• Journal of Acupuncture Research
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• v.19 no.2
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• pp.65-77
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• 2002

Objective : This study was undertaken to determine which compound of Bee Venom for herb-acupuncture has cytotoxicity on mouse mast cell line. Methods : We compared crude bee venom and its compounds such as melittin, mast cell degranulating peptide (MCD peptide), apamin with control groups on cytotoxicity by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Results & Conclusion : 1. Crude bee venom showed significant cytotoxic effect(p<0.01) in 1 hour treatment with $1{\mu}g/m{\ell}$ in comparison with control group in 1 hour treatment with low concentration of $10-4{\mu}g/m{\ell}$, $10-3{\mu}g/m{\ell}$, $10-2{\mu}g/m{\ell}$, $10-1{\mu}g/m{\ell}$ and $1{\mu}g/m{\ell}$, but it showed no significant cytotoxic effect in 6 hours treatment. 2. Melittin group showed no significant cytotoxic effect in comparison with control group in 1 and 6 hours treatment with low concentration of $10-4{\mu}g/m{\ell}$, $10-3{\mu}g/m{\ell}$, $10-2{\mu}g/m{\ell}$, $10-1{\mu}g/m{\ell}$ and $1{\mu}g/m{\ell}$. 3. MCD peptide and Apamin group showed no significant cytotoxic effect in comparison with control group in 1 and 6 hours treatment with low concentration of $10-4{\mu}g/m{\ell}$ $10-3{\mu}g/m{\ell}$, $10-2{\mu}g/m{\ell}$, $10-1{\mu}g/m{\ell}$ and $1{\mu}g/m{\ell}$. 4. Crude bee venom showed significant cytotoxic effect(p<0.01) in 1 and 6 hours treatment in comparison with control group in 1 and 6 hours treatment with high concentration of $10{\mu}g/m{\ell}$, $20{\mu}g/m{\ell}$ and $102{\mu}g/m{\ell}$. 5. Melittin group showed significant cytotoxic effect(p<0.01) in 1 hour treatment in comparison with control group in 1 hour treatment with high concentration of $10{\mu}g/m{\ell}$ but it showed no significant cytotoxic effect in 6 hours treatment. 6. Crude bee venom and its compounds have more cytotoxic effect in 1 hour treatment than in 6 hours treatment. It means cytotoxicity tends to decrease according to the treatment time.

• Journal of Pharmacopuncture
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• v.14 no.4
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• pp.23-32
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• 2011

Objectives: This study was performed to examine the antigenic potential of pure melittin (Sweet Bee Venom - SBV) extracted from the bee venom by utilizing protein isolation method of gel filtration. Methods: All experiments were conducted at Biotoxtech (Chungwon, Korea), authorized a non-clinical studies institution, under the regulations of Good Laboratory Practice (GLP). Antigenic potential of SBV was examined by active systemic anaphylaxis (ASA) and passive cutaneous anaphylaxis (PCA) in guinea pigs. SBV was subcutaneously administered at 0.07 and 0.28mg/kg and also as a suspension with adjuvant (Freund's complete adjuvant: FCA). Ovalbumin (OVA) as a suspension with adjuvant was used to induce positive control response ($5mg/m{\ell}$-FCA). Results: 1. In the ASA test, experimental groups showed some symptoms of anaphylaxis like piloerection, hyperpnea and staggering gait. 2. In the PCA test, low dosage group did not show any antibody responses, whereas high dosage group showed positive responses. 3. In the weight measurement and clinical observation, experimental groups didn't show any significant changes compared with control group. 4. In the autopsy of body, the abnormalities of lung were detected in the corpse. This means that the cause of death may induced anaphylactic shock. Conclusions: Above findings suggested that SBV had antigenic potential in guinea pig. Further studies on the subject should be conducted to yield more concrete evidences.

