Cytotoxic constituents of propolis inducing anticancer effects: a review

Objectives Propolis is a honeybee product used extensively in traditional medicine for its antioxidant, anti-inflammatory, immunomodulatory and anticancer effects. Propolis exhibits a broad spectrum of biological activities because it is a complex mixture of natural substances. In this review, the antitumour effects of propolis extracts and its constituents (e. g. flavonoids, terpenes and caffeic acid phenethyl ester) are discussed.Key findings The effect of propolis on experimental carcinogenesis is discussed, as well as its possible mechanisms of action against tumours, involving apoptosis, cell cycle arrest and interference on metabolic pathways. Propolis seems to be efficient against different tumour cells both in vitro and in vivo, which suggests its potential in the development of new anticancer drugs.Summary Propolis extracts may be important economically and would allow a relatively inexpensive cancer treatment. Preclinical investigations are needed to further elucidate the benefits of propolis and its antitumour properties.

The effects of propolis on antibody production by laying hens

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effect of narcosis with CO2 on the ovarian development in queens of Apis mellifera (Hymenoptera, Apini)

Exposure of 1, 4, 7 & 10 day-old virgin queens of Apis mellifera Linne 1758 for 1 min to CO2 accelerated their ovarian development, having a similar effect as mating on the initial formation of the ovarian follicles. In 3 day-old queens the exposure to CO2 enhanced the initial stage of germ cell differentiation into oocytes and nurse cells in the ovarioles, a developmental stage only seen in 5 day-old untreated virgin queens, the age at which they are ready to mate. In 10 day-old untreated virgin queens, some regions of the ovarioles presented tissue disorganization and many cells with pycnotic nuclei. However, narcotized virgin queens of the same age did not present such ovary degeneration. Conversely, they showed nitid follicle formation, arising in the ovarioles' initial differentiation between nurse and oocytic chambers, although still without vitellogenesis. The accelerative effect of CO2 is limited to the ages near to those proper for mating, since 15 and 18 day-old treated virgin queens presented ovaries with extensive regions of high tissue disorganization and a great number of cells with pycnotic nuclei. According to the results, the narcosis presented three levels of effect on the ovary of honeybee virgin queens: 1) accelerated the germ cell differentiation, 2) preserved the tissue integrity even after the queen mating period and 3) stimulated the initial differentiation of a vitellarium. This later condition was only seen in untreated queens after mating. All these effects are not maintained if the queen is kept virgin beyond 15 days old.

Enzymatic activity of hypopharyngeal gland extractsfrom workers of Apis mellifera (Hymenoptera, Apidae, Apinae)

This research presents a comparative study of enzymatic activity of the hypopharyngeal gland extracts from workers of Apis mellifera in three physiologic stages: newly emerged, nurse and forager workers, with the objective of contributing to the comprehension of the gland function. In order to determinate the enzymes present in the extracts, the Api Zym kit (Bio Merieux) was used to test the activity of 19 different enzymes. The enzymes found in larger amounts only in the hypopharyngeal glands from certain individuals were the following: in newly emerged workers, the N-acetyl-double down arrow-glucosaminidase that may be digesting the chitin of some food ingested by the bee; in forager workers, the acid phosphatase that is likely acting in authophagic processes, the a-glucosidase, in the processing of nectar into honey, and the double down arrow-glucosidases, in the pollen digestion.

Alterations induced by the juvenile hormone in glandular cells of the Apis mellifera venom gland: Applications on newly emerged workers (Hymenoptera, Apidae)

Histological and histochemical analyses were carried out in order to evaluate the influence of the topical application of a synthetic juvenile hormone on the secretory cycle and degeneration of the venom gland of Apis mellifera. Newly emerged workers received the topical application of synthetic hormone and the results were compared to the normal development of the secretory cycle in virgin and mated queens. The first worker group received the juvenile hormone diluted in hexane (2 mu g/mu L), the second received only mu L of hexane, and the third did not receive any kind of application. After the application the workers were returned to the colony and collected at the ages of 14 and 25 days of adult life. The groups with virgin queens and the other with mated queens, did not receive the treatment. The results show that the individuals treated with juvenile hormone and with pure hexane presented differences in the histological and cytochemical aspects of the secretory cells of the venom gland. The data indicate that both the juvenile hormone and hexane accelerate the activity of the secretory cycle and the degeneration of the venom gland; however, the juvenile hormone proved to be more effective than hexane. (c) 2006 Elsevier Ltd. All rights reserved.

