Behavioral responses of male lobsters to Ecdysones

Four ecdysones with molting hormone activity, β‐ecdysone, inokosterone, ponasterone A, and cyasterone were tested for their capacity to elicit behavioral responses, especially sexual responses in the male lobster (Homarus americanus). Of these ecdysones, only β‐ecdysone elicited a low key alerting response. However, no particular type of behavior, such as feeding, aggressive or sexual behavior could be observed. A role of β‐ecdysone as a crustacean sex pheromone is not supported by these experiments and the general validity of this notion is discussed.     

Concentration‐dependent chemosensory responses towards pheromones are influenced by receiver attributes in Asian elephants

The physical structure of a signal is not sufficient to determine its meaning. For chemical signals between conspecifics, this concept is termed “pheromonal parsimony.” The function of a compound depends not only on its molecular structure but also on its context, which can include signal concentration and various receiver attributes. We sought to investigate the contextual flexibility of chemosensory responses through bioassays with Asian elephant (Elephas maximus) sex pheromones of various concentrations (frontalin, from males, and (Z)‐7‐dodecenyl acetate [Z7‐12:Ac], from females). We hypothesized that elephants would respond stronger to higher concentrations, especially towards the opposite‐sex pheromone, and that receiver age and sexual experience would modify responses. We examined responses of 73 captive elephants to both compounds. Pheromone concentration impacted the rate of chemosensory response, which was further modified by the sex, age and/or sexual experience of the receiver. Response rates increased with concentration for each compound across both sexes. Experience shaped male responses with older, physiologically primed males responding more often. The interaction between experience and age affected female response to frontalin, but not to Z7‐12:Ac. Furthermore, response thresholds were modified by sexual experience in most cases: experienced animals generally had lower thresholds than inexperienced animals. Elephants responded to each solution according to its perceived relevance, including concentration. These results also indicate that receiver attributes (e.g., sex, age and sexual experience) may modify seemingly fixed chemosensory responses and further emphasize the flexibility of vertebrate communication systems.     

Behavioral responses of maleHeliothis armigera (Lepidoptera: Noctuidae) moths in a flight tunnel to combinations of components identified from female sex pheromone glands

Six compounds were identified from gland extracts of the cotton bollworm, Heliothis armigera(Hubner): (Z)-11-hexadecenal (Z11-16:Ald), (Z)-9-hexa-decenal (Z9-16:Ald), hexadecanal, (Z)-11-hexadecenol (Z11-16:OH), (Z)-7-hexadecenal (Z7-16:Ald), and (Z)-9-tetradecenal (Z9-14:Ald). Each of the compounds that were identified was examined for its ability to elicit sexual responses from male moths in a flight tunnel. Males flew upwind to Z11-16:Ald alone, but greater levels of copulatory responses were evoked with the addition of 2.5% Z9-16:Ald to the Z11-16:Ald. Addition of hexadecanal to the binary mixture had no effect in raising the behavioral response of the males in the flight tunnel. The effect of Z7-16:Ald on male flight depended on the loading. The addition of 1% of this component to 2 mg of the binary mixture reduced levels of copulatory response, but the same addition (1 %) to 10 g of the binary mixture increased copulatory response. The addition of 79-14:Ald or Z11-16:OH to the binary mixture reduced behavioral responses of males. High loadings of the binary mixture (200–2000 g) were better than a low loading (10 g) in eliciting response of males.Contribution from the Agricultural Research Organization, The Volcani Center, Bet Dagan, Israel. No. 2455-E, 1988 series.     

A Diterpene Endoperoxide from Microtoena insuavis （Hance） Prain ex Dunn

A novel endoperoxlde diterpene, 7a-hydroxy-abieta-8（14）-en-18-oi 9α,13α-endoperoxide （compound 1）, was isolated from the stems of Microtoena insuavis （Hance） Prain ex Dunn, along with 4,4＇-dlhydroxytruxillic acid （compound 2）, gallic acid （compound 3）, ellaglc acid （compound 4）, 3-O-methylellaglc acid 3＇-O-α- rhamnopyranoslde （compound 5）, 3＂＇-O-methylcrenatoslde （compound 6）, crenatoslde （compound 7）, aptgenin （compound 8）, succinic acid （compound 9）, β-sitosterot （compound 10）, and β-daucosterol （compound 11）. The structures of these compounds were elucidated on the basis of spectral evidence.     

