Putrescine은 일반적으로 미생물의 활동에 의해 발생되며, 신선함의 척도로서 사용된다. 그러나 동결건조된 로열젤리에 대한 putrescine 의 분석법은 아직 확립되지 않은 실정이다. 본 연구에서는 C18 컬럼을 이용하여 동결건조 로열젤리 내 putrescine을 분석하기 위한 UPLC 분석 법을 확립하고자 하였다. 새롭게 확립된 분석법은 7분 이내에 putrescine을 분석 가능하였으며, 이러한 분석법을 검증하기 위해 특이성, 직선성, 정밀성, 정확성, 정량한계, 정성한계 등을 평가하였다. 본 연구를 통해 동결건조 로열젤리의 신선한 정도를 평가하기 위한 분석법을 제공하였으 며, 추후 안전성의 척도에 대한 자료로서 활용 가능할 것으로 기대되어진다.

Royal jelly (RJ) is a gelatinous substance that bees produce to feed bees and queen bees. It’s frequently sold as a dietary supplement to treat a variety of physical ailments and chronic diseases. While it has long been used in traditional medicine, its applications in Western medicine remain controversial. The inhibitory effect of royal jelly on osteoarthritis was investigated in primary cultured rat cartilage cells and monosodium-iodoacetate (MIA)-induced arthritis rat model 10-hydroxy-2-decenoic acid (10-HAD) is the main fatty acid present in RJ. Among the criteria for RJ quality analysis, 10-HAD content has been proposed as a freshness parameter. We investigated the effect of RJ on the improvement of osteoarthritis on SD rats and they were divided into five groups. In this study, we examined the effect of enzymatic royal jelly (ERJ) administration on osteoarthritis. To determine the antiinflammatory effects of RJ, tumor necrosis factor alpha (TNF-α) and Interleukin-6 (IL-6) expression were measured after lipopolysaccharide (LPS) activation in RAW 264.7 cells. In in vivo animal study, osteoarthritis was induced by intra-articular injection of MIA into knee joints of rats. As a results, ERJ showed that TNF-α and IL-6 levels were decreased by ERJ treatment in a dosedependent manner. In conclusion, ERJ extract was able to inhibit articular cartilage degeneration by preventing extracellular matrix degradation and cartilage cell damage. It was considered that ERJ extract may be a potential therapeutic treatment for degenerative osteoarthritis.

Royal jelly (RJ) is a well-known functional and medicinal food for human health promotion. Major royal jelly proteins (MRJPs), which are the major protein components in RJ, exhibit antimicrobial activities. However, the identities of the MRJPs of RJ responsible for its antioxidant effects have remained unclear. Here, we report that honeybee (Apis cerana) MRJP 2 (AcMRJP2) acts as an antimicrobial and antioxidant agent in RJ. Using recombinant AcMRJP2, which was produced in baculovirus-infected insect cells, we established the antimicrobial and antioxidant roles of MRJP 2. AcMRJP2 bound to the surfaces of bacteria, fungi, and yeast, which then induced structural damage in the microbial cell walls and led to a broad spectrum of antimicrobial activities. AcMRJP2 protected mammalian and insect cells via the direct shielding of the cell against oxidative stress, which led to reduced levels of caspase-3 activity and oxidative stress-induced cell apoptosis, followed by increased cell viability. Moreover, AcMRJP2 exhibited DNA protection activity against reactive oxygen species (ROS). Our data indicate that AcMRJP2 could play a crucial role as an antimicrobial agent and antioxidant in RJ, suggesting that MRJP 2 is a component responsible for the antimicrobial and antioxidant activities of RJ.

The hypopharyngeal gland (HPG) of the honeybee worker produces royal jelly (RJ) and has a developmental cycle closely related to the division of labor. In this study, we investigated to compare the HPG acini diameter of differently aged worker bees with high royal jelly producing colony (HRC) or less producing colony (LRC). Additionally, we also evaluated whether the fresh weight of the head is a reliable indicator of the developmental status of HPG. The HRC showed a significantly higher RJ production about two-times as compared with those of the LRC. We measured the HG-diameters on days 1, 3, 6, 9, 12, 15. The microscopic analysis revealed that the acini size of the HRC was significantly larger than the LRC. In addition, the acini diameter of HRC was 15% longer than the LRC on the first day after emerging. It was shown that the fastest development during 3 days which is preparing for nurse the brood. The HPG acini diameters increased in both colonies in a similar fashion until day 12 and then decreased. We also compared the fresh head weight of the experimental colonies, differences were similar to the development of HPG. Therefore, high royal jelly production may have a positive correlation between HPG acini size and the fresh head weight.

