Baculoviruses as vectors for foreign gene expression in insect cells

Recombinant DNA technology has enabled the expression of a large number of eukaryotic genes in vitro. There has been growing interest in recent years in the development of baculovirus expression vectors as an easily manipulated gene expression system. Both virus and insect cell culture are relatively easy to handle, and biologically active proteins have been produced abundantly from a variety of eukaryotic, prokaryotic and viral genes. This review describes the molecular biology of baculoviruses and some of the current applications of the baculovirus expression system in insect cells. Specific emphasis is placed on those features relevant to the use of this system in pesticide research.     

The kinetics of insecticide action. Part IV: the in-vivo distribution of pyrethroid insecticides during insect poisoning

The distribution of pyrethroids in insects has been studied using a combination of mathematical modelling and experimental observation. This approach has resulted in the formulation of a physiological model of the pharmaco-kinetics of cypermethrin applied topically to larvae of Spodoptera littoralis. Development of this model from simpler two- and three-compartment models is described. Simple models, whilst capable of complex behaviour, consider only the average rates and magnitudes of pharmaco-kinetic processes over whole animals, and cannot account for differences in concentration of insecticide between individual tissues. This can be achieved by using physiological models, but these require more experimental information for their validation. Moreover, unless simplifying assumptions are made, analytical solutions are not feasible for the large number of equations necessary to define such models. The modelling studies prompted an investigation of (1) in-vivo binding of insecticide to insect tissues, (2) the sizes of body compartments, and (3) the factors which affect the distribution of toxicant between these compartments. Binding has a marked effect on pharmacokinetic profiles and may result in oscillatory behaviour. During poisoning, the total bound cypermethrin increases proportionally to the cube root of the elapsed time. This results in a rapid rate of increase over early elapsed times (< 3h) which slows to approach a more linear form thereafter. Average sizes for the body compartments of larvae of Spodoptera littoralis Boisd, and the steady-state distribution of cypermethrin in these compartments are described. Although the haemolymph, which acts as the main distributive phase during poisoning, forms the largest compartment by volume, it has a low affinity for cypermethrin and distribution reaches steady state within 5 min after topical application. The nerve cord (the target tissue), which is the smallest compartment, has the highest steady-state concentration of cypermethrin. The distribution of cypermethrin in larval tissues is related to the ratios of tissue dry matter to water content.     

Review—mechanism of action of herbicidal and fungicidal compounds on cell membranes

The eukaryotic cell membrane is thought to consist of a mobile bilayer of phospholipid, sometimes intercalated with sterols, in which peripheral and transmembrane proteins are embedded. This provides a model whereby the mode of action of many fungicidal and herbicidal compounds can be rationalised and understood. Some compounds, such as the polyene antibiotics, steroidal saponins and certain phytoalexins, induce membrane malfunction by direct insertion, often complexing with vital components. Others, such as paraquat and the nitrodiphenyl ether herbicides, cause membrane damage by inducing lipid peroxidations. Furthermore, there are many compounds, including the sterol-biosynthesis-inhibiting fungicides and the aryloxyphenoxypropionate and cyclohexanedione herbicides, which interfere with the biosynthesis of membrane components. Because membrane structure is fundamentally similar in eukaryotes, it is important to discover the reasons for any differential toxicity displayed by these compounds and, if necessary, to find ways of optimising desirable patterns of selectivity.     

Studies into the action of the diphenyl ether herbicides acifluorfen and oxyfluorfen. Part I: Activation by light and oxygen in leaf tissue

The effect of acifluorfen and oxyfluorfen on chlorophyll bleaching, lipid peroxidation and photosynthesis in pea leaf discs was studied. Both her- bicides induced light-dependent bleaching and lipid peroxidation, the level of damage being greater at higher light intensities. Photosynthetic carbon dioxide fixation was only partially inhibited in treated leaf discs incubated in darkness, thus indicating that these herbicides did not inhibit photo- synthesis as a primary mode of action. Leaf discs maintained in darkness showed no visible signs of injury, and light-dependent herbicide-induced damage was reduced by incubating discs under nitrogen, orpre-incubating them with the electron-transport inhibitor monuron. It is suggested that acifluorfen and oxyfluorfen are activated by a light-dependent process, which requires photosynthetic electron transport.     

Permethrin treatments in canadian forests. Part 1: Impact on stream fish

The effects of permethrin on native and caged fish when aerially applied directly to forest streams at dosages between 8.8 and 70 g ha^?1 were studied between 1976 and 1981. None of the applications caused mortality of caged or native fish in streams. Trout and salmon diets were altered by the treatments due to effects on fish food organisms. The duration of the effects varied from several months to over a year with increasing dosage. Reductions in salmonid growth rates and reductions in fish densities in treated areas, presumably due to emigration, were documented following severe impacts on aquatic invertebrates in salmon nursery streams. Growth rates and population densities both recovered within four months after treatment.     

Permethrin treatments in canadian forests. Part 2: Impact on stream invertebrates

A series of experiments was conducted between 1977 and 1981 to evaluate the effects of aerially applied permethrin on forest stream invertebrates. All permethrin applications to forest streams resulted in large drift increases, and most produced measurable reductions in benthos density. Recovery of benthic fauna following the various permethrin applications was apparent from 1 to 18 months post-spray. In double-application experiments, the second treatments reduced benthos density to a point at which recovery of numbers was slower than after the impact resulting from a single application of the same dosage.     

Some fluorine-containing pheromone analogues

Two analogues of the aphid alarm pheromone (E)-β-farnesene, a trifluorofarnesene and a difluoro-1-norfarnesene, were found to be highly active and were more readily detectable than the parent compound. For (Z)-hexadec-11-enal, a component of some lepidopteran sex attractant pheromones, replacement of the carbonyl oxygen with a difluoromethyl group to give a difluoroheptadecadiene resulted in loss of activity. A trifluoroacetoxyhexadecanolide was a more volatile analogue of the mosquito oviposition pheromone (?)-(5R,6S)-6-acetoxy-5-hexadecanolide and was highly active.     

Vapour pressure of permethrin

The vapour pressures of the separate isomers of permethrin have been determined over a range of temperatures by two laboratories using different versions of the gas saturation method. A statistical analysis of the sets of data from the two laboratories shows no significant differences for the cis isomers but a small difference in the 20°C extrapolated values for the trans isomer exists. However, this difference is discounted as arising from experimental error and the mean values of the estimated vapour pressures at 20°C are given as 2.5 μPa (cis) and 1.5 μPa (trans).     

The pyrethrins and related compounds. Part XXVIII: Alkenyl- and alkynyl-substituted benzyl esters

Over 100 benzyl esters of pyrethroidal acids were synthesised and tested for insecticidal activity to establish detailed structure–activity relationships in compounds with side-chains similar to those in the natural pyrethrins. Alkenyl, and corresponding alkynyl, side-chains were effective, both at the 3- and 4-positions, as were side-chains with extended substitution in either E or Z forms. A cyano group at the α-position increases activity if the side-chain is at C-3, but lowers it drastically if the substituent is at C-4. Similarly, methyl groups at C-2 and/or C-6 may increase activity whether the unsaturated side-chain is at C-3 or C-4, but only in the absence of an α-cyano group.