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Insect venom hypersensitivity: experience in a clinical immunology/allergy service in Singapore.

INTRODUCTION: To study the profile of patients with allergy to the venom of insect stings. METHODS: 31 consecutive cases referred to our clinical immunology/allergy outpatient service from June 1, 1998 to June 30, 2002 were reviewed. RESULTS: These patients comprised 3.5 percent of 889 cases referred during the study period. Their mean age was 28.8 +/- 10.5 (range 19-57) years and the majority were males (90.3 percent). Of these, 20 (64.5 percent) were Chinese, four (12.9 percent) were Malays and seven (22.6 percent) were of other races. 19 patients (61.3 percent) were men from the uniformed services including 12 (63.2 percent) full-time National Servicemen. 71 percent (22 patients) were stung for the first time. Urticaria (22 cases, 71.0 percent), dyspnoea (13, 41.9 percent), angioedema (12, 38.7 percent) and syncope (ten, 32.3 percent) were the most common manifestations of insect allergy. Anaphylaxis occurred in 22 (71.0 percent) cases, constituting 30.1 percent of all cases of anaphylaxis referred to our service during the study period. Although the causative insect was identified as honeybee (12, 38.7 percent), ant (four, 12.9 percent), wasp (three, 9.7 percent), and fire ant (two, 6.5 percent) by the majority of patients, ten (32.2 percent) patients were unable to identify the causative insect. The two patients stung by fire ants were Americans working in Singapore who had been stung while in the United States. Among those with anaphylaxis, honeybee, wasp and fire ant venom, for which specific immunotherapy is available, were identified as the cause in 40.9 percent, 4.5 percent, and 4.5 percent, respectively. CONCLUSION: Insect venom hypersensitivity made up 3.5 percent of allergy/immunology referrals and 32.8 percent of cases of anaphylaxis referred to our institution. The majority were military servicemen who developed allergic reactions during the course of duty. The inability to identify the causative insect in 50 percent with sting anaphylaxis limits the role of specific immunotherapy in our patients.

Adult↗

[Allergic reactions to insect bites].

An allergic reaction to stings by insects can occur within minutes and may be fatal. The history serves to assess the severity of the reaction and may aid in the identification of the insect involved. The diagnosis is confirmed by a skin test and the presence of insect venom specific IgE. Minor allergic reactions can be treated with drugs, the more severe forms require hyposensitization therapy with pure insect venom, especially when the risk of life threatening reaction due to reexposure to insect sting is high. A protection against insect stings can be achieved within days. During the venom immunotherapy an antibody rise occurs and is followed by a suppression of specific IgE production later on. Allergy against hymenoptera venom is a suitable model for hyposensitization treatment.

Ant Venoms↗

Perception and use of insect repellent among soldiers in the Singapore Armed Forces.

The Singapore Armed Forces currently uses the 75% DEET formulation insect repellent. To study the perception, use, and acceptability of this insect repellent, a survey was carried out on servicemen who had participated in field exercises. Although over 80% of the servicemen knew the proper use of the insect repellent and brought along the army-issued repellent in the field, less than half used this repellent frequently while on exercise. Eighty-three percent felt that the army repellent was only effective sometimes and that it lasted for 4 hours or less. Skin irritation was a common side effect when using the repellent. About 70.4% had used commercial insect repellents and perceived them to be more long lasting and with fewer side effects. The results from our study indicate that the army-issued insect repellent currently used is not totally acceptable to our servicemen. Thus, there is a pressing need to develop or acquire a better insect repellent that is more or just as effective, has fewer side effects in terms of smell and skin irritation, and with an improved mode of delivery, such as aerosol or pump spray.

Adolescent↗

Detectors for polarized skylight in insects: a survey of ommatidial specializations in the dorsal rim area of the compound eye.

Apart from the sun, the polarization pattern of the sky offers insects a reference for visual compass orientation. Using behavioral experiments, it has been shown in a few insect species (field crickets, honey bees, desert ants, and house flies) that the detection of the oscillation plane of polarized skylight is mediated exclusively by a group of specialized ommatidia situated at the dorsal rim of the compound eye (dorsal rim area). The dorsal rim ommatidia of these species share a number physiological properties that make them especially suitable for polarization vision: each ommatidium contains two sets of homochromatic, strongly polarization-sensitive photoreceptors with orthogonally-arranged analyzer orientations. The physiological specialization of the dorsal rim area goes along with characteristic changes in ommatidial structure, providing actual anatomical hallmarks of polarized skylight detection, that are readily detectable in histological sections of compound eyes. The presence of anatomically specialized dorsal rim ommatidia in many other insect species belonging to a wide range of different orders indicates that polarized skylight detection is a common visual function in insects. However, fine-structural disparities in the design of dorsal rim ommatidia of different insect groups indicate that polarization vision arose polyphyletically in the insects.

