Insect growth regulatory activity of naturally occurring quinones and their derivatives in Dysdercus koenigii Fabr. (Hem., Pyrrhocoridae)

Plumbagin, a naphthoquinone occurring in the plants belonging to Plumbago sp. is known to have insect growth disrupting activities. In the present study, a comparative evaluation of bioactivities of related naphthoquinones, juglone and menadione and benzoquinones, 2,6-dimethylbenzoquinone and 2,3,6-trimethylbenzoquinone along with 2,6-dimethylhydroquinone have been undertaken using Dysdercus koenigii . The LD₅₀ values of the compounds showed wide variation. Although all the test compounds disrupted the normal growth at sublethal doses, 2,6-dimethylbenzoquinone and menadione showed activity at low doses.     

Mutagenicity of natural naphthoquinones and benzoquinones in the Salmonella/microsome test

The mutagenicities of naturally occurring naphthoquinones and benzoquinones were tested by the pre-incubation method with Salmonella typhimurium strains TA98, TA100 and TA2637, which all contain plasmid pKM101. 6 of the 16 naphthoquinones tested, i.e., plumbagin, naphthazarin, 2-hydroxy-naphthoquinone, vitamin K3 (menadione), juglone and 7-methyljuglone, were mutagenic to strain TA2637 with metabolic activation. Except for juglone and 7-methyl-juglone, these compounds also had slight mutagenic effects on strain TA98 with S9 mix. All the mutagenic naphthoquinones contain one or two hydroxyl and/or methyl substituents. The naphthoquinone mompain, which has four hydroxyl groups, was not mutagenic. Unsubstituted beta-naphthoquinone, naphthoquinones with a prenyl side chain and all bi-naphthoquinone derivatives tested were non-mutagenic. None of the 13 benzoquinones examined was mutagenic to any of the strains used with or without metabolic activation. These results show that natural naphthoquinones are mutagenic when they have only one or two hydroxyl and/or methyl substituents.     

1,4-Naphthoquinones as inducers of oxidative damage and stress signaling in HaCaT human keratinocytes

Selected biological effects of 1,4-naphthoquinone, menadione (2-methyl-1,4-naphthoquinone) and structurally related quinones from natural sources - the 5-hydroxy-naphthoquinones juglone, plumbagin and the 2-hydroxy-naphthoquinones lawsone and lapachol - were studied in human keratinocytes (HaCaT). 1,4-naphthoquinone and menadione as well as juglone and plumbagin were highly cytotoxic, strongly induced reactive oxygen species (ROS) formation and depleted cellular glutathione. Moreover, they induced oxidative DNA base damage and accumulation of DNA strand breaks, as demonstrated in an alkaline DNA unwinding assay. Neither lawsone nor lapachol (up to 100 μM) were active in any of these assays. Cytotoxic and oxidative action was paralleled by stimulation of stress signaling: all tested quinones except lawsone and lapachol strongly induced phosphorylation of the epidermal growth factor receptor (EGFR) and the related ErbB2 receptor tyrosine kinase. EGFR activation by plumbagin, juglone and menadione was attenuated by a superoxide dismutase mimetic, indicating that ROS-related mechanisms contribute to EGFR activation by these naphthoquinones.     

Effect of Artemisia annua oil on development and reproduction of Dysdercus koenigii F. (Hem., Pyrrhocoridae)

The effect of topical application of Artemisia annua oil on the development and reproduction of Dysdercus koenigii was studied. On the basis of 48 h mortality, the LD₅₀ for fifth instar nymphs was 0.48 μl/nymph. The treated nymphs showed 2 days delay in development, and also produced intermediates. The treatment decreased haemolymph protein concentration and disturbed its electrophoretic protein pattern. The adults which emerged from treated nymphs showed poor ovary development. There was a greater median neurosecretory activity in the treated insects compared with control insects till day 6 of adult life.     

Toxicity of plumbagin and juglone to the eggs of the cotton stainer,Dysdercus koenigii

Toxicity of two plumbaginoids viz., plumbagin and juglone to the eggs of the cotton stainer,Dysdercus koenigii was studied by a residual film technique. Of these two, plumbagin showed the higher toxicity against different-age eggs with LC₅₀ ranging from 0.0044 to 0.0066%. Eggs showed low susceptibility in the middle of embryogenesis. The toxicity of plumbaginoids, especially plumbagin, is discussed in relation to mode of action and prospects of its use as an ovicide in control of the insect.     

