Royal jelly as a novel bio-stimulant for asymbiotic seed germination andprotocorm development in Phalaenopsis

Objective: In this study the effect of Royal jelly on the growth of two important members of Bacteroides spp.; Bacteroides fragilis and Bacteroides thetaiotaomicron, was evaluated. Also the physicochemical properties and cytotoxicity effects of Royal jelly on Caco-2 cell line as gastrointestinal epithelial cell model, assessed.
 Methods : Bacteria, Bacteroides fragilis and Bacteroides thetaiotaomicron were grown on brain heart infusion (BHI) broth medium supplemented with Royal jelly in 3 different concentrations (2.5, 5 and 10% v/v), both of the bacteria (1.5×108 cfu/mL) were inoculated to BHI broth contained Royal jelly in anaerobic condition. To calculate the bacterial optical density (OD), the absorbance was measured at 600 nm after an overnight. Also Caco-2 cells, was used to study the effects of Royal jelly on epithelial cell viability, and the Physicochemical properties consist of total proteins, polysaccharides, phenolic compounds, total lipids, ash and moisture by UV-VIS spectrophotometric and gravimetric methods were evaluated .
 Results: The growth of B. fragillis and B. thetaiotaomicron were increased by Royal jelly (2.5, 5 and 10% v/v concentrations) and the results indicated that Royal jelly increased the growth of bacteria in a dose dependent manner (p<0.001). In addition MTT assay showed more than 95% viability of Caco-2 cells treated with Royal jelly. The Iranian Royal jelly sample contains 59.01% water, 11.57% proteins, 12% lipids, 12.63 % polysaccharide and 5% mineral.
 Conclusion: The present study showed that Royal jelly has a potential effect in the preserving gut microbiota and it is suggested that Royal jelly as a complementary and alternative medicine can be used to treatment diseases are associated with gut microbiota- host interactions and immune regulating. Although we need to expand our knowledge by designing clinical trials to confirm the therapeutic effects of Royal jelly on gut microbiota modulation as a barrier function.

Synergistic effect of royal jelly in combination with glycerol and dimethyl sulfoxide on cryoprotection of Romanov ram sperm

BackgroundOne 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.Principal FindingsWe 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.Conclusions/SignificanceThese 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.

Background: Banana (Musa spp.) is one of the most consumable fruits and cultivated around the globe. It contains high nutritional value as well as the high demand of the market. The microbes are the main problem for the propagation of banana plants in tissue culture. Chitosan is one of the best substances for the eradication of contamination and also growth stimulators of banana plants. This study is based on the micro-propagation of the bantala variety of Musa species and free from microbe infection. Methods: The rhizome and sucker as explants of Musa cv. Bantala. The different combination concentrations of 6-Benzylaminopurine (BAP), indole-3-acetic acid (IAA) and chitosan (CS) were tried in Murashige and Skoog medium for in vitro response of plants, shoot initiation and shoot proliferation. The formation of rooting was used as the half-strength Murashige and Skoog (MS) medium with indole-3-butyric acid (IBA) and chitosan (CS). Result: The best response, shoot initiation and shoot proliferation were observed at 6-Benzylaminopurine (5.0 mg/L)+indole-3-acetic acid (0.5 mg/L)+chitosan (25 mg/L) and 6-Benzylaminopurine (4.0 mg/L)+indole-3-acetic acid (0.5 mg/L)+chatoyant (25 mg/L) in both of rhizome and sucker respectively. The maximum root formations were observed in the medium containing half-strength Murashige and Skoog medium+1.0 mg/L indole-3-butyric acid+25 mg/L chitosan in the rhizome and 0.8 mg/L indole-3-butyric acid+25 mg/L chitosan in the sucker. The successful survival rate of sucker and rhizome under the acclimatization condition was recorded as 90% and 88% compared with control as 66% and 63% respectively. This standardized protocol might be useful for the mass production of bantala variety as well as other cultivars of banana plants.

