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Multiple hypotheses have been put forth to understand why defense chemistry in individual plants is so diverse. A major challenge has been teasing apart the importance of concentration vs. composition of defense compounds and resolving the mechanisms of diversity effects that determine plant resistance against herbivores. Accordingly, we first outline nonexclusive mechanisms by which phytochemical diversity may increase toxicity of a mixture compared to the average effect of each compound alone. We then leveraged independent in vitro, in vivo transgenic, and organismal experiments to test the effect of equimolar concentrations of purified milkweed toxins in isolation vs. mixtures on the specialist and sequestering monarch butterfly. We show that cardenolide toxin mixtures from milkweed plants enhance resistance against this herbivore compared to equal concentrations of single compounds. In mixtures, highly potent toxins dominated the inhibition of the monarch's target enzyme (Na+/K+-ATPase) in vitro, revealing toxin-specific affinity for the adapted enzyme in the absence of other physiological adaptations of the monarch. Mixtures also caused increased mortality in CRISPR-edited adult Drosophila melanogaster with the monarch enzyme in vivo, whereas wild-type flies showed lower survival regardless of mixture type. Finally, although experimentally administered mixtures were not more toxic to monarch caterpillars than single compounds overall, increasing caterpillar sequestration from mixtures resulted in an increasing burden for growth compared to single compounds. Phytochemical diversity likely provides an economical plant defense by acting on multiple aspects of herbivore physiology and may be particularly effective against sequestering specialist herbivores. [ABSTRACT FROM AUTHOR]

Theories have been widely proposed and tested for impacts of soil nitrogen (N) on phytochemical defenses. Among the hundreds of distinct cardenolide toxins produced by milkweeds (Asclepias spp.), few contain N, yet these appear to be the most toxic against specialist herbivores. Because N- and non-N-cardenolides coexist in milkweed leaves and likely have distinct biosynthesis, they present an opportunity to address hypotheses about drivers of toxin expression. We tested effects of soil N and herbivore-damage on cardenolide profiles of two milkweed species differing in life-history strategies (Asclepias syriaca and A. curassavica), and the toxicity of their leaves. In particular leaf extracts were tested against the target enzymes (Na+/K+-ATPase extracted from neural tissue) from both monarch butterflies (Danaus plexippus) as well as less cardenolide-resistant queen butterflies, D. gilippus. Increasing soil N enhanced biomass of Asclepias syriaca but had weak effects on cardenolides, including causing a significant reduction in the N-cardenolide labriformin; feeding by monarch caterpillars strongly induced N-cardenolides (labriformin), its precursors, and total cardenolides. Conversely, soil N had little impact on A. curassavica biomass, but was the primary driver of increasing N-cardenolides (voruscharin, uscharin and their precursors); caterpillar induction was weak. Butterfly enzyme assays revealed damage-induced cardenolides substantially increased toxicity of both milkweeds to both butterflies, swamping out effects of soil N on cardenolide concentration and composition. Although these two milkweed species differentially responded to soil N with allocation to growth and specific cardenolides, leaf toxicity to butterfly Na+/K+-ATPases was primarily driven by herbivore-induced defense. Thus, both biotic and abiotic factors shape the composition of phytochemical defense expression, and their relative importance may be dictated by plant life-history differences. [ABSTRACT FROM AUTHOR]

Background: Poekilocerus pictus (Fabricius 1771), a painted grasshopper, sequesters cardenolides from its food plant, the Apple of Sodom or Aak, Calotropis procera (Aiton) W.T. Aiton (Family-Asclepiadaceae). In our present investigation, we were able to isolate Pseudomonas aeruginosa KRK6 from the intestine of Poekilocerus pictus responsible for the biotransformation of cardenolides. Methods: Pseudomonas aeruginosa KRK6 was grown in methanolic extracts of Calotropis procera and the modified cardenolides were detected by Liquid Chromatography-Mass Spectrometry (LCMS) and also used to induce apoptosis in cancer cells (MCF-7 cells and T-47 D). Result: The modified cardenolides CPMEP6 was found to induce apoptosis in human breast adenocarcinoma cells (MCF cells-IC50= 6.31±0.4 µg/mL, T-47D cells-IC50= 10.1±1.02 µg/mL). Phosphatidylserine exposure and DNA fragmentation suggested apoptosis in treated cancer cells. CPMEP6 induced apoptosis in cancer cells via the mitochondrial pathway by down-regulating BCL-2 protein expression and up-regulating BAX protein expression. [ABSTRACT FROM AUTHOR]

Since ancient times, people have used medicinal plants to treat various diseases. Nerium oleander is a toxic plant due to the presence of various cardiac glycosides. Oleander, at a dose of 50 mg kg-1 inhibits Na+/K+ ATPase pump in mice. Despite being toxic, it is used in traditional medicine. Almost all of the parts of oleander plant contain various phytochemicals such as phenols, tannins, flavonoids, coumarins, sterols, triterpenes, alkaloids, and different types of cardenolides. Oleander extract has neuroprotective, sedative, and anticonvulsant properties, especially against ischemic stroke. Cardenolides like nerizoside, neritaloside, odoroside-H, and neridiginoside offer advantages when used as CNS depressants. CNS depressant action of oleander is primarily due to a rise in GABA levels. At low to moderate dose, it shows depressant activity but in high doses, body became motionless inducing tremors and lethargy. Oleanderʼs dual role of toxic and therapeutic activity raises concerns about its application. Yet due to presence of vast constituents, it can be used for modern medicine. Cardenolides showing antidepressant activity on mice. [Display omitted] • Nerium oleander is considered to be a poisonous plant. • But it contains various phytochemicals, including antioxidants, anticancer agents, CNS depressants, and anticonvulsants. • This review aims to identify the therapeutic benefits of the plant and its toxicity. [ABSTRACT FROM AUTHOR]

Plants in the genus Erysimum produce both glucosinolates and cardenolides as a defense mechanism against herbivory. Two natural isolates of Erysimum cheiranthoides (wormseed wallflower) differed in their glucosinolate content, cardenolide content, and their resistance to Myzus persicae (green peach aphid), a broad generalist herbivore. Both classes of defensive metabolites were produced constitutively and were not further induced by aphid feeding. To investigate the relative importance of glucosinolates and cardenolides in E. cheiranthoides defense, we generated an improved genome assembly, genetic map, and segregating F2 population. The genotypic and phenotypic analysis of the F2 plants identified quantitative trait loci, which affected glucosinolates and cardenolides, but not the aphid resistance. The abundance of most glucosinolates and cardenolides was positively correlated in the F2 population, indicating that similar processes regulate their biosynthesis and accumulation. Aphid reproduction was positively correlated with glucosinolate content. Although the overall cardenolide content had little effect on aphid growth and survival, there was a negative correlation between aphid reproduction and helveticoside abundance. However, this variation in defensive metabolites could not explain the differences in aphid growth on the two parental lines, suggesting that processes other than the abundance of glucosinolates and cardenolides have a predominant effect on aphid resistance in E. cheiranthoides. [ABSTRACT FROM AUTHOR]

