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Properties of rabbit erythrocytes treated with phospholipase A2 from bee venom.

In isotonic saline-buffered conditions, phospholipase A2 from bee venom cleaves up to 65% of the phosphatidylcholine of the rabbit RBC membrane without causing significant hemolysis; however, the volume and the osmotic fragility of the treated RBC is modified. The osmotic behaviour of PLA-treated RBC after reincubation in autologous plasma suggests an heterogeneity in the enzymatic attack among the RBC population. In vivo RBC survival is strongly impaired by PLA treatment.

Animals↗

Modulation of spike frequencies by varying the ambient magnetic field and magnetite candidates in bees (Apis mellifera).

1. Spontaneous activity was recorded from neurons in the second abdominal ganglion of bees. 2. Thirty per cent intensity modulations of the horizontal component of the background magnetic field provoked changes in the firing pattern of single neurons. 3. Two classes of neurons were distinguished and confirmed by statistical analysis. 4. Electron dense material in hairs and in or near the cutex may be single domain (SD) and superparamagnetic (SPM) magnetite. 5. A hypothesis is proposed for magnetoreception. Magnetite would act as an amplifier of the external magnetic induction changes. 6. The amplified magnetic field would influence neuronal elements only in restricted regions near the magnetite.

Abdomen↗

Evidence for light-induced release of Ca2+ from intracellular stores in bee photoreceptors.

In the drone retina light elicits an increase in the extracellular Ca2+ concentration ([Ca]o). After withdrawal of extracellular Ca2+ and addition of 1 mM EGTA (ethyleneglycol-bis (beta-aminoethyl ether) N,N,N',N'-tetraacetic acid) a short light flash still caused an increase in [Ca]o as measured with Ca2(+)-sensitive microelectrodes. This increase vanished after a few flashes. When [Ca]o was reduced to 10(-5) M Ca2+ (no EGTA) the rise in Cao disappeared after several light flashes. Subsequent stimulation with a dim steady light produced a decrease in [Ca]o and caused a recovery of the increase in [Ca]o elicited by a test flash. The results show that bee photoreceptors have a light-depletable intracellular Ca2+ store which can be reloaded by dim continuous lights.

Animals↗

Dufour's gland triglycerides from Anthophora, emphoropsis (Anthophoridae) and Megachile (Megachilidae) bees (Hymenoptera: Apoidea).

Lipids secreted from the Dufour's glands of the bees Anthophora centriformis, Anthophora californica tarsata, Emphoropsis miserabilis, Megachile (Delomegachile) gemula, M. (Phaenosarus) fortis, M. (Argyropile) parallela, and M. (Melanosarus) xylocopoides primarily consist of short chain (C2-C20) fatty acid triglycerides. Nearly 95% of the triglycerides from Anthophora and Emphoropsis have one palmitoyl-, stearoyl- or arachidoyl moiety, plus two shorter acids (C2-C16). Virtually all triglycerides (98%) from the four Megachile species possess only short-chain fatty acids (C2-C16), often with one like pair of acids.

Animals↗

Purity control of different bee venom melittin preparations by capillary zone electrophoresis.

A rapid capillary zone electrophoresis method was developed for the determination of melittin during its purification from bee venom by column chromatography. Low-pH buffer was selected to decrease the adsorption of the analytes on capillary wall. The average relative standard deviations of the migration time of melittin and of the peak area were 1.2 and 2.3%, respectively.

Bee Venoms↗

Habituation and dishabituation of exploratory and appetitive responses in the honey bee (Apis mellifera L.).

Sucrose-induced antennal and proboscis motor responses of the honey bee were comparatively studied using muscle responses. The activity of the proboscis muscle responses was correlated with sucrose stimulus concentration, but that of antennal muscle responses was not. Both responses habituated following repetitive stimulation with a low sucrose concentration and dishabituated upon stimulation with a high sucrose concentration. Despite the different response properties of the responses, their habituation kinetics were indistinguishable. The absence of sensory adaptation was confirmed by taste hair recordings. The potential for habituation was related to the functions of the responses studied. The exploratory antennal response used for continuous sampling of the environment habituated to a smaller extent than the appetitive proboscis response.