Plumbagin and juglone induce caspase-3-dependent apoptosis involving the mitochondria through ROS generation in human peripheral blood lymphocytes

The phytochemicals plumbagin and juglone have recently been gaining importance because of their various pharmacological activities. In this study, these compounds are shown to induce concentration- and time-dependent toxicity in human peripheral blood lymphocytes via the apoptotic pathway. Flow cytometry data revealed the occurrence of about 28% early apoptotic cells after 6 h exposure to 10 μM plumbagin and 35% late apoptotic cells and about 43% sub-G1 population after 24 h. The cytotoxic effect of plumbagin was at least twofold higher than that of juglone as evidenced by the IC₅₀ value for cytotoxicity. Characteristic apoptotic features such as chromatin condensation and apoptotic body formation were observed through TEM, and membrane blebbing and cell surface smoothening were seen in SEM studies. Generation of ROS was evidenced through the HPLC analysis of superoxide-specific 2-OH-E+ formation. In addition, a decrease in GSH levels parallel to ROS production was observed. Reversal of apoptosis in both NAC- and Tempol-pretreated cells indicates the involvement of both ROS generation and GSH depletion in plumbagin- and juglone-induced apoptosis. The mechanistic pathway involves a decrease in MMP; alterations in the levels of Bcl-2, Bax, and cytosolic cytochrome c; and PARP-1 cleavage subsequent to caspase-3 activation.     

Benzoquinone levels as a function of age and gender of the red flour beetle, Tribolium castaneum

Liquid chromatography with both photodiode array and electrochemical detection was used to analyze as a function of age and gender the levels of two ρ-quinones, methyl-1,4-benzoquinone (MBQ) and ethyl-1,4-benzoquinone (EBQ), which are found in defensive secretions of the red flour beetle, Tribolium castaneum. We developed a method to simultaneously analyze quinones and hydroquinones excreted from or in homogenates of individual beetles. The major components present in beetle extracts were the benzoquinones and not free or conjugated forms of the hydroquinones. Greater than 95% of the quinone/hydroquinone mixture in extracts was present in the oxidized form. Because of their lability, however, the quinones were quantified indirectly as their hydroquinone derivatives after extraction in dilute acid supplemented with ascorbic acid as a reducing agent. Comparisons of whole body rinses and homogenates revealed that rinses recovered only up to 60% of the total quinones that were extracted after homogenization. The levels recovered also depended on the age and sex of the individual beetles sampled. ρ-Benzoquinones in both male and female beetles increased after adult eclosion and cuticle sclerotization for 40–50 days and then remained at their highest levels (15–21 μg MBQ and 22–32 μg EBQ per beetle) through 80 days posteclosion. Virgin females that were collected 40–80 days after eclosion contained approximately 40% more of these compounds than males of the same age. The build-up of ρ-benzoquinones subsequent to cuticle sclerotization apparently reflects the need for an adequate cuticular barrier for self-protection from these defensive compounds.     

Synthesis and SAR study of novel anticancer and antimicrobial naphthoquinone amide derivatives

A series of novel naphthoquinone amide derivatives of the bioactive quinones, plumbagin, juglone, menadione and lawsone, with various amino acids were synthesized. The compounds were characterized by ¹H NMR, ¹³C NMR, Mass, IR and elemental analysis. All the compounds were evaluated for their anticancer activity against HeLa and SAS cancer cell lines and 3D-QSAR indicated the presence of electron donating group near sulphur enhanced the activity against HeLa cells. Among the derivatives synthesized, compounds 11f, 10a, 10b and 10g were the most active with IC₅₀ values of 16, 12, 14 and 24.5 μM, respectively. The analogues were also screened for antimicrobial activity against two human bacterial pathogens, the Gram-positive Methicillin resistant Staphylococcus aureus (MRSA) and the Gram-negative Pseudomonas aeruginosa and a human yeast pathogen, Fluconazole resistant Candida albicans (FRCA). Among the synthesized compounds, 8g, 10g and 11g exhibited maximum antibacterial activity towards MRSA and antifungal activity against FRCA in well diffusion method.     