Phytochemical is a collective term for plant chemicals with varied structure and function. The most common sources of phytochemicals are fruits, vegetables, whole grains, nuts and seeds, and other plant foods. Calotropin is a pharmacologically active compound isolated from milkweed plants like Calotropis procera, Calotropis gigantea, and Asclepias curassavica that belong to the Asclepiadaceae family which is used for medicinal purposes in many Asian countries. Calotropin is identified as a highly potent cardenolide that has a similar chemical structure to cardiac glycosides (such as digoxin and digitoxin). Among cardenolides, calotropin is identified as the most promising agent. Calotropin has cytotoxic and anti-tumor impacts, with cancers of the breast, colon, lung, and leukemia malignancies exhibiting the most significant effects. The effects of calotropin on cancer have been extensively studied in preclinical pharmacological studies in vitro using cancer cell lines and in vivo in experimental animal models that have targeted antitumor mechanisms and anticancer signaling pathways. During ancient times, calotropin was utilized in various techniques. A macerated bark extract is frequently utilized for de-hearing hides and tanning. Calotropin is a particularly effective abortifacient or interceptive agent in females. Cardenolide calotropin is poisonous. This critique focused on its chemistry and therapeutic activity in various cancer cells. [ABSTRACT FROM AUTHOR]

IntroductionThe Nerium oleander plant contains cardenolides that may cause human poisoning when ingested. A long-standing belief holds that it is possible to be poisoned by eating hot dogs or other foods cooked on Nerium oleander branch skewers. Oleandrin levels in frankfurters cooked on fresh and dry Nerium oleander skewers were measured.MethodsHot dogs were cooked separately on either dried or fresh oleander branch skewers using a disposable charcoal grill. The hot dogs were then frozen and transported to an analytical laboratory where oleandrin content was measured via liquid chromatography/mass spectroscopy (LC/MS).ResultsThe oleandrin content of hot dogs cooked on dried and fresh skewers did not exceed 343 ng and 701 ng, respectively.ConclusionHot dogs cooked on Nerium oleander skewers contain a negligible amount of oleandrin with respect to that sufficient to cause human poisoning. Reports of poisonings occurring in this manner are most likely the result of an urban myth.

The presence of secondary metabolites in flower nectar can mediate interactions between plants, pollinators, herbivores, and microbes. Milkweeds range in concentrations of cardenolides in flower nectar from ~ 1 to 100 ng μl–1. Using three different behavioral assays with bumblebees Bombus impatiens, we examined the impacts of the commercially available cardenolide ouabain at the range of concentrations at which cardenolides naturally occur in milkweeds. We show that after four days of exposure bees in consumption assays drank more of a nectar solution with a low ouabain concentration of 10 ng μl–1 than the control sucrose nectar, and over the course of the experiment bees consumed less of the 100 ng μl–1 ouabain solution than the control. Bee activity levels in Petri dish arena assays were not impacted by ouabain consumption, even at the highest concentrations; however, in free‐flying choice assays, bees preferentially visited artificial flowers containing 10 ng μl–1 ouabain more than flowers with sucrose control, or flowers with 100 ng μl–1 of ouabain. We therefore conclude that cardenolides may provide plants advantages to pollination at the low end of the naturally occurring range of concentrations for cardenolides, but may be costly to plants at the high end of the range. This research highlights that secondary metabolites in nectar may be under selection to be maintained at low concentrations, and species with high concentrations of secondary metabolites may be under conflicting selection pressures to maintain high circulating levels to deter herbivores at the cost of reduced pollination. [ABSTRACT FROM AUTHOR]

Cardiac glycosides (CGs) are a class of bioactive organic compounds well-known for their application in treating heart disease despite a narrow therapeutic window. Considerable evidence has demonstrated the potential to repurpose CGs for cancer treatment. Chemical modification of these CGs has been utilized in attempts to increase their anti-cancer properties; however, this has met limited success as their mechanism of action is still speculative. Recent studies have identified the DNA damage response (DDR) pathway as a target of CGs. DDR serves to coordinate numerous cellular pathways to initiate cell cycle arrest, promote DNA repair, regulate replication fork firing and protection, or induce apoptosis to avoid the survival of cells with DNA damage or cells carrying mutations. Understanding the modus operandi of cardiac glycosides will provide critical information to better address improvements in potency, reduced toxicity, and the potential to overcome drug resistance. This review summarizes recent scientific findings of the molecular mechanisms of cardiac glycosides affecting the DDR signaling pathway in cancer therapeutics from 2010 to 2022. We focus on the structural and functional differences of CGs toward identifying the critical features for DDR targeting of these agents. [ABSTRACT FROM AUTHOR]

Plant toxicity shapes the dietary choices of herbivores. Especially when herbivores sequester plant toxins, they may experience a trade-off between gaining protection from natural enemies and avoiding toxicity. The availability of toxins for sequestration may additionally trade off with the nutritional quality of a potential food source for sequestering herbivores. We hypothesized that diet mixing might allow a sequestering herbivore to balance nutrition and defence (via sequestration of plant toxins). Accordingly, here we address diet mixing and sequestration of large milkweed bugs (Oncopeltus fasciatus) when they have differential access to toxins (cardenolides) in their diet. In the absence of toxins from a preferred food (milkweed seeds), large milkweed bugs fed on nutritionally adequate non-toxic seeds, but supplemented their diet by feeding on nutritionally poor, but cardenolide-rich milkweed leaf and stem tissues. This dietary shift corresponded to reduced insect growth but facilitated sequestration of defensive toxins. Plant production of cardenolides was also substantially induced by bug feeding on leaf and stem tissues, perhaps benefitting this cardenolide-resistant herbivore. Thus, sequestration appears to drive diet mixing in this toxic plant generalist, even at the cost of feeding on nutritionally poor plant tissue. [ABSTRACT FROM AUTHOR]