Morphological characterization of cell death during the ovary differentiation in worker honey bee

Cell death that occurs during ovary differentiation in the honeybee worker's larval development accounts for ovariole reabsorption. From a morphological standpoint, three modes of death were detected. Germinative cells in the ovarioles die by an apoptotic-like process, whereas the somatic cells die by an autophagic process, type 11 cell death; and during pupation, stromatic and ovarian capsular cells die through cytoplasmic disintegration, releasing their components into the hemolymph. These modes of cell death are in part determined by the pattern of tissue organization within which the cell occurs. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Structure determination of hydroxytrypargine: A new tetrahydro-beta-carboline toxin from the venom of the spider Parawixia bistriata

A new, highly active tetrahydro-p-carboline toxin from the spider Parawixia bistriata, the most-common species of social spider occurring in Brazil, was isolated. The new toxin was identified as 1,2,3,4-tetrahydro-6-hydroxy-beta-carboline (= N-[3-(2,3,4,9-tetrahydro-6-hydroxy-1H-pyrido[3,4-b]indol-1-yl)propyl]guanidine; 3). This type of alkaloid, not common among spider toxins, was found to be the most-potent constituent of the spider's chemical weaponry to kill prey. When P bistriata catch arthropods in their web, they apparently attack their prey in groups of many individuals injecting their venoms. In vivo toxicity assays with 3 demonstrated a potent lethal effect to honeybees, giving rise to clear neurotoxic effects (paralysis) before death. The compound's toxicity (LD50 value) was determined to be ca. 8 ng/g of honeybee. The investigation of the pharmacological properties and neurotoxic actions of 3 may be used in the future for the development of new drugs to be applied for pest control in agriculture.

The venomous secrets of the web droplets from the viscid spiral of the orb-weaver spider Nephila clavipes (Araneae, Tetragnatidae)

The capture web of N. clavipes presents viscous droplets, which play important roles in web mechanics and prey capture. By using scanning and transmission electron microscopy, it was demonstrated that the web droplets are constituted of different chemical environments, provided by the existence both of an aqueous and a lipid layer, which, in turn, present a suspension of tenths of vesicles containing polypeptides and/or tipids. GC/EI-MS Analysis of the contents of these vesicles led to the identification of some saturated fatty acids, such as decanoic acid, undecanoic acid, dodecanoic acid, tetradecanoic acid, octadecanoic acid, and icosanoic acid, while other components were unsaturated fatty acids, such as (Z)-tetradec-9-enoic acid, (Z)-octadec-9-enoic acid, and (Z)-icosa-11-enoic acid; and polyunsaturated fatty acids like (9Z,12Z)-octadeca-9,12-dienoic acid, (9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid, and (11Z,14Z)-icosa-11,14-dienoic acid. Toxic proteins such as calcium-activated proteinase and metalloproteinase jararhagin-like precursor were also identified by using a proteomic approach, indicating the possible involvement of these enzymes in the pre-digestion of spiders' preys web-captured. Apparently, the mixture of fatty acids are relatively toxic to insects by topical application (LD50 64.3 +/- 7.6 ng mg(-1) honeybee), while the proteins alone present no topical effect; however, when injected into the prey-insects, these proteins presented a moderate toxicity (LD50 40.3 +/- 4.8 ng mg(-1) honeybee); the mixture of fatty acids and proteins is very toxic to the preys captured by the web droplets of the viscid spiral of Nephila clavipes when topically applied on them (LD50 14.3 +/- 1.8ng mg(-1) honeybee).

Programmed cell death in the larval salivary glands of Apis mellifera (Hymenoptera, Apidae)

