3-amino-1,2,4-triazole inhibits macrophage NO synthase.

Murine macrophages activated by interferon-gamma and lipopolysaccharide become leishmanicidal through a process involving L-arginine-derived nitrogen oxidation products. Both nitrite secretion and parasite killing by activated macrophages were inhibited by 3-amino-1,2,4-triazole as well as the related compound, 3-amino-1,2,4-triazine. Moreover, NO synthase activity in cytosolic extracts of activated cells was inhibited by both compounds. 4-amino-1,2,4-triazole, an isomer of 3-amino-1,2,4-triazole, was without effect. Our results suggest that besides its known inhibitory effect on catalases and peroxidases, 3-amino-1,2,4-triazole is an inhibitor of NO synthase. The resemblance between the tautomeric form of 3-amino-1,2,4-triazole and the guanidino group of L-arginine, the natural substrate for NO synthase, might be responsible for the observed inhibition.     

Histidine sensitive variant of the blue-green alga Nostoc muscorum: response to corepressors of histidine biosynthesis.

Summary A model has been proposed to account for growth inhibition by L-histidine in a variant strain of Nostoc muscorum. This strain has been characterized for its response to 3-amino-1,2,4-triazole and 1,2,4-triazole-3-alanine known to act as false corepressors of the histidine biosynthesis genes. The histidine sensitive strain retained its sensitivity to triazole alanine while the inhibitory effects of aminotriazole were much reduced indicating a change in regulation of his genes. The probable interactions between nif and his genes in cyanobacteria (blue-green algae) have been discussed.     

Further characterization of the two catalases inEscherichia coli

Crude extracts of wild-typeEscherichia coli contain two catalase species that separate on native polyacrylamide gels. The slow-migrating enzyme (HPII) has two pH optima of activity (at pH 6.8 and 10.5), is activated at 70°C, is sensitive to inhibitory by 3-amino-1,2,4-triazole and has Km values of 18.2 mM at pH 6.8 and of 10 mM at pH 10.5. The fast-migrating enzyme has a single pH optimum of 6.8 and is composed of two isozymes (HPI-A and HPI-B). Its activity is labile at 70°C, it is relatively resistant to inhibition by 3-amino-1,2,4-triazole and has a Km value of 3.7 mM.     

Metabolic Response to Treatment with Cold,Paraquat, or 3-Amino-1,2,4-triazole in Leaves of Winter Wheat

We treated leaves of winter wheat (Triticum aestivum L.) with cold, paraquat, or 3-amino-1,2,4-triazole and compared the responses. We assayed the activities of glucose-6-phosphate dehydrogenase, catalase, dehydroascorbate reductase and ascorbate free radical reductase and levels of hydrogen peroxide, glucose-6-phosphate, fructose-6-phosphate, ascorbate, dehydroascorbate, reduced and oxidized glutathione. With any of the three treatments, contents of cellular peroxides and hexose phosphates were raised. The content of ascorbate was lowered markedly by paraquat treatment, which produces active oxygen species, whereas such a decrease did not occur in other two treatments. When the plants were treated with 3-amino-1,2,4-triazole, which is a specific inhibitor of catalase, the content of oxidized glutathione increased severalfold. The glucose-6-phosphate dehydrogenase activity increased with all three treatments, but it decreased after glyphosate treatment, which does not stimulate the formation of peroxides. The activities of catalase and dehydroascorbate reductase were increased by the treatment of cold and paraquat, while 3-amino-1,2,4-triazole did not affect the dehydroascorbate reductase activity. The activity of ascorbate free radical reductase increased after treatment by paraquat only.     

Effect of cobalt chloride and 3-amino-1,2,4-triazole on the induction of cytochrome P-450 synthesis by phenobarbitone in rat liver

Administration of cobalt chloride and 3-amino-1,2,4-triazole leads to a suppression of phenobarbitone-mediated increase in total cytochrome P-450 as well as cytochrome P-450b contents of the liver. This suppression is due to a decrease in the content of the protein species which is the result of a decrease in its rate of synthesis as measured in vivo and in vitro. Cobalt chloride as well as 3-amino-1,2,4-triazole treatments lead to a decrease in the translatability of cytochrome P-450b RNA without affecting total protein synthesis. It is a possibility that a small pool of heme regulates the RNA levels for the cytochrome P-450 species.     