Introduction:The present research work was undertaken with a view to developing a suitable protocol forin vitroplant regeneration of economically important plant (Glycine max) (Bangladesh Agricultural Research Institute BARI- 5) variety,viaboth direct and indirect organogenesis fromin vitrogrown seedlings.Methods:For micropropagation explants were cultured on MS and half strength Murashige and Skoog (MS) medium supplemented with various plant growth regulators (cytokinins and auxins). In the present study for inducting of callus, among 3 different hormone combinations, the suitable medium was 3.32 mg/L 2, 4-D containing MS medium and the callus was deep green in color. Different type of media like MS, 1/2 MS and MS with different (6-Benzyl Amino Purine) BAP concentration was used for seed germination ofGlycine max. 100% of seed germination was observed in MS +1 mg/L BAP containing the medium.Results:In the present investigation, different concentration of cytokinins and auxins{BAP, 2, 4-D, and Naphthalene Acetic Acid (NAA)} were used individually or in combinations with MS medium to observe their effect on multiple shoot regeneration from the cotyledonary nodal segment. 100% shoot formation from cotyledonary nodal segment was recorded in 1.5 mg/L BAP and 0.15 mg/L BAP + 0.025 mg/L NAA containing MS medium, the best number of shoot was 10.9±2.0 found in MS + 1.5 mg/L BAP containing medium and highest length of shoot was 2 cm recorded in 1.5 mg/L BAP + 0.3 mg/L (different concentrations of Giberrellic acid) GA3containing MS medium. In addition, for root inductionin vitroraised well developed and elongated shoots were excised and cultured on MS and 1/2 MS medium supplemented with various concentration of Indole-3-Butyric acid (IBA). It was observed that MS medium containing 0.1 mg/L IBA and 1/2 MS medium containing 0.25 mg/L IBA was optimal for root induction. In which 100% shoots rooted well within 13 days of culture. The highest average number of roots per shoot was 6 recorded in MS +0.5 mg/L IBA containing the medium and highest average length of root was 8 cm recorded in 0.1 mg/L IBA containing MS medium.Conclusion:The most effective surface sterilization treatment for explants ofGlycine maxhas been found in 0.1% HgCl2solution for 15 minutes.

Aim:This study examined the healing potential of royal jelly on the acetic acid induced wounds healing in male rat’s gastric mucosa.Background:Scientific reports suggest that, bee products can help in the wounds healing.Methods:96 adult male Wistar rats were divided into in 4 groups as follows: control, omeprazole 20 mg/kg, and royal jelly 50 and 200 mg/kg). Wound was induced in stomach mucosa of each rat with 100% acetic acid. Samples groups received omeprazole or royal jelly from 1st to 14th day after acetic ulcer induction. Gastric ulcer healing and histopathological parameters were evaluated on 4, 7, 10, 15th days after ulceration. Both descriptive and statistical analyses were used. P <0.05 was considered as significant. Results:The royal jelly administration significantly reduced the depth of lesion in comparison with the control group (p<0.05) and attuned histopathological changes in the treatment groups. The largest healing effect was demonstrated with royal jelly on 10th treatment day, at a higher concentration (200 mg/kg). Conclusion:These findings supported that royal jelly had effectively contributed to the wound healing, valid gastroprotective activity, and can be used for peptic ulcer therapy.

ABSTRACTIntroduction:Royal jelly (RJ) has been consumed as food or as a supplement because of its high nutritional and medicinal values. A fresh harvested RJ is yellowish to whitish in color and contains proteins, free amino acids, lipids, vitamins, and sugar. Without proper storage conditions, such as at 4°C, the color of RJ changes to much darker yellow and produces a rancid smell. To prolong its shelf life, RJ is usually mixed with honey. Alginate, a natural and edible polymer derived from seaweed, is commonly used to encapsulate drugs and food due to its ability to form gels by reacting with divalent cations. However, there is a lack of research on the microencapsulation of RJ in alginate using electrospray. The electrospray technique has the advantage in producing consistent size and shape of alginate microbeads under optimum parameters.Aim:This research aimed to optimize electrospray-operating parameters in producing alginate–RJ microbeads.Materials and Methods:Optimization of alginate–RJ microbeads electrospray parameters was carried out using 24 factorial design with three center points (19 runs). The studied parameters were flow rate, high voltage, nozzle size, and tip-to-collector distance, whereas the responses were particle size, particle size distribution, and sphericity factor. The responses of each run were analyzed using Design-Expert software.Results:Nozzle size is a significant parameter that influences the particle size. Flow rate is a significant parameter influencing the sphericity factor.Conclusion:Screening of the electrospray-operating parameters paves the way in determining the significant parameters and their design space to produce consistent alginate–RJ microbeads.