The morphological and histochemical features of degeneration in honeybee (Apis mellifera) salivary glands were investigated in 5th instar larvae and in the pre-pupal period. The distribution and activity patterns of acid phosphatase enzyme were also analysed. As a routine, the larval salivary glands were fixed and processed for light microscopy and transmission electron microscopy. Tissue sections were subsequently stained with haematoxylin-eosin, bromophenol blue, silver, or a variant of the critical electrolyte concentration (CEC) method. Ultrathin sections were contrasted with uranyl acetate and lead citrate. Glands were processed for the histochemical and cytochemical localization of acid phosphatase, as well as biochemical assay to detect its activity pattern. Acid phosphatase activity was histochemically detected in all the salivary glands analysed. The cytochemical results showed acid phosphatase in vesicles, Golgi apparatus and lysosomes during the secretory phase and, additionally, in autophagic structures and luminal secretion during the degenerative phase. These findings were in agreement with the biochemical assay. At the end of the 5th instar, the glandular cells had a vacuolated cytoplasm and pyknotic nuclei, and epithelial cells were shed into the glandular lumen. The transition phase from the 5th instar to the pre-pupal period was characterized by intense vacuolation of the basal cytoplasm and release of parts of the cytoplasm into the lumen by apical blebbing; these blebs contained cytoplasmic RNA, rough endoplasmic reticule and, occasionally, nuclear material. In the pre-pupal phase, the glandular epithelium showed progressive degeneration so that at the end of this phase only nuclei and remnants of the cytoplasm were observed. The nuclei were pyknotic, with peripheral chromatin and blebs. The gland remained in the haemolymph and was recycled during metamorphosis. The programmed cell death in this gland represented a morphological form intermediate between apoptosis and autophagy.

Análise morfológica da glândula de veneno de Apis mellifera L. (Hymenoptera: Apidae) em populações de Mato Grosso do Sul

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Analysis of Mortality in Africanized Honey Bee Colonies with High Levels of Infestation by Varroa destructor

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

A morphological view of the relationship between indirect flight muscle maturation and the flying needs of two species of advanced eusocial bees

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cytoskeletal organization of bee ovarian follicles during oogenesis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Bee Visitors of Quaresmeira Flowers (Tibouchina granulosa Cogn.) in the Region of Dourados (MS-Brasil)

This research was developed in the center-urban area at the city of Dourados (MS), in Tibouchina granulosa trees, a plant popularly known as quaresmeira in Brazil. The floral visitors of this species were recorded in three daily periods: 7 - 8 a.m., 1 - 2 p.m. and 4 - 5 p. m. Bee visitors of those plants were collected with an entomological net when they landed on the flowers. The bees were anesthetized in a closed camera containing etila acetate, conserved in Dietrich ' s fixative and, soon afterwards, transferred to 70% ethyl alcohol for subsequent identification. Almost 300 specimens of Africanized Apis mellifera, Trigona spinipes and Tetragonisca angustula were collected. The presence of these three species of bees in the flowers of T. granulosa suggests that they can be considered the main floral visitors of that vegetal species. T. angustula workers did not visit the flowers of T. granulosa between 7 and 8 a.m., but they were the main floral visitors from 4 to 5 p.m. The workers of T. spinipes presented a very aggressive behavior against the Africanized A. mellifera workers on the quaresmeira flowers, defending the food sources with strength and efficiency and provoking a drastic reduction in the number of honeybee visits between 1 and 2 p.m. Nevertheless, they did not interfere in the opportunistic activity of foraging by T. angustula workers, mainly between 4 and 5 p.m. The most important result of this research was the detection of the Competitive Exclusion Principle between A. mellifera and T. spinipes species in relation to the foraging behavior, when they exploit the floral resources of T. granulosa.

Profiling the proteome complement of the secretion from hypopharyngeal gland of Africanized nurse-honeybees (Apis mellifera L.)

The protein complement of the secretion from hypopharyngeal gland of nurse-bees (Apis mellifera L.) was partially identified by using a combination of 2D-PAGE, peptide sequencing by MALDI-PSD/MS and a protein engine identification tool applied to the honeybee genome. The proteins identified were compared to those proteins already identified in the proteome complement of the royal jelly of the honey bees. The 2D gel electrophoresis demonstrated this protein complement is constituted of 61 different polypepides, from which 34 were identified as follows: 27 proteins belonged to MRJPs family, 5 proteins were related to the metabolism of carbohydrates and to the oxido-reduction metabolism of energetic Substrates, I protein was related to the accumulation of iron in honeybee bodies and I protein may be a regulator of MRJP-1 oligomerization. The proteins directly involved with the carbohydrates and energetic metabolisms were: alpha glucosidase, glucose oxidase and alpha amylase, whose are members of the same family of enzymes, catalyzing the hydrolysis of the glucosidic linkages of starch; alcohol dehydrogenase and aldehyde dehydrogenase, whose are constituents of the energetic metabolism. The results of the present manuscript support the hypothesis that the most of these proteins are produced in the hypoharyngeal gland of nurse-bees and secreted into the RJ. (C) 2004 Elsevier Ltd. All rights reserved.