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PubMed1981

Bee venom immunotherapy: clinical and immunologic observations.

In the present study the authors evaluated the usefulness of pure venom (PV) in the diagnosis and prevention of bee allergy in a group of 25 patients. In addition, they followed changes in specific IgE and IgG serum levels during the course of immunotherapy. They found excellent correlation between the clinical history and skin test reactivity to PV. The RAST values correlated well with skin test but were somewhat less specific. They used a rather moderate schedule of desensitization by which a 100 mcg maintenance dose was attained by the 11th week. This schedule resulted in minimal side effects and was extremely effective. None of the live insect challenged patients had major reactions. Successful immunization was not associated by a decrease in specific IgE level. In contrast, most patients exhibited an increase in specific serum IgG levels. An increase in IgG level did not prove to be an absolute indication of successful immunization.

Bee Venoms↗
PubMed1997

Antennal lobe neurons of the honey bee, Apis mellifera, express a D2-like dopamine receptor in vitro.

We have used the D2-specific dopamine receptor ligand spiperone [N-(p-aminophenethyl) spiperone; NAPS] coupled to the fluorophore 7-nitrobenz-2-oxa-1,3-diazole-4-yl (NBD) to visualize dopamine receptors expressed in vitro by neurons of the primary antennosensory centers (antennal lobes) of the brain of the honey bee, Apis mellifera. Changes in the percentage of antennal lobe neurons exhibiting spiperone binding sites over time in culture and at different stages of metamorphic adult development have been investigated. Neurons obtained from animals at all stages of development exhibited spiperone binding sites, but only after 2 days or more in vitro. The percentage of antennal lobe neurons in vitro expressing spiperone binding sites increased significantly with the development of the antennal lobe neuropil. Fluorescently labelled spiperone (120 nM) could be displaced effectively by 1 mM dopamine but not by the same concentration of tyramine, octopamine, or serotonin. In addition, the D2 antagonist spiperone and the D2/D1 antagonist fluphenazine were more effective at displacing the fluorescent ligand than the D1-specific antagonist SCH23390. Our results indicate that Apis antennal lobe neurons in culture express a dopamine receptor and that this receptor is more likely to be D2-like than D1-like in nature. The receptor is expressed early in the metamorphic adult development of the antennal lobe neuropil of the brain.

Animals↗
PubMed1998

Antigen-independent suppression of the IgE immune response to bee venom phospholipase A2 by maternally derived monoclonal IgG antibodies.

The IgE immune response to ovalbumin in rats can be suppressed by prior immunization of the dams. The results reported in this paper extend this observation to include a different antigen and another species, namely the IgE immune response to bee venom phospholipase A2 (PLA2) in CBA/J mice. The degree of suppression seemed to depend on the amount of IgG antibodies transferred to the offspring. Moreover, we found that the maternally mediated suppression of the IgE response could be achieved in a completely antigen-free system in which exogenous monoclonal anti-PLA2 IgG antibodies were transferred from the dams to the offspring. The following results were obtained: (i) The IgE suppression by monoclonal IgG antibodies was induced as efficiently with one single anti-PLA2 IgG1 antibody as with a mixture of ten antibodies (nine IgG1, one IgG2b). (ii) Even after several immunizations up to an age of 6 months with a dose of PLA2 that normally induces IgE production, none of the F1 mice developed an IgE response. (iii) This long-lasting suppression was observed in mice which were first immunized at an age of 4 weeks (i.e. when low amounts of maternally derived monoclonal IgG were still present), as well as in mice which were first immunized at an age of 8 weeks, when no such maternal antibodies could be detected in their sera. The corresponding IgG responses showed, compared to normal mice, a transient enhancement in the maternally influenced mice. It is concluded that the immunological experience of the mother is of particular importance for the isotype regulation in the newborns, especially with respect to the ability to elicit an IgE response. The possible implications for the development of allergic diseases in humans are discussed.

Animals↗
PubMed2002

Cloning and characterization of Acmar1, a mariner-like element in the asiatic honey bee, Apis cerana japonica (Hymenoptera, Apocrita).