Optimizing culture conditions lead to the improvement of oocyte developmental competence and additives with anti-oxidative activity in culture media improved embryonic development. Royal jelly (RJ) is a product from the cephalic glands of nurse bees that has considerable health effects. The aim of this study was to investigate the effect of different concentrations of RJ on the maturation, cleavage, and blastocyst rates and gene expression in the oocyte and cumulus cells during in vitro maturation (IVM) of sheep oocyte. IVM of oocyte was performed in the presence of control (RJ0), 2.5 (RJ2.5), 5 (RJ5), 10 (RJ10), 20 (RJ20), and 40 (RJ40) mg/mL of RJ. Following the maturation period, parthenogenetic activation was carried out in two treatment groups (RJ0 and RJ10) and embryonic development was examined three and eight days thereafter. Moreover, the relative expression of BCL2 and BAX in oocyte as well as BCL2, BAX, HAS2, PTGS2, and STAR in cumulus cells were assessed. The results indicated that the addition of 10 mg/mL of RJ (90 ± 4.51%) to the maturation medium linearly increased the oocyte maturation rate compared to the control group (57 ± 2.42%), then it remained constant to the RJ40 (93 ± 3.10%) group. The higher RJ concentrations were associated with increased (p < 0.01) cleavage (53.3 ± 1.55% to 82.3 ± 2.82%) and blastocyst rate (15.5 ± 1.16% to 33.8 ± 3.09%) from the RJ0 to the RJ10 group. The relative mRNA expression of BCL2 and BAX in the oocyte was higher at RJ10. In cumulus cells, the expression of BCL2 was not affected, but that of BAX decreased, and expression of HAS2, PTGS2, and STAR were increased following the addition of RJ to the maturation media. In conclusion, the addition of 10 mg/mL of RJ to maturation medium improved blastocyst formation and decreased the apoptotic incidence in sheep cumulus cells and the oocyte during the in vitro development.

Bee products as valuable nutritional ingredients: Determination of broad free amino acid profiles in bee pollen, royal jelly, and propolis

By damaging the liver, hepatitis B can result in acute and chronic diseases, such as cirrhosis or hepatocellular carcinoma. Viable treatments for such diseases using natural products and determinative biomarkers have been proposed but require evaluation to improve their effects. Therefore, this study aims to examine how effectively a specific natural product (namely, royal jelly) protects the body from the copy number of the virus, as well as TLR1 to TLR9 gene expressions. The effectiveness of royal jelly was tested by giving it (orally) to 30 hepatitis B patients for one month. HBV copy number and mRNA levels of TLRs were explored using Real Time PCR technique, and liver enzymes were evaluated too. Orally treatment with royal jelly led to a significant decrease in HBV-DNA copy number, down-regulation of TLR2 and TLR8, and up-regulation of TLR3. However, mRNA levels of the TLRs were not altered in the female, while TLR1, TLR2, and TLR5 were significantly decreased in the male participants. It seems that royal jelly has anti-viral and anti-inflammatory roles in the in vivo conditions in a dependent manner in TLR3, TLR2, and TLR8. Therefore, it can be suggested as a safe complementary agent for patients with hepatitis B.

Antioxidative action of royal jelly in the yeast cell

The article reviewed the foreign and Russian research of the changes in quality of royal jelly (Apis mellifera L.) caused by paratypic factors. The main quality standards of royal jelly are the total content of protein, sugars and 10-HDA concentration. The total protein and 10-HDA content determine the therapeutic properties of royal jelly. In 12 countries standards for royal jelly have been developed, these criteria do not differ much, but in Russia this standard regulates the product with no regard to modern methods of evaluation and quality standard. The quality of royal jelly is influenced by many factors. Honey bees of different breed origin produce royal jelly with different content of total protein and 10-HDA. Dark honey bees produce royal jelly with an average 0.3% higher concentration of protein and 10 - HDA. A negative correlation between the total gain of royal jelly and 10-HDA concentration has been determined (r=-0.77, p<0.05). The quality_of melliferous plants have a direct influence on the change in the total amount of sugars, protein and 10-hDa in royal jelly. By getting royal jelly from rape and lime bee pastures its quality criteria are higher, compared to apiaries with deadnettle plants. When plants don’t distill nectar, artificial carbohydrate-protein feedings have to be applied. It raises the total sugar content in royal jelly, reducing the quality. During the storage of royal jelly amino acid content varies. According to storage method this index ranges from 20% to 50%.