Regulation of Crustacean Molting: A Multi-Hormonal System

SYNOPSIS. In order to increase in size, arthropods must firstmolt (shed) their confining exoskeleton. This molting processis under the immediate control of the steroid molting hormone20-hydroxyecdysone (20-HE). Both the initial rise in circulatinghormone concentration and a coordinated decline are necessaryfor successful molting. Synthesis and/or release of ecdysone,the precursor to 20-HE, is regulated by the neuropeptide molt-inhibitinghormone (MIH). We have determined the primary amino acid sequenceof MIH in the lobster, Homarus americanus. This peptide hasa high degree of identity with the lobster hyperglycemic hormone.Another endocrine factor that appears to be involved in moltingis the juvenile hormone-like terpenoid methyl farnesoate (MF).We have characterized hemolymph MF binding proteins during themolt cycle. In addition, recent data indicate that MF may stimulatethe secretion of 20-HE in vivo and in vitro.     

Photoperiodic regulation of seasonal responses in Indian weaver bird (Ploceus philippinus)

Photoperiod (=day length) is the vital factor for the regulation of behavioral and physiological activities in many avian species. This study investigated the seasonal cycles of testicular growth and secondary sexual characteristics of Indian weaver bird under natural day length (NDL) and the effects of duration and intensity of light on photoperiodic induction. In the first experiment, groups of birds (n = 7 each) were exposed to under NDL in April 2008 and May 2009 for 8 and 12 months, respectively. In second and third experiment, birds (n = 6 each group) were exposed to different photoperiods (11.5L:12.5D, 12L:12D, 13L:11D, and 15L:9D) at the same (500 lux) light intensity, and to 13L:11D at different light intensities (10-, 50-, 500-, and 800-lux). Observations on testis size, molt, and plumage score were recorded 2-week (molt and plumage) or at 4-week intervals (testes). Both the NDL groups showed similar seasonal cycles of testicular growth-regression and secondary sexual characteristics. Second and third experiments suggest that the photoperiodic induction was depending upon duration and intensity of the light. Birds showed testicular growth-regression cycle followed by molt and plumage color change only under 13L:11D and 15L:9D and only 500- and 800-lux under 13L:11D photoperiod but not under 11.5L:12.5D and 12L:12D and 10- and 50-lux light intensities. Pre- and post-nuptial molting on body feathers were progressed with gonadal stimulation–maturation and regression cycle under 13L:11D and 15L:9D. Results under different light–dark cycles suggest that day length of about 12 h or more and above the threshold level of light intensity are essential for the induction of photoperiodic responses.     

Water-Soluble Constituents of Cudrania tricuspidata （Carr.） Bur.

In order to find new structural and biologically active compounds, the constituents of the bark of Cudrania tricuspidata （Carr.） Bur. were investigated and a new 6-p-hydroxybenzyltaxifolin glucoside, named tricusposide （compound 1）, together with 16 known compounds, was isolated by solvent partition, macroporous adsorption resin AB-8, silica gel, Sephadex LH-20 chromatography. Using spectroscopic methods, the structures of the compounds were elucidated as 6-p-hydroxybenzyl taxifolin-7-O-β-D-glucoside （compound 1）, dihydroquerctin-7-O-β-D-glucoside （compound 2）, dihydrokaempferol-3-O-β-D-glucoside （compound 3）, dihydroquercetin （compound 4）, peonoside （compound 5）, sphaerobioside （compound 6）, quercimeritrin （compound 7）, genistein （compound 8）, aromadendrin （compound 9）, kaempferol （compound 10）, genistin （compound 11）, 3,4-dihydroxystyryl alcohol （compound 12）, sucrose （compound 13）, 1,3,5,6-tetrahydroxyxanthone （compound 14）, gericudranin E （compound 15）, gericudranin C （compound 16）, and orobol （compound 17）. Compounds 2-6, 8, 9, 12-14, and 17 were isolated from this genus for the first time.     