A mariner-like element was cloned from the genome of the Asiatic honey bee, Apis cerana japonica (Hymenoptera, Apocrita). The (composite) clone, named Acmar1, was 1,378 bp long, and encoded 336 amino acids corresponding to a transposase-like putative polypeptide in a single open reading frame. The D,D(34)D motif, the catalytic domain of the mariner transposase, was present, although there was a deletion of five amino acid residues within it as compared with the active transposase in Drosophila mauritiana. Nineteen-bp-long imperfect inverted terminal repeat-like sequences flanked by TA dinucleotides, the typical target site for mariner insertion, were observed. Southern blot analysis using a fragment covering two-thirds of the Acmar1 transposase coding sequence as a probe indicated the presence of multiple Acmar1-like elements in the genome. Maximum-parsimony phylogenetic analysis based on the transposase amino acid sequences of insect mariner-like elements revealed that Acmar1 is a member of the mellifera subfamily.

Amino Acid Sequence↗
PubMed2004

Antiarthritic effect of bee venom: inhibition of inflammation mediator generation by suppression of NF-kappaB through interaction with the p50 subunit.

OBJECTIVE: To investigate the molecular mechanisms of the antiarthritic effects of bee venom (BV) and melittin (a major component of BV) in a murine macrophage cell line (Raw 264.7) and in synoviocytes obtained from patients with rheumatoid arthritis. METHODS: We evaluated the antiarthritic effects of BV in a rat model of carrageenan-induced acute edema in the paw and in a rat model of chronic adjuvant-induced arthritis. The inhibitory effects of BV and melittin on inflammatory gene expression were measured by Western blotting, and the generation of prostaglandin E(2) (PGE(2)) and nitric oxide (NO) and the intracellular calcium level were assayed. NF-kappaB DNA binding and transcriptional activity were determined by gel mobility shift assay or by luciferase assay. Direct binding of BV and melittin to the p50 subunit of NF-kappaB was determined with a surface plasmon resonance analyzer. RESULTS: BV (0.8 and 1.6 mug/kg) reduced the effects of carrageenan- and adjuvant-induced arthritis. This reducing effect was consistent with the inhibitory effects of BV (0.5, 1, and 5 mug/ml) and melittin (5 and 10 mug/ml) on lipopolysaccharide (LPS; 1 mug/ml)-induced expression of cyclooxygenase 2, cytosolic phospholipase A(2), inducible NO synthase, generation of PGE(2) and NO, and the intracellular calcium level. BV and melittin prevented LPS-induced transcriptional and DNA binding activity of NF-kappaB via the inhibition of IkappaB release and p50 translocation. BV (affinity [K(d)] = 4.6 x 10(-6)M) and melittin (K(d) = 1.2 x 10(-8)M) bound directly to p50. CONCLUSION: Target inactivation of NF-kappaB by directly binding to the p50 subunit is an important mechanism of the antiarthritic effects of BV.

Animals↗
PubMed2004

From the linden flower to linden honey--volatile constituents of linden nectar, the extract of bee-stomach and ripe honey.

Honey is produced by honeybees (Apis mellifera), which collect nectar from flowers, digest it in their bodies, and deposit it in honeycombs, where it develops into ripe honey. We studied the evolution of the volatile constituents from the nectar of linden blossoms (Tilia cordata) to honey via the 'intermediate' honeybee. The sampling of the contents of the honey stomach or honey sack of the bee is unique. Extracts were prepared from nectar, from the liquid of the honey stomach, and from ripe honey. The chemistry is extremely complex, and compounds spanning from monoterpenes (hydrocarbons, ethers, aldehydes, acids, and bifunctional derivatives), isoprenoids, aromatic compounds (phenylpropanoids, phenols), and products degraded from fatty acids to alkaloids, were identified. Some compounds definitely stem from the plants, whereas other interesting constituents can be attributed to animal origin. Two derivatives of decanoic acid, 9-oxodec-2-enoic acid (12) and 9-hydroxydec-2-enoic acid, identified in the honey are known to be constituents of the so-called 'Queen's pheromone'. Two metabolites of these acids were identified in the extract of the honey stomach: 8-oxononanal (10), a new natural product, and 8-oxononanol (11). There structures were confirmed by synthesis. Nectar and honey stomach contain many aldehydes, which, due to the highly oxidative atmosphere in the honeycomb, are found as corresponding acids in the honey. Two acids were newly identified as 4-isopropenylcyclohexa-1,3-diene-1-carboxylic acid (14) and 4-(1-hydroxy-1-methylethyl)-cyclohexa-1,3-diene-1-carboxylic acid (15).