Animals↗

Peripheral and central structures involved in insect gustation.

Studies in insect gustation have a long history in general physiology, particularly with work on fly labellar and tarsal sensilla and in the general field of insect-plant interactions, where work on immature Lepidoptera and chrysomelid beetles has been prominent. Much more emphasis has been placed on the physiological characteristics of the sensory cells than on the central cellular mechanisms of taste processing. This is due to the fairly direct access for physiological experimentation presented by many taste sensilla and to the obvious importance of tastants in insect feeding and oviposition behaviour. In some of the insect models used for gustatory studies, advances have been made in understanding the basic morphology of the central neuropils involved in the first stages of taste processing. There is much less known about the physiology of interneurons involved. In this review, we concentrate on four insect models (Manduca sexta, Drosophila melanogaster, Neobellieria bullata (and other large flies), and Apis mellifera) to summarize morphological knowledge of peripheral and central aspects of insect gustation. Our views of current interpretations of available data are discussed and some important areas for future research are highlighted.

Animals↗

Mono- and dibasic proteolytic cleavage sites in insect neuroendocrine peptide precursors.

Regulatory peptides are synthesized as part of larger precursors that are subsequently processed into the active substances. After cleavage of the signal peptide, further proteolytic processing occurs predominantly at basic amino acid residues. Rules have been proposed in order to predict which putative proteolytic processing sites are actually used, but these rules have been established for vertebrate peptide precursors and it is unclear whether they are also valid for insects. The aim of this paper is to establish the validity of these rules to predict proteolytic cleavage sites at basic amino acids in insect neuropeptide precursors. Rules describing the cleavage of mono- and dibasic potential processing sites in insect neuropeptide precursors are summarized below. Lys-Arg pairs not followed by an aliphatic or basic amino acid residue are virtually always cleaved in insect regulatory peptide precursors, but cleavages of Lys-Arg pairs followed by either an aliphatic or a basic amino acid residue are ambiguous, as is processing at Arg-Arg pairs. Processing at Arg-Lys pairs has so far not been demonstrated in insects and processing at Lys-Lys pairs appears very rare. Processing at single Arg residues occurs only when there is a basic amino acid residue in position -4, -6, or -8, usually an Arg, but Lys or His residues work also. Although the current number of such sites is too limited to draw definitive conclusions, it seems plausible that cleavage at these sites is inhibited by the presence of aliphatic residues in the +1 position. However, cleavage at single Arg residues is ambiguous. When several potential cleavage sites overlap the one most easily cleaved appears to be processed. It cannot be excluded that some of the rules formulated here will prove less than universal, as only a limited number of cleavage sites have so far been identified. It is likely that, as in vertebrates, ambiguous processing sites exist to allow differential cleavage of the same precursor by different convertases and it seems possible that the precursors of allatostatins and PBAN are differentially cleaved in different cell types. Arch. Insect Biochem. Physiol. 43:49-63, 2000.

Amino Acid Sequence↗

Odour plumes and odour-mediated flight in insects.

Flying insects often follow odour plumes to find resources. Some insects may employ an 'aim-and-shoot' strategy using mechanoreceptors before flight to determine wind direction. Once airborne, insects must use optomotor anemotaxis to set a course upwind. This mechanism uses a visual appraisal of how wind modifies the insect's path. A straight upwind course yields a front-to-rear image flow directly below the insect. Details of this process in male moths flying to female pheromone have emerged mainly from wind tunnel studies. Loss of the pheromone triggers 'casting', or wide lateral excursions without upwind progress, whereas contact with a plume usually induces a zigzag path upwind. The temporally regular counterturns in casting and zigzagging seem to be generated by a central programme. Brief contact with a filament of odour induces a heading towards upwind, and an optimal rate of encounter promotes a rapid, straight upwind course. Other insects, such as parasitoid wasps seeking a host and tsetse flies seeking a blood meal, seem not to have a temporally regular pattern of counter-turns and often fly straight upwind. The availability of visual cues from the odour source itself, the aerial distribution of odour set by turbulent diffusion, and light and wind levels all influence the success of these manoeuvres.

Animals↗

Insect--plant adaptations.