Major royal jelly proteins (MRJPs), important protein components of bee royal jelly (RJ) and exclusive nourishments for queen, exhibit various biological and pharmacological activities. RJ is one of the most studied bee products, but the crucial roles for MRJP2 as an antimicrobial and antioxidant agents remain largely unknown. Here we demonstrated the antimicrobial and antioxidant functions of the Asiatic honeybee (Apis cerna) MRJP2 (AcMRJP2). Recombinant AcMRJP2 of approximately 53 kDa was expressed in baculovirus-infected insect cells, and it exhibited antimicrobial activity against bacteria, fungi, and yeast via binding to microbial surfaces and inducing structural damage in microbial cell walls. AcMRJP2 protected mammalian and insect cells against oxidative damage through shielding of cell membranes. Interestingly, AcMRJP2 exhibited DNA protection activity and DPPH radical-scavenging activity. Altogether, our data demonstrated that AcMRJP2 functions as antimicrobial and antioxidant agents.

Major royal jelly proteins (MRJPs) are important protein components of bee royal jelly (RJ) and exhibit various biologicaland pharmacological activities. The antimicrobial activities of royalisins and the jelleines contained within MRJP 1 andMRJP 2 in RJ have been elucidated. However, the antimicrobial effects of other bee RJ MRJPs remain largely unknown.In this study, we demonstrated that the Asiatic honeybee (Apis cerana) MRJP 4 (AcMRJP4) exhibits antimicrobial activitiesagainst bacteria, fungi, and yeast. Recombinant AcMRJP4 was expressed as a 63-kDa protein in baculovirus-infected insectcells. However, some of the recombinant AcMRJP4 proteins were cleaved into two fragments (i.e., 48-kDa (AcMRJP4-48)and 15-kDa (AcMRJP4-15) proteins) by the proteolytic cleavage of the C-terminus of the recombinant AcMRJP4. Interestingly,AcMRJP4, AcMRJP4-48, and AcMRJP4-15 exhibited antimicrobial activities, with AcMRJP4-15 exhibiting the highestantimicrobial activity, followed by AcMRJP4. AcMRJP4-15, which is a hydrophilic peptide with 88 amino acid residuesthat contains a high content of Asn and positively charged amino acids, induced structural damage in the cell walls ofthe assayed bacteria, fungi, and yeast. Altogether, our data demonstrated that AcMRJP4 functions as an antimicrobial agent.

Royal jelly (RJ) is one of the most attractive functional foods that have been a commercial product, especially in dietetics and cosmetics in many countries. However, RJ has been evoked with dermatitis, acute asthma and anaphylaxis because of major RJ proteins. Therefore, to access water soluble royal jelly (WSRJ) that removed allergy-induced proteins as an effective whitening agent for cosmetics and potential external treatment for topical use, we investigated its ability to inhibit melanin biosynthesis. B16F1 cells were treated with 10 nM α-melanocyte-stimulating hormone (α -MSH) for 48hr, and then were treated with various doses of WSRJ for 36hr. WSRJ (1-10ug/ml) inhibited direct tyrosinase activity and cellular tyrosinase activity, which lead to the decrease of melanin synthesis in α-MSH stimulated B16F1 melanoma cells. In addition, we examined RT-PCR and Western blotting for melanogenesis-related genes such as tyrosinase, tyrosinase related protein 1 (TRP-1) and 2. WSRJ suppressed mRNA and protein expression of tyrosinase, tyrosinase related protein (TRP)-1 and TRP-2 in α-MSH stimulated B16F1 cells, and similar to positive control, arbutin. Our findings suggest that WSRJ induced the downregulation of melanogenesis by inhibiting tyrosinase, TRP-1 and 2 activations. It may serve as a new candidate in the new skin-whitening agents.

Royal jelly (RJ) is exclusive food that is secreted from the hypopharyngeal and mandibular glands of worker honeybees, and it is well known to be a necessary for the growth of the queen honeybee Although fresh royal jelly have been demonstrated to enhance wound healing, the wound healing effects of water soluble royal jelly (WSRJ) have not been elucidated. We investigated whether WSRJ promotes the migration, attachment, and proliferation of human dermal fibroblasts (HDFs) during in vitro wound healing. HDFs were treated with 1-5ug/ml WSRJ and RJ for up to 24hr following wound formation. Cell migration was assessed by measuring recovery from wound margin, while cell attachment and proliferation were determined by MTT assay. By observing the numbers of cell attached, we confirmed that not only WSRJ but also RJ did not affect on the initial cell adhesion. WSRJ (5 ug/ml) enhanced cell migration rate approximately 84.3% in HDFs at 24hr, whereas RJ (5 ug/ml) increased cell migration rate 71.3% in HDFs at 24hr, which is similar to cell migration rate of WSRJ 1 ug/ml (73.7%). In cell proliferation assays, WSRJ induced an increase in the number of HDFs, compared with control and RJ. In conclusion, WSRJ promotes cell migration with increased cell proliferation in an in vitro wound healing model.