Adaptation, Physiological↗

Evidence for peripherally antinociceptive action of propofol in rats: behavioral and spinal neuronal responses to subcutaneous bee venom.

In the present study, behavioral and in vivo electrophysiological methods were used to examine the peripheral effects of propofol on tonic ongoing pain-related responses produced by subcutaneous bee venom-induced inflammatory pain state. Local administration of 0.5 microg propofol produced significant suppression of the well-established ongoing pain responses in both conscious rats and dorsal horn nociceptive neurons. The locally antinociceptive action of propofol is not caused by systemic effect, because contralateral administration of the same dose of drug did not produce any effect. This result indicates that besides central actions, propofol has peripherally antinociceptive action as well.

Analgesics↗

The anti-inflammatory effect of peripheral bee venom stimulation is mediated by central muscarinic type 2 receptors and activation of sympathetic preganglionic neurons.

The anti-inflammatory effect (AI) induced by peripheral injection of diluted bee venom (dBV) involves activation of spinal cord circuits and is mediated by catecholamine release from adrenal medulla, but the precise neuronal mechanisms involved are not fully understood. In a recent study, we demonstrated that an increase in spinal acetylcholine is involved in mediating the anti-inflammatory effect of dBV and that this mediation also involves adrenomedullary activation. The present study utilized the mouse air pouch inflammation model to evaluate the involvement of spinal acetylcholine receptors and sympathetic preganglionic neurons (SPNs) in dBV's anti-inflammatory effect (dBVAI). Intrathecal (IT) pretreatment with atropine (muscarinic cholinergic antagonist) but not hexamethonium (nicotinic cholinergic antagonist) significantly suppressed dBVAI on zymosan-evoked leukocyte migration. Subsequent experiments showed that IT pretreatment with methoctramine (a muscarinic receptor type 2; M(2) antagonist), but not pirenzepine (an M(1) antagonist) or 4-DAMP (an M(3) antagonist), suppressed the dBVAI. In addition, dBV stimulation specifically increased Fos expression in SPNs of the T7-T11, but not the T1-T6 or T12-L2 spinal cord segments, in animals with zymosan-induced inflammation. Moreover, IT methoctramine pretreatment suppressed this dBV-induced Fos expression specifically in SPNs of T7-T11 level. Peripheral sympathetic denervation using 6-hydroxydopamine (6-OHDA) treatment (which spares sympathetic adrenal medullary innervation) did not alter dBVAI. Collectively these results indicate that dBV stimulation leads to spinal cord acetylcholine release that in turn acts on spinal M(2) receptors, which via a hypothesized disinhibition mechanism activates SPNs that project to the adrenal medulla. This activation ultimately leads to the release of adrenal catecholamines that contribute to dBVAI.

Animals↗

Antinociceptive effect and the mechanism of bee venom acupuncture (Apipuncture) on inflammatory pain in the rat model of collagen-induced arthritis: Mediation by alpha2-Adrenoceptors.

The antinociceptive effect and the mechanism of bee venom acupuncture (BVA) on inflammatory pain, especially in the rat model of collagen-induced arthritis (CIA), have not yet been fully studied. This study was designed to investigate the antinociceptive effect and its mu-opioid and alpha2-adrenergic mechanism of BVA in the CIA rat model. To induce CIA, male Sprague-Dawley rats were immunized with bovine type II collagen emulsified in Freund's incomplete adjuvant followed by a booster injection 14 days later. The antinociceptive effect was evaluated by tail flick latency (TFL). After induction of arthritis, the inflammatory pain threshold decreased as time passed, and there was no big change of the pain threshold after 3 weeks. Three weeks after the first immunization, BVA (0.25 mg/kg) injected into the Zusanli acupoint (ST36) showed the antinociceptive effect. Furthermore, the antinociceptive effect of BVA was blocked by yohimbine (alpha2-adrenergic receptor antagonist, 2 mg/kg, i.p) pretreatment, but not by naloxone (mu-opioid receptor antagonist, 2 mg/kg, i.p.) pretreatment. These results suggest that BVA can relieve inflammatory pain in CIA and the antinociceptive effect of BVA can be mediated by alpha2-adrenergic receptor.

