Estudios organolépticos, fisicoquímicos, microbiológicos e interacción con excipientes farmacéuticos de un extracto purificado de cera de Apis mellifera / Organoleptic, physicochemical and microbiological studies and its interaction with pharmaceutical excipients of a purified extract from Apis mellifera wax

El D-002, ingrediente activo antioxidante extraído de la cera de abejas Apis mellifera, fue caracterizado desde el punto de vista físicoquímico, de igual forma se analizó su interacción con excipientes de interés farmacéutico. El D-002 es un polvo fluido inodoro de color blanco a crema, con pérdidas por secado £ 1 por ciento; es insoluble en agua y etanol, y muy ligeramente soluble en otros disolventes orgánicos. Su composición, determinada por cromatografía de gases, fue: 1-tetracosanol (6-15 por ciento), 1-hexacosanol (7-20 por ciento), 1-octacosanol (12-20 por ciento), 1-triacontanol (25-35 por ciento) 1-dotriacontanol (18-25 por ciento) y 1-tetratriacontanol (£ 7,5 por ciento), para una pureza ³ 85 por ciento. Fue estable durante 5 años en la zona climática IV y su análisis por calorimetría diferencial de barrido mostró 2 transiciones de fusión a 59,0 y 81,1 ºC sin descomposición, una alta estabilidad térmica hasta 200 ºC, así como la ausencia de interacciones con lactosa, almidón, croscarmelosa sódica, polivinil pirrolidona, celulosa microcristalina y estearato de magnesio, lo que posibilita el empleo de estos excipientes en la formulación de las tabletas(AU)

The D002, an antioxidant active ingredient extracted from the Apis mellifera bees wax was characterized from the physicochemical point of view analyzing its interaction with excipients of pharmaceutical interest. The D-002 is a creamy white odourless fluid powder with losses by £ 1 percent dry; it is water and ethanol insoluble and very slightly soluble in other organic solvents. Its composition, determined by gas chromatography was: 1-tetracosanol (6-15 percent), 1-hexacosanol (7-20 percent), 1-octacosanol (12-20 percent), 1-triacontanol (25-35 percent, 1-dotriacontanol (18-25 percent) and 1-tetratriacontaol (£ 7,5 percent) for ³ 85 percent of purity. It remained stable during 5 years in the IV climatic zone and its analysis by differential scanning calorimetry showed 2 fusion transitions at 59,0 and 81,1 ºC. without decomposition, a high thermal stability up to 200 ºC, as well as a lack of interactions with lactose, starch, sodium croscarmelosa, polyvinylpyrrolidone, microcrystalline cellulose, and magnesium stearate allowing the use of these excipients in tablets formula(AU)

Estudios organolépticos, fisicoquímicos, microbiológicos e interacción con excipientes farmacéuticos de un extracto purificado de cera de Apis mellifera / Organoleptic, physicochemical and microbiological studies and its interaction with pharmaceutical excipients of a purified extract from Apis mellifera wax

El D-002, ingrediente activo antioxidante extraído de la cera de abejas Apis mellifera, fue caracterizado desde el punto de vista físicoquímico, de igual forma se analizó su interacción con excipientes de interés farmacéutico. El D-002 es un polvo fluido inodoro de color blanco a crema, con pérdidas por secado £ 1 por ciento; es insoluble en agua y etanol, y muy ligeramente soluble en otros disolventes orgánicos. Su composición, determinada por cromatografía de gases, fue: 1-tetracosanol (6-15 por ciento), 1-hexacosanol (7-20 por ciento), 1-octacosanol (12-20 por ciento), 1-triacontanol (25-35 por ciento) 1-dotriacontanol (18-25 por ciento) y 1-tetratriacontanol (£ 7,5 por ciento), para una pureza ³ 85 por ciento. Fue estable durante 5 años en la zona climática IV y su análisis por calorimetría diferencial de barrido mostró 2 transiciones de fusión a 59,0 y 81,1 ºC sin descomposición, una alta estabilidad térmica hasta 200 ºC, así como la ausencia de interacciones con lactosa, almidón, croscarmelosa sódica, polivinil pirrolidona, celulosa microcristalina y estearato de magnesio, lo que posibilita el empleo de estos excipientes en la formulación de las tabletas