The adaptation of insects to plants probably commenced in the early Permian period, though most current associations will be more recent. A major burst of adaptation must have followed the rise of the Angiosperms in the Cretaceous period, though some particular associations are as recent as this century. Living plants form a large proportion of the potential food in most habitats, though insects have had to overcome certain general hurdles to live and feed on them. Insects affect the reproduction and survival of plants, and thus the diversity of plant secondary chemicals may have evolved as a response. Where an insect species has a significant effect on a plant species that is its only host, coevolution may be envisaged. A spectacular example is provided by Heliconius butterflies and passion flower vines, studied by L.E. Gilbert and others. But such cases may be likened to 'vortices in the evolutionary stream': most plant species are influenced by a range of phytophagous insects so that selection will be for general defences--a situation termed diffuse coevolution. Evidence is presented on recent host-plant shifts to illustrate both the restrictions and the flexibility in current insect-plant associations.

Adaptation, Biological↗

Pharmacologic characterization of muscarinic receptors of insect brains.

Muscarinic receptors in brain membranes from honey bees, houseflies, and the American cockroach were identified by their specific binding of the non-selective muscarinic receptor antagonist [3H]quinuclidinyl benzilate ([3H]QNB) and the displacement of this binding by agonists as well as subtype-selective antagonists, using filtration assays. The binding parameters, obtained from Scatchard analysis, indicated that insect muscarinic receptors, like those of mammalian brains, had high affinities for [3H]QNB (KD = 0.47 nM in honey bees, 0.17 nM in houseflies and 0.13 nM in the cockroach). However, the receptor concentration was low (108, 64.7, and 108 fmol/mg protein for the three species, respectively). The association and dissociation rates of [3H]QNB binding to honey bee brain membranes, sensitivity of [3H]QNB binding to muscarinic agonists, and high affinity for atropine were also features generally similar to muscarinic receptors of mammalian brains. In order to further characterize the three insect brain muscarinic receptors, the displacement of [3H]QNB binding by subtype-selective antagonists was studied. The rank order of potency of pirenzepine (PZ), the M1 selective antagonist, 11-[2-[dimethylamino)-methyl)1-piperidinyl)acetyl)-5,11- dihydro-6H-pyrido(2,3-b)-(1,4)-benzodiazepin-6 one (AF-DX 116), the M2-selective antagonist, and 4-DAMP (4-diphenylacetoxy-N-methylpiperidine methiodide) the M3-selective antagonist, was also the same as that of mammalian brains, i.e., 4-DAMP greater than PZ greater than AF-DX 116. The three insect brain receptors had 27-50-fold lower affinity for PZ (Ki 484-900 nM) than did the mammalian brain receptor (Ki 16 nM), but similar to that reported for the muscarinic receptor subtype cloned from Drosophila. Also, the affinity of insect receptors for 4-DAMP (Ki 18.9-56.6 nM) was much lower than that of the M3 receptor, which predominates in rat submaxillary gland (Ki of 0.37 nM on [3H]QNB binding). These drug specificities of muscarinic receptors of brains from three insect species suggest that insect brains may be predominantly of a unique subtype that is close to, though significantly different from, the mammalian M3 subtype.

Animals↗

Antioxidant systems in insects.

Insects possess a suite of antioxidant enzymes and small molecular weight antioxidants that may form a concatenated response to an onslaught of dietary and endogenously produced oxidants. Antioxidant enzymes such as superoxide dismutase, catalase, glutathione transferase, and glutathione reductase have been characterized in insects. Water-soluble and lipid-soluble antioxidants such as ascorbate, glutathione, tocopherols, and carotenoids have not been well studied in insects but may play very important antioxidant roles. Additionally, the peritrophic matrix and trehalose may possess important antioxidant functions in insects. The enzymatic recycling of ascorbate, first noted in green plants, may also exist in insects. A greater understanding of these antioxidant systems may provide greater understanding about the ecological relationships of insects with their hosts.

Animals↗

Cytochromes P450 of insects: the tip of the iceberg.

The cytochrome P450-dependent monooxygenases are an extremely important metabolic system involved in the metabolism of endogenous compounds and xenobiotics. Collectively, P450 monooxygenases can metabolize numerous substrates and carry out multiple oxidative reactions. The large number of substrates metabolized is due to the plethora of P450 isoforms and to the broad substrate specificity of some isoforms. Monooxygenases of insects have several functional roles, including growth, development, feeding and protection against xenobiotics, including resistance to pesticides and tolerance to plant toxins. This review begins with background information about P450s and their evolution, followed by a discussion of the extraordinary diversity of insect P450s. Given the enormous interest in studying individual P450s, we then provide a synopsis of the different methods that have been used in their isolation and the substrates that are known to be metabolized. We conclude by summarizing the lessons we have learned from the study of individual insect P450s, including their roles in insecticide resistance, plant-insect interactions and insect physiology. However, these studies are just the 'tip of the iceberg'. Our knowledge continues to expand at a rapid pace, suggesting that the next decade will outpace the last in terms of improving our understanding of the cytochromes P450 of insects.