Plant secondary metabolites that defend leaves from herbivores also occur in floral nectar. While specialist herbivores often have adaptations providing resistance to these compounds in leaves, many social insect pollinators are generalists, and therefore are not expected to be as resistant to such compounds. The milkweeds, Asclepias spp., contain toxic cardenolides in all tissues including floral nectar. We compared the concentrations and identities of cardenolides between tissues of the North American common milkweed Asclepias syriaca, and then studied the effect of the predominant cardenolide in nectar, glycosylated aspecioside, on an abundant pollinator. We show that a generalist bumblebee, Bombus impatiens, a common pollinator in eastern North America, consumes less nectar with experimental addition of ouabain (a standard cardenolide derived from Apocynacid plants native to east Africa) but not with addition of glycosylated aspecioside from milkweeds. At a concentration matching that of the maximum in the natural range, both cardenolides reduced activity levels of bees after four days of consumption, demonstrating toxicity despite variation in behavioral deterrence (i.e., consumption). In vitro enzymatic assays of Na+/K+-ATPase, the target site of cardenolides, showed lower toxicity of the milkweed cardenolide than ouabain for B. impatiens, indicating that the lower deterrence may be due to greater tolerance to glycosylated aspecioside. In contrast, there was no difference between the two cardenolides in toxicity to the Na+/K+-ATPase from a control insect, the fruit fly Drosophila melanogaster. Accordingly, this work reveals that even generalist pollinators such as B. impatiens may have adaptations to reduce the toxicity of specific plant secondary metabolites that occur in nectar, despite visiting flowers from a wide variety of plants over the colony's lifespan. [ABSTRACT FROM AUTHOR]

Phytochemical investigation of the aerial parts and latex of Pergularia tomentosa from Jordan, utilizing column chromatography accompanied with NMR (1 & 2D), IR, UV, and HR-ESI-Mass spectroscopy have led to the isolation and characterization of two new compounds, pergularol (1) and 3-O-acetyl-28-hydroxytaraxasterol (2) for the first time along with other ten known compounds including linoleic acid (3), 3'-didehydroafroside (4), apigenin (5), β-sitosteryl glucoside (6), luteolin (7), apigenin 7-(6''-crotonoyl)glucoside (8), 3'-O-β-glucopyranosylcalotropin (9), apigenin-7-O-β-D-glycoside (10), pergularine A (11), taraxasterol (12). Compound (8) is isolated for the first time from Pergularia genus. [ABSTRACT FROM AUTHOR]

Plants in the genus Erysimum produce both glucosinolates and cardenolides as a defense mechanism against herbivory. Two natural isolates of Erysimum cheiranthoides (wormseed wallflower) differed in their glucosinolate content, cardenolide content, and their resistance to Myzus persicae (green peach aphid), a broad generalist herbivore. Both classes of defensive metabolites were produced constitutively and were not further induced by aphid feeding. To investigate the relative importance of glucosinolates and cardenolides in E. cheiranthoides defense, we generated an improved genome assembly, genetic map, and segregating F2 population. The genotypic and phenotypic analysis of the F2 plants identified quantitative trait loci, which affected glucosinolates and cardenolides, but not the aphid resistance. The abundance of most glucosinolates and cardenolides was positively correlated in the F2 population, indicating that similar processes regulate their biosynthesis and accumulation. Aphid reproduction was positively correlated with glucosinolate content. Although the overall cardenolide content had little effect on aphid growth and survival, there was a negative correlation between aphid reproduction and helveticoside abundance. However, this variation in defensive metabolites could not explain the differences in aphid growth on the two parental lines, suggesting that processes other than the abundance of glucosinolates and cardenolides have a predominant effect on aphid resistance in E. cheiranthoides.

As a general rule, plants defend against herbivores with multiple traits. The defense synergy hypothesis posits that some traits are more effective when co‐expressed with others compared to their independent efficacy. However, this hypothesis has rarely been tested outside of phytochemical mixtures, and seldom under field conditions. We tested for synergies between multiple defense traits of common milkweed (Asclepias syriaca) by assaying the performance of two specialist chewing herbivores on plants in natural populations. We employed regression and a novel application of random forests to identify synergies and antagonisms between defense traits. We found the first direct empirical evidence for two previously hypothesized defense synergies in milkweed (latex by secondary metabolites, latex by trichomes) and identified numerous other potential synergies and antagonisms. Our strongest evidence for a defense synergy was between leaf mass per area and low nitrogen content; given that these "leaf economic" traits typically covary in milkweed, a defense synergy could reinforce their co‐expression. We report that each of the plant defense traits showed context‐dependent effects on herbivores, and increased trait expression could well be beneficial to herbivores for some ranges of observed expression. The novel methods and findings presented here complement more mechanistic approaches to the study of plant defense diversity and provide some of the best evidence to date that multiple classes of plant defense synergize in their impact on insects. Plant defense synergies against highly specialized herbivores, as shown here, are consistent with ongoing reciprocal evolution between these antagonists. [ABSTRACT FROM AUTHOR]

In the struggle against dietary toxins, insects are known to employ target site insensitivity, metabolic detoxification, and transporters that shunt away toxins. Specialized insects across six taxonomic orders feeding on cardenolide-containing plants have convergently evolved target site insensitivity via specific amino acid substitutions in the Na/K-ATPase. Nonetheless, in vitro pharmacological experiments have suggested a role for multidrug transporters (Mdrs) and organic anion transporting polypeptides (Oatps), which may provide a basal level of protection in both specialized and non-adapted insects. Because the genes coding for these proteins are evolutionarily conserved and in vivo genetic evidence in support of this hypothesis is lacking, here we used wildtype and mutant Drosophila melanogaster (Drosophila) in capillary feeder (CAFE) assays to quantify toxicity of three chemically diverse, medically relevant cardenolides. We examined multiple components of fitness, including mortality, longevity, and LD50, and found that, while the three cardenolides each stimulated feeding (i.e., no deterrence to the toxin), all decreased lifespan, with the most apolar cardenolide having the lowest LD50 value. Flies showed a clear non-monotonic dose response and experienced high levels of toxicity at the cardenolide concentration found in plants. At this concentration, both Mdr and Oatp knockout mutant flies died more rapidly than wildtype flies, and the mutants also experienced more adverse neurological effects on high-cardenolide-level diets. Our study further establishes Drosophila as a model for the study of cardenolide pharmacology and solidifies support for the hypothesis that multidrug and organic anion transporters are key players in insect protection against dietary cardenolides.