The D002, an antioxidant active ingredient extracted from the Apis mellifera bees wax was characterized from the physicochemical point of view analyzing its interaction with excipients of pharmaceutical interest. The D-002 is a creamy white odourless fluid powder with losses by £ 1 percent dry; it is water and ethanol insoluble and very slightly soluble in other organic solvents. Its composition, determined by gas chromatography was: 1-tetracosanol (6-15 percent), 1-hexacosanol (7-20 percent), 1-octacosanol (12-20 percent), 1-triacontanol (25-35 percent, 1-dotriacontanol (18-25 percent) and 1-tetratriacontaol (£ 7,5 percent) for ³ 85 percent of purity. It remained stable during 5 years in the IV climatic zone and its analysis by differential scanning calorimetry showed 2 fusion transitions at 59,0 and 81,1 ºC. without decomposition, a high thermal stability up to 200 ºC, as well as a lack of interactions with lactose, starch, sodium croscarmelosa, polyvinylpyrrolidone, microcrystalline cellulose, and magnesium stearate allowing the use of these excipients in tablets formula

Efectos del extracto de alcoholes purificados de la cera de abejas (Apis mellifera) en dos modelos de analgesia / Effect of purified alcohols extract from the bee wax (Apis mellifera) in two analgesia models

El D-002 es una mezcla de 6 alcoholes alifàticos de alto peso molecular purificada de la cera de abejas (Apis mellifera). Este estudio tuvo como objetivo investigar los efectos del tratamiento por vía oral con D-002 sobre las contorsiones abdominales inducidas por àcido acético y en el modelo del plato caliente en ratones. Los animales se distribuyeron aleatoriamente en 5 grupos (10-20 animales/grupo): uno control que recibió el vehículo goma acacia/H2O, tres tratados con D-002 (25, 125 y 250 mg/kg) y uno con aspirina (modelo de contorsiones abdominales) o morfina (plato caliente). El D-002 (25-250 mg/kg) inhibió significativamente las contorsiones inducidas por àido acético en un 44,5; 44,8 y 47,1 por ciento respectivamente; sin embargo, no modificó la latencia de la respuesta en el modelo del plato caliente. Estos resultados muestran que el tratamiento por vía oral con D-002 (25-250 mg/kg) es capaz de inhibir de forma moderada las contorsiones abdominales por àcido acético sin afectar la respuesta al plato caliente. Esto sugiere que el D-002 ejerce una acción analgésica periférica pero no a nivel central.

The D-002 is a mixture of 6 high molecular weight aliphatic acids purified from bee wax (Apis mellifera). The aim of present study was to research the effects of an oral treatment using D-002 on the acetic acid- induced abdominal writhings and in the hot plate model in mice. Animals were randomized distributed to 5 groups (10-20 animals/group): a control one received the Gum Arabic vehicle/H(2)0, three received D-002 (25, 125 and 250 mg/kg), and another received aspirin (abdominal contortions model) or morphine (hot plate). The D-002 inhibited the above mentioned writhings in a 44,5; 44,8 and 47,1 respectively; however, not modified the response latency in the hot plate model. These results demonstrate that the D-002 (25-250 mg/kg) oral treatment may to inhibit in a moderate way the above mentioned writhings without to affect the response to hot plate. It suggests that the D-002 exerts a peripheral analgesic action but not a central level.

