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PubMedDate not supplied

Effect of Apis mellifera propolis from two Brazilian regions on caries development in desalivated rats.

The purpose of the present study was to evaluate the effect of Apis mellifera propolis collected from two regions of Brazil on caries development in desalivated rats. Ethanolic extracts of propolis (EEP) were prepared from crude propolis samples collected in Minas Gerais state (MG), southeastern Brazil, and Rio Grande do Sul state (RS), southern Brazil. The flavonoid composition of EEP was analyzed by high-performance thin-layer chromatography (HPTLC) and reversed-phase high-performance liquid chromatography (HPLC). For the animal study, 30 specific pathogen-free Wistar rats were infected with Streptococcus sobrinus 6715 and surgically desalivated. The rats were randomly divided into three groups which were treated with 80% ethanol (control), EEP from MG and EEP from RS. The animals were placed in a König-Höfer programmed feeder and received 17 meals of diet 2000 daily at hourly intervals. The solutions were applied on the rat molars (25 microl on molars of each quadrant) twice a day, by using graduate syringes. After 3 weeks, the animals were killed by CO(2) asphyxiation. For microbial assessment, the left jaw was removed and sonicated in 154 mM NaCl solution. Dental caries was evaluated according to Larson's modification of Keyes' system. The HPTLC patterns and HPLC profiles demonstrated that both quality and quantity of flavonoid aglycones of EEP from MG were different compared to EEP from RS. In general, it is apparent that EEP from RS contained the highest concentrations of pinocembrin, chrysin, acacetin and galangin. The group of animals treated with EEP from RS showed the lowest smooth-surface and sulcal caries scores as well as less caries severity in smooth-surface and sulcal lesions, and these data were statistically different when compared with the control group. The group treated with EEP from MG only demonstrated a significant difference in the severity of sulcal lesions when compared to the control group. The percentage of S. sobrinus was lower in the groups treated with EEP, but did not differ statistically from the control group. The results showed that the cariostatic effect of propolis depends on its composition, and consequently the region of collection of propolis samples.

Analysis of Variance↗
PubMedDate not supplied

Effects of Apis mellifera propolis on the activities of streptococcal glucosyltransferases in solution and adsorbed onto saliva-coated hydroxyapatite.

Propolis, a resinous hive product collected by Apis mellifera bees, has been used for thousands of years in folk medicine. Ethanolic extracts of propolis (EEP) have been shown to inhibit the activity of a mixture of crude glucosyltransferase (Gtf) enzymes in solution. These enzymes synthesize glucans from sucrose, which are important for the formation of pathogenic dental plaque. In the present study, the effects of propolis from two different regions of Brazil on the activity of separate, purified Gtf enzymes in solution and on the surface of saliva-coated hydroxyapatite (sHA) beads were evaluated. The EEP from Minas Gerais (MG; Southeastern Brazil) and Rio Grande do Sul (RS; Southern Brazil) were tested for their ability to inhibit the enzymes GtfB (synthesis of insoluble glucan), GtfC (insoluble/soluble glucan) and GtfD (soluble glucan). The effects of propolis on Gtf from Streptococcus sanguis (soluble glucan synthesis) was also explored. The EEP from both regions effectively inhibited the activity of all Gtfs in solution (75-95%) and on the surface of sHA beads (45-95%) at concentrations between 0.75 and 3.0 mg of propolis/ml. However, the two samples of propolis showed different levels of inhibition on each of the enzymes tested. In general, EEP RS demonstrated a significantly higher inhibitory activity on GtfB and C activities (both solution and surface assays) than EEP MG at concentrations between 0.047 and 0.187 mg/ml (p<0.05). EEP MG, on the other hand, exhibited a greater inhibitory effect on the activities of surface GtfD (at 0.375, 0.75 and 1.5 mg/ml) and S. sanguis Gtf (at 1.5 and 3.0 mg/ml; p<0.05). These data indicate that EEP is a potent inhibitor of Gtf enzymes in solution and adsorbed on an experimental pellicle; however, its effect on Gtf activity is variable depending on the geographical origin of the propolis samples. There is a need to identify the active compounds of propolis.