Insects are incapable of biosynthesising sterols de novo so they need to obtain them from their diets or, in certain cases, from symbiotic microorganisms. Sterols serve a structural role in cellular membranes and act as precursors for signalling molecules and defence compounds. Many phytophagous insects dealkylate phytosterols to yield primarily cholesterol, which is also the main sterol that carnivorous and omnivorous insects obtain in their diets. Some phytophagous species have secondarily lost the capacity to dealkylate and consequently use phytosterols for structural and functional roles. The polyhydroxylated steroid hormones of insects, the ecdysteroids, are derived from cholesterol (or phytosterols in non-dealkylating phytophagous species) and regulate many crucial aspects of insect development and reproduction by means of precisely regulated titres resulting from controlled synthesis, storage and further metabolism/excretion. Ecdysteroids differ significantly from vertebrate steroid hormones in their chemical, biochemical and biological properties. Defensive steroids (cardenolides, bufadienolides, cucurbitacins and ecdysteroids) can be accumulated from host plants or biosynthesised within the insect, depending on species, stored in significant amounts in the insect and released when it is attacked. Other allelochemical steroids serve as pheromones. Vertebrate-type steroids have also been conclusively identified from insect sources, but debate continues about their significance. Side chain dealkylation of phytosterols, ecdysteroid metabolism and ecdysteroid mode of action are targets of potential insect control strategies., (© 2024. All color figures to remain in the eBook and to be printed in color.)

Bioactivity-guided isolation of the CHCl3-soluble fraction of the roots of Calotropis gigantea was carried out to obtain a new cardenolide glycoside, caloside G. Its absolute structure was elucidated based on NMR and ECD spectroscopic data interpretation. Caloside G showed noteworthy cytotoxicity against the PANC-1 human pancreatic and HeLa human cervical carcinoma cell lines, with the submicromolar IC50 values of 0.038 and 0.09 µM, respectively. [ABSTRACT FROM AUTHOR]

[Display omitted] • Three new steroidal sapogenins (1-3) were isolated from the roots of Cynanchum otophyllum. • Compound 1 harbouring a rare beta -OH at the C-8 position in the cardenolide skeleton. • Compound 1 showed the strongest cytotoxicity with IC 50 of 6.86–22.09 μM. Three new steroidal sapogenins, 3 β ,8 β ,14 β -trihydroxycarda-5,20(22)-dienolide (1), (20 R)-12 β ,20-epoxy-3 β ,8 β ,14 β -trihydroxy-17 α -pregn-5-en-17-yl 4-hydroxy-3-methoxybenzoate (2), and 3,20-dioxo-8 β ,14 β ,17 β -trihydroxy-17 α -pregn-5-en-12 β -yl 4-hydroxybenzoate (3), along with four known C 21 -steroidal sapogenins (4-7) were isolated from the hydrolysate of Cynanchum otophyllum roots. Among them, compound 1 harbouring a rare β- OH at the C-8 position in the cardenolide skeleton, was identified for the first time in the genus Cynanchum. Their structures were elucidated via extensive spectroscopic analysis, including UV, IR, HR-ESI-MS, 1D and 2D NMR. Compounds 1-7 were evaluated for their cytotoxic activities against four cancer cell lines (MCF-7, H1299, HeLa, and HepG2). Compound 1 exhibited significant inhibitory effects on the growth of the four cell lines, exhibiting IC 50 values ranging from 6.86 to 22.09 μM. [ABSTRACT FROM AUTHOR]

The crested rat, Lophiomys imhausi , is the only mammal known to sequester plant toxins. Found in eastern Africa, this large rodent is thought to defend against predation by coating specialized hairs along its sides with cardenolide toxins from the poison arrow tree, Acokanthera schimperi. To better understand the ecology of this unusual poisonous mammal, we used camera traps, livetrapping, and captive behavioral observations, to study L. imhausi in central Kenya. Although crested rats were rarely detected with camera traps, 25 individuals were caught in live traps, with estimated densities of up to 15 rats/km2 at one of nine trapping sites. Trapping records and behavioral observations suggest that L. imhausi live in male–female pairs, with juveniles that might exhibit delayed dispersal. We observed chewing of A. schimperi and/or anointing in 10 of 22 individuals, confirming the previous poison sequestration observation. We monitored crested rat activity using cameras and found that chewing on A. schimperi and cardenolide exposure had no effect on feeding, movement, or total activity. One crested rat also fed on milkweed (Gomphocarpus physocarpus ; Gentaniales: Apocynaceae), but did not anoint with this cardenolide containing plant. This observation, combined with L. imhausi 's selective use of A. schimperi , suggests the potential for use of alternative poison sources. This research provides novel insight into the ecology of L. imhausi , while also suggesting that more field observations, feeding trials, and chemical analyses are needed to understand their behavior and physiology. Furthermore, their complex social interactions, slow life history, and fragmented populations suggest that L. imhausi could be at risk of decline. [ABSTRACT FROM AUTHOR]

Erysimum cheiranthoides L (Brassicaceae; wormseed wallflower) accumulates not only glucosinolates, which are characteristic of the Brassicaceae, but also abundant and diverse cardenolides. These steroid toxins, primarily glycosylated forms of digitoxigenin, cannogenol, and strophanthidin, inhibit the function of essential Na+/K+-ATPases in animal cells. We screened a population of 659 ethylmethanesulfonate-mutagenized E. cheiranthoides plants to identify isolates with altered cardenolide profiles. One mutant line exhibited 66% lower cardenolide content, resulting from greatly decreased cannogenol and strophanthidin glycosides, partially compensated for by increases in digitoxigenin glycosides. This phenotype was likely caused by a single-locus recessive mutation, as evidenced by a wildtype phenotype of F1 plants from a backcross, a 3:1 wildtype:mutant segregation in the F2 generation, and genetic mapping of the altered cardenolide phenotype to one position in the genome. The mutation created a more even cardenolide distribution, decreased the average cardenolide polarity, but did not impact most glucosinolates. Growth of generalist herbivores from two feeding guilds, Myzus persicae Sulzer (Hemiptera: Aphididae; green peach aphid) and Trichoplusia ni Hübner (Lepidoptera: Noctuidae; cabbage looper), was decreased on the mutant line compared to wildtype. Both herbivores accumulated cardenolides in proportion to the plant content, with T. ni accumulating higher total concentrations than M. persicae. Helveticoside, a relatively abundant cardenolide in E. cheiranthoides, was not detected in M. persicae feeding on these plants. Our results support the hypothesis that increased digitoxigenin glycosides provide improved protection against M. persicae and T. ni, despite an overall decrease in cardenolide content of the mutant line. [ABSTRACT FROM AUTHOR]

