Lifespan-Extending Effects of Royal Jelly and Its Related Substances on the Nematode Caenorhabditis elegans

One of the most important challenges in the study of aging is to discover compounds with longevity-promoting activities and to unravel their underlying mechanisms. Royal jelly (RJ) has been reported to possess diverse beneficial properties. Furthermore, protease-treated RJ (pRJ) has additional pharmacological activities. Exactly how RJ and pRJ exert these effects and which of their components are responsible for these effects are largely unknown. The evolutionarily conserved mechanisms that control longevity have been indicated. The purpose of the present study was to determine whether RJ and its related substances exert a lifespan-extending function in the nematode Caenorhabditis elegans and to gain insights into the active agents in RJ and their mechanism of action.We found that both RJ and pRJ extended the lifespan of C. elegans. The lifespan-extending activity of pRJ was enhanced by Octadecyl-silica column chromatography (pRJ-Fraction 5). pRJ-Fr.5 increased the animals' lifespan in part by acting through the FOXO transcription factor DAF-16, the activation of which is known to promote longevity in C. elegans by reducing insulin/IGF-1 signaling (IIS). pRJ-Fr.5 reduced the expression of ins-9, one of the insulin-like peptide genes. Moreover, pRJ-Fr.5 and reduced IIS shared some common features in terms of their effects on gene expression, such as the up-regulation of dod-3 and the down-regulation of dod-19, dao-4 and fkb-4. 10-Hydroxy-2-decenoic acid (10-HDA), which was present at high concentrations in pRJ-Fr.5, increased lifespan independently of DAF-16 activity.These results demonstrate that RJ and its related substances extend lifespan in C. elegans, suggesting that RJ may contain longevity-promoting factors. Further analysis and characterization of the lifespan-extending agents in RJ and pRJ may broaden our understanding of the gene network involved in longevity regulation in diverse species and may lead to the development of nutraceutical interventions in the aging process

The effect of post-translational modifications on the antimicrobial activity of apalbumin2.

This study illustrates multifunctionality of proteins of honeybee royal jelly (RJ) and how their neofunctionalization result from various PTMs of maternal proteins. Major proteins of RJ, designated as apalbumins belong to a protein family consisting of nine members with Mr of 49-87 kDa and they are accompanied by high number of minority homologs derived from maternal apalbumins. In spite of many data on diversity of apalbumins, the molecular study of their individual minority homologous is still missing. This work is a contribution to functional proteomics of second most abundant protein of RJ apalbumin2 (Mr 52.7 kDa). We have purified a minority protein from RJ; named as apalbumin2a, differ from apalbumin2 in Mr (48.6 kDa), in N-terminal amino acids sequences - ENSPRN and in N-linked glycans. Characterization of apalbumin2a by LC-MALDI TOF/TOF MS revealed that it is a minority homolog of the major basic royal jelly protein, apalbumin2, carrying two fully occupied N-glycosylation sites, one with high-mannose structure, HexNAc2Hex9, and another carrying complex type antennary structures, HexNAc4Hex3 and HexNAc5Hex4. We have found that apalbumin2a inhibit growth of Paenibacillus larvae. The obtained data call attention to functional plasticity of RJ proteins with potential impact on functional proteomics in medicine

THE QUALITY OF HONEY FOR BEES AND MAN

Honey is used both as food ingredient and in a variety of treatments for diverse ailments. For this reason, consumer expectation of its quality and purity is particularly high. The aim of the BEE SHOP Honey Department was the evaluation of honey quality and authenticity, through the development of new instrument for the verification of the botanical origin and the presence of impurities. Honey quality can be important also for the colony itself and therefore the Honey Department focused on the physiological properties of honey which can be beneficial for honeybees and for the prevention of bee diseases. Testing the origin of honeybees is also important to enforce the EC directive for organic beekeeping, since the use of regional bee strains is an important quality criterion for organic honey. To identify the origin of the honeybee the BEE SHOP Honey Department has developed a DNA based diagnostic tool. Through the analysis of honey and nectar samples by HPLC-MS-MS methods, suitable markers were detected for Robinia, Tilia, Citrus, Eucalyptus and chestnut unifloral honeys, and the following phytochemicals have been proposed for the determination of honey floral origins: myricetin, tricetin and luteolin for Eucalyptushoney, kynurenic acid related compounds for chestnut honey, terpenoids for Linden honey, hesperetin for Citrus honey, kaempferol rhamnosides for Acacia (Robinia) honey. Two novel protocols were introduced for the evaluation of honey quality: DRIFTS (Diffuse Reflectance Infrared Fourier Transform Spectroscopy) and HR-NMR (High Resolution Nuclear Magnetic Resonance). These techniques, coupled with appropriate multivariate statistical analysis, demonstrated to be suitable for the verification of botanical origin and the detection of honey adulteration by sugar syrups. The HR-NMR method seems to be suitable also for a quantitative determination of the adulteration levels. A new sensitive enzyme-linked immunoassay (ELISA) for the quantitative determination of apalbumin1a major compound of the royal jelly (RJ) proteins in honey has been developed. This protein can be used as a marker for honey quality because its concentration varies with the botanical origin. The highest content was determined in chestnut honey, in comparison with acacia and rape honey, while the lowest amount was detected in honey obtained supplying bee colony with saccharose syrup. The antimicrobial potential of honey based on proteins of honeybee origin was tested by microtiter based assays. The inhibition of P. larvae growth was observed in the protein fraction of cherry and rape honeys and honeydew. Moreover, a protein fraction corresponding to apalbumin2a has been identified in honey. This protein, purified from RJ, had specific antibiotic properties against P. larvae. The anti quorum-sensing (QS) activities of honeys with different floral origin have been evaluated, using bacterial strains in which quorum-sensing activated the pigment violacein. 29 honey samples inhibited QS even at the lowest concentration. The anti-QS activity was concentration-dependent and relied on the floral origin. Among all honeys, chestnut and linden samples were the strongest quorum-sensing inhibitors (QSI)