Simultaneous choice bioassays accompanied by physiological changes identify civetone and decanoic acid as pheromone candidates for giant pandas

Chemicals present in urine are thought to play an important role in mate identification in the solitary giant panda (Ailuropoda melanoleuca). During the breeding season, females will deposit chemical signals to advertise sexual receptivity to potential mates. The goal of this study was to determine if specific volatile compounds found in female urine could be considered as pheromones that elicit behavioral and physiological responses in males. Experimental simultaneous choice trials were conducted with captive male giant pandas (n = 3) housed at Memphis Zoo, San Diego Zoo, and Zoo Atlanta. Octanoic acid, 1H-pyrrole-2-carboxaldehyde, decanoic acid, and civetone were selected as stimuli because previous studies reported their elevation in urine during the breeding season. Male interest was determined by a behavioral preference toward these volatile compounds diluted in synthetic urine compared with nontreated synthetic urine. Male urine samples were collected 1 week prior, during, and 1 week after the experimental period to assess changes in urinary semiochemical composition and urinary androgen concentrations. No significant differences in investigation response (p = .395) or flehmen response (p = .600) were found when stimuli were compared; however, decanoic acid and civetone elicited a behavioral preference over the control (response ratio > 0.5). The relative abundance of 16 compounds identified in male urine was significantly elevated (p < .05) above baseline values after the males were exposed to the stimuli. Androgen levels were significantly elevated (p < .05) in one male after exposure to 1H-pyrrole-2-carboxaldehyde, decanoic acid, and civetone. These data suggested that civetone and decanoic acid in female urine may motivate sexual responses in males.     

Tyrosinase inhibitory components from Aloe vera and their antiviral activity

A new compound, 9-dihydroxyl-2'-O-(Z)-cinnamoyl-7-methoxy-aloesin (1), and eight known compounds (2–9) were isolated from Aloe vera. Their structures were elucidated using 1D/2D nuclear magnetic resonance and mass spectra. Compound 9 exhibited reversible competitive inhibitory activity against the enzyme tyrosinase, with an IC₅₀ value of 9.8?±?0.9?µM. A molecular simulation revealed that compound 9 interacts via hydrogen bonding with residues His244, Thr261, and Val283 of tyrosinase. Additionally, compounds 3 and 7 were shown by half-leaf assays to exhibit inhibitory activity towards Pepper mild mottle virus.     

RESPONSE OF PARASITOID MICROPLITIS MEDIATOR TO PLANT VOLATILES IN AN OLFACTOMETER*

Abstract The behavioral responses of Microplitis mediator were measured in a four‐armed olfactometer. Leaves extract of the suitable host plant of its insect host, cotton, elicited higher olfactory responses; while leaves extract of the less suitable host plant of its insect host, tomato or tobacco, elicited moderate responses. Volatiles from the least suitable host plant of its insect host, hot pepper, elicited the lowest responses. The different preferences to four plant leaves extracts suggested that M. mediator could discriminate between suitable and less suitable host plants of its insect host by semiochemicals. Green leaf volatiles (GLVs), cis‐3‐hexen‐1‐yl acetate, hexanal, and nonan‐1‐ol, are the most attractive compound among the 7 tested compounds. The parasitoids preferred host‐damaged plant to undamaged plant, suggesting that they were able to distinguish between host and non‐host of their insect host through varying chemical blends. Experience with cotton leaves extract prior to experiment enhanced their responses to the same odor.     

