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Rhodnius prolixus: salivary antihemostatic components decrease with Trypanosoma rangeli infection.

Feeding behavior and some antihemostatic properties of the saliva of Rhodnius prolixus infected with Trypanosoma rangeli were studied and compared with those of uninfected controls. Insects with salivary gland infection pierced host skin more often and drew less blood and at a lower rate than controls when feeding on a rabbit. None of these differences was observed when feeding was performed through a membrane feeder. Salivary gland homogenates from infected insects, at 30 days after feeding/infection, had a significantly lower amount of total protein/salivary gland pair and less anticoagulant activity than controls. Also, infected salivary glands exhibited significantly reduced apyrase activities and reactive nitrogen groups when compared to controls. It is concluded that salivary infection of T. rangeli impairs the ability of the vector to locate blood vessels by affecting salivary antihemostatic properties, thus enhancing the possibility of intradermal inoculation of parasites into the mammalian host.

Animals

Large-scale propagation of insect cells.

Cultured insect cells have many uses in agriculture and medicine. They can be used in the diagnosis and isolation of a number of viruses infecting both animals and plants and for the laboratory study of these viruses. Large-volume culture of insect cells has been envisioned as a way of producing viruses for use in controlling insect pests and for the production of viral antigens for vaccine preparations. Recently they have become a potentially valuable way of producing a variety of proteins for human and veterinary medicine using the genetically engineered baculovirus expression vectors. The development of satisfactory cell lines and culture methods has proceeded at a slow, irregular pace, inhibited by the lack of knowledge of the physiology of the insect, its small size, and often by the lack of consistent, adequate support for the necessary developmental research. However, now that the basic culture systems are available, cell lines have been developed or can easily be developed for most needs. Suitable media are available and recent developments in refining existing media formations have resulted in low-cost media containing little protein to interfere with down-stream processing of cellular metabolites. Future developments are likely to further improve the media formulations and lower the cost. Technical problems relating to oxygen demand and cell fragility that inhibited the continued development of large-volume culture systems beyond the laboratory a few years ago now appear to be solved or at least are solvable. The successful culture of the Spodoptera cells in bioreactors of 40-liter capacity indicates that means of producing insect cells or their metabolic products on a commercial scale can be made economically feasible.

Animals

Insect prothoracicotropic hormone: a new member of the vertebrate growth factor superfamily.

Prothoracicotropic hormone (PTTH) is a brain neurosecretory protein that controls insect development. PTTH of the silkmoth Bombyx mori is a homodimeric protein, the subunit of which consists of 109 amino acids. Clear-cut sequence similarity to any other proteins has not been observed. By disulfide-bond pattern analysis and modeling of the PTTH structure based on the known three-dimensional (3D) structures of growth factor family with cystine-knot motif, we propose that the PTTH protomer adopts the fold unique to the structural superfamily of the growth factors, beta-nerve growth factor (beta-NGF), transforming growth factor-beta 2 (TGF-beta 2), and platelet-derived growth factor-BB (PDGF-BB). The insect neurohormone PTTH appears to be a member of the growth factor superfamily, sharing a common ancestral gene with the three vertebrate growth factors, beta-NGF, TGF-beta 2 and PDGF-BB.

Amino Acid Sequence

Glutamate receptor inhibitors as potential insecticides.

Philanthotoxin (PhTX) is a neurotoxic constituent of the paralytic venom of the digger wasp, Philanthus triangulum. PhTX inhibits glutamate receptors of insect muscles mostly as a channel blocker, thereby producing muscle paralysis. Since glutamate receptor blockers may be of value as selective insect control agents, numerous derivatives of PhTX were synthesized and tested for their potencies as inhibitors of insect skeletal muscle glutamate receptors. Structure-activity relationship studies revealed that shortening the polyamine chain length reduced potency, and quaternarization of the nitrogen destroyed it. The potency was increased by a bulky anchoring group with moderate hydrophobicity at the end of the polyamine chain. The conversion of the tryosyl moiety to 3,5-diiodo-tyrosyl also increased potency and so did lengthening the butyryl chain from 4 to 10 carbons. Not only did PhTXs inhibit different subtypes of glutamate receptors, including the mammalian N-methyl-D-aspartate receptor, but also nicotinic receptors of insects and vertebrates. Because of this low selectively, and the hydrophilicity of the derivatives tested, which interferes with their penetration to the target receptor, these compounds cannot be used as insecticides. Nevertheless, the insect skeletal muscle glutamate receptor is a viable target for selective insecticides and major changes in PhTX structure may possibly produce derivatives that can be potential insecticides.