Enhancement of Bacillus thuringiensis with monosodium glutamate against larvae of obliquebanded leafroller (Lep.:Tortricidae)

Abstract:  Monosodium glutamate, a taste enhancer widely used in food industry, was tested in the laboratory to determine its phagostimulatory effects on larvae of Choristoneura rosaceana (Harris). Larvae fed apple leaves treated with 50–700  μ g/l monosodium glutamate increased leaf tissue consumption by approximately 40%. The stimulatory effect of monosodium glutamate (at 675  μ g/l concentration) was maintained throughout 10 days of continuous exposure. Adding 675  μ g/l monosodium glutamate to commercial formulation of Bacillus thuringiensis ssp. kurstaki, (DiPel^®2X DF), lowered LC₅₀ from 450 to 150  μ g/l (P < 0.05, Lethal Ratio Significance Test), indicating good potential for monosodium glutamate to enhance B. thuringiensis -based formulations.     

Interaction of benzoquinones with mitochondria interferes with oxidative phosphorylation characteristics

Studies with four benzoquinones, viz. juglone, embelin, maesaquinone and maesanin, on rat liver mitochondria oxidative phosphorylation have been carried out. Three of the benzoquinones are uncouplers in the order juglone greater than maesoquinone greater than embelin, while maesanin is an inhibitor of electron transport and oxidative phosphorylation.     

Occurrence, biosynthesis and function of isoprenoid quinones

Isoprenoid quinones are one of the most important groups of compounds occurring in membranes of living organisms. These compounds are composed of a hydrophilic head group and an apolar isoprenoid side chain, giving the molecules a lipid-soluble character. Isoprenoid quinones function mainly as electron and proton carriers in photosynthetic and respiratory electron transport chains and these compounds show also additional functions, such as antioxidant function. Most of naturally occurring isoprenoid quinones belong to naphthoquinones or evolutionary younger benzoquinones. Among benzoquinones, the most widespread and important are ubiquinones and plastoquinones. Menaquinones, belonging to naphthoquinones, function in respiratory and photosynthetic electron transport chains of bacteria. Phylloquinone K₁, a phytyl naphthoquinone, functions in the photosynthetic electron transport in photosystem I. Ubiquinones participate in respiratory chains of eukaryotic mitochondria and some bacteria. Plastoquinones are components of photosynthetic electron transport chains of cyanobacteria and plant chloroplasts. Biosynthetic pathway of isoprenoid quinones has been described, as well as their additional, recently recognized, diverse functions in bacterial, plant and animal metabolism.     

Bioactivity of two major constituents isolated from the essential oil of Artemisia judaica L

The essential oil of Artemisia judaica L., grown on Sinai Peninsula of Egypt, was extracted via hydrodistillation. Chromatographic separation on repeated silica gel columns led to isolate two compounds namely piperitone and trans-ethyl cinnamate. Insecticidal, antifeedant and antifungal properties of the isolated compounds were examined. Both compounds showed pronounced insecticidal and antifeedant activity against the third instar larvae of Spodoptera littoralis (Boisd). trans-Ethyl cinnamate (LD₅₀ = 0.37 μg/larva) was more toxic than piperitone (LD₅₀ = 0.68 μg/larva). The two isolated compounds revealed antifeedant activity in a concentration dependent manner, with complete feeding inhibition at a concentration of 1000 μg/ml. When tested for antifungal activity against four plant pathogenic fungi, the isolated compounds exhibited a moderate to high activity.     

Cytotoxicity Mechanism of Two Naphthoquinones (Menadione and Plumbagin) in Saccharomyces cerevisiae

Background

Quinones are compounds extensively used in studies of oxidative stress due to their role in plants as chemicals for defense. These compounds are of great interest for pharmacologists and scientists, in general, because several cancer chemotherapeutic agents contain the quinone nucleus. However, due to differences in structures and diverse pharmacological effects, the exact toxicity mechanisms exerted by quinones are far from elucidatation.

