Monique S. J. Simmonds

MOLECULES OF INTEREST ROSMARINIC ACID

Rosmarinic acid is an ester of caffeic acid and 3,4-dihydroxyphenyllactic acid. It is commonly found in species of the Boraginaceae and the subfamily Nepetoideae of the Lamiaceae. However, it is also found in species of other higher plant families and in some fern and hornwort species. Rosmarinic acid has a number of interesting biological activities, e.g. antiviral, antibacterial, antiinflammatory and antioxidant. The presence of rosmarinic acid in medicinal plants, herbs and spices has beneficial and health promoting effects. In plants, rosmarinic acid is supposed to act as a preformed constitutively accumulated defence compound. The biosynthesis of rosmarinic acid starts with the amino acids L-phenylalanine and L-tyrosine. All eight enzymes involved in the biosynthesis are known and characterised and cDNAs of several of the involved genes have been isolated. Plant cell cultures, e.g. from Coleus blumei or Salvia officinalis, accumulate rosmarinic acid in amounts much higher than in the plant itself (up to 36% of the cell dry weight). For this reason a biotechnological production of rosmarinic acid with plant cell cultures has been proposed.

Flavonoid-insect interactions: recent advances in our knowledge.

Recent contributions to the role of phenolics, especially flavonoids, in different aspects of insect-plant interactions are reviewed, including data on the effects of rutin on the feeding behaviour of a range of noctuid larvae.

Importance of flavonoids in insect-plant interactions : feeding and oviposition

Jeffrey Harborne and colleagues have been responsible for collating the majority of data on the role of flavonoids in insect plant interactions. This article examines some of this information and assesses our knowledge about the role flavonoids play in insect feeding and oviposition behaviour. It is clear that insects can discriminate among flavonoids and that these compounds can modulate the feeding and oviposition behaviour of insects, but further work is required to understand the neural mechanisms associated with these behavioural responses. Despite the wealth of data about the diversity of flavonoids in plants, very few of these compounds have been tested against insects and their role in the evolution of host range in insect--plant interactions has yet to be determined.

Antifeedant effects of azadirachtin and structurally related compounds on lepidopterous larvae.

The antifeedant activity of azadirachtin, azadirachtin‐derivatives and related limonoids was assessed in choice and no‐choice bioassays against four species of Lepidoptera: Spodoptera littoralis, Spodoptera frugiperda, Heliothis virescens and Heliothis armigera. The choice bioassay showed that the feeding behaviour of S. littoralis was affected by more of the compounds than that of either S. frugiperda or H. virescens. H. armigera was the least affected. Azadirachtin and dihydroazadirachtin were the most potent of the 40 compounds tested. The results showed that hydrogenation of the C‐22,23 double bond did not decrease antifeedant activity and the nature of the substitutes at C‐1, C‐3 and C‐11 were important. Molecules with bulky substitutes at either C‐22 or C‐23 were usually ineffective antifeedants as were compounds lacking an epoxide. Compounds recorded as active antifeedants in the choice bioassay were not always as active in the no‐choice test. The value of the bioassays in assessing the mode of action of the compounds is discussed.

Actions of azadirachtin, a plant allelochemical, against insects

Investigations of the antifeedant mode of action of azadirachtin and four synthetic analogues, 22,23-dihydroazadirachtin, 3-tigloylazadirachtol, 11-methoxydihydroazadirachtin and 22,23-bromoethoxydihydroazadirachtin have revealed that both polyphagous and oligophagous insects are behaviourally responsive to azadirachtin, with the most responsive species being able to differentiate extremely small changes in the parent molecule. In Lepidoptera the antifeedant response is correlated also with increased neural activity of the chemoreceptors. When locusts are treated on crop plants, the antifeedant and physiological actions of azadirachtin and analogues work in concert and result in feeding deterrence, growth and moulting aberrations and mortality with the same order of potency as for antifeedancy. Specific binding studies using [3H]dihydroazadirachtin carried out on locust testes and Spodoptera Sf9 cells have shown that the competitive binding of the different analogues of azadirachtin to these binding sites occurs in a similar order of potency to that found with antifeedant and IGR bioassays. This suggests a causal link between specific binding to membrane proteins and the ability of the molecule to exert biological effects.

In vitro and In vivo evaluation of betulinic acid as an antimalarial

The lupane‐type triterpene betulinic acid was isolated from an ethanol extract of the root bark of the Tanzanian tree Uapaca nitida Müll‐Arg. (Euphorbiaceæ). The in vitro antiplasmodial IC50 values of betulinic acid against chloroquine resistant (K1) and sensitive (T9‐96) Plasmodium falciparum were found to be 19.6 µg/mL and 25.9 µg/mL, respectively. The in vitro activities of several related triterpenes were also evaluated. Betulin was found to be inactive at 500 µg/mL for both K1 and T9‐96. Ursolic acid exhibited IC50 values of 36.5 µg/mL and 28 µg/mL, and oleanolic acid exhibited IC50 values of 88.8 µg/mL and 70.6 µg/mL against K1 and T9‐96, respectively. When betulinic acid was tested for in vivo activity in a murine malaria model (P. berghei) the top dosage employed of 250 mg/kg/day was ineffective at reducing parasitaemia and exhibited some toxicity. Betulinic acid has not previously been evaluated for in vivo activity. This is believed to be the first compound to be isolated from U. nitida. Copyright

