O. ochengi microfilaria, adults, and C. elegans wild-type worms with LC(50) values of 0.06 mg/ml, 0.09 mg/ml after 24h and 0.44 mg/ml after 48 h, respectively. Remarkably, the efficacy of the plant extract was not significantly altered in the ivermectin- and levamisole-resistant C. elegans mutant strains
lev-1(
e211),
glc-2(
ok1047),
lev-9(
x16) and
avr-14(
ad1302),
avr-15(
ad1051),
glc-1(
pk54). The albendazole resistant strain
ben-1(
e1880) exhibited a moderate increase of the LC(50) value to 1.5mg/ml after 48 h. These results are in good accordance with the use of A. leiocarpus extract against nematode infections by traditional healers, herdsmen and pastoralists. Moreover, the data indicate that the plant extract could be used to treat nematode infections even in cases of drug resistance towards established anthelmintic drugs.
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[
J Helminthol,
2014]
The effect of three phenols (ellagic, gentisic and gallic acids) from the axlewood tree Anogeissus leiocarpus on Onchocerca ochengi and drug-resistant strains of Caenorhabditis elegans, a model organism for research on nematode parasites, is investigated. Worms were incubated in different concentrations of phenols and their survival was monitored after 48h. Among the three acids, ellagic acid strongly affected the survival of O. ochengi microfilariae, O. ochengi adults, a wild-type C. elegans and anthelmintic-resistant strains of C. elegans, namely albendazole (CB3474), levamisole (CB211, ZZ16) and ivermectin (VC722, DA1316), with LC50 values ranging from 0.03mm to 0.96mm. These results indicate that the binding of ellagic acid in the worm differs from that of resistant strains of C. elegans. The efficacy of both gallic and gentisic acids was not significantly changed in resistant strains of C. elegans treated with levamisole (ZZ16, LC50=9.98mm, with gallic acid), albendazole (CB3474, LC50=7.81mm, with gentisic acid) and ivermectin (DA1316, LC50=10.62mm, with gentisic acid). The efficacy of these three pure compounds is in accordance with the use of A. leiocarpus from its locality of origin. The in vivo toxicity data reveal that the thresholds are up to 200 times higher than the determined LC50 values. Thus, ellagic acid could be a potential option for the treatment of nematode infections, even in cases of drug resistance towards established anthelmintic drugs.
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[
Biosci Biotechnol Biochem,
2016]
We compared the growth inhibitory effects of all aldohexose stereoisomers against the model animal Caenorhabditis elegans. Among the tested compounds, the rare sugars d-allose (d-All), d-talose (d-Tal), and l-idose (l-Ido) showed considerable growth inhibition under both monoxenic and axenic culture conditions. 6-Deoxy-d-All had no effect on growth, which suggests that C6-phosphorylation by hexokinase is essential for inhibition by d-All.
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[
Bioorg Med Chem Lett,
2016]
Biological activities of unusual monosaccharides (rare sugars) have largely remained unstudied until recently. We compared the growth inhibitory effects of aldohexose stereoisomers against the animal model Caenorhabditis elegans cultured in monoxenic conditions with Escherichia coli as food. Among these stereoisomers, the rare sugar d-arabinose (d-Ara) showed particularly strong growth inhibition. The IC50 value for d-Ara was estimated to be 7.5mM, which surpassed that of the potent glycolytic inhibitor 2-deoxy-d-glucose (19.5mM) used as a positive control. The inhibitory effect of d-Ara was also observed in animals cultured in axenic conditions using a chemically defined medium; this excluded the possible influence of E. coli. To our knowledge, this is the first report of biological activity of d-Ara. The d-Ara-induced inhibition was recovered by adding either d-ribose or d-fructose, but not d-glucose. These findings suggest that the inhibition could be induced by multiple mechanisms, for example, disturbance of d-ribose and d-fructose metabolism.
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[
Bioorg Med Chem Lett,
2019]
The biological activities of deoxy sugars (deoxy monosaccharides) have remained largely unstudied until recently. We compared the growth inhibition by all 1-deoxyketohexoses using the animal model Caenorhabditis elegans. Among the eight stereoisomers, 1-deoxy-d-allulose (1d-d-Alu) showed particularly strong growth inhibition. The 50% inhibition of growth (GI<sub>50</sub>) concentration by 1d-d-Alu was estimated to be 5.4mM, which is approximately 10 times lower than that of d-allulose (52.7mM), and even lower than that of the potent glycolytic inhibitor, 2-deoxy-d-glucose (19.5mM), implying that 1d-d-Alu has a strong growth inhibition. In contrast, 5-deoxy- and 6-deoxy-d-allulose showed no growth inhibition of C. elegans. The inhibition by 1d-d-Alu was alleviated by the addition of d-ribose or d-fructose. Our findings suggest that 1d-d-Alu-mediated growth inhibition could be induced by the imbalance in d-ribose metabolism. To our knowledge, this is the first report of biological activity of 1d-d-Alu which may be considered as an antimetabolite drug candidate.
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[
Biochim Biophys Acta Proteins Proteom,
2020]
d-Aspartate oxidase (DDO) is a flavin adenine dinucleotide (FAD)-containing flavoprotein that stereospecifically acts on acidic D-amino acids (i.e., free d-aspartate and D-glutamate). Mammalian DDO, which exhibits higher activity toward d-aspartate than D-glutamate, is presumed to regulate levels of d-aspartate in the body and is not thought to degrade D-glutamate in vivo. By contrast, three DDO isoforms are present in the nematode Caenorhabditis elegans, DDO-1, DDO-2, and DDO-3, all of which exhibit substantial activity toward D-glutamate as well as d-aspartate. In this study, we optimized the Escherichia coli culture conditions for production of recombinant C. elegans DDO-1, purified the protein, and showed that it is a flavoprotein with a noncovalently but tightly attached FAD. Furthermore, C. elegans DDO-1, but not mammalian (rat) DDO, efficiently and selectively degraded D-glutamate in addition to d-aspartate, even in the presence of various other amino acids. Thus, C. elegans DDO-1 might be a useful tool for determining these acidic D-amino acids in biological samples.
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[
J Appl Glycosci (1999),
2019]
D-Allose (D-All), C-3 epimer of D-glucose, is a rare sugar known to suppress reactive oxygen species generation and prevent hypertension. We previously reported that D-allulose, a structural isomer of D-All, prolongs the lifespan of the nematode Caenorhabditis elegans. Thus, D-All was predicted to affect longevity. In this study, we provide the first empirical evidence that D-All extends the lifespan of C. elegans. Lifespan assays revealed that a lifespan extension was induced by 28 mM D-All. In particular, a lifespan extension of 23.8 % was achieved (p< 0.0001). We further revealed that the effects of D-All on lifespan were dependent on the insulin gene
daf-16 and the longevity gene
sir-2.1, indicating a distinct mechanism from those of other hexoses, such as D-allulose, with previously reported antiaging effects.