This study aimed to evaluate the effect of substrates in the emergence and early development of Baru seedlings. The experiment was performed in a greenhouse in a completely randomized design with five treatments (substrates) and four replications. The substrates were: S1) Oxisol; S2) Oxisol + washed sand (1: 1 v: v); S3) washed sand; S4)&nbsp; Oxisol + washed sand + manure (1: 1: 1 v: v: v); S5) Oxisol + sand washed + poultry litter (1: 1: 1 v: v: v). The emergence, emergence speed index, seedling height (SH), stem diameter (SD), root length, shoot dry mass (SDM), root dry mass (RDM), SH/SD and SDM/RDM ratios and the Dickson quality index were evaluated. The emergence and early development of seedlings were influenced by the type of substrate. The Oxisol substrate is the most efficient for the early development of Baru seedlings. Key words: Dipteryx alata Vog, emergence, germination, reforestation.

Oral administration of royal jelly (RJ) promotes the wound healing in diabetic mice. Concerns have arisen about the efficacy of RJ on wound healing process of normal skin cells. In this study, a wound was created by scratching normal human dermal fibroblasts, one of major cells involved wound healing process. 0.1, 1.0 or 5 ug/ml RJ were promptly treated up to 48 hrs, and migration was analyzed by the closure of wound margin. Furthermore, altered levels of lipids, which is recently reported to be responsible for wound healing process, were analyzed by HPTLC. Migration of fibroblasts was peaked up to ~24hr after wounding. RJ treatment significantly accelerated migration of fibroblasts in a dose dependent manner at 8hr. Although RJ accelerated migration of fibroblasts at either 20 hr or 24hr, the efficacy of RJ was less portent than at 8hr. Among various lipid classes of fibroblasts, the levels of triglycerol and cholesterol were significantly decreased with 5 ug/ml RJ. In spite of a dose‐dependent increase of sphinganine, no altered level of sphingosine, ceramides and glucosylceramide was noted with any concentration of RJ. We demonstrate that RJ enhances migration of fibroblasts with altered levels of various lipids in the wound healing process.This work is supported by Biogreen21 grant (20090801‐060‐001‐001‐02‐00) of Rural Development Administration

An efficient cryopreservation protocol was developed for mature seeds of Oncidium flexuosum Sims. Seed morphology, protocorm formation, and early seedling development were also assessed. The effects of phloroglucinol and Supercool X-1000® as cryoprotectant additives in the vitrification solution were investigated. Dehydration using the plant vitrification solution 2 (PVS2) for 60 and 120 min prior to immersion in liquid nitrogen promoted the highest frequency of in vitro seed germination 6 weeks following culture on half-strength Murashige and Skoog (½ MS) medium. Mature seeds submitted to vitrification for 120 min in PVS2 and 1 % phloroglucinol at 0 °C enhanced germination by 68 %, whereas in PVS2 and 1 % Supercool X-1000® germination was just moderately enhanced (26 %). In vitro-germinating seedlings developed healthy shoots and roots without the use of plant growth regulators. After 6 months of growth, there were no differences between in vitro- and ex vitro-grown seedlings for various phenotypic characteristics, including shoot length, number of leaves, number and length of roots, and fresh and dry weight. Seedlings were transferred to greenhouse conditions and successfully acclimatized, further developing into normal plants with over 90 % survival. Comparative analysis of seedlings from control and vitrified seeds using flow cytometry indicated that no change in ploidy levels occurred as a result of cryopreservation, therefore maintaining seedlings genetic stability. In this study, vitrification with PVS2 for 120 min with the addition of 1 % phloroglucinol offers a simple, safe, and feasible protocol for cryopreservation of O. flexuosum mature seeds.