Methodology/Principal Findings

Using Saccharomyces cerevisiae, we evaluated the main mechanisms of toxicity of two naphthoquinones, menadione and plumbagin, by determining tolerance and oxidative stress biomarkers such as GSH and GSSG, lipid peroxidation levels, as well as aconitase activity. The importance of glutathione transferases (GST) in quinone detoxification was also addressed. The GSSG/GSH ratio showed that menadione seemed to exert its toxicity mainly through the generation of ROS while plumbagin acted as an electrophile reacting with GSH. However, the results showed that, even by different pathways, both drugs were capable of generating oxidative stress through their toxic effects. Our results showed that the control strain, BY4741, and the glutathione transferase deficient strains (gtt1Δ and gtt2Δ) were sensitive to both compounds. With respect to the role of GST isoforms in cellular protection against quinone toxicity, we observed that the Gtt2 deficient strain was unable to overcome lipid peroxidation, even after a plumbagin pre-treatment, indicating that this treatment did not improve tolerance when compared with the wild type strain. Cross-tolerance experiments confirmed distinct cytotoxicity mechanisms for these naphthoquinones since only a pre-treatment with menadione was able to induce acquisition of tolerance against stress with plumbagin.

Conclusions/Significance

These results suggest different responses to menadione and plumbagin which could be due to the fact that these compounds use different mechanisms to exert their toxicity. In addition, the Gtt2 isoform seemed to act as a general protective factor involved in quinone detoxification.     

Glasshouse tests of quinones, polyhydroxybenzenes and related compounds against potato common scab

Forty quinones, polyhydroxybenzenes and related compounds were tested for possible control of potato common scab, caused by Streptomyces scabies. They were incorporated, at 50 μg/g or less, into scab-infected soil in which potato plants were then grown in the glasshouse. Quintozene at 50/μ g/g was used as standard. The chemicals varied very widely in their effects on scab, and in their phytotoxicity, as measured by decrease in yield of tubers. Corresponding 1,4-benzoquinones and hydroquinones behaved similarly. The unsubstituted and the phenyl-, chloro-,2,5- and 2,6-dichloro-com-pounds were as effective as quintozene; with the exception of chloro-hydroquinone, they did not decrease yield. Chloranil and chloroneb (the dimethylether of 2,5-dichlorohydroquinone) were much less effective, and chloroneb decreased yield as well. 4-terf-butyl catechol was more effective than any other compound tested, including quintozene, and at suitable rates (25 or 12-5 μg/g) did not decrease yield. Other hydroxybenzenes and some 1,4-naphthaquinones (including dichlone) and anthraquinones were almost ineffective.     

Effects of azadirachtin and of simpler epoxy-alcohols on survival and behaviour of Galleria mellonella (Lepidoptera)

Abstract:  Investigations into the toxicity of three simpler molecules based on the epoxy-alcohol fragment of azadirachtin have revealed insecticidal activity on the greater wax moth Galleria mellonella L. larvae. The simpler epoxy-alcohols doses giving 50% mortalities (LD₅₀) for G. mellonella larvae were in the increasing order from glycidol (0.022 mg/g), 4,5-epoxy-2-pentanol (0.068 mg/g) and finally, glycerol diglycidyl ether (0.147 mg/g). The three epoxy-alcohols exhibited higher insecticidal activity when compared with the commercial neem product for which the dose giving 50% mortalities was 10.6 mg/g and to azadirachtin that killed the larvae only by injection (dose of 0.20 mg/g of larvae body weight). Our results confirm the importance of the epoxy-alcohol junction between the two parts of the azadirachtin molecule for the biological activity. Other effects of the epoxy-alcohols tested were blackening of larvae and morphological deformities of some adults hatching. In future, the molecules should be complexified (degree of ramification, length of chain and presence of bulky ramified substituent) to obtain an insecticide as toxic for insects only and environmentally safe as azadirachtin but more stable, and their physiological activities on insect's tissues and cells should be studied.     