The significance of blends of two sex pheromone components on the behavior of maleSpodoptera litura F. (Lepidoptera: Noctuidae)

Behavioral responses of maleSpodoptera litura to the female sex pheromone components, (Z, E)-9, 11-tetradecadienyl acetate (compound A) and (Z, E)-9, 12-tetradecadienyl acetate (compound B), and mixtures of compounds A and B were analyzed in the laboratory. Male orientation flight in a wind tunnel was induced by a lure dispenser onto which 1.1×10⁻³ ng of a mixture (A∶B=10∶1) was absorbed, while 1 ng of compound A was required to induced the same level of behavior. The blend ratio was important for trap catch and orientation flight of males and a 10∶1 mixture of compounds A and B was most effective. A mixture (10∶1) was more effective than compound A alone in induction of movement, walking, and flying in resting males in a glass tube. These results indicated that 2 sex pheromone components act as a set from the initial to final steps of male attraction to female.     

多炔类化合物对亚洲玉米螟产卵驱避作用及玉米螟的触角电位反应

以茵陈二炔为结构母体,人工合成了11个多炔类化合物。采用蜡纸接卵法,测定了11 个化合物对亚洲玉米螟Ostrinia furnacalis的产卵驱避活性。结果表明: 当处理浓度为20 μg/cm²时,受试的化合物对亚洲玉米螟的产卵行为具有一定程度的驱避作用,其中化合物9（1-苯基-4-甲基-丁二炔）对亚洲玉米螟产卵驱避作用明显, 调查处理后的3天、4天、 5天和6天,其产卵驱避率分别为85.71%、80.00%、61.27%和62.51%。触角电位测定表明,受试的11个化合物对亚洲玉米螟成虫触角感受器具有刺激作用,其中化合物6和化合物9能强烈地刺激产生高振幅的动作电位。10 mg/mＬ浓度处理,测得化合物6触角电位相对值, 雌、雄虫分别为68.22% 和106.60%,化合物9分别为199.19% 和220.60%。经回归分析所测11个化合物的产卵驱避活性与触角电位反应相对值,两者呈现一定程度上的相关性。还讨论了合成的多炔类化合物对亚洲玉米螟可能的作用靶标和结构与活性间的关系。     

Association between olfactory sensitivity and behavioral responses of Drosophila suzukii to naturally occurring volatile compounds

Drosophila suzukii Matsumura (Diptera: Drosophilidae) is an invasive, destructive crop pest that originated in South East Asia. D. suzukii recently invaded Western countries and is threatening both European and American fruit industries. It is extremely attracted to otherwise undamaged, ripening fruits, unlike most other Drosophila species that attack only decaying or rotten fruits. Recent studies on different insect species showed that several naturally occurring compounds of easy market availability showing deterrent action may be used to supplement mass catches with food traps. Based on these considerations, the aim of the present work was to test the effects of some natural compounds (alone or in the mixture) on the olfactory system of the D. suzukii and the behavioral responses evoked. We measured by electroantennogram (EAG) recordings, the olfactory sensitivity of antennae to increasing concentrations of eugenol, vanillin, menthol, cis-jasmone; eugenol + vanillin, +menthol, +cis-jasmone; vanillin + menthol, +cis-jasmone. In addition, the behavioral responses to the same compounds and mixtures were evaluated. Our electrophysiological results show a dose-response relationship between the EAG amplitudes and the increasing concentrations of the olfactory compound. The behavioral results show that the number of laid eggs is significantly different between the standard diet and the standard diet + natural compound. These results underline a specificity in the olfactory sensitivity and in the ovipositing behavior of D. suzukii females; also, they could be valuable for the identification of key chemicals aimed at the future development of strategies in the management and control of this harmful insect for crops.     

The metabolism of gamma-aminobutyric acid (GABA) in the lobster nervous system--uptake of GABA in the nerve-muscle preparations