Efectos del extracto de alcoholes purificados de la cera de abejas (Apis mellifera) en dos modelos de analgesia / Efectos del extracto de alcoholes purificados de la cera de abejas (Apis mellifera) en dos modelos de analgesia / Effect of purified alcohols extract from the bee wax (Apis mellifera) in two analgesia models / Effect of purified alcohols extract from the bee wax (Apis mellifera) in two analgesia models

El D-002 es una mezcla de 6 alcoholes alifàticos de alto peso molecular purificada de la cera de abejas (Apis mellifera). Este estudio tuvo como objetivo investigar los efectos del tratamiento por vía oral con D-002 sobre las contorsiones abdominales inducidas por àcido acético y en el modelo del plato caliente en ratones. Los animales se distribuyeron aleatoriamente en 5 grupos (10-20 animales/grupo): uno control que recibió el vehículo goma acacia/H2O, tres tratados con D-002 (25, 125 y 250 mg/kg) y uno con aspirina (modelo de contorsiones abdominales) o morfina (plato caliente). El D-002 (25-250 mg/kg) inhibió significativamente las contorsiones inducidas por àido acético en un 44,5; 44,8 y 47,1 por ciento respectivamente; sin embargo, no modificó la latencia de la respuesta en el modelo del plato caliente. Estos resultados muestran que el tratamiento por vía oral con D-002 (25-250 mg/kg) es capaz de inhibir de forma moderada las contorsiones abdominales por àcido acético sin afectar la respuesta al plato caliente. Esto sugiere que el D-002 ejerce una acción analgésica periférica pero no a nivel central(AU)

El D-002 es una mezcla de 6 alcoholes alifàticos de alto peso molecular purificada de la cera de abejas (Apis mellifera). Este estudio tuvo como objetivo investigar los efectos del tratamiento por vía oral con D-002 sobre las contorsiones abdominales inducidas por àcido acético y en el modelo del plato caliente en ratones. Los animales se distribuyeron aleatoriamente en 5 grupos (10-20 animales/grupo): uno control que recibió el vehículo goma acacia/H2O, tres tratados con D-002 (25, 125 y 250 mg/kg) y uno con aspirina (modelo de contorsiones abdominales) o morfina (plato caliente). El D-002 (25-250 mg/kg) inhibió significativamente las contorsiones inducidas por àido acético en un 44,5; 44,8 y 47,1 por ciento respectivamente; sin embargo, no modificó la latencia de la respuesta en el modelo del plato caliente. Estos resultados muestran que el tratamiento por vía oral con D-002 (25-250 mg/kg) es capaz de inhibir de forma moderada las contorsiones abdominales por àcido acético sin afectar la respuesta al plato caliente. Esto sugiere que el D-002 ejerce una acción analgésica periférica pero no a nivel central(AU)

The D-002 is a mixture of 6 high molecular weight aliphatic acids purified from bee wax (Apis mellifera). The aim of present study was to research the effects of an oral treatment using D-002 on the acetic acid- induced abdominal writhings and in the hot plate model in mice. Animals were randomized distributed to 5 groups (10-20 animals/group): a control one received the Gum Arabic vehicle/H(2)0, three received D-002 (25, 125 and 250 mg/kg), and another received aspirin (abdominal contortions model) or morphine (hot plate). The D-002 inhibited the above mentioned writhings in a 44,5; 44,8 and 47,1 respectively; however, not modified the response latency in the hot plate model. These results demonstrate that the D-002 (25-250 mg/kg) oral treatment may to inhibit in a moderate way the above mentioned writhings without to affect the response to hot plate. It suggests that the D-002 exerts a peripheral analgesic action but not a central level(AU)

The D-002 is a mixture of 6 high molecular weight aliphatic acids purified from bee wax (Apis mellifera). The aim of present study was to research the effects of an oral treatment using D-002 on the acetic acid- induced abdominal writhings and in the hot plate model in mice. Animals were randomized distributed to 5 groups (10-20 animals/group): a control one received the Gum Arabic vehicle/H(2)0, three received D-002 (25, 125 and 250 mg/kg), and another received aspirin (abdominal contortions model) or morphine (hot plate). The D-002 inhibited the above mentioned writhings in a 44,5; 44,8 and 47,1 respectively; however, not modified the response latency in the hot plate model. These results demonstrate that the D-002 (25-250 mg/kg) oral treatment may to inhibit in a moderate way the above mentioned writhings without to affect the response to hot plate. It suggests that the D-002 exerts a peripheral analgesic action but not a central level(AU)