Bacterial Proteins↗
PubMed2002

Behavioral studies of learning in the Africanized honey bee (Apis mellifera L.).

Experiments on basic classical conditioning phenomena in adult and young Africanized honey bees (Apis mellifera L.) are described. Phenomena include conditioning to various stimuli, extinction (both unpaired and CS only), conditioned inhibition, color and odor discrimination. In addition to work on basic phenomena, experiments on practical applications of conditioning methodology are illustrated with studies demonstrating the effects of insecticides on learning and the reaction of bees to consumer products. Electron microscope photos are presented of Africanized workers, drones, and queen bees. Possible sub-species differences between Africanized and European bees are discussed.

Animals↗
PubMed2002

The water-soluble fraction (<10 kD) of bee venom (Apis mellifera) produces inhibitory effect on apical transporters in renal proximal tubule cells.

Human envenomation caused by bee stings has been reported to cause acute renal failure and the pathogenetic mechanisms of these renal functional changes are still unclear. Bee venom is also a complex mixture of enzymes and proteins. Thus, this study was conducted to examine the effects of bee venom (BV, Apis mellifera) fractions on apical transporters' activity and its related signal pathways in primary cultured renal proximal tubule cells. Whole BV was extracted into three fractions according to solubility [a water-soluble fraction (BVA), an ethylacetate-soluble fraction (BVE), and a hexane-soluble fraction (BVH)]. BVA fraction was further separated to three portions according to molecular weights: BF1 (>20 kD), BF2 (10-20 kD), and BF3 (<10 kD). Each fraction was treated to the PTCs to the ratio of BV (1 microg/ml). BVA (930 ng/ml) significantly decreased cell viability, but BVH (27 ng/ml) and BVE (43 ng/ml) did not. BF3 (710 ng/ml) among BVA fractions predominantly decreased cell viability and inhibited alpha-methyl-D-glucopyranoside (alpha-MG), phosphate (Pi), and Na(+) uptake. In addition, BF3 increased [(3)H] arachidonic acid release, lipid peroxide formation, and Ca(2+) uptake. These effects of BF3 were blocked by mepacrine and AACOCF(3) (phospholipase A(2) inhibitors) or N-acetylcysteine, vitamin C, and vitamin E (antioxidants). In conclusion, BF3 (<10 kD) among BV fractions is the most effective portion in BV-induced inhibition of alpha-MG, P(i), and Na(+) uptake and these effects of BF3 are associated with phospholipase A(2)-oxidative stress-Ca(2+) signal cascade in the primary cultured rabbit renal proximal tubule cells.

Animals↗
PubMed1977

Detoxified hymenoptera venoms: preliminary studies of in vitro cytotoxicity and antigenicity of Apis mellifera 'venomoid'.

The preparation of detoxified venom (venomoid) of Apis mellifera is described. The venomoid obtained by formaldehyde detoxification retained significant antigenicity. Radioallergosorbent test (RAST) inhibition studies indicated a loss of the affinity of the venomoid for the venom-specific human IgE antibody. In rabbits the venomoid was capable of producing precipitating antibodies which cross-reacted with the original venom. Cytotoxic activity of venomoid as measured by nonimmunological histamine release and hemolytic activity with human cells was not detectable in vitro. Further studies seem warranted for the establishment of a venomoid as a diagnostic and/or therapeutic agent in the management of the hypersensitivity reactions to venom.

Animals↗
PubMed1998

Mechanism of action of honey bee (Apis mellifera L.) venom on different types of muscles.