Purpose: Angiogenesis related abnormalities underlie several life-threatening disorders. Despite approved therapies, scientists have yet to develop highly efficient, low cost approaches with minimal side effects. Methods: We evaluated the antiangiogenic activity of 50% hydroalcoholic extracts of Pergularia tomentosa L. root and aerial parts along with their EtOAc and water fractions, in vivo and in vitro. Transgenic zebrafish line Tg(fli1:EGFP) was used for in vivo assay and human umbilical vein endothelial cell (HUVEC) migration test along with possibility of tube formation were performed as in vitro tests. Furthermore, microvasculature in chicken chorioallantoic membrane (CAM) was assessed under P. tomentosa treatment. The fractionation of the 50% hydroalcoholic extracts was led to the identification of the best active fraction in this study. The metabolite profiling of the active fraction was also carried out using LC-HRESIMS analysis. Results: Pergularia tomentosa markedly inhibited intersegmental vessel (ISV) formation at 48 h post-fertilization (hpf) embryos in zebrafish. The water fraction of root hydroalcoholic extract (PtR2), showed strong antiangiogenic effect with minimal adverse viability impacts. Over 80% of embryos showed more than 50% inhibition in their ISV development at 20 and 40 μg/mL. PtR2 at 20 μg/mL substantially reduced human umbilical vein endothelial cell (HUVEC) migration up to 40%, considerable destruction of the formed tubes in the tube formation and microvasculature in CAM assays. Immunocytochemistry showed a marked reduction in vascular endothelial cadherin (VE-cadherin) abundance at cell junctions concurrent with substantial reduction of phospho-Akt (p-Akt) and β-catenin protein expressions. Phytochemical profile of PtR2 showed a rich source of cardenolide structures, including ghalakinoside, calactin and calotropin derivatives. Conclusion: Thus, the P. tomentosa cardenolide-rich fraction (PtR2) may hold a considerable promise for an antiangiogenic impact by impairment of endothelial cell (EC) migration and viability. [ABSTRACT FROM AUTHOR]

Calotropis gigantea produces specialized secondary metabolites known as cardenolides, which have anticancer and antimalarial properties. Although transcriptomic studies have been conducted in other cardenolide-producing species, no nuclear genome assembly for an Asterid cardenolide-producing species has been reported to date. A high-quality de novo assembly was generated for C. gigantea, representing 157,284,427 bp with an N50 scaffold size of 805,959 bp, for which quality assessments indicated a near complete representation of the genic space. Transcriptome data in the form of RNA-sequencing libraries from a developmental tissue series was generated to aid the annotation and construction of a gene expression atlas. Using an ab initio and evidence-driven gene annotation pipeline, 18,197 high-confidence genes were annotated. Homologous and syntenic relationships between C. gigantea and other species within the Apocynaceae family confirmed previously identified evolutionary relationships, and suggest the emergence or loss of the specialized cardenolide metabolites after the divergence of the Apocynaceae subfamilies. The C. gigantea genome assembly, annotation, and RNA-sequencing data provide a novel resource to study the cardenolide biosynthesis pathway, especially for understanding the evolutionary origin of cardenolides and the engineering of cardenolide production in heterologous organisms for existing and novel pharmaceutical applications.

Background: Pergularia tomentosa L. has several traditional uses and biological activities which need to be more investigated. Objective: In this study, some ecological and phytochemical properties of Pergularia tomentosa were assessed. The flavonoids and cardenolides in the aqueous extract of leaves were identified with LCMS techniques. Methods: Some quantity characteristics of P. tomentosa were measured. Extraction was performed according to the traditional method (aqueous extract at room temperature). Metabolite profiling was conducted using untargeted liquid chromatography coupled to high resolution mass spectrometry (LCeESI-OrbitrapeMS). Data were analyzed by Xcalibur version 2.1. Results: Results showed that length of roots, leaves and fruits and the height of aerial parts were 23-30 cm, 2.5-3.2 cm, 4.9- 5.4 cm and 40-58.00 cm, respectively. Physiography and canopy cover were two important variables on these properties. We also identified 19 natural products in aqueous extract, 15 phenolics and flavonoids and 4 cardenolides. Three flavonoids (8, 11 and 14) and one cardenolide (17) had been previously isolated in P. tomentosa and 3 cardenolides (5, 18 and 19) has been reported for the first time in this plant. Conclusion: Results indicated that the leaves of P. tomentosa are a rich source of flavonoids and cardenolides. These findings suggest that leaves of this plant can be a good source of natural antioxidant and antitumor compounds. However, this plant needs special conservational efforts. Therefore, planting this plant under the canopy or along the borders of waterways can help in an appropriate habitat for its conservation and reclamation.

Milkweeds (Asclepias) are used in wide-ranging studies including floral development, pollination biology, plant-insect interactions and co-evolution, secondary metabolite chemistry, and rapid diversification. We present a transcriptome and draft nuclear genome assembly of the common milkweed, Asclepias syriaca. This reconstruction of the nuclear genome is augmented by linkage group information, adding to existing chloroplast and mitochondrial genomic resources for this member of the Apocynaceae subfamily Asclepiadoideae. The genome was sequenced to 80.4× depth and the draft assembly contains 54,266 scaffolds ≥1 kbp, with N50 = 3,415 bp, representing 37% (156.6 Mbp) of the estimated 420 Mbp genome. A total of 14,474 protein-coding genes were identified based on transcript evidence, closely related proteins, and ab initio models, and 95% of genes were annotated. A large proportion of gene space is represented in the assembly, with 96.7% of Asclepias transcripts, 88.4% of transcripts from the related genus Calotropis, and 90.6% of proteins from Coffea mapping to the assembly. Scaffolds covering 75 Mbp of the Asclepias assembly formed 11 linkage groups. Comparisons of these groups with pseudochromosomes in Coffea found that six chromosomes show consistent stability in gene content, while one may have a long history of fragmentation and rearrangement. The progesterone 5β-reductase gene family, a key component of cardenolide production, is likely reduced in Asclepias relative to other Apocynaceae. The genome and transcriptome of common milkweed provide a rich resource for future studies of the ecology and evolution of a charismatic plant family.