Contenido microbiológico cultivable del tracto intestinal y polen almacenado de Apis mellifera (Hymenoptera: Apidae) / Cultured Microbiological Content of the Intestinal Tract and Stored Pollen of Apis mellifera (Hymenoptera: Apidae)

Se caracterizaron los microorganismos cultivables asociados con Apis mellifera. Las muestras fueron tomadas a partir de polen almacenado (joven y maduro) y transportado en corbículas y tracto digestivo de las abejas (forrajeras y recién nacidas). Se aislaron bacterias pertenecientes a los géneros Pseudomonas, Streptococcus, Micrococcus, Lactobacillus, Klebsiella, Proteus, Yersinia y Arthrobacter y hongos de los géneros Rhizopus, Alternaria y Epicoccum. De acuerdo a sus propiedades bioquímicas, algunas de estas bacterias pueden estar involucradas en la degradación de los compuestos de la capa externa del polen y son adquiridas por las abejas a través del alimento y contacto con otros individuos de la colmena. La presencia de los hongos se explica por su amplia distribución en el ambiente, ya que los tres géneros se encuentran comúnmente en el suelo y en las plantas que las abejas pueden seleccionar como fuente de alimento.

Z-band proteins in the flight muscle and leg muscle of the honeybee.

Monoclonal antibodies (mAb's) have been raised against proteins in preparations of Z-discs isolated from honeybee fibrillar flight muscle. These antibodies have identified four Z-disc antigens on immunoblots of honeybee fibrillar proteins. Antibody alpha binds to the 90-100 kD protein, alpha-actinin; mAb P interacts with the protein, projectin, an extremely large polypeptide (greater than 600kD) found in the connecting filaments which link thick filaments to the Z-band in insect asynchronous flight muscle. Two other mAb's recognize previously uncharacterized insect Z-band proteins. Monoclonal antibody Z(400) binds to a pair of proteins with molecular masses near 400 kD and 600 kD. Antibody Z(175) recognizes two components, 158 kD and 175 kD, that are not only immunologically similar but have nearly identical peptide maps. Indirect immunofluorescence microscopy studies show that the proteins recognized by mAb's alpha, Z(175) and Z(400) are located at the Z-band, while the mAb P antigen is found on either side of it. Three of the four antibodies we have obtained recognize leg muscle proteins. Monoclonal antibodies alpha and P comigrate on SDS gels with analogous components from flight muscle. Only the smaller of the two proteins identified in flight muscle by mAb Z(400) is found in leg muscle, however. Furthermore, no Z(175) antigens have been detected in the non-fibrillar tissue by either monoclonal or polyclonal antibodies. Immunofluorescence microscopy studies localize the alpha and Z(400) antigens at the Z-line in leg muscle fibrils. Surprisingly, however, mAb P binds within the A-bands of synchronous fibres, not between the A- and Z-bands as in asynchronous fibrillar muscle.

Apidaecins: antibacterial peptides from honeybees.

Although insects lack the basic entities of the vertebrate immune system, such as lymphocytes and immunoglobulins, they have developed alternative defence mechanisms against infections. Different types of peptide factors, exhibiting bactericidal activity, have been detected in some insect species. These humoral factors are induced upon infection. The present report describes the discovery of the apidaecins, isolated from lymph fluid of the honeybee (Apis mellifera). The apidaecins represent a new family of inducible peptide antibiotics with the following basic structure: GNNRP(V/I)YIPQPRPPHPR(L/I). These heat-stable, non-helical peptides are active against a wide range of plant-associated bacteria and some human pathogens, through a bacteriostatic rather than a lytic process. Chemically synthesized apidaecins display the same bactericidal activity as their natural counterparts. While only active antibacterial peptides are detectable in adult honeybee lymph, bee larvae contain considerable amounts of inactive precursor molecules.

Antibodies to the mammalian adhesion molecule J1/tenascin and its carbohydrate epitope L2/HNK-1 label the receptor lymph cavities of insect sensilla.