Animals↗
PubMed2006

The molecular mechanism of bee venom phospholipase A2 inactivation by bolinaquinone.

The molecular basis of the inactivation of bee venom PLA2 by the marine natural product bolinaquinone (BLQ) was studied by several spectral techniques (CD, fluorescence, and NMR spectroscopy, mass spectrometry), biomimetic reactions, and molecular modeling. Our data suggest competitive inhibition based on a BLQ-PLA2 noncovalent molecular recognition. However, BLQ is also able to react selectively with Lys133 through conjugate addition followed by a beta elimination. The biological implications of both the covalent and noncovalent molecular events are discussed.

Bee Venoms↗
PubMed2005

Octopamine-like immunoreactivity in the honey bee and cockroach: comparable organization in the brain and subesophageal ganglion.

A serum raised against octopamine reveals in cockroaches and honey bees structurally comparable systems of perikarya and their extensive yet discrete systems of arborizations in neuropils. Numerous and prominent clusters of lateral cell bodies in the brain as well as many midline perikarya provide octopamine-like immunoreactive processes to circumscribed regions of the subesophageal ganglion, antennal lobe glomeruli, optic neuropils, and neuropils of the protocerebrum. There is dense octopaminergic innervation in the protocerebral bridge and ellipsoid body of the central complex. The antennal lobes are supplied by at least three octopamine-immunoreactive neurons. In contrast, the mushroom bodies show the fewest immunoreactive elements. In Apis a single axon supplies sparse immunoreactive processes to the calyces' basal ring, collar, and lip. A diffuse arrangement of immunoreactive processes invades all zones of the mushroom body calyces in Periplaneta. These processes derive from an ascending axon ascribed to a dorsal unpaired median neuron at the maxillary segment of the subesophageal ganglion. In both taxa octopamine-immunoreactive processes invade only the gamma lobes of the mushroom bodies, omitting their other divisions. The present observations are discussed with respect to possible roles of octopamine in sensory integration and association.

Animals↗
PubMed1989

Sensory pathways and motoneurons of the proboscis reflex in the suboesophageal ganglion of the honey bee.

A morphological description is given for the motoneurons underlying the proboscis extension reflex in the honey bee (Apis mellifera). Sensory neurons from the mandibles and labium were examined and their projection areas were related to those of the motoneurons. Motoneurons are located in the ventral part of the suboesophageal ganglion, have their somata in the hemiganglion ipsilateral to the filled nerve, and send prominent branches through identified ventral commissures into the contralateral hemiganglion. Motoneurons to the same muscle have parallel-running projections into the contralateral hemiganglion, often parallel-running neurites and show vast areas of dendritic overlap. Arborizations of motoneurons are not restricted to their neuromere of origin but invade adjacent neuromeres, resulting in regions of dendritic overlap of motoneurons to different muscles. Sensory fibers from the mouthparts terminate in medio- and mediolateral parts of the suboesophageal ganglion, where their arborizations overlap with projections from motoneurons. A few sensory fibers descend in ventral parts of the cervical connective. Although sensory neurons from the mandible are restricted to the ipsilateral hemiganglion, those of the labium also show contralateral branches. These observations are discussed in the context of the suboesophageal ganglion as a ganglion composed of the embryonic fusion of three single neuromeres.

Animals↗
PubMed1995

Carbohydrate-dependent, HLA class II-restricted, human T cell response to the bee venom allergen phospholipase A2 in allergic patients.