Milkweeds (Asclepias) are used in wide-ranging studies including floral development, pollination biology, plant-insect interactions and co-evolution, secondary metabolite chemistry, and rapid diversification. We present a transcriptome and draft nuclear genome assembly of the common milkweed, Asclepias syriaca. This reconstruction of the nuclear genome is augmented by linkage group information, adding to existing chloroplast and mitochondrial genomic resources for this member of the Apocynaceae subfamily Asclepiadoideae. The genome was sequenced to 80.4× depth and the draft assembly contains 54,266 scaffolds ≥1 kbp, with N50 = 3,415 bp, representing 37% (156.6 Mbp) of the estimated 420 Mbp genome. A total of 14,474 protein-coding genes were identified based on transcript evidence, closely related proteins, and ab initio models, and 95% of genes were annotated. A large proportion of gene space is represented in the assembly, with 96.7% of Asclepias transcripts, 88.4% of transcripts from the related genus Calotropis, and 90.6% of proteins from Coffea mapping to the assembly. Scaffolds covering 75 Mbp of the Asclepias assembly formed 11 linkage groups. Comparisons of these groups with pseudochromosomes in Coffea found that six chromosomes show consistent stability in gene content, while one may have a long history of fragmentation and rearrangement. The progesterone 5β-reductase gene family, a key component of cardenolide production, is likely reduced in Asclepias relative to other Apocynaceae. The genome and transcriptome of common milkweed provide a rich resource for future studies of the ecology and evolution of a charismatic plant family. [ABSTRACT FROM AUTHOR]

The repeated evolutionary specialization of distantly related insects to cardenolide-containing host plants provides a stunning example of parallel adaptation. Hundreds of herbivorous insect species have independently evolved insensitivity to cardenolides, which are potent inhibitors of the alpha-subunit of Na+,K+-ATPase (ATPa). Previous studies investigating ATPa-mediated cardenolide insensitivity in five insect orders have revealed remarkably high levels of parallelism in the evolution of this trait, including the frequent occurrence of parallel amino acid substitutions at two sites and recurrent episodes of duplication followed by neo-functionalization. Here we add data for a sixth insect order, Orthoptera, which includes an ancient group of highly aposematic cardenolide-sequestering grasshoppers in the family Pyrgomorphidae. We find that Orthopterans exhibit largely predictable patterns of evolution of insensitivity established by sampling other insect orders. Taken together the data lend further support to the proposal that negative pleiotropic constraints are a key determinant in the evolution of cardenolide insensitivity in insects. Furthermore, analysis of our expanded taxonomic survey implicates positive selection acting on site 111 of cardenolide-sequestering species with a single-copy of ATPa, and sites 115, 118 and 122 in lineages with neo-functionalized duplicate copies, all of which are sites of frequent parallel amino acid substitution. [ABSTRACT FROM AUTHOR]

Natural products possess a significant role in anticancer therapy and many currently-used anticancer drugs are of natural origin. Cerberin (CR), a cardenolide isolated from the fruit kernel of Cerbera odollam, was found to potently inhibit cancer cell growth (GI50 values < 90 nM), colony formation and migration. Significant G2/M cell cycle arrest preceded time- and dose-dependent apoptosis-induction in human cancer cell lines corroborated by dose-and time-dependent PARP cleavage and caspase 3/7 activation, in addition to reduced Bcl-2 and Mcl-1 expression. CR potently inhibited PI3K/AKT/mTOR signalling depleting polo-like kinase 1 (PLK-1), c-Myc and STAT-3 expression. Additionally, CR significantly increased the generation of reactive oxygen species (ROS) producing DNA double strand breaks. Preliminary in silico biopharmaceutical assessment of CR predicted >60% bioavailability and rapid absorption; doses of 1-10 mg/kg CR were predicted to maintain efficacious unbound plasma concentrations (>GI50 value). CR's potent and selective anti-tumour activity, and its targeting of key signalling mechanisms pertinent to tumourigenesis support further preclinical evaluation of this cardiac glycoside. [ABSTRACT FROM AUTHOR]

A major prediction of coevolutionary theory is that plants may target particular herbivores with secondary compounds that are selectively defensive. The highly specialized monarch butterfly (Danaus plexippus) copes well with cardiac glycosides (inhibitors of animal Na+/K+-ATPases) from its milkweed host plants, but selective inhibition of its Na+/K+-ATPase by different compounds has not been previously tested. We applied 17 cardiac glycosides to the D. plexippus-Na+/K+-ATPase and to the more susceptible Na+/K+-ATPases of two non-adapted insects (Euploea core and Schistocerca gregaria). Structural features (e.g., sugar residues) predicted in vitro inhibitory activity and comparison of insect Na+/K+-ATPases revealed that the monarch has evolved a highly resistant enzyme overall. Nonetheless, we found evidence for relative selectivity of individual cardiac glycosides reaching from 4- to 94-fold differences of inhibition between non-adapted Na+/K+-ATPase and D. plexippus-Na+/K+-ATPase. This toxin receptor specificity suggests a mechanism how plants could target herbivores selectively and thus provides a strong basis for pairwise coevolutionary interactions between plants and herbivorous insects.

A phytochemical investigation of the whole plants of Adonis multiflora Nishikawa & Koki Ito. resulted in the isolation and identification of two new cardenolides—adonioside A (1) and adonioside B (6)—as well as four known cardenolides: tupichinolide (2) oleandrine (3), cryptostigmin II (4), and cymarin (5). Their structures were elucidated on the basis of NMR, MS, and IR spectroscopic analyses. Compounds 1, 2, 5, and 6 showed significant cytotoxicity against six human cancer cell lines (HCT-116, HepG2, HeLa, SK-OV-3, and SK-MEL-5, and SK-BR-3).

Graphical abstract Highlights • SpHL-GEVPG showed good stability over 30 days at 4 °C. • SpHL-GEVPG treatment showed higher cytotoxicity than doxorubicin treatment against MDA-MB-231 and SKBR3. • SpHL-GEVPG treatment at 27 nM was able to inhibit approximately 75% of formation of A549 lung cancer cells colonies. • SpHL-GEVPG showed to be 5 – 10 times more potent than paclitaxel in A549 lung tumor bearing-Balb/C nude female mice. Abstract Cancer is an important public health problem, being one of the leading causes of death worldwide. Most antineoplastic agents cause severe toxic effects and some types of cancer do not respond or are resistant to the existing pharmacotherapy, necessitating the research and development of new therapeutic strategies. Cardenolides have shown significant antitumor activity due to their ability to inhibit the Na+K+ATPase enzyme, and the expression of this enzyme is increased in tumor cells. Glucoevatromonoside containing peracetylated glucose hydroxyl groups (GEVPG) is a cardenolide derivative that has low solubility in aqueous media, which constitutes a barrier to its potential biological applications. In this context, the use of liposomes represents a promising strategy to deliver GEVPG, thus allowing its intravenous administration. In this study, long-circulating and fusogenic liposomes containing GEVPG (SpHL-GEVPG) were developed, and their chemical and physicochemical properties were evaluated. SpHL-GEVPG presented adequate properties, including a mean diameter of 182.2 ± 2.7 nm, a polydispersity index equal to 0.36 ± 0.03, a zeta potential of –2.37 ± 0.31 mV, and a GEVPG entrapment of 0.38 ± 0.04 mg/mL. Moreover, this formulation showed a good stability after having been stored for 30 days at 4 °C. The cytotoxic studies against breast (MDA-MB-231, MCF-7, and SKBR-3) and lung (A549) cancer cell lines demonstrated that SpHL-GEVPG treatment significantly reduced the cell viability. In addition, the SpHL-GEVPG formulation presented a good selectivity toward these cancer cells. The evaluation of the therapeutic efficacy of the treatment with SpHL-GEVPG showed a potent anticancer effect in an A549 human lung cancer xenograft model. SpHL-GEVPG administered at doses of 1.0 and 2.0 mg/kg (i.v.) induced antitumor effect comparable to paclitaxel given at dose of 10 mg/kg (i.v.) to mice. Therefore, the results of the present work indicate the potential applicability of SpHL-GEVPG as a new anticancer formulation. [ABSTRACT FROM AUTHOR]