1. The effect of crude honeybee (Apis mellifera) venom on the skeletal, smooth as well as cardiac muscles were studied in this investigation. 2. Perfusion of gastrocnemius-sciatic nerve preparation of frogs with 1 microgram/ml venom solution has weakened the mechanical contraction of the muscle without recovery. Blocking of nicotinic receptors with 3 micrograms/ml flaxedil before bee venom application sustained normal contraction of gastrocnemius muscle. 3. The electrical activity of duodenum rabbits was recorded before and after the application of 1 microgram/ml venom solution. The venom has depressed the amplitude of the muscle contraction after 15 min pretreatment with atropine nearly abolished the depressor effect of the venom on smooth muscle. 4. In concentrations from 0.5-2 micrograms/ml, bee venom caused decrease of heart rate of isolated perfused toad heart. This bradycardia was accompanied by elongation in the P-R interval. A gradual and progressive increase in the R-wave amplitude reflected a positive inotropism of the venom. Application of 5 micrograms/ml verapamil, a calcium channels blocking agent, abolished the noticed effect of the venom. 5. Marked electrocardiographic changes were produced within minutes of the venom application on the isolated perfused hearts, like marked injury current (elevation or depression of the S-T segment), atrioventricular conduction disturbances and sinus arrhythmias. Atropine and nicotine could decrease the toxic effect of the venom on the myocardium. 6. Results of the present work lead to the suggestion that bee venom is mediated through the peripheral cholinergic neurotransmitter system. General neurotoxicity of an inhibitory nature involving the autonomic as well as neuromuscular system are established as a result of the venom, meanwhile a direct effect on the myocardium membrane stabilization has been suggested.

Animals↗
PubMed2005

Web GIS in practice III: creating a simple interactive map of England's Strategic Health Authorities using Google Maps API, Google Earth KML, and MSN Virtual Earth Map Control.

This eye-opener article aims at introducing the health GIS community to the emerging online consumer geoinformatics services from Google and Microsoft (MSN), and their potential utility in creating custom online interactive health maps. Using the programmable interfaces provided by Google and MSN, we created three interactive demonstrator maps of England's Strategic Health Authorities. These can be browsed online at http://www.healthcybermap.org/GoogleMapsAPI/ - Google Maps API (Application Programming Interface) version, http://www.healthcybermap.org/GoogleEarthKML/ - Google Earth KML (Keyhole Markup Language) version, and http://www.healthcybermap.org/MSNVirtualEarth/ - MSN Virtual Earth Map Control version. Google and MSN's worldwide distribution of "free" geospatial tools, imagery, and maps is to be commended as a significant step towards the ultimate "wikification" of maps and GIS. A discussion is provided of these emerging online mapping trends, their expected future implications and development directions, and associated individual privacy, national security and copyrights issues. Although ESRI have announced their planned response to Google (and MSN), it remains to be seen how their envisaged plans will materialize and compare to the offerings from Google and MSN, and also how Google and MSN mapping tools will further evolve in the near future.

Editorial↗
PubMed2005

The remarkable conservation of corticotropin-releasing hormone (CRH)-binding protein in the honeybee (Apis mellifera) dates the CRH system to a common ancestor of insects and vertebrates.

CRH-binding protein (CRH-BP) is a key factor in the regulation of CRH signaling; it modulates the bioactivity and bioavailability of CRH and its related peptides. The conservation of CRH-BP throughout vertebrates was only recently demonstrated. Here we report the presence of CRH-BP in the honeybee (Apis mellifera) and other insects. Honeybee CRH-BP resembles previously characterized vertebrate CRH-BP sequences with respect to conserved cysteine residues, gene organization, and overall sequence identity. Phylogenetic analyses confirm the unambiguous orthology of insect and vertebrate CRH-BP sequences. Soon after their discovery, it was noted that insect diuretic hormone-I (DH-I) and its receptor share similarities with the vertebrate CRH family and their receptors. Despite these similarities, demonstration of common ancestry of DH-I and the vertebrate CRH family is still speculative: the mature neuropeptides are short, and their genes differ substantially with regard to the number of coding exons. Moreover, DH and CRH receptors belong to the much larger family of G protein-coupled receptors. In contrast, the unique and conspicuous features of CRH-BP greatly facilitate the establishment of orthology over much larger evolutionary distances. The identification of CRH-BP in insects clearly indicates that this gene predates vertebrates by at least several hundred million years. Moreover, our findings imply that a CRH system is shared by insects and vertebrates alike and, consequently, that it has been present at least since the common ancestor to both phylogenetic lines of proto- and deuterostomians.

Amino Acid Sequence↗
PubMed2003

Hepatic biotransformation of the new calcium-mimetic agent, RWJ-68025, in the rat and in man--API-MS/MS identification of metabolites.