The T cell-independent antibody response to polysaccharide antigen (Ag) is believed to result from their inability to bind major histocompatibility complex (MHC) restriction elements. However, recent studies using glycosylated analogues of known immunogenic peptides revealed that glycopeptides can interact with MHC molecules and are able to elicit specific T cell responses in experimental animals. This raises questions about the possible role which carbohydrates can play in T cell responses following natural exposure to glycoprotein antigens. Analyzing the fine specificity of the human T cell response against the major bee venom allergen phospholipase A2 (PLA), a 16-20-kDa protein glycosylated at a single site (Asn13), we have identified several T cell clones which proliferate in response to the glycoprotein but not to its non-glycosylated variants. Neither the carbohydrate moiety alone nor the combination of carbohydrate and non-glycosylated protein could substitute for the intact glycoprotein. Antibody directed against the carbohydrate moiety inhibited Ag-induced proliferation of these clones whereas control clones with known peptide specificity were not affected, providing additional evidence for the involvement of carbohydrates in T cell recognition. Moreover, peripheral blood mononuclear cells of two individuals from whom glycosylation-dependent T cell clones have been isolated showed significantly higher proliferation in response to glycosylated compared to non-glycosylated Ag, suggesting that glycosylation can contribute in some cases extensively to the immunogenicity of a glycoprotein Ag. Thus, this report shows that glycosylation-dependent Ag recognition by T cells can also occur following natural exposure to a glycoprotein.

Allergens↗
PubMed1991

Developmental study of afferented and deafferented bee antennal lobes.

The role of antennal sensory projections on the ontogeny of the bee antennal lobe was analyzed using both light and transmission electron microscopy. Normal and deafferented developing antennal lobes were examined. The results obtained show that (1) initiation of synaptogenesis in the antennal lobe is independent of the arrival of sensory inputs; (2) sensory inputs are necessary for setting up the glomerular antennal lobe organization; (3) regressive events, such as the reduction of synapse density, occur during the development of the antennal lobe; and (4) glomeruli formation appears as related to glia development.

Animals↗
PubMed1999

Phylogeny of the bee genus Halictus (Hymenoptera: halictidae) based on parsimony and likelihood analyses of nuclear EF-1alpha sequence data.

We investigated higher-level phylogenetic relationships within the genus Halictus based on parsimony and maximum likelihood (ML) analysis of elongation factor-1alpha DNA sequence data. Our data set includes 41 OTUs representing 35 species of halictine bees from a diverse sample of outgroup genera and from the three widely recognized subgenera of Halictus (Halictus s.s., Seladonia, and Vestitohalictus). We analyzed 1513 total aligned nucleotide sites spanning three exons and two introns. Equal-weights parsimony analysis of the overall data set yielded 144 equally parsimonious trees. Major conclusions supported in this analysis (and in all subsequent analyses) included the following: (1) Thrincohalictus is the sister group to Halictus s.l., (2) Halictus s.l. is monophyletic, (3) Vestitohalictus renders Seladonia paraphyletic but together Seladonia + Vestitohalictus is monophyletic, (4) Michener's Groups 1 and 3 are monophyletic, and (5) Michener's Group 1 renders Group 2 paraphyletic. In order to resolve basal relationships within Halictus we applied various weighting schemes under parsimony (successive approximations character weighting and implied weights) and employed ML under 17 models of sequence evolution. Weighted parsimony yielded conflicting results but, in general, supported the hypothesis that Seladonia + Vestitohalictus is sister to Michener's Group 3 and renders Halictus s.s. paraphyletic. ML analyses using the GTR model with site-specific rates supported an alternative hypothesis: Seladonia + Vestitohalictus is sister to Halictus s.s. We mapped social behavior onto trees obtained under ML and parsimony in order to reconstruct the likely historical pattern of social evolution. Our results are unambiguous: the ancestral state for the genus Halictus is eusociality. Reversal to solitary behavior has occurred at least four times among the species included in our analysis.

Animals↗
PubMed2000

Analysis of the complete genome sequence of acute bee paralysis virus shows that it belongs to the novel group of insect-infecting RNA viruses.