At least twelve plant families contain species that synthesize cardiac glycosides as defense against herbivory. These inhibitors of animal Na+, K+-ATPases also have medical uses in treating congestive heart failure and other diseases. However, despite extensive ecological research and centuries of use in both traditional and modern medicine, the complete cardiac glycoside biosynthesis pathway has yet to be elucidated in any plant species. To a large extent, this research deficit results from the fact that cardiac glycosides are produced exclusively by non-model plant species such as Digitalis that have not been amenable to the development of mutagenesis, cloning, and genetic mapping approaches. Recent advances in genome sequencing, transcript profiling, plant transformation, transient expression assays, and plant metabolite analysis have provided new opportunities for the investigation and elucidation of cardiac glycoside biosynthesis pathways. The genetic tools that have been developed for Brassicaceae, in particular Arabidopsis thaliana, may be directly applicable to Erysimum, a Brassicaceae genus that characteristically produces cardiac glycosides as defensive metabolites. We propose that Erysimum cheiranthoides (wormseed wallflower), a rapid-cycling, self-pollinating species with a relatively small, diploid genome, would be a suitable model system to advance research on the biosynthesis of cardiac glycosides in plants. [ABSTRACT FROM AUTHOR]

Abstract Cardenolides are plant-derived toxic substances. Their cytotoxicity and the underlying mechanistic signaling axes have been extensively documented, but only a few anti-viral activities of cardenolides and the associated signaling pathways have been reported. Previously, we reported that a variety of cardenolides impart anti-transmissible gastroenteritis coronavirus (TGEV) activity in swine testicular (ST) cells, through targeting of the cell membrane sodium/potassium pump, Na+/K+-ATPase. Herein, we further explore the potential signaling cascades associated with this anti-TGEV activity in ST cells. Ouabain, a representative cardenolide, was found to potently diminish TGEV titers and inhibit the TGEV-induced production of IL-6 in a dose dependent manner, with 50% inhibitory concentrations of 37 nM and 23 nM respectively. By pharmacological inhibition and gene silencing, we demonstrated that PI3K_PDK1_RSK2 signaling was induced in TGEV-infected ST cells, and ouabain imparted a degree of anti-TGEV activity via further augmentation of this existing PI3K_PDK1 axis signaling, in a manner dependent upon its association with the Na+/K+-ATPase. Finally, inhibition of PI3K by LY294002 or PDK1 by BX795 antagonized the anti-viral activity of ouabain and restored the TGEV virus titer and yields. This finding is the first report of a PI3K_PDK1 signaling axis further induced by ouabain and implicated in the suppression of TGEV activity and replication; greatly illuminates the underlying mechanism of cardenolide toxicity; and is expected to result in one or more anti-viral applications for the cardenolides in the future. Graphical abstract Unlabelled Image Highlights • Ouabain eliminated TGEV titers and inhibited viral replication. • Ouabain diminished TGEV induced IL-6 production. • Ouabain enhanced PI3K or PDK1 activation induced by TGEV via Na+/K+-ATPase. • PI3K or PDK1 inhibition antagonized the anti-TGEV activity of ouabain. • Ouabain augmented the PI3K_PDK1 axis signaling that inhibited TGEV activity. [ABSTRACT FROM AUTHOR]

A major prediction of coevolutionary theory is that plants may target particular herbivores with secondary compounds that are selectively defensive. The highly specialized monarch butterfly (Danaus plexippus) copes well with cardiac glycosides (inhibitors of animal Na+/K+-ATPases) from its milkweed host plants, but selective inhibition of its Na+/K+-ATPase by different compounds has not been previously tested. We applied 17 cardiac glycosides to the D. plexippus -Na+/K+-ATPase and to the more susceptible Na+/K+-ATPases of two non-adapted insects (Euploea core and Schistocerca gregaria). Structural features (e.g., sugar residues) predicted in vitro inhibitory activity and comparison of insect Na+/K+-ATPases revealed that the monarch has evolved a highly resistant enzyme overall. Nonetheless, we found evidence for relative selectivity of individual cardiac glycosides reaching from 4- to 94-fold differences of inhibition between non-adapted Na+/K+-ATPase and D. plexippus -Na+/K+-ATPase. This toxin receptor specificity suggests a mechanism how plants could target herbivores selectively and thus provides a strong basis for pairwise coevolutionary interactions between plants and herbivorous insects. [ABSTRACT FROM AUTHOR]

Cerbera odollam is a plant species of the Apocynaceae family. It is often dubbed the 'suicide tree' due to its strong cardiotoxic effects, which make it a suitable means to attempt suicide. The plant grows in wet areas in South India, Madagascar, and Southeast Asia; and its common names include Pong-Pong and Othalanga. The poison rich part of the plant is the kernel which is present at the core of its fruit. The bioactive toxin in the plant is cerberin, which is a cardiac glycoside of the cardenolide class. Cerberin has a mechanism of action similar to digoxin; hence, Cerbera odollam toxicity manifests similar to acute digoxin poisoning. Ingestion of its kernel causes nausea, vomiting, hyperkalemia, thrombocytopenia, and ECG abnormalities. Exposure to high doses of Cerbera odollam carries the highest risk of mortality. Initial management includes supportive therapy and administration of atropine followed by temporary pacemaker insertion. Administration of digoxin immune Fab may be considered in severe cases, although efficacy is variable and data limited to isolated case reports. [ABSTRACT FROM AUTHOR]