The in-vitro biotransformation of a new calcium-mimetic agent and benzenemethanamine analogue, RWJ-68025, was studied after incubation with rat and human hepatic S9 fractions in the presence of an NADPH-generating system. Unchanged RWJ-68025 (44-48% of the sample) plus 12 metabolites were profiled, quantified, and tentatively identified on the basis of API (ionspray)-MS and MS/MS data, and ethyl derivatization for phenolic and carboxylic metabolites. Four metabolic pathways for RWJ-68025 were proposed: pathway 1, O-demethylation; pathway 2, phenyl oxidation; pathway 3, methyl oxidation; and pathway 4, N-dealkylation/acetylation. Pathway 1 formed a major metabolite, O-desmethyl-RWJ-68025 (M1; RWJ-68311; 26% in rat; 16% in human fraction). Pathway 2 produced one major (M2; 12-17% in rat and human fraction) and two minor phenolic metabolites (M4 and M5; all <1% in both species), and in conjunction with step 1, formed hydroxy-M1 (M3; 4-5% in both species). Pathways 3 and 4 formed seven minor oxidized metabolites (M6-M12). RWJ-68025 was extensively metabolized in the rat and human hepatic S9 fractions.

Animals↗
PubMed1977

Occurrence of intercellular bridges between follicle epithelial cells in the ovary of Apis mellifica queens.

Hitherto unknown intercellular bridges or fusomes between the follicle epithelial cells investing the oocytes of Apis mellifica queens have been observed both with light and electron microscopy. Usually each follicle cell has 2-3 intercellular bridges. In surfacial paraffin sections, the intercellular bridges can be seen to connect a series of follicle cells which may be branching. The intercellular bridges lie close to the egg cortex and this position is relatively constant. The width of the fusomal ring canal varies in different developmental stages. In stages 3 and 4 of oogenesis, which are the main vitellogenic stages, the intercellular bridges measure 0-5 micron, while in stages 1 and 2 they have a diameter ranging from 1-5 to 3-5 micron. In these stages the intercellular bridges are provided with numerous transverse microfilaments which disappear later. The fusomal lips are thickened and consist of electron-dense material and an additional layer of less electron-dense material both inside and outside. Ribosomes flow across the bridge. The intercellular bridges may serve to synchronize the differentiation and functional activity of the follicle epithelium during the course of oogenesis.

Animals↗
PubMed2005

Honeybee (Apis mellifera) vision can discriminate between and recognise images of human faces.

Recognising individuals using facial cues is an important ability. There is evidence that the mammalian brain may have specialised neural circuitry for face recognition tasks, although some recent work questions these findings. Thus, to understand if recognising human faces does require species-specific neural processing, it is important to know if non-human animals might be able to solve this difficult spatial task. Honeybees (Apis mellifera) were tested to evaluate whether an animal with no evolutionary history for discriminating between humanoid faces may be able to learn this task. Using differential conditioning, individual bees were trained to visit target face stimuli and to avoid similar distractor stimuli from a standard face recognition test used in human psychology. Performance was evaluated in non-rewarded trials and bees discriminated the target face from a similar distractor with greater than 80% accuracy. When novel distractors were used, bees also demonstrated a high level of choices for the target face, indicating an ability for face recognition. When the stimuli were rotated by 180 degrees there was a large drop in performance, indicating a possible disruption to configural type visual processing.

Animals↗
PubMed1995

Motor pattern of the stinging response in the honeybee Apis mellifera

In the stinging response of the worker honeybee (Apis mellifera), rhythmic movements of the lancets on the stylet are produced by alternating contractions of a set of stinging muscles (a protractor, M198, and a retractor, M199) on each side during co-contraction of the frucula muscles (M197s) on both sides. In this study, stinging movements were elicited by tactile stimulation to the sternum in isolated abdomens, in intact animals and in preparations in which the connectives between the sixth and terminal abdominal ganglia were cut. There was a close relationship among the following three temporal variables of stinging motoneurone pattern: the interval between successive bursts of a stinging muscle, the duration of a burst and the time lag between the bursts of homologous stinging muscles on both sides. All of these variables increased linearly as the sting was inserted deeper into a soft object and the tension on the lancets increased. When sensory nerves from the proprioceptors (campaniform sensilla on the tapering sting shaft and hair plates at the basal cuticular plate) were cut on both sides, the relative timing of bursts of homologous stinging muscles on both sides and antagonistic stinging muscles on each side became more variable. When a proprioceptive input was removed from one side during penetration of the sting, the frequency of the bursts of stinging muscles was higher and the duration of bursts was shorter on the cut side than on the intact side; nevertheless, a sting muscle was still activated out of phase with its antagonistic muscle on the ipsilateral side and its homologous muscle on the contralateral side. These results suggest that the motor pattern driving the rhythmic movements of stinging muscles is produced by a central pattern generator consisting of a pair of oscillators located in the terminal abdominal ganglion and that the precise timing of the motor pattern in a hemiganglion is controlled mainly by proprioceptive inputs on its own side.