The lack of information on the mechanism of inactivation of the crustacean neuromuscular inhibitory transmitter compound prompted a study of the disposition of radioactive γ-aminobutyric acid (GABA) in lobster nerve-muscle preparations. A specific GABA transport system was found. Radioactive GABA was concentrated by the tissues to levels several times those in the medium, and net uptake could be demonstrated. The process was dependent on sodium ions in the medium; neither lithium nor choline could substitute for sodium. Incubations with increasing GABA concentrations indicated that uptake was a saturable mechanism with an apparent K_m of 5.8 × 10⁻⁵m . Of many compounds tested, only desmethylimipramine, chlopromazine (and several related compounds), and certain close structural analogues (guanidinoacetic acid, β-guani-dinopropionic acid and,β-hydroxy-GAB A) were effective inhibitors of uptake. The inhibition with all these compounds, however, was at high concentrations (5 × 10⁻⁴ to 10⁻³m ) which limited their usefulness for physiological studies. A separate uptake mechanism for glutamate was found in the lobster nerve-muscle preparations. This process was not described in detail, but certain properties are similar to those of the GABA transport system. The cellular location of the GABA uptake system remains unknown. By analogy with noradrenaline inactivation, however, it is postulated that uptake could serve to terminate the physiological actions of GABA by rapidly removing it from its sites of action in synaptic clefts.     

Design and synthesis of N-alkyl-N′-substituted 2,4-dioxo-3,4-dihydropyrimidin-1-diacylhydrazine derivatives as ecdysone receptor agonist

Based on the discovery of thymine as an ecdysteroid agonist, a series of 1,4-disubstituted diacylhydrazine derivatives with a thymine moiety were designed and synthesized. The activities of these compounds against Spodoptera litura (Fabricius) were evaluated by the insect immersion method. Results showed that compound 2h with an N-cyclohexylmethyl substituent exhibits the most potent agonist activity with a median lethal concentration of 23.21 μg/mL. This compound also caused malformation of molting larvae and adults. Compound 2h was further demonstrated as an ecdysteroid agonist by reporter gene assay on the Spodoptera frugiperda cell line (Sf9 cells). A molecular docking study indicated that hydrophobic interactions and the formation of hydrogen bonds between the compounds and the ecdysone receptor play critical roles in promoting the binding affinity of the compound. The structure of compound 2h may serve as a favorable template for the development of new ecdysteroid agonists with a pyrimidinedione moiety.     

Microbial Reactions on 7α‐Hydroxyfrullanolide and Evaluation of Biotransformed Products for Antibacterial Activity

7α‐Hydroxyfrullanolide ( 1 ), a known sesquiterpenoid, was isolated from Sphaeranthus indicus using an antibacterial‐activity‐directed fractionation method. This compound had exhibited a significant antibacterial activity against Gram‐positive bacteria. Chemical and microbial reactions were performed to prepare eight different analogues of compound 1 in order to evaluate these newly synthesized compounds for antibacterial activity. These compounds were 1β,7α‐dihydroxyfrullanolide ( 2 ), 7α‐hydroxy‐1‐oxofrullanolide ( 3 ), 4,5‐dihydro‐7α‐hydroxyfrullanolide ( 4 ), 11,13‐dihydro‐7α‐hydroxyfrullanolide ( 5 ), 13‐acetyl‐7α‐hydroxyfrullanolide ( 6 ), 2α,7α‐dihydroxysphaerantholide ( 7 ), 4α,5α‐epoxy‐7α‐hydroxyfrullanolide ( 8 ), and 4β,5β‐epoxy‐7α‐hydroxyfrullanolide ( 9 ). Microbial reactions on 1 using whole‐cell cultures of Cunninghamella echinulata and Curvularia lunata yielded compounds 2 – 4 . Incubation of compound 1 with the liquid cultures of Apsergillus niger and Rhizopus circinans yielded metabolites 5 – 7 , while 8 and 9 were prepared by carrying out an epoxidation reaction on 1 using meta‐chloroperbenzoic acid (mCPBA). Structures of compounds 2 – 9 were elucidated with the aid of extensive NMR spectral studies. Compounds 2 – 4 were found to be new metabolites. Compounds 1 – 9 were evaluated for antibacterial activity and found to exhibit a wide range of bioactivities. Antibacterial‐activity data of 1 – 9 suggested that the bioactivity of 1 is largely due to the presence of C(4)?C(5), C(11)?C(13), and a γ‐lactone moiety.     