Abstract: Antipredator defense of herbivorous insects often relies on the potential toxicity of defensive chemicals sequestered from their host plants. The colorful Lygaeinae (Heteroptera: Lygaeidae) store a concentrated mixture of toxic cardenolides (cardiac glycosides) in specialized storage compartments of the bugs' integument, from which they are released upon attack. Larvae and adults of the large milkweed bug Oncopeltus fasciatus (Heteroptera: Lygaeinae) are specialized to feed on cardenolide‐containing milkweeds in the plant genus Asclepias and display a conspicuous red and black colorations. To investigate whether O. fasciatus gained improved protection by feeding on a toxic host plant (Asclepias syriaca), compared to a nontoxic alternative (sunflower seeds), we fed nymphs and adults of O. fasciatus to the golden orb‐weaver Nephila senegalensis. While visually oriented vertebrates, such as avian predators, have been intensively investigated for their reaction to defensive compounds and aposematic coloration, less attention has been paid to invertebrate predators. Their different perceptual abilities can provide important opportunities for testing hypotheses on warning coloration and chemical defenses. The predation trials showed that the bugs fed on Asclepias were significantly less likely to be killed than the bugs reared on a cardenolide‐free diet. This suggests that sequestered cardenolides in O. fasciatus nymphs and adults represent a significant fitness advantage on an individual level against this invertebrate predator. Yet, when testing for avoidance learning in the spiders, negative experience did not change the way how similar prey was attacked at the next encounter. In this case, visual or chemical aposematism thus does not seem to matter for predator learning. [ABSTRACT FROM AUTHOR]

Abstract: While it is well established that climate change affects species distributions and abundances, the impacts of climate change on species interactions has not been extensively studied. This is particularly important for specialists whose interactions are tightly linked, such as between the monarch butterfly (Danaus plexippus) and the plant genus Asclepias, on which it depends. We used open‐top chambers (OTCs) to increase temperatures in experimental plots and placed either nonnative Asclepias curassavica or native A. incarnata in each plot along with monarch larvae. We found, under current climatic conditions, adult monarchs had higher survival and mass when feeding on A. curassavica. However, under future conditions, monarchs fared much worse on A. curassavica. The decrease in adult survival and mass was associated with increasing cardenolide concentrations under warmer temperatures. Increased temperatures alone reduced monarch forewing length. Cardenolide concentrations in A. curassavica may have transitioned from beneficial to detrimental as temperature increased. Thus, the increasing cardenolide concentrations may have pushed the larvae over a tipping point into an ecological trap; whereby past environmental cues associated with increased fitness give misleading information. Given the ubiquity of specialist plant–herbivore interactions, the potential for such ecological traps to emerge as temperatures increase may have far‐reaching consequences. [ABSTRACT FROM AUTHOR]

Premise of the Study: Pachypodium (Apocynaceae) is a genus of iconic stem‐succulent and poisonous plants endemic to Madagascar and southern Africa. We tested hypotheses about the mode of action and macroevolution of toxicity in this group. We further hypothesized that while monarch butterflies are highly resistant to cardenolide toxins (a type of cardiac glycoside) from American Asclepias, they may be negatively affected by Pachypodium defenses, which evolved independently. Methods: We grew 16 of 21 known Pachypodium spp. and quantified putative cardenolides by HPLC and also by inhibition of animal Na+/K+‐ATPase (the physiological target of cardiac glycosides) using an in vitro assay. Pachypodium extracts were tested against monarch caterpillars in a feeding bioassay. We also tested four Asclepias spp. and five Pachypodium spp. extracts, contrasting inhibition of the cardenolide‐sensitive porcine Na+/K+‐ATPase to the monarch's resistant form. Key Results: We found evidence for low cardenolides by HPLC, but substantial toxicity when extracts were assayed on Na+/K+‐ATPases. Toxicity showed phylogenetic signal, and taller species showed greater toxicity (this was marginal after phylogenetic correction). Application of Pachypodium extracts to milkweed leaves reduced monarch growth, and this was predicted by inhibition of the sensitive Na+/K+‐ATPase in phylogenetic analyses. Asclepias extracts were 100‐fold less potent against the monarch compared to the porcine Na+/K+‐ATPase, but this difference was absent for Pachypodium extracts. Conclusions: Pachypodium contains potent toxicity capable of inhibiting sensitive and cardenolide‐adapted Na+/K+‐ATPases. Given the monarch's sensitivity to Pachypodium, we suggest that these plants contain novel cardiac glycosides or other compounds that facilitate toxicity by binding to Na+/K+‐ATPases. [ABSTRACT FROM AUTHOR]

A series of naturally occurring cardenolides that exhibit potent anti-transmissible gastroenteritis virus (TGEV) activity in swine testicular (ST) cells has been identified. In an immunofluorescence assay, these cardenolides were found to diminish the expressions of TGEV nucleocapsid and spike protein, which was used as an indication for viral replication; block TGEV infection induced apoptosis and cytopathic effects; and impart the same trend of inhibitory activity against Na + /K + -ATPase as for anti-TGEV activity. The viral titer inhibition was found to take place in a dose-dependent manner. Knocking down expression of Na + /K + -ATPase, the cellular receptor of cardenolides, in ST cells was found to significantly impair the susceptibility of ST cells to TGEV infectivity. Thus, we have identified Na + /K + -ATPase as an anti-viral drug target and its antagonists, cardenolides, a novel class of anti- TGEV agents. [ABSTRACT FROM AUTHOR]

Prey have evolved a number of defenses against predation, and predators have developed means of countering these protective measures. Although caterpillars of the monarch butterfly, Danaus plexippus L., are defended by cardenolides sequestered from their host plants, the Chinese mantid Tenodera sinensis Saussure guts the caterpillar before consuming the rest of the body. We hypothesized that this gutting behavior might be driven by the heterogeneous quality of prey tissue with respect to toxicity and/or nutrients. We conducted behavioral trials in which mantids were offered cardenolide-containing and cardenolide-free D. plexippus caterpillars and butterflies. In addition, we fed mantids starved and unstarved D. plexippus caterpillars from each cardenolide treatment and nontoxic Ostrinia nubilalis Hü bner caterpillars. These trials were coupled with elemental analysis of the gut and body tissues of both D. plexippus caterpillars and corn borers. Cardenolides did not affect mantid behavior: mantids gutted both cardenolide-containing and cardenolide-free caterpillars. In contrast, mantids consumed both O. nubilalis and starved D. plexippus caterpillars entirely. Danaus plexippus body tissue has a lower C:N ratio than their gut contents, while O. nubilalis have similar ratios; gutting may reflect the mantid's ability to regulate nutrient uptake. Our results suggest that post-capture prey processing by mantids is likely driven by a sophisticated assessment of resource quality. [ABSTRACT FROM AUTHOR]