Journal Article↗
PubMed1999

Mechanisms of thermal stability during flight in the honeybee apis mellifera

Thermoregulation of the thorax allows honeybees (Apis mellifera) to maintain the flight muscle temperatures necessary to meet the power requirements for flight and to remain active outside the hive across a wide range of air temperatures (Ta). To determine the heat-exchange pathways through which flying honeybees achieve thermal stability, we measured body temperatures and rates of carbon dioxide production and water vapor loss between Ta values of 21 and 45 degrees C for honeybees flying in a respirometry chamber. Body temperatures were not significantly affected by continuous flight duration in the respirometer, indicating that flying bees were at thermal equilibrium. Thorax temperatures (Tth) during flight were relatively stable, with a slope of Tth on Ta of 0.39. Metabolic heat production, calculated from rates of carbon dioxide production, decreased linearly by 43 % as Ta rose from 21 to 45 degrees C. Evaporative heat loss increased nonlinearly by over sevenfold, with evaporation rising rapidly at Ta values above 33 degrees C. At Ta values above 43 degrees C, head temperature dropped below Ta by approximately 1-2 degrees C, indicating that substantial evaporation from the head was occurring at very high Ta values. The water flux of flying honeybees was positive at Ta values below 31 degrees C, but increasingly negative at higher Ta values. At all Ta values, flying honeybees experienced a net radiative heat loss. Since the honeybees were in thermal equilibrium, convective heat loss was calculated as the amount of heat necessary to balance metabolic heat gain against evaporative and radiative heat loss. Convective heat loss decreased strongly as Ta rose because of the decrease in the elevation of body temperature above Ta rather than the variation in the convection coefficient. In conclusion, variation in metabolic heat production is the dominant mechanism of maintaining thermal stability during flight between Ta values of 21 and 33 degrees C, but variations in metabolic heat production and evaporative heat loss are equally important to the prevention of overheating during flight at Ta values between 33 and 45 degrees C.

Journal Article↗
PubMed2002

Discrimination of closed shapes by two species of bee, Apis mellifera and Megachile rotundata.

In the present study, the performance of two bee species, the honeybee Apis mellifera and the leaf-cutter bee Megachile rotundata, in discriminating among various closed (convex) shapes was examined systematically for the first time. Bees were trained to each of five different shapes, a disc, a square, a diamond and two different triangles, all of the same area, using fresh bees in each experiment. In subsequent tests, the trained bees were given a choice between the learned shape and each of the other four shapes. Two sets of experiments were conducted with both species. In the first, solid black shapes were presented against a white background, thus providing a high luminance contrast. In the second, the shapes carried a random black-and-white pattern and were presented 5 cm in front of a similar pattern, thus producing motion contrast, rather than luminance contrast, against the background. The results obtained with the solid shapes reveal that both bee species accomplish the discrimination, although the performance of the honeybee is significantly better than that of the leaf-cutter bee. Furthermore, the effectiveness of the various shapes differs between the two species. However, in neither species is the discrimination performance correlated with the amount of overlap of the black areas contained in the various pairs of shapes, suggesting that, in our experiments, shape discrimination is not based on a template-matching process. We propose that it is based on the use of local parameters situated at the outline of the shape, such as the position of angles or acute points and, in particular, the position and orientation of edges. This conclusion is supported by the finding that bees of both species accomplish the discrimination even with the patterned shapes. These shapes are visible only because of the discontinuity of the speed of image motion perceived at the edge between the shape and the background.