Animals↗

Alteration of pyruvate metabolism in African trypanosomes during differentiation from bloodstream into insect forms.

In the presence of glucose and ample oxygen, insect form African trypanosomes release pyruvate more than 100-fold more slowly than do bloodstream forms. This rate decrease could not be accounted for simply by an increased mitochondrial pyruvate oxidation rate as inhibiting mitochondrial respiration increases pyruvate efflux to rates only 2-3% of that observed for bloodstream form trypanosomes. Alternatively, decreased pyruvate efflux from insect form trypanosomes could not be accounted for by decreased pyruvate transporter activity, which, surprisingly, was nearly as high in insect form trypanosomes as reported by us earlier for bloodstream forms (J.P. Barnard, B. Reynafarje, and P.L. Pedersen (1993) J. Biol. Chem. 268, 3654-3661). Rather, the low pyruvate efflux rate appears to be due primarily to reduced levels of the enzyme pyruvate kinase, which, in contrast to conclusions of an earlier study, is readily detected in insect form trypanosomes in the absence of added activators at an activity level about 4% of that found in bloodstream forms. Insect form pyruvate kinase seems to be located in the cytosol and exhibits kinetic profiles and constants nearly identical to those reported by us earlier for the bloodstream form enzyme (J.P. Barnard, and P.L. Pedersen (1988) Mol. Biochem. Parasitol. 31, 141-148). It is suggested that the reduced levels of pyruvate kinase, and hence the reduced pyruvate efflux rates, in insect form trypanosomes result from down regulation of the gene encoding the cytosolic enzyme.

Animals↗

Trypanosomatidae: Phytomonas detection in plants and phytophagous insects by PCR amplification of a genus-specific sequence of the spliced leader gene.

In this paper we describe a method for the detection of Phytomonas spp. from plants and phytophagous insects using the PCR technique by targeting a genus-specific sequence of the spliced leader (SL) gene. PCR amplification of DNA from 48 plant and insect isolates previously classified as Phytomonas by morphological, biochemical, and molecular criteria resulted in all cases in a 100-bp fragment that hybridized with the Phytomonas-specific spliced leader-derived probe SL3'. Moreover, this Phytomonas-specific PCR could also detect Phytomonas spp. in crude preparations of naturally infected plants and insects. This method shows no reaction with any other trypanosomatid genera or with plant and insect host DNA, revealing it to be able to detect Phytomonas spp. from fruit, latex, or phloem of various host plants as well as from salivary glands and digestive tubes of several species of insect hosts. Results demonstrated that SLPCR is a simple, fast, specific, and sensitive method that can be applied to the diagnosis of Phytomonas among cultured trypanosomatids and directly in plants and putative vector insects. Therefore, the method was shown to be a very specific and sensitive tool for diagnosis of Phytomonas without the need for isolation, culture, and DNA extraction of flagellates, a feature that is very convenient for practical and epidemiological purposes.

Animals↗

Differential budding efficiencies of human T-cell leukemia virus type I (HTLV-I) Gag and Gag-Pro polyproteins from insect and mammalian cells.

In this study, we examined the ability of human T-cell leukemia virus type I (HTLV-I) Gag and Gag-Pro to assemble immature virus-like particles (VLPs) and bud from insect and mammalian cells. Transmission electron microscopy of insect cells infected with a recombinant baculovirus carrying the entire gag gene revealed that Pr53(Gag) is targeted to the plasma membrane, where it extensively accumulates and forms electron-dense evaginations. However, no particles could be detected either inside the cells or in the culture supernatants. With the Gag-Pro-expressing construct, we observed HTLV-I-specific cytoplasmic proteolysis of the Gag precursor, but again no particle released in the culture supernatants. Transmission electron microscopic analysis of insect cells expressing Gag-Pro polyprotein revealed large vacuoles in the cytoplasm and no budding particles at the plasma membrane. In contrast, human immunodeficiency virus type 1 Gag polyprotein expressed in insect cells is able to release VLPs. These data showed that unlike other retroviruses, Pr53(Gag) is unable to be released as immature VLPs from insect cells. To determine whether the block in particle budding and release is due to an intrinsic property of Pr53(Gag) or the absence of essential cellular factors in insect cells, we expressed Gag and Gag-Pro polyproteins in human 293 cells. The results indicate that Pr53(Gag) and p24 capsid are released within particles into the culture supernatants of human 293 cells. We found that the myristylation of the N-terminal glycine residue is essential for Gag release. Altogether, these results strongly suggest that the proper assembly of HTLV-I particles is dependent on mammalian host cell factors.