Bacterial ectosymbionts in cuticular organs chemically protect a beetle during molting stages

In invertebrates, the cuticle is the first and major protective barrier against predators and pathogen infections. While immune responses and behavioral defenses are also known to be important for insect protection, the potential of cuticle-associated microbial symbionts to aid in preventing pathogen entry during molting and throughout larval development remains unexplored. Here, we show that bacterial symbionts of the beetle Lagria villosa inhabit unusual dorsal invaginations of the insect cuticle, which remain open to the outer surface and persist throughout larval development. This specialized location enables the release of several symbiont cells and the associated protective compounds during molting. This facilitates ectosymbiont maintenance and extended defense during larval development against antagonistic fungi. One Burkholderia strain, which produces the antifungal compound lagriamide, dominates the community across all life stages, and removal of the community significantly impairs the survival probability of young larvae when exposed to different pathogenic fungi. We localize both the dominant bacterial strain and lagriamide on the surface of eggs, larvae, pupae, and on the inner surface of the molted cuticle (exuvia), supporting extended protection. These results highlight adaptations for effective defense of immature insects by cuticle-associated ectosymbionts, a potentially key advantage for a ground-dwelling insect when confronting pathogenic microbes. Subject terms: Antibiotics, Zoology, Microbial ecology     

The contribution of olfactory receptor neurons to the perception of pheromone component ratios in male redbanded leafroller moths

Summary (Z)-11-tetradecenyl acetate (Z-11, 14:AC) must be in a 1009 ratio with (E)-11-tetradecenyl acetate (E-11,14:AC) to produce maximal wing fanning and attraction in male redbanded leafrollers. Earlier electrophysiological studies had indicated that mixtures of these pheromone components elicited responses from olfactory receptor neurons that appeared to differ from those expected on the basis of the responses to the individual components. Here we evaluate whether the behavioral sensitivity to particular ratios of Z- and E-11,14:AC has a correlate in the response properties of olfactory receptor neurons.The stimuli included the ratios of Z- and E-11, 14:AC used in earlier behavioral work plus several different mixtures of the seven components found in the pheromone blend, and equivalent amounts of the individual components. These stimuli were presented over a range of intensities to individual trichoid sensilla on the male antenna. In common with earlier results, the receptor neuron with the larger amplitude action potential responded most strongly to Z-11,14:AC, whereas the companion receptor neuron in the sensillum responded most strongly to E-11,14:AC. In contrast with earlier results, each receptor neuron responded exclusively to its own most effective stimulus, without regard to the presence of any other compound. They failed to respond uniquely to any of the other five compounds in the female pheromone blend, or to any of the tested combinations of these compounds. These minor components also failed to modulate the responses elicited in receptor neurons by appropriate ratios of Z- and E-11,14:AC. Thus, the responses of the two types of olfactory receptor neurons found in trichoid sensilla failed to show an optimum at the pheromone ratio known to elicit peak behavioral activity.Abbreviation RBLR redbanded leafroller moth     

Design,Synthesis, Cytotoxic Screening and Molecular Docking Studies of Novel Hybrid Thiosemicarbazone Derivatives as Anticancer Agents

Thiosemicarbazones have been the focus of scientists owing to their broad clinical anticancer range. Herein, A Series of new thiosemicarbazone derivatives 5 – 9 were synthesized and confirmed through the use of different spectroscopic techniques along with elemental analysis. The in vitro cytotoxic activity of compounds 5 – 9 against MCF-7 and A549 cell lines and normal breast cells were assessed. Several compounds were found to be active. The most active compound 7 caused MCF-7 cell cycle arrest at G1/ S phases; and induced apoptosis at the pre-G1 phase. The apoptosis-inducing activity of compound 7 was proofed by the elevation of caspase 3/7 activity and also by up-regulation of the expression of Bax and p53 proteins together with the down-regulation of the expression of the Bcl-2 protein. It also had a strong inhibitory effect topoisomerase IIβ enzyme. Molecular Docking study revealed that the synthesized compounds had good docking scores compared to the standard drug Etoposide towards the topoisomerase IIβ protein (3QX3). Overall, these findings confirmed that the new thiosemicarbazone derivatives could aid in the development of promising cancer drug candidates.