Role of haem in the synthesis and assembly of cytochrome P-450

By using 3-amino-1,2,4-triazole, an inhibitor of haem synthesis, and 2-allyl-2-isopropylacetamide, a drug that degrades the haem moiety of cytochrome P-450, the involvement of haem in cytochrome P-450 synthesis and assembly was investigated. Phenobarbital was used to stimulate apo-(cytochrome P-450) synthesis. Degradation of preformed cytochrome P-450 haem does not result in a concomitant release of the apoprotein from the endoplasmic reticulum. The availability of haem for cytochrome P-450 synthesis in the normal animal is not rate-limiting. Prolonged inhibition of haem synthesis in vivo decreases the rate of apo-(cytochrome P-450) synthesis, although this effect is not discernible under conditions of short-term inhibition of haem synthesis. Under the former conditions exogenous haemin is able to counteract the decrease in the rate of apoprotein synthesis. In animals receiving successive injections of phenobarbital plus 3-amino-1,2,4-triazole, compared with those receiving phenobarbital only, the holo-(cytochrome P-450) content measured spectrally shows a greater decrease than could be accounted for by the decrease in the content of the total apoprotein. In addition to less haem being available under these conditions, the free apoprotein appears to have undergone some modification, such that its haem-binding capacity is considerably decreased. This particular effect could be due to a direct interaction of 3-amino-1,2,4-triazole or its metabolites with cytochrome P-450 rather than a consequence of haem deficiency. Apo-(cytochrome P-450) is capable of binding to the endoplasmic reticulum in a form and at a site, which can be reconstituted with haemin to yield the functional protein.     

丁氟螨酯对二斑叶螨生长发育的影响

[目的]丁氟螨酯是一种新型酰基乙腈类非内吸性杀螨剂,对害螨的各个螨态都有很高活性,具有较高的应用价值.本文评价了丁氟螨酯对二斑叶螨生长发育的影响,以期为合理用药和二斑叶螨的综合防治提供理论依据.[方法]采用浸叶法测定丁氟螨酯对二斑叶螨成螨与卵的致死中浓度、雌成螨产卵量、各螨态存活率以及各发育历期的影响.[结果]经丁氟螨...     

Effect of cannibalism on predation,oviposition and longevity of the predacious mite,Agistemus exsertus Gonzalez (Acari: Stigmaeidae)

Cannibalism (intraspecific predation) on conspecific eggs was investigated in the predatory stigmaeid mite, Agistemus exsertus Gonzalez in the absence of eggs of Tetranychus urticae Koch (no-choice tests) and presence of three densities of prey eggs simultaneously (choice tests) in the laboratory. Data show that cannibalism occurs in immatures and adult females of the predator, which successfully developed and reproduced on conspecific eggs as an alternative prey in the absence of prey eggs. In no-choice tests, cannibalism rate on conspecific eggs by A. exsertus stages was significantly lower than the predation rate on T. urticae eggs. The predatory mite exhibited a marked decline in oviposition rate when preyed on conspecific eggs compared with feeding on prey eggs. The developmental duration and longevity of A. exsertus females were significantly longer 1.9 and 1.7 times, respectively, when fed on conspecific eggs than feeding on T. urticae eggs. The propensity of the predator towards cannibalism depends on the prey density, when T. urticae eggs and conspecific eggs are present simultaneously. Provision of increased densities of prey eggs significantly decreased cannibalism and predation by A. exsertus stages, which fed generally less on conspecific eggs than on T. urticae eggs in choice tests. The oviposition rate of the predator increased significantly, as the egg density of the prey increased. The developmental period and longevity of A. exsertus females showed significantly gradual shortness with increasing egg density of the prey.     

Lethal activity of beauvericin,a Beauveria bassiana mycotoxin,against the two‐spotted spider mites,Tetranychus urticae Koch

The entomopathogenic fungi Beauveria bassiana (Balsamo) Vuillemin is known to produce a broad range of secondary metabolites. Beauvericin, a cyclic hexadepsipeptide, is the best known mycotoxin produced by B. bassiana; however, reports discussing the insecticidal activity of beauvericin per se are limited. In this study, we assessed the lethal activity of beauvericin against Tetranychus urticae Koch (Acarina: Tetranychidae). In addition, screening for suitable application of the mycotoxin against T. urticae on greenhouse strawberries is discussed. Beauvericin was able to control successfully T. urticae where concentrations of 10, 100 and 1,000 µg/g recorded mortalities of 84%, 100% and 100%, respectively, against motile stages. Furthermore, beauvericin inhibited egg hatching up to 83.3%, 69.3% and 53.3%, respectively, using the same concentrations under laboratory conditions. Under greenhouse conditions, the efficacy recorded was 52.6%, 85.7%, 72.4% and 72.4% at 1, 3, 7 and 10 days post‐inoculation, respectively. Beauvericin was efficacious under greenhouse conditions since the application increased strawberry yields while showing no phytotoxicity and ecotoxicological risk. Resistance to beauvericin was not detected initially at the unselected strain of T. urticae. Yet, the laboratory selection of populations of T. urticae exposed to beauvericin resulted in relatively resistant T. urticae strain that displayed no cross‐resistance to cyflumetofen and bifenazate. The acaricidal activity of beauvericin documented in this study would increase the efficacy of integrated pest management strategies.     