Acupuncture Points↗

Glutathione S-transferase in the insect Apis mellifera macedonica kinetic characteristics and effect of stress on the expression of GST isoenzymes in the adult worker bee.

The glutathione S-transferase present in the adult worker bee Apis mellifera macedonica was purified and analyzed for its physicochemical and kinetic properties. The enzyme is heterodimeric with subunit molecular masses of 29 and 25 kDa, respectively. Two main isoenzymes with distinct kinetic properties are present, with isoelectric points of 7.40 for the alkaline and 4.58 for the acidic forms, respectively. The two enzymes are induced independently by factors such as insecticide treatments and environmental conditions, including low temperatures or starvation.

Animals↗

Sex determination: balancing selection in the honey bee.

Sequences of alleles of the honey bee's primary sex-determining gene have extremely high diversity, with many amino acid variants, suggesting that different alleles of this gene have been maintained in populations for very long evolutionary times.

Alleles↗

Unilateral subcutaneous bee venom but not formalin injection causes contralateral hypersensitized wind-up and after-discharge of the spinal withdrawal reflex in anesthetized spinal rats.

This study aimed to investigate the effect of tonic nociception on spinal withdrawal reflexes including (1) long lasting spontaneous responses elicited by subcutaneous (s.c.) administration of formalin (2.5%, 50 microl) and bee venom (BV, 0.2 mg/50 microl) into the hind paw and (2) corresponding ipsilateral (primary) and contralateral (secondary) hypersensitivity to noxious pinch and repetitive supra-threshold (1.5 x T) electrical stimuli at different frequencies (3 Hz: wind-up; 20 Hz: after-discharge) in anesthetized spinal rats. Spinal withdrawal reflexes were studied by simultaneously assessing single motor units (SMUs) electromyographic (EMG) activities from the bilateral medial gastrocnemius (MG) muscles. Subcutaneous formalin-induced persistent spontaneous SMU EMG responses were in typical biphasic manner with an apparent silent period (about 13-18 min), but in contrast, BV elicited monophasic long lasting (about 1 h) SMU EMG responses without any resting state. The mechanically and electrically evoked responsiveness of SMUs were enhanced significantly by ipsilateral BV injection, whereas enhanced electrically, but not mechanically, evoked responses (including wind-up and after-discharge) were found at the non-injection site of the contralateral hind paw. However, s.c. administration of formalin was only able to establish ipsilateral hypersensitivity of the SMUs to repeated electrical, not mechanical, stimulation. Neither mechanically nor electrically evoked contralateral hypersensitivity of the SMUs was found during the ipsilateral formalin-induced nociception. For pharmacological intervention, intrathecal administration of the non-N-methyl-d-aspartate (non-NMDA) receptor antagonist CNQX (40 nmol/10 microl), but not the non-competitive NMDA receptor antagonist MK-801 (40 nmol/10 microl), significantly depressed BV-induced contralateral hypersensitivity of the SMUs to repeated 3 Hz (wind-up) and 20 Hz (after-discharge) frequencies of electrical stimulation. Using the extracellular SMU recording technique, we found that s.c. administration of formalin and BV shows a significant difference in long lasting spontaneous firing of SMUs. This is consistent with previous observations in animal behavioral studies. Additionally, contralateral electrically evoked hypersensitivity of the SMUs was found only following BV injection, not in the formalin test. The maintenance and development of BV-induced contralateral hypersensitivity of the spinal withdrawal reflex to noxious electrical stimulation indeed depend on different central pharmacological receptors. The spinal non-NMDA, but not the NMDA, receptors may play important role in BV-induced contralateral central hyperexcitability and sensitization.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

Pivotal involvement of neurogenic mechanism in subcutaneous bee venom-induced inflammation and allodynia in unanesthetized conscious rats.