Antifeeding activity of limonoids from Khaya senegalensis (Meliaceae)

Abstract: Fifteen B,D-secolimonoids of mexicanolide, rearranged phragmalin, methyl angolensate and glycoside types have been isolated from ether and acetone extracts of the stem bark of Khaya senegalensis (Desr.). The antifeedant activity of the isolated compounds was assessed by conventional choice leaf disc method on the third-instar larvae of Spodoptera littoralis (Boisd.). Khayalactol, 1- O -acetylkhayanolide A, 2-hydroxyseneganolide, khayanolide A, khayanolide D and methyl angolensate displayed strong antifeedant activity at 1000  μ g/ml with antifeedant percentages of 83.8, 61.9, 60.1, 59.5, 57.1 and 55.7, respectively. 2-Hydroxyseneganolide and khayanolide D showed high activity at 500, 300 and 200  μ g/ml while 1- O -acetylkhayanolide A was the only compound that revealed antifeedant activity at a concentration as low as 100  μ g/ml. Antifeedant activity was dose-dependent in some of the isolated compounds. Correlation between antifeedant activity of the isolated compounds and chemical structure was discussed.     

Antitermitic quinones from Diospyros sylvatica

Six quinones were isolated from the chloroform extract of the roots of Diospyros sylvatica and identified as 2-methyl-anthraquinone, plumbagin, diosindigo, diospyrin, isodiospyrin and microphyllone. The effect of the root extract on the orientation and survival of the subterranean termite, Odontotermes obesus was tested. In addition, four of these quinones were tested on the survival of the subterranean termite. In a direct-choice experiment, exposure to an extract-treated filter disc had a significantly repellent effect over the solvent-treated filter disc. The no-choice experiment revealed the toxic property of the extract as well as the tested quinones and showed high mortality of the O. obesus workers after 48 h on forced exposure. The major termiticidal components identified were plumbagin, isodiospyrin and microphyllone while diospyrin was not toxic to termites at the concentration tested. All the quinones are reported for the first time from D. sylvatica.     

Regulation of 1-dehydrogenation and 20 beta-reduction of ketosteroids by microorganisms]

Regulation of two microbiological processes--1-dehydrogenation and 20 beta-reduction of ketosteroids--was studied using several exogeneous quinones which (a) stimulated 1-dehydrogenation and (b) inhibited the accompanying reaction, the reduction of 20-ketogroup of the steroid molecule. The least active compounds were benzoquinones. The best regulators are 1,4-naphthoquinone, 1,2-naphthoquinone, and 7-methoxy-1,2-naphthoquinone. Possible mechanism of the action of naphthoquinones on Mycobacterium globiforme is discussed.     

The w/w+ SMART assay of Drosophila melanogaster detects the genotoxic effects of reactive oxygen species inducing compounds

The somatic mutation and recombination w/w+ eye assay has been used for genotoxic evaluation of a broad number of chemicals with different action mechanisms yielding high values of sensitivity, specificity and accuracy. The aim of this work was to determine the utility of this assay in the evaluation of reactive oxygen species inducers. For this, we have tested eight compounds: diquat, paraquat, menadione, juglone, plumbagin, streptonigrin, tert-butyl hydroperoxide and 4-nitroquinoline 1-oxide, using the Drosophila Oregon K strain which had previously shown advantageous conditions to test this type of compounds. Diquat was the only chemical for which the results were clearly negative, probably because its high toxicity, whereas indications of a marginal genotoxicity raised for menadione. The remaining compounds were evaluated as positives. The conclusion of these experiments is that the w/w+ assay is capable to detect genotoxic effects induced by compounds that generate reactive oxygen species through different action mechanisms.     

The w/w SMART assay of Drosophila melanogaster detects the genotoxic effects of reactive oxygen species inducing compounds

The somatic mutation and recombination w/w⁺ eye assay has been used for genotoxic evaluation of a broad number of chemicals with different action mechanisms yielding high values of sensitivity, specificity and accuracy. The aim of this work was to determine the utility of this assay in the evaluation of reactive oxygen species inducers. For this, we have tested eight compounds: diquat, paraquat, menadione, juglone, plumbagin, streptonigrin, tert-butyl hydroperoxide and 4-nitroquinoline 1-oxide, using the Drosophila Oregon K strain which had previously shown advantageous conditions to test this type of compounds. Diquat was the only chemical for which the results were clearly negative, probably because its high toxicity, whereas indications of a marginal genotoxicity rised for menadione. The remaining compounds were evaluated as positives. The conclusion of these experiments is that the w/w⁺ assay is capable to detect genotoxic effects induced by compounds that generate reactive oxygen species through different action mechanisms.