Animals

Isolation and characterization of the Saccharomyces cerevisiae LPP1 gene encoding a Mg2+-independent phosphatidate phosphatase.

The DPP1-encoded diacylglycerol pyrophosphate (DGPP) phosphatase enzyme accounts for half of the Mg2+-independent phosphatidate (PA) phosphatase activity in Saccharomyces cerevisiae. The LPP1 (lipid phosphate phosphatase) gene encodes a protein that contains a novel phosphatase sequence motif found in DGPP phosphatase and in the mouse Mg2+-independent PA phosphatase. A genomic copy of the S. cerevisiae LPP1 gene was isolated and was used to construct lpp1Delta and lpp1Delta dpp1Delta mutants. A multicopy plasmid containing the LPP1 gene directed a 12.9-fold overexpression of Mg2+-independent PA phosphatase activity in the S. cerevisiae lpp1Delta dpp1Delta double mutant. The heterologous expression of the S. cerevisiae LPP1 gene in Sf-9 insect cells resulted in a 715-fold overexpression of Mg2+-independent PA phosphatase activity relative to control insect cells. The Mg2+-independent PA phosphatase activity encoded by the LPP1 gene was associated with the membrane fraction of the cell. The LPP1 gene product also exhibited lyso-PA phosphatase and DGPP phosphatase activities. The order of substrate preference was PA > lyso-PA > DGPP. Like the dpp1Delta mutant, the lpp1Delta mutant and the lpp1Delta dpp1Delta double mutant were viable and did not exhibit obvious growth defects. Biochemical analyses of lpp1Delta, dpp1Delta, and lpp1Delta dpp1Delta mutants showed that the LPP1 and DPP1 gene products encoded nearly all of the Mg2+-independent PA phosphatase and lyso-PA phosphatase activities and all of the DGPP phosphatase activity in S. cerevisiae. Moreover, the analyses of the mutants showed that the LPP1 and DPP1 gene products played a role in the regulation of phospholipid metabolism and the cellular levels of phosphatidylinositol and PA.

Animals

Hormonal control of transmitter plasticity in insect peptidergic neurons. II. Steroid control of the up-regulation of bursicon expression.

Each abdominal ganglion of the central nervous system of the tobacco hawkmoth, Manduca sexta contains four individually identified lateral neurosecretory cells (LNCs) that undergo a postembryonic transmitter switch in vivo. In the embryonic and caterpillar stages, the primary LNC transmitter is cardioacceleratory peptide 2 (CAP2), a myoregulatory peptide. During metamorphosis, these cells stop expressing CAP2 and instead produce bursicon, a classic insect peptide hormone responsible for cuticular tanning. We have previously reported that this transmitter plasticity is under the control of the insect steroid hormone 20-hydroxyecdysone (20-HE), which surges twice during the last larval instar. In that report we showed that the CAP2 decline is indirectly regulated by the first 20-HE rise, the commitment pulse (CP). Here we provide evidence that the rise in bursicon levels in the LNCs is directly triggered by the second 20-HE surge, the prepupal peak (PP). We performed several experimental manipulations that exposed LNCs to the PP without the CP; cells treated in this manner exhibited a significant rise in bursicon content. In contrast, bursicon levels remained unchanged in those LNCs exposed only to the CP. Exposure to the PP triggered a precocious increase in bursicon expression in LNCs from the penultimate larval stage. Increased bursicon levels in the LNCs were also induced by direct infusion of 20-HE. Taken together, the results of these experiments suggest that the rise in bursicon in the LNCs during metamorphosis is due to the direct action of the PP on the LNCs. Thus, the two 20-HE surges combine to regulate the CAP2-to-bursicon switch in the LNCs, the first acting indirectly to cause a decline in CAP2 levels and the second triggering a rise in bursicon expression, possibly by a direct action on the LNCs.