A comparison of dietary selection behaviour in larval Locusta migratoria and Spodoptera littoralis

ABSTRACT. . Final instar nymphs of the oligophagous acridid Locusta migratoria (L.) and larvae of the polyphagous noctuid Spodoptera littoralis (Boisduval) were fed for 4, 8 or 12 h, the conditioning period, on one of four artificial diets. Of these, diet PC contained 20% protein and 10% digestible carbohydrate; another, P, contained 20% protein but with the digestible carbohydrate component replaced by cellulose; a third, C, had the protein component substituted by cellulose, and the fourth, O, had both protein and digestible carbohydrate replaced. After this conditioning period, insects were given a choice of two diets, P and C, and hence an opportunity to select for the nutrients, if any, which were lacking in their previous food. Amounts eaten and selection behaviour were then recorded in detail for a total of 9 h. This paper deals with total amounts of diet eaten during the conditioning and choice periods. Spodoptera larvae were more sensitive than the locusts to being fed a nutritionally inadequate conditioning diet, and ate only small quantities of the P, C and O diets as compared with the PC diet, irrespective of the duration of conditioning. Locusts, on the other hand, when restricted to the P diet continued to eat relatively large amounts of it throughout a 12 h conditioning period. Those nymphs fed the C diet ingested large quantities (more than of the PC diet) up until 8 h, after which intake fell. When offered a choice, both species selected for the nutrients missing from the conditioning diet, even if the conditioning period had been as short as 4 h. During the first hour of choice locusts selected the P diet if they had been previously fed C and the C diet if previously fed P. Those deprived of both nutrients increased consumption of both P and C diets. Spodoptera larvae were more sensitive to prior deprivation of digestible carbohydrate than of protein. During the first hour of choice they selected the C diet if previously fed P or O but did not choose the P diet if previously fed C. In the subsequent 8 h of choice, however, a strong selection for the P diet after previous deprivation became apparent. In the locust, the selection for nutrients missing from the conditioning diet continued for the following 8 h of choice but became masked by a tendency, shown by all nymphs, to select C over P. The functional significance and possible physiological basis of all these responses is discussed.

Behavioral and electrophysiological study of antifeedant mechanisms associated with polyhydroxy alkaloids

Eleven polyhydroxy alkaloids of plant origin were tested for anti-feedant effects against larvae of the lepidopteransSpodoptera littoralis, Spodoptera frugiperda, Heliothis virescens, andHelicoverpa armigera. Data from behavioral and electrophysiological investigations were correlated to reveal information on the mode of action of the antifeedants. The pyrrolidine DMDP was an effective antifeedant for all four species, whereas the piperidines fagomine and XZ-1 and the pyrrolizidine alexine were all ineffective as antifeedants. The activity of the pyrrolidines CYB-3 and DAB-1, the piperidines DNJ, DMJ, and BR1, and the bicyclic octahydroindolizine castanospermine varied among species. The investigation focuses on the structural similarities between some of the alkaloids and some common phagostimulatory sugars and illustrates a neural interaction involving the neurons that are differentially responsive to alkaloids and sugars. InS. littoralis, the neurons responding specifically to the alkaloids DMDP, DAB-1, and castanospermine and to the sugars fructose, sucrose, and glucose are more active when the compounds are applied singly than when an alkaloid and a sugar are applied together. The implications for the occurrence and functioning of different sugar receptor sites are discussed.

Omic techniques in systems biology approaches to traditional Chinese medicine research: Present and future ,

Omic techniques have become key tools in the development of systems biology. As the holistic approaches underlying the practice of traditional Chinese medicine (TCM) and new tendencies in Western medicine towards personalised medicine require in-depth knowledge of mechanisms of action and active compounds, the use of omic techniques is crucial for understanding and interpretation of TCM development, especially in view of its expansion in Western countries. In this short review, omic applications in TCM research are reviewed which has allowed some speculation regarding future perspectives for these approaches in TCM modernisation and standardisation. Guidelines for good practice for the application of omics in TCM research are also proposed.

Variation in chemosensitivity and the control of dietary selection behaviour in the locust

Investigations into the behavioural and underlying physiological mechanisms of dietary selection are presented for the locust, Locusta migratoria. Locusts were fed for 4, 8 or 12 h on one of four chemically defined artificial diets: diet PC, which was nutritionally complete; diet P, containing no digestible carbohydrate; diet C, containing no protein; and diet O, which lacked both protein and digestible carbohydrate. Following this pretreatment, the locusts were provided with both the P and the C diet in a choice test. Detailed analyses of selection behaviour indicated that diets lacking a nutrient for which the insect was deficient were either rejected before a meal was initiated, or, if feeding commenced, eaten in meals of only short duration, while those containing the appropriate nutrients were accepted more readily and eaten in longer meals. Electrophysiological studies showed that this behaviour was paralleled by nutrient-specific changes in gustatory responsiveness. Locusts pretreated for 4h on C diet had increased gustatory responsiveness to stimulation with an amino acid mix, but not to sucrose, while insects fed on P diet showed increased responsiveness to stimulation with sucrose, but not to the amino acid mix. This result is consistent with earlier experiments in which levels of blood nutrients were shown to modulate taste responsiveness in the locust.