The complete genome sequence of acute bee paralysis virus (ABPV) was determined. The 9470 nucleotide, polyadenylated RNA genome encoded two open reading frames (ORF1 and ORF2), which were separated by 184 nucleotides. The deduced amino acid sequence of the 5' ORF1 (nucleotides 605 to 6325) showed significant similarity to the RNA-dependent RNA polymerase, helicase, and protease domains of viruses from the picornavirus, comovirus, calicivirus, and sequivirus families, as well as to a novel group of insect-infecting RNA viruses. The 3' ORF2 (nucleotides 6509-9253) was proposed as encoding a capsid polyprotein with three major structural proteins (35, 33, and 24 kDa) and a minor protein (9.4 kDa). This was confirmed by N-terminal sequence analysis of two of these proteins. The overall genome structure of ABPV showed similarities to those of Drosophila C virus, Plautia stali intestine virus, Rhopalosiphum padi virus, and Himetobi P virus, which have been classified into a novel group of picorna-like insect-infecting RNA viruses called cricket paralysis-like viruses. It is suggested that ABPV belongs to the cricket paralysis-like viruses.

Amino Acid Sequence↗
PubMed1983

Caste-specific maturation of the endocrine system in the female honey bee larva.

The endocrine system of female honey bee larvae has been studied through postembryonic development with histological and autoradiographic techniques. During larval development, brain and retrocerebral complex proceed from immature cells to an active endocrine system. Caste-specific retardation occurs in the worker during this process. In the developing queen, the differentiation of the neurosecretory cells (NSC) and the outgrowth of their axons occurs from the second instar onward and is nearly completed in the fourth, whereas in the worker larva these processes are delayed by more than one instar. In the queen, RNA synthesis starts in the NSC at the end of the third instar and in the worker at the fifth instar. Stainable neurosecretory material is present only in fifth instar queen larvae. The queen's corpora cardiaca become active at the end of the fourth, those of the worker in the fifth instar. In the corpora allata (CA), nuclei undergo several phases of endomitosis. These phases of polyploidization end at the beginning (queen) or at the end (worker) of the fifth instar respectively. CA volume in the queen is twice that of a worker at its height at the end of larval development. these data demonstrate a caste-specific maturation of the endocrine organs which results in differences in hormone titres.

Animals↗
PubMed1982

Antennal hygroreceptors of the honey bee, Apis mellifera L.

Antennal hygroreceptors of the honey bee, Apis mellifera L., have been investigated electrophysiologically and the sensillum containing these receptors with SEM. Moist and dry hygroreceptors have been identified along with a thermal receptor in a specialized coeloconic sensillum. This sensillum comprises a cuticular, shallow depression (diameter; 4 micrometer) having a central opening (1.4-1.5 micrometer) and a mushroom-shaped protrusion (1.4-1.5 micrometer) from the opening. The head of the protrusion is irregular in shape and is not perforated. This sensillum has been thus far referred to as a "sensillum campaniformium" (Dietz and Humphreys 1971), henceforth, it is referred to as a coelocapitular sensillum. The responses of both moist and dry hygroreceptors are of a phasic-tonic manner. Both receptors are antagonistic with respect to their responses to humidity; one responds with an increase in impulse frequency to rising humidity, the other to falling humidity. The humidity-response relationship is independent of stimulus flux.

Animals↗
PubMed1984

The first optic ganglion of the bee. V. Structural and functional characterization of centrifugally arranged interneurones.

The organization, characterization and connectivity patterns of four different interneurone types were studied with the use of Golgi light- and electron-microscopic techniques. All four cell types originate in the outer chiasma; they have an efferent end-branch in the lamina and an afferent one terminating in the distal region of the second optic ganglion, the medulla. These interneurones are referred to as: (i) Garland-cell: The efferent fibre has on its tangential branch numerous centripetal side branches, so-called "garlands", which synapse with first- and second-order visual cells. (ii) Y-cell: The lamina branch bifurcates before entering the lamina. It innervates two neighbouring cartridges. Synaptic contacts were seen in two different laminar strata where bottle-brush-like collaterals occurred. (iii) Single bottle-brush cell: The efferent part has only one centrifugal branch, which can be compared morphologically and in terms of synaptology with those of the Y-cell. (iv) Triptych-cell: The lamina component innervates three neighbouring cartridges at three different laminar layers interconnecting different first- and second-order visual neurones. The present study provides some essential qualitative and quantitative fine-structural information, which - when compared with adequate physiological data - may lead to a better understanding of the function of the first visual information-processing centre of the bee.