We compared the grafting success in total of 107 rearing Apis carana queens cells, to which we grafted 540 larvae. The wax for cups we prepared from A. mellifera and A. cerana wax. The A. cerana wax cups were found that artificial queen cell cups with the internal diameter of 8.0 mm at the mouth and 8.0 mm depth were highly preferred by the bees for rearing of queens from the grafted larvae. From the 210 grafted larvae into A. mellifera wax bees accepted 30 queens cells, only (16.67 %) ; A. cerana wax bees accepted 59 queens cells (33 %) ; plastic cup bees accepted 18 queens cells, only (10 %). In the preference test the grafting success in the A. cerana wax cups were better than in the A. mellifera wax and plastic cup. The results show better acceptance of larvae grafted into the pure A. cerana wax cups for rearing A. cerana queen. A new method for rearing honey bees, A. cerana, in vivo was developed and the effects of royal jelly from A. mellifera. We used royal jelly diluted 50:50 with sterile water (The royal jelly is kept frozen until used). A small amount of royal jelly is placed at the center of each cell cup. Young A. cerana larvae were transferred into the queen cups containing ± 10 ㎍ of the Royal jelly from A. mellifera and A. cerana. The average rates of acceptance were affected significantly due to the royal jelly source in the queen cell cups. It is so workable first to produce pure A. cerana wax for making the queen cups before a beekeeper starts with grafting.

This study was conducted to evaluate whether \"Royal jelly\" (RJ) added to Tris-buffer dilute contributed to supporting post-thaw viability and longevity of frozen canine spermatozoa. Two Japanese spitzs (2 to 4 years of age) were used as a semen donor. Semen was collected by manual masturbation and separated into 3 fractions. Only the sperm-rich fraction having sperm motility of more than 70%, containing sperm concentration of 2~410 cells/ml and having dead or abnormal spermatozoa of less than 15% was used for the experiment. Each ejaculated semen was centrifuged at 400 g for 5 min and then diluted in a Tris-buffer supplemented with 20 ml egg yolk (Ext I), 4% glycero1 and 1% Equex STM Paste (Ext II) or g1ycero1, Equex STM paste and RJ of various concentrations (Ext II-RJ). After freezing and thawing, viability of spermatozoa in Ext II -RJ containing 1% RJ immediately after thawing (67.59.6) was significantly lower than that of Ext II , Ext II -RJ containing 0.01 or 0.1% RJ (77.512.5, 78.78.2 and 80.06.3). However, Ext II-RJ containing 0.1% RJ yielded higher viability than Ext II, Ext II-RJ containing 0.01% at or 1% 1 h after thawing (69.58.1 vs. 55.012.9, 57.59.6 and 41.512.6; P<0.05). At 1 h after thawing, the viability of spermatozoa thawed in 7 (68.812.5) was significantly higher than that of spermatozoa thawed in 38 (48.816.3), although there was no difference in the viability between both groups immediately after thawing (77.59.6 and 81.38.1). Post-thaw viability and longevity of post-thaw spermatozoa in Ext II-RJ containing 0.1% RJ was higher in those in Ext II at 1 h (65.012.9 vs. 42.512.6), 2 h (52.512.6 vs. 27.517.1) and 3 h (40.014.1 vs. 20.012.1) after thawing. These results indicated that addition of 0.1% af to Tris-buffer enhanced post-thaw viability and longevity of canine spermatozoa and this additive can be used for increasing the possibility of collision between spermatozoa and ova during insemination.emination.

A fine granule was prepared using freeze-dried royal jelly. For its preparation, which depended on operational parameters like its glucose-to-total sugar content ratio (X1,0-100%), ethanol concentration (X2,75-95%) and sprayed ethanol solution content (X3,8-12%) using freeze-dried royal jelly, the response surface methodology was used to monitor the optimum conditions for the yield, the fragmentation rate with shaking, and the organoleptic properties. The maximum yield was 89.99% with a glucose–to-total sugar content ratio of 59.30%, an ethanol concentration of 88.64%, and a sprayed ethanol solution content of 11.83%. The minimum fragmentation rate by shaking was 0.82% at the glucose–to-total-sugar content ratio of 22.35%, the ethanol concentration of 77.21%, and the sprayed ethanol solution content of 10.59%. The sensory score for the overall palatability of the organoleptic properties was 7.45 at the glucose–to-total-sugar content ratio of 31.81%, the ethanol concentration of 93.96%, and the sprayed ethanol solution content of 10.51%.