Adaptation of acaricide stress facilitates Tetranychus urticae expanding against Tetranychus cinnabarinus in China

The two‐spotted spider mite, Tetranychus urticae, and the carmine spider mite, Tetranychus cinnabarinus, are invasive and native species in China, respectively. Compared with T. cinnabarinus, T. urticae has expanded into most parts of China and has become the dominant species of spider mite since 1983, when it was first reported in China. However, the mechanism of the demographic conversion has not been illuminated. In this study, one T. urticae field population and one T. cinnabarinus field population were isolated from the same plant in the same field, and the toxicological characteristics were compared between these two species. Laboratory bioassays demonstrated that T. urticae was more tolerant to commonly used acaricides than T. cinnabarinus. The activities of detoxification enzymes were significantly greater in T. urticae, and the fold changes of enzymes activities in T. urticae were also greater following exposure to acaricides. Furthermore, more metabolism‐related genes were upregulated at a basal level, and more genes were induced in T. urticae following exposure to acaricides. The comparison of proteins and genes between both species led credence to the hypothesis that T. urticae was more resistant to acaricides, which was the reason explaining the expansion of invasive T. urticae against native T. cinnabarinus. Laboratory simulation experiments demonstrated that following the application of acaricides, the composition of a mixed T. urticae/T. cinnabarinus population would change from a T. cinnabarinus‐dominant to a T. urticae‐dominant population. This study not only reveals that T. urticae possesses stronger detoxification capacity than its sibling species T. cinnabarinus, which facilitated its persistent expansion in China, but also points to the need to accurately identify Tetranychus species and to develop species‐specific management strategies for these pests.     

INHIBITORY EFFECT OF AMITROLE ON SCENEDESMUS QUADRICAUDA

The multiplication of the alga Scenedesmus quadricauda is inhibited by Amitrole (3-amino-1,2,4-triazole). This inhibition is reversed by addition of adenine to the growth medium. An electron-microscopic study has been made of the cytological changes which accompany the inhibition by Amitrole and the subsequent reversal. In the presence of Amitrole the cells accumulate a large quantity of starch which is lost very rapidly as soon as the Amitrole block is released and cell multiplication is resumed. ³Present address: Department of Biology and Botany, University of British Columbia, Vancouver, Canada.     

Intracellular location of cytochrome oxidase active in vivo in the cyanophytes,Synechocystis sp. PCC6714 andAnacystis nidulans Tx20 and R2, grown under various conditions

Summary Intracellular location of cytochrome oxidase (cytoxidase) active in vivo was studied cytochemically with four strains of cyanophytes, using 3,3-diaminobenzidine (DAB) oxidation. DAB was oxidized in the dark bySynechocystis sp. PCC6714 and two strains ofAnacystis nidulans (Tx20 and R2) grown under photoautotrophic, heterotrophic and salt-stressed conditions, respectively. Electron microscopic observations showed that DAB-oxidation in the dark occurred in the thylakoids, but was insignificant on or around the cytoplasmic membrane. However, deposition of DAB-oxidation product around the cytoplasmic membrane was observed with cells of the thylakoid-less cyanophyteGlaeobacter violaceus ATCC29082. All DAB oxidations observed with the four strains were inhibited completely by cyanide, the inhibitor of cyt-oxidase, but not by 3-amino-1,2,4-triazole, the inhibitor of peroxidase. The results show that (1) DAB was oxidized by the cyt-oxidase functioning in the respiratory system, and that (2) cyt-oxidase in thylakoids was active in vivo.Abbreviations AT 3-amino-1,2,4-triazole - Cyt oxidase cytochrome oxidase - ETS electron transport system - DAB 3,3-diaminobenzidine     