Animals↗
PubMed2004

Localization of alpha-glucosidases I, II, and III in organs of European honeybees, Apis mellifera L., and the origin of alpha-glucosidase in honey.

Three kinds of alpha-glucosidases, I, II, and III, were purified from European honeybees, Apis mellifera L. In addition, an alpha-glucosidase was also purified from honey. Some properties, including the substrate specificity of honey alpha-glucosidase, were almost the same as those of alpha-glucosidase III. Specific antisera against the alpha-glucosidases were prepared to examine the localization of alpha-glucosidases in the organs of honeybees. It was immunologically confirmed for the first time that alpha-glucosidase I was present in ventriculus, and alpha-glucosidase II, in ventriculus and haemolymph. alpha-Glucosidase III, which became apparent to be honey alpha-glucosidase, was present in the hypopharyngeal gland, from which the enzyme may be secreted into nectar gathered by honeybees. Honey may be finally made up through the process whereby sucrose in nectar, in which glucose and fructose also are naturally contained, is hydrolyzed by secreted alpha-glucosidase III.

Amino Acid Sequence↗
PubMed2005

Preparation of trehalase inhibitor validoxylamine A by biocatalyzed hydrolysis of validamycin A with honeybee (Apis cerana Fabr.) beta-glucosidase.

Validoxylamine A is structurally similar to trehalose and acts a potent competitive inhibitor of trehalase. It has recently been receiving increased attention as a potential material for the development of new insecticides or drugs. In this study, beta-glucosidase extracted from honeybees (Apis cerana Fabr.) was used as a catalyst to produce validoxylamine A through enzymatic hydrolysis of validamycin A. Beta-glucosidase was separated and purified from honeybees, and its characteristics were examined. The results showed that beta-glucosidase was stable across a range of temperatures from 30 to 40 degrees C and across a relatively wide range of pH values from 5.0 to 7.5. Investigation of the biocatalyzed hydrolysis process from validamycin A to validoxylamine A with beta-glucosidase revealed that both the substrate (validamycin A) and the product (validoxylamine A) inhibited beta-glucosidase activity. The inhibition constant of the substrate Kis value was 5.01 mM, and that of the product Kip value was 1.32 mM. This product inhibition was competitive.

Animals↗
PubMed2005

Spontaneous recovery from extinction depends on the reconsolidation of the acquisition memory in an appetitive learning paradigm in the honeybee (Apis mellifera).

Memory retrieval initiates two consolidation processes: consolidation of an extinction memory and reconsolidation of the acquisition memory. The strength of the consolidation processes depends on both the strength of the acquisition memory and the strength of retrieval trials and is correlated with its sensitivity to inhibition. We demonstrate that in the honeybee (Apis mellifera), memory retrieval of a consolidated appetitive olfactory memory leads to both consolidation processes, depending on the number of retrieval trials. Spontaneous recovery from extinction is induced by many (five), but not by few (one and two), retrieval trials. Spontaneous recovery is blocked by emetine, an inhibitor of protein synthesis. We conclude that reconsolidation of the acquisition memory underlies spontaneous recovery.

Animals↗
PubMed2005

Focal and temporal release of glutamate in the mushroom bodies improves olfactory memory in Apis mellifera.

In contrast to vertebrates, the role of the neurotransmitter glutamate in learning and memory in insects has hardly been investigated. The reason is that a pharmacological characterization of insect glutamate receptors is still missing; furthermore, it is difficult to locally restrict pharmacological interventions. In this study, we overcome these problems by using locally and temporally defined photo-uncaging of glutamate to study its role in olfactory learning and memory formation in the honeybee, Apis mellifera. Uncaging glutamate in the mushroom bodies immediately after a weak training protocol induced a higher memory rate 2 d after training, mimicking the effect of a strong training protocol. Glutamate release before training does not facilitate memory formation, suggesting that glutamate mediates processes triggered by training and required for memory formation. Uncaging glutamate in the antennal lobes shows no effect on memory formation. These results provide the first direct evidence for a temporally and locally restricted function of glutamate in memory formation in honeybees and insects.

Animals↗