The bee venom (BV) model is a valid inflammatory pain model in animals and has been extended to human studies using its principle protein, mellitin. After subcutaneous (s.c.) injection of BV, long-lasting spontaneous nociception followed by thermal hyperalgesia, static allodynia, and local inflammatory response (edema) can be observed in rats. We hypothesize that (1) neurogenic components may contribute to the BV-induced inflammatory response and (2) static and dynamic mechanical allodynia may exist simultaneously in the BV model. Using different approaches including sciatic nerve transection (SCT), L4-L6 dorsal rhizotomy (DRT) and local treatment of the sciatic nerve with capsaicin, we found that SCT, DRT, and local capsaicin onto the sciatic nerve produced a significant inhibition of the BV-induced increase in volume of the injected paw, with a stronger effect of the SCT and the local capsaicin treatments than that of the DRT treatment. Static and dynamic mechanical allodynia in the BV test was assessed by measuring the paw withdrawal mechanical threshold and the paw withdrawal latency before and after the BV injection, respectively. Local capsaicin onto the sciatic nerve produced a significant inhibition of the BV-induced decrease in the paw withdrawal mechanical threshold, but not the paw withdrawal latency, of the injected paw. These findings suggest that neurogenic components, via dorsal root reflex and axon reflex mechanisms, are probably involved in the maintenance and the development of the BV-induced inflammation. In addition, the capsaicin-sensitive primary afferents may play differential roles in the development of the BV-induced static and dynamic mechanical allodynia.

Analysis of Variance↗

Cross-presentation of a CMV pp65 epitope by human dendritic cells using bee venom PLA2 as a membrane-binding vector.

We have used bee venom phospholipase A2 as a vector to load human dendritic cells ex vivo with a major histocompatibility complex (MHC) class I-restricted epitope fused to its C-terminus. The fusion protein bound to human monocyte-derived dendritic cells and was internalized into early endosomes. In vitro immunization experiments showed that these dendritic cells were able to generate specific CD8 T cell lines against the epitope carried by the fusion protein. Cross-presentation did not require proteasome, transporter associated with antigen processing, or endosome proteases, but required newly synthesized MHC molecules. Comparison of the antigen presentation pathway observed in this study to that followed by other toxins used as vectors is discussed.

Antigen Presentation↗

Candida riodocensis and Candida cellae, two new yeast species from the Starmerella clade associated with solitary bees in the Atlantic rain forest of Brazil.

Two new ascomycetous yeast species belonging to the Starmerella clade were discovered in nests of two solitary bee species in the Atlantic rain forest of Brazil. Candida riodocensis was isolated from pollen-nectar provisions, larvae and fecal pellets of nests of Megachile sp., and Candida cellae was found in pollen-nectar provisions of Centris tarsata. Analysis of the sequences of the D1/D2 large-subunit ribosomal DNA showed that C. riodocensis is phylogenetically related to C. batistae, and the closest relative of C. cellae is C. etchellsii. The type strains are C. riodocensis UFMG-MG02 (=CBS 10087(T) = NRRL Y-27859(T)) and C. cellae UFMG-PC04 (=CBS 10086(T) = NRRL Y-27860(T)).

Animals↗

Properties of the various Botmar1 transcripts in imagoes of the bumble bee, Bombus terrestris (Hymenoptera: Apidae).

Botmar1 elements are mariner-like elements (MLEs), class II transposable elements that occur in the genome of the bumble bee, Bombus terrestris. Each haploid B. terrestris genome contains about 230 Botmar1, consisting entirely of 1.3-kb and 0.85-kb elements. During their evolution in the B. terrestris genome, two Botmar1 lineages have been differentiated in terms of their nucleic acid sequences and the differences found in their 5' untranslated regions suggest that they could be transcribed differently in B. terrestris. Here, we show that small amounts of Botmar1 mRNA occur in RNA extracts purified from B. terrestris imagoes. This indicates that the Botmar1 transcription is either weak in imagoes, or is restricted to very few cells. The cloning of several mRNAs reveals that only lineage-2 Botmar1 elements are transcribed. This transcription is specific, and uses cardinal initiators and terminators of eukaryotic elements in the Botmar1 elements. The intrastrand stem-loop folds in the mRNA theoretically synthesized by elements of the first lineage suggest that mRNA maintenance in cells might be self-regulated by RNA interference.

3' Untranslated Regions↗