Enhanced cellular membrane contrast in a marine alga by Osmium-azole complexes

Summary Addition of certain heterocyclic nitrogen-carbon compounds to standard osmium tetroxide solutions used as secondary fixative resulted in an enhanced general membrane contrast in cells of the marine algaEmiliania huxleyi (Lohmann) Kamptner.Ultrastructural cell morphology and the contrast distribution were compared between cells treated according to a standard secondary fixation procedure and cells post-fixed when above mentioned heterocyclic compounds were introduced; in both cases some of the ultrathin sections were post-stained.Different compounds were tested: 1,2,4-triazole (TRA), 3-amino-1,2,4-triazole (A-TRA), 5-amino-tetrazole (A-TEA) and 2,4,6-tri-amino-1,3,5-triazine (melamine).The results were interpreted to indicate the possible bonding types arising from interaction of the heterocyclic compounds with osmium tetroxide and with membrane constituents.Interpretations were partly inspired by considerations from coordination chemistry.All above tests which did not include post-staining of thin sections could be performed at alkaline pH, and consequently calcified structures were preserved.The enhanced osmium accumulation at membranes was verified with X-ray microanalysis, which also showed that in the cases where membranes were visibly contrasted, localization of probable sites of intracellular non-crystalline calcium was facilitated.     

Host plant-mediated interactions between Tetranychus urticae and Eotetranychus carpini borealis (Acari: Tetranychidae)

Competitive interactions between Tetranychus urticae and Eotetranychus carpini borealis were investigated under laboratory and greenhouse conditions. When placed first to colonize red raspberry leaves, T. urticae caused extinction of E. carpini borealis populations. E. carpini borealis had detrimental effects on T. urticae but did not cause T. urticae population extinction. The area used by E. carpini borealis for egg deposition decreased with an increase in the number of T. urticae eggs. Red raspberry showed an induced response to T. urticae and E. carpini borealis feeding after two weeks of infestation. Population growth of each species was reduced on plants that were previously infested by conspecifics but inducible response by one species had no effects on the other species.     

The Predatory Mite Phytoseiulus persimilis Adjusts Patch-leaving to Own and Progeny Prey Needs

Integration of optimal foraging and optimal oviposition theories suggests that predator females should adjust patch leaving to own and progeny prey needs to maximize current and future reproductive success. We tested this hypothesis in the predatory mite Phytoseiulus persimilis and its patchily distributed prey, the two-spotted spider mite Tetranychus urticae. In three separate experiments we assessed (1) the minimum number of prey needed to complete juvenile development, (2) the minimum number of prey needed to produce an egg, and (3) the ratio between eggs laid and spider mites left when a gravid P. persimilis female leaves a patch. Experiments (1) and (2) were the pre-requirements to assess the fitness costs associated with staying or leaving a prey patch. Immature P. persimilis needed at least 7 and on average 14±3.6 (SD) T. urticae eggs to reach adulthood. Gravid females needed at least 5 and on average 8.5±3.1 (SD) T. urticae eggs to produce an egg. Most females left the initial patch before spider mite extinction, leaving prey for progeny to develop to adulthood. Females placed in a low density patch left 5.6±6.1 (SD) eggs per egg laid, whereas those placed in a high density patch left 15.8±13.7 (SD) eggs per egg laid. The three experiments in concert suggest that gravid P. persimilis females are able to balance the trade off between optimal foraging and optimal oviposition and adjust patch-leaving to own and progeny prey needs.     

Influence of substituents on the structures of hybrid complexes constructed from tetranuclear copper(I) 1,2,4-triazolate clusters and octamolybdates

By changing the substituents on 1,2,4-triazole ring, six novel organic-inorganic hybrid complexes constructed from tetranuclear copper(I) 1,2,4-triazolate clusters and octamolybdates, [{Cu₄(L)_x}Mo₈O₂₆] (L = 3,5-diamino-1,2,4-triazole (datrz) and x = 4 for 1; L = 3-amino-1,2,4-triazole (3atrz) and x = 4 for 2; L = 3,5-dimethyl-1,2,4-triazole (dmtrz) and x = 4 for 3; L = 3,5-dimethyl-4-amino-1,2,4-triazole (dmatrz) and x = 6 for 4; L = 3,5-diethyl-4-amino-1,2,4-triazole (deatrz) and x = 4 for 5; L = 3,5-di(n-propyl)-4-amino-1,2,4-triazole (dpatrz) and x = 3 for 6), were obtained. The tetranuclear Cu(I) cluster in compound 1 acts as charge-compensating unit, which is the first polynuclear metal 1,2,4-triazole structure only with N1, N2 bridging mode. Compounds 2, 4, 5 and 6 are of polymeric 1D chains and 3 is of a 2D layer structure. In 2, three distinct Cu(I)-coordination geometries, distorted tetrahedral, T-shaped and V-shaped linear Cu(I), are observed in the same structure. The first extended hybrid structure constructed by δ-octamolybdates is founded in 4. A novel [Mo₈O₂₆]⁴⁻ anion is found in 5, which contains only three crystallographically independent Mo atoms. In compounds 5 and 6, terminal oxo groups of octamolybdate cluster act as μ₃-oxo bridges to link the copper(I) coordination complexes; such an unusual linking manner is unique in the coordination chemistry of octamolybdates with transition metal fragments. The influences of substituent on the structures of the tetranuclear units are also discussed in details.     