Data from light- and electronimmunocytochemistry gave evidence that the antibodies to the mammalian adhesion molecule J1/tenascin and its carbohydrate structure L2/HNK-1 react with immunoreactive structures present in the inner and outer receptor lymph cavities of antennal sensilla of the honey bee. Immunoreactivity was additionally present in the cytoplasm of the enveloping cells surrounding the receptor lymph cavities. Cell contacts between enveloping cells and between dendrites and enveloping cells were never observed to be antigen positive.

Isolation and partial characterization of insulin of the honeybee (Apis mellifica).

In the honeybee (Apis mellifica), insulin-like material was partially purified with acid ethanol extractions by a classic method for recovering insulin and following gel filtration on a Sephadex G-50 column. The preparations were characterized by their ability to cross-react with porcine insulin antibodies. Insulin-like biological activity was demonstrated using the insulin bioassay. Stimulation of glucose oxidation or lipogenesis was measured by isolated rat adipocytes. Insulin seems to be more widespread in invertebrates than was previously assumed.

Structure of a protein catalyzing the formation of 11 cis-retinal in the visual cycle of invertebrate eyes.

A pigment made up of a protein able to bind retinal as well as retinol is described. The molecule consists of a dimer with a molecular weight of 50,000 which binds one molecule of retinal. The binding site for retinal is a Schiff base buried in the interior of the protein. Retinol is probably bound to the protein in the same site as for retinal, although not covalently, as suggested by the absorbance spectra. The protein, extracted from honeybee retina, is involved in visual pigment metabolism, and its structure may elucidate the mechanism of the stereospecific photoisomerization of all trans-retinal to 11-cis-retinal.

Is it possible to use the honey bee adult as a bioindicator for the detection of pesticide residues in plants?

Pesticide residues are usually determined by physical, chemical and biological methods. The simplicity and adaptability of bioassay methods have won their acceptance in the field of residue analysis. Theoretically, any organism that is susceptible to a pesticide may be used for its bioassay in any environmental sample. This means that such organism may serve as a bioindicator for the detection of certain pollutants. The susceptibility of honey bees (Apis melifera L.) to many insecticides commonly used in crop protection led to an attempt to use it as a bioindicator for the determination of residues of some insecticides in plant materials, as well as to detect toxicity hazards to honey bees of some commonly used insecticides. Results of this work which have been recently published may suggest "Yes" to answer the question posed in the title of this subject.

Iron-containing cells in the honey-bee (Apis mellifera). I. Adult morphology and physiology.

Particulate iron was found within the trophocytes of the fat body of the adult honey-bee. These iron granules differed in their structure and composition from iron granules found in other biological systems. The granules had an average diameter of 0.32 +/- 0.07 micron and were composed of iron, calcium and phosphorus in a non-crystalline arrangement. The granules were apparently randomly distributed within the cytoplasm of the cells, and were not associated with any particular cellular organelle. Electron microscopy revealed the presence of cell junctions between the trophocytes. In tissues treated with colloidal lanthanum, 20-nm gaps were seen between the outer leaflets of the cells forming the cell junction. Physiological studies showed that these cells are electrically coupled, but the coupling ratio is low, as a result of extensive coupling to many cells.

Insect optic lobe neurons identifiable with monoclonal antibodies to GABA.

Five monoclonal antibodies against GABA were tested on glutaraldehyde fixed sections of optic lobes of three insect species, blowflies, houseflies and worker bees. The specificity of these antibodies was analyzed in several tests and compared with commercially available anti-GABA antiserum. A very large number of GABA-like immunoreactive neurons innervate all the neuropil regions of these optic lobes. Immunoreactive processes are found in different layers of the neuropils. The immunoreactive neurons are amacrines and columnar or noncolumnar neurons connecting the optic lobe neuropils. In addition some large immunoreactive neurons connect the optic lobes with centers of the brain. Some neuron types could be matched with neurons previously identified with other methods. The connections of a few of these neuron types are partly known from electron microscopy or electrophysiology and a possible role of GABA in certain neural circuits can be discussed.