Animals↗

Polyploidy in insect evolution.

Of all living organisms insects are the group with the highest number of existing species. It is, of course, true that a fraction of the total number of insects has been cytologically studied. Polyploid forms are rare exceptions among them. Polyploidy in insects is always associated with the parthenogenetic mode of reproduction. The cytologically verified cases are described. As for the geographic distribution of polyploid insects, they have successfully colonized vast land areas. Their distributions are, in general, northern and montane. The polyploid races are in general far more widespread than their diploid bisexual ancestors. The possible models of origin of polyploid insects are covered as well as data on their gaenetic variability. There are apparent environmental correlations in the distribution of certain forms. Most polyploid insects have life cycles extending over two (or more) years. They are also in general flightless forms. Hypotheses on the relation between heterozygosity in polyploids as well as the consequences of mutations in polyploid lineages are also presented.

Animals↗

Synthesis and metabolism of vertebrate-type steroids by tissues of insects: a critical evaluation.

This review covers the synthesis and the metabolism of vertebrate-type steroids (progesterone, testosterone, estradiol, corticosteroids) by insect tissues and discusses the significance of the reactions for insect physiology. Biosynthesis of vertebrate-type steroids from cholesterol hitherto has been demonstrated in only two insect species, i.e. the water beetle Acilius sulcatus (Coleoptera) and the tobacco hornworm Manduca sexta (Lepidoptera). In Acilius, steroid synthesis is associated with exosecretion (chemical defense). Nothing, however, is known about a physiological role of the C21 steroid conjugate present in ovaries and eggs of Manduca. No synthesis of vertebrate-type steroids was observed in any other insect investigated to date. Most metabolic conversions of steroids by insects concerned oxidoreduction of oxygen groups (hydroxysteroid dehydrogenase activity) and (polar and apolar) conjugate formation. All important enzymatic steps involved in synthesis and catabolism, as known from studies with tissues of vertebrates, were not, or hardly observed. The conclusion is drawn that typical vertebrate-type (C21, C19 and C18) steroids probably do not act as physiologically active substances in insects.

Animals↗

Review and patents and literature. The use of insect cell cultures for recombinant protein synthesis: Engineering aspects.

The use of the insect cell/baculovirus expression system for producing recombinant proteins of bacterial, plant, insect, and mammalian origin has become widespread. The popularity of this eukaryotic expression system is due to many factors, including (1) potentially high protein expression levels, (2) ease and speed of genetic engineering, (3) ability to accommodate large DNA inserts, (4) protein processing similar to higher eukaryotic cells (e.g., mammalia cells), and (5) ease of insect cell growth (e.g., suspension growth). The following review of the literature discusses two engineering aspects of recombinant protein synthesis by insect cell cultures: bioreactor scale-up and insect cell line selection. Following this review patent abstracts and additional literature pertaining to expression of recombinant proteins in insect cell culture are listed.

Animals↗

Extensive sequence conservation among insect, nematode, and vertebrate vitellogenins reveals ancient common ancestry.

The eggs of most oviparous animals are provisioned with a class of protein called vitellogenin (Vg) which is stored as the major component of yolk. Until recently, deduced amino acid sequences were available only from vertebrate and nematode Vgs, which proved to be homologous. The sequences of several insect Vgs are now known, but early attempts at pairwise alignments with vertebrate and nematode Vgs have been problematic, leading to conflicting conclusions about how closely insect Vgs are related to the others. In this paper we demonstrate that insect Vg sequences can be confidently aligned with one another along their entire lengths and with multiple vertebrate and nematode Vg sequences along most of their spans. Although divergence is high, conservation among insect, vertebrate, and nematode Vg sequences is widespread with a preponderance of glycine, proline, and cysteine residues among strictly conserved amino acids, establishing conclusively that Vgs from the three phyla are homologous. Areas of least-certain alignment are primarily in and around insect and vertebrate polyserine domains which are not homologous. Phylogenetic reconstructions of Vgs based on sequence identities indicate that the insect lineage is the most diverged and that the mammalian serum protein, apolipoprotein B-100, arose from a Vg ancestor after the nematode/vertebrate divergence.

Amino Acid Sequence↗