Animals

Insecticidal toxins from the bacterium Photorhabdus luminescens.

Transgenic plants expressing Bacillus thuringiensis (Bt) toxins are currently being deployed for insect control. In response to concerns about Bt resistance, we investigated a toxin secreted by a different bacterium Photorhabdus luminescens, which lives in the gut of entomophagous nematodes. In insects infected by the nematode, the bacteria are released into the insect hemocoel; the insect dies and the nematodes and bacteria replicate in the cadaver. The toxin consists of a series of four native complexes encoded by toxin complex loci tca, tcb, tcc, and tcd. Both tca and tcd encode complexes with high oral toxicity to Manduca sexta and therefore they represent potential alternatives to Bt for transgenic deployment.

Amino Acid Sequence

Expression of the mosquitocidal toxins of Bacillus sphaericus and Bacillus thuringiensis subsp. israelensis by recombinant Caulobacter crescentus, a vehicle for biological control of aquatic insect larvae.

In the quest for effective control of mosquitoes, attention has turned increasingly to strains of the bacteria Bacillus sphaericus and Bacillus thuringiensis subsp. israelensis, which produce potent toxins with specific mosquitocidal activities. However, sedimentation of the bacterial spores limits the duration of effective control after field application of these bacilli. We describe here the cloning of genes encoding the 51.4- and 41.9-kDa toxins from B. sphaericus 2297, the 100-kDa toxin from B. sphaericus SSII-1, and the 130-kDa toxin from B. thuringiensis subsp. israelensis into the broad-host-range plasmid pRK248 and the transfer of these genes for expression in Caulobacter crescentus CB15. The recombinant C. crescentus cells were shown to be toxic to mosquito larvae. Caulobacter species are ubiquitous microorganisms residing in the upper regions of aquatic environments and therefore provide the potential for prolonged control by maintaining mosquitocidal toxins in larval feeding zones.

Animals

[Physical preservation of food].

Physical preservation procedures are mostly aimed at prolonging the durability of foods by slowing down or repressing the spoilage mechanisms by the alteration of relevant parameters. Of these mechanisms of microbiological, enzymatic, chemical and mechanical type, especially the first-named are dealt with. Growth and multiplication of micro-organisms can be influenced by temperature, water activity and high-energy radiation. Depending on the susceptibility to deterioration of the products and the intensity of the processes based on these parameters, different degrees of durability are obtained. Micro-organisms are killed by the application of high temperatures in conjunction with the treatment times required in a given case. Relatively mild treatments affect only the vegetative forms (pasteurization), whereas more aggressive treatments are required to kill spores as well (sterilization). Recontamination of the products which were subjected to the two procedures must be avoided. Pasteurized foods require additional cold storage to prevent the spores from sprouting. Low temperatures reduce and repress the growth of micro-organisms (refrigeration) or prevent any activity of the micro-organisms (deep-freezing). The latter deprives the micro-organisms of the water they need for their growth. The same principle applies to another method of preservation where so much water is extracted from the product that the residual humidity no longer allows the micro-organisms to be active (drying). Ionising radiation can be used to reduce a potential risk to hygiene, to influence physiological processes, to control insects and to lengthen the durability of fresh food in the short term. Finally optimum product quality can be maintained by combining various procedures. If such methods cope with controlling the microbiological situation, attention must nevertheless be given to the other spoilage mechanisms as well, as they may prove to be limiting factors with respect to product quality.

Bacteria