Structure of thick filaments from insect flight muscle.

The supramolecular organization of thick (myosin) filaments isolated from insect flight muscle was studied using negative staining and shadowing techniques. The electron microscopical findings favour the two-stranded arrangement of double cross-bridges rather than a four- or six-stranded structures of single cross-bridges. The thick filament backbone consists of 12-subfilaments of myosin rods with a diameter of about 4 nm in agreement with the X-ray data. Furthermore, about 4 nm stripping was observed on the filament shaft, resembling to the structure of the light meromyosin paracrystals with a periodicity of 4.8 nm. The existence of a hinge region at the origin of the projections and between the tail and myosin heads are confirmed. According to the morphological observations the stalks of the projections can be characterized with flexible properties, as well. At high level of the supramolecular organization, the myosin filaments, the connecting filaments and the Z-filaments are systematically interconnected after a remarkable myosin filament branching (trifurcation) at the Z line level, which supports the view that a continuous longitudinal filament network constitutes the structure of the myofibrils.

Lipophorin of the larval honeybee, Apis mellifera L.

Most insects have a major lipoprotein species in the blood (hemolymph) that serves to transport fat from the midgut to the storage depots in fat body cells and from the fat body to peripheral tissues. The generic name lipophorin is used for this lipoprotein. In larvae of the honeybee, Apis mellifera, a lipophorin has been found with properties that correlate well with those of the only other lipophorin reported for an immature insect, that of the tobacco hornworm, Manduca sexta. The honeybee lipophorin (Mr = 530,000) has a density of 1.13 g/ml, contains approximately 41% lipid and 59% protein, and contains two apoproteins, apoLp-I, Mr = 250,000 and apoLp-II, Mr = 80,000, both of which are glycosylated. The lipids consist predominantly of polar lipids, of which phospholipids and diacylglycerols represent 60% of the total. When the intact lipophorin is treated with trypsin, apoLp-I is rapidly proteolyzed, while apoLp-II is resistant, indicating a difference in exposure of the two apoproteins to the aqueous environment. Honeybee apoLp-II cross-reacts with antibodies to M. sexta apoLp-II, but not to anti-M. sexta apoLp-I. No cross-reactivity of honeybee apoLp-I to anti-M. sexta apoLp-I was observed.

Sclerotin and lipid in the waterproofing of the insect cuticle.

In all the cuticles studied waterproofing is effected by extracuticular material, a mixture of sclerotin precursors and lipids, exuded from the tubular filaments of the pore canals. In Rhodnius larval abdomen it is a layer of thickness similar to the outer epicuticle, believed to be composed of 'sclerotin' and wax, in Schistocerca larval sternal cuticle and in Carausius sternal cuticle it is similar. In Tenebrio adult sternal cuticle of the abdomen, in both the extracuticular exudation and the contents of the distal endings of the tubular filaments, the wax component is obscured by hard 'sclerotin'. In Manduca larva a very thin layer of 'sclerotin' and wax is covered by an irregular wax layer, average 0.75 micron, twice the thickness of the inner epicuticle. In Periplaneta and Blattella the abdominal cuticle is covered by a soft waxy layer, often about 1 micron thick, which is mixed with argentaffin material. Below this is a very thin waterproof layer of wax and 'sclerotin' continuous with the contents of the tubular filaments, which is readily removed by adsorptive dusts. In Apis adult abdominal terga free wax plus sclerotin precursors form a thin layer which is known to be removed by adsorptive dusts. In Calliphora larva there is a very thin layer of the usual mixed wax and sclerotin and below this a thick (0.5 micron) layer, lipid staining and strongly osmiophil, likewise extracuticular and exuded from the epicuticular channels. This material (which is often called 'outer epicuticle') has the same staining and resistance properties as the true outer epicuticle on which it rests. In the abdomen of Calliphora adult the waterproofing wax-sclerotin mixture forms a thin layer over the entire cuticle including the surface of the microtrichia. There is also a thin detachable layer of free wax on the surface.