Regioselective synthesis of 6-aryl-benzo[h][1,2,4]-triazolo[5,1-b]quinazoline-7,8-diones as potent antitumoral agents

Three-component coupling of aldehyde, 2-hydroxy-1,4-naphthoquinone and 3-amino-1,2,4-triazole has been achieved using a catalytic amount of sulfamic acid under solvent free conditions to produce a novel series of 6-aryl-benzo[h][1,2,4]-triazolo[5,1-b]quinazoline-7,8-dione derivatives in good yields and with high regioselectivity. These compounds are found to exhibit potent antitumoral properties.     

Behavioral responses to prey density by three acarine predator species with different degrees of polyphagy

Behavioral responses by three acarine predators, Phytoseiulus persimilis, Typhlodromus occidentalis, and Amblyseius andersoni (Acari: Phytoseiidae), to different egg and webbing densities of the spider mite Tetranychus urticae (Acari: Tetranychidae) on rose leaflets were studied in the laboratory. Prey patches were delineated by T. urticae webbing and associated kairomones, which elicit turning back responses in predators near the patch edge. Only the presence of webbing affected predator behavior; increased webbing density did not increase patch time. Patch time increased with increased T. urticae egg density in the oligophagous P. persimilis, but was density independent in the polyphagous species T. occidentalis and A. andersoni. Patch time in all three species was more strongly correlated with the number of prey encounters and attacks than with the actual prey number present in the patch. Patch time was determined by (a) the turning back response near the patch edge; this response decayed through time and eventually led to the abandonment of the patch, and (b) encounters with, and attacks upon, prey eggs; these prolonged patch time by both an increment of time spent in handling or rejecting prey and an increment of time spent searching between two successive prey encounters or attacks. Although searching efficiency was independent of prey density in all three species, the predation rate by P. persimilis decreased with prey density because its searching activity (i.e. proportion of total patch time spent in searching) decreased with prey density. Predation rates by T. occidentalis and A. andersoni decreased with prey density because their searching activity and success ratio both decreased with prey density. The data were tested against models of predator foraging responses to prey density. The effects of the degree of polyphagy on predator foraging behavior were also discussed.     

Cytochemische Darstellung von Peroxisomen in Pilzzellen

Zusammenfassung Mit Hilfe der cytochemischen Methode vonFahimi (1968) wurde in den Zellen der PilzeNeurospora crassa, Rhizopus nigricans undSaccbaromyces cerevisiae das Enzym Katalase in distinkten Granula nachgewiesen. Die Spezifität der Färbereaktion wurde durch ihre Hemmbarkeit mit 3-Amino-1,2,4-triazol gezeigt.

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Cytochemical localization of peroxisomes in fungal cells

Summary In the cells of the fungiNeurospora crassa, Rhizopus nigricans, andSaccharomyces cerevisiae catalase was detected in distinct granula with the cytochemical procedure ofFahimi (1968). The specificity of the staining reaction was demonstrated by the inhibitory action of 3-amino-1,2,4-triazole.

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Frau Prof. Dr. B.Haccius danke ich für die Überlassung der Pilzkulturen.     

Predation,oviposition and conversion rates of the predacious mite,Neoseiulus californicus (McGregor) consuming different densities of Tetranychus urticae Koch,Bemisia tabaci (Genn.) and Thrips tabaci Lind.

Rates of prey consumption, egg production and prey conversion by the predacious mite, Neoseiulus californicus (McGregor) were estimated at different densities of Tetranychus urticae Koch, Bemisia tabaci (Genn.) and Thrips tabaci Lind. in the laboratory. N. californicus females functionally and numerically responded to the increasing densities of T. urticae nymphs, B. tabaci nymphs and T. tabaci larvae showing Holling’s type II. The maximum mean predation and oviposition rates by the predator females devouring T. urticae, B. tabaci and T. tabaci occurred at 15, 10 and 10 prey individuals/day, respectively, followed by the plateau levels at higher prey densities. N. californicus females exhibited the highest feeding and oviposition on T. urticae nymphs, followed by T. tabaci larvae and B. tabaci nymphs. The predator females showed the highest efficiency in converting the prey into egg progeny at 5 individuals/day of the previous prey species, respectively. T. urticae was the most favourable for N. californicus females, followed by T. tabaci and B. tabaci.