Animals↗
PubMed1975

The retina-lamina projection in the visual system of the bee, Apis mellifera.

Single Golgi impregnated visual cells and their axons were treated from the retina to the first synaptic layer (lamina) in serial electron microscopic sections. This analysis of the retina-lamina projection was undertaken in the upper dorso-median eye region which is known to be involved in the perception of polarized light. For identification of individual visual cells and their fibres a numbering system was used which relates the number of each of the nine visual cells within one retinula to the transverse axis of the rhabdom (TRA) (Fig. 1). Because of the twist of the retinula along its course to the basement membrane (Fig. 6), individual visual cells change their position relative to any eye-constant co-ordinate system. Each axon bundle originating from one 9-celled retinula performs a 180 degrees-rotation before entering the lamina (Fig. 2). The direction of rotation (clockwise or counter-clockwise), which may differ even between adjacent bundles, is related to the two mirror-image types of rhabdoms in the corresponding retinulae and is opposite to the direction of rhabdom twist. Thus, even in small groups of the in total 5500 ommatidia in the eye of the bee, two types of retinulae exist which can be characterized by the geometry of the rhabdoms as well as by the direction of rotation of the retinulae and the axon bundles (Fig. 1). Visual cell numbers 1, 2, and 9, the microvilli of which are oriented in the direction of TRA, form three long visual fibres terminating in the second synaptic layer (medulla). In cross sections of laminar pseudocartridges they appear as the smallest fibre profiles arranged in a symmetrical line of the pseudocartridge bundle (=the transverse axis of the pseudocartridge; TPA) (Fig. 4). The remaining six fibres (cell numbers 3-8) only project to the lamina (short visual fibres; svf's). Two of them (cell numbers 5 and 6), which are the largest cells in the proximal retinula and have their microvilli perpendicularly arranged to TRA (Fig. 1), give rise to the two thickest axons of the underlaying pseudocartridge. In cross sections, t he connecting line of these two axons is orthogonally oriented to TPA (Fig. 5). A model was developed, in which all long visual fibres originate from ultraviolet receptors and in which the polarization sensitivity of the basal ninth cell is enhanced by the twist of the rhabdom. Finally, this model is discussed in light of behavioral experiments revealing the ultraviolet receptors as the only cells involved in the detection of polarized light.

Animals↗
PubMed1975

The first optic ganglion of the bee. I. Correlation between visual cell types and their terminals in the lamina and medulla.

Each visual unit (ommatidium) of the compound eye of the honey bee contains nine retinula cells, six of which end as axons in the first synaptic ganglion, the lamina, and three in the second optic ganglion, the medulla. A technique allowing light- and electron microscopy to be performed on the same silver-impregnated sections has made it possible to follow all types of retinula axons of one ommatidium to their terminals in order to study the shape of the terminal branches with their position in the cartridge. 1. The axons of retinula cells 1-6 (numbered according to Menzel and Snyder, 1974) end as three different types of short visual fibres (svf) in the lamina; the axons of retinula cells 7-9 run through the lamina to terminate in the medulla and are known as long visual fibres (lvf). Retinula cells of each type are identified by the location of their cell bodies and by the direction of their microvilli. The retinula cells 1 and 4 (group I according to Gribakin, 1967) end as svf type 1 with three tassel-like branches in stratum B of the first synaptic region. The pair of cells 3, 6 and the pair 2, 5 (group II) end in the first synaptic region in stratum A. Cells 3 and 6 have forked endings, svf type 2, whereas cells 2 and 5 have tapered endings, svf type 3. The remaining retinula cells 7, 8 and 9 have long fibres. Nos. 7 and 8 (group III) have tapered endings and are termed lvf types 1 and 2, respectively. The 9th cell is the lvf type 3 with a highly branched ending. 2. The nine axons in the bundle from one ommatidium have relative positions which do not change from the proximal retina to the monopolar cell body layer. 3. By following silver-stained retinula cells and their corresponding axons, it is possible to describe mirror-image arrangements of fibres in the axon bundles in different parts of the eye. This correlation of numbered retinula cells with specific axon types, together with the highly organized pattern in an axon bundle, allows the correlation between histological and physiological findings on polarization and colour perception.

Animals↗