• Journal of Microbiology
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• v.34 no.3
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• pp.274-278
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• 1996

Two peptides, MA-inv AND MA-ME, with the sequence 10-17 of maganin 2 at their-N-termini were designed and synthesized. The peptides had higher antifungal activities against Aspergilus fumigatus without hemolytic activities. The minimal inhibition concentratory (MIC) values of both peptides against A. fumigatus were 5 .mu.g/ml, whereas those of the native peptides, magainin 2 and melittin, were 10.mu.g/ml. At 3 .mu.g/ml, MA-inv and MA-ME inhibited the mycelium growth of A. fumigatus by 94.6% and 97.3% respectively, whereas magainin 2 and melittin inhibited by 62.2% and 32.4, respectively. MA-inv showed up to 80% inhibition of (1, 3)-.betha.-D-glucan synthase activity of A. fumigatus. The peptides also showed up to 80% inhibition of (1, 3)-.betha.-D glucan synthase activity of A. fumigatus. The peptides also showed antifungal activities for other fungi of Aspergillus sp. However, the antibiotic activities of MA-ME against Escherichia coli, Bacillus subtilis and Fusarium oxysporum were more effective than those of MA-inv, suggesting that the C-terminal sequences of MA-inv and MA-ME may also influence their antibiotic activities. These results suggest that the N-terminal sequence of the designed peptides, KKFGKAFV, is important for their antifungal activities against A. fumigatus and their C- terminal sequences are related to the organism selectivity.

• Analytical Science and Technology
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• v.27 no.6
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• pp.277-283
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• 2014

The effects of concentration, pressure, and molecular sige on removing allergenic substance (phospholipase $A_2$) in bee venom by ultrafiltration were investigated. The membrane pore sizes were selected based on the molecular weight of the main compounds. The conditions of concentration and pressure were selected randomly. As results, we obtained the optimum condition (1 mg/mL, 20 psi, 10,000 dalton) for removing $PLA_2$ at constant concentration of melittin and apamin and confirmed the separation results by HPLC and SDS-PAGE.

• The Korean Journal of Physiology and Pharmacology
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• v.12 no.6
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• pp.343-347
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• 2008

This study was carried out to investigate the wound healing effect of caffeic acid in skin-incised mice. Caffeic acid showed significant effects on anti-inflammatory activity and wound healing, such as myeloperoxidase activity, lipid peroxidation, phospholipase $A_2$ activity and collagen-like polymer synthesis, in incised-wound tissue. On the other hand, it significantly stimulated collagen-like polymer synthesis in NIH 3T3 fibroblast cells, while inhibited both silica-induced reactive oxygen species generation and melittin-induced arachidonic acid release and $PGE_2$ production in Raw 264.7 cells, and histamine release in RBL 2H3 cells stimulated by melittin or arachidonic acid. Therefore, caffeic acid appears to have a potent antioxidant and anti-inflammatory effect in cell culture system, which may be related to wound healing in skin-incised mice.

• Journal of Pharmacopuncture
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• v.12 no.2
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• pp.85-90
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• 2009

Objectives : Sweet bee venom(SBV) is pure melittin, the main component of bee venom, made by removing another components through gel filtration chromatography and propionic acid/urea polyacrylamide gel electrophoresis. In the using SBV, 2 patients were experienced anaphylaxis by SBV. So, we reported the process of anaphylaxis and consider these situation. Methods : We observed 2 patients suffered from anaphylaxis after treated with SBV in the Korean Medical Hospital, Sangji University. Results : Though SBV was removed allergen from bee venom, it is not possible to complete prevention of anaphylactic shock in the clinics. So, Korean medical doctor using BV or SBV must be prepare the system consider a countermeasure by anaphylaxis.

• 한국약용작물학회:학술대회논문집
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• 2008.11a
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• pp.451-452
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• 2008

• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.04a
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• pp.281-281
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• 1996

The immunoassay method is frequently used for the identification and quantitation of ultratrace amount of bioactive substances. Homogeneous liposome immunoassays, which can avoid the use of radioisotopes and separation steps, have recently been reported in many publications. Cytolysin-mediated liposome immunoassay using melittin ever been studied but showed limited applications. Here, we designed a homogeneous liposome immunoassay using Clostridium perfringens phospholipase C (PLC), an enzyme which catalyzes the hydrolysis of phosphatidylcholine in biological membranes, as a cytolysin.

• Proceedings of the Korean Magnetic Resonance Society Conference
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• 1999.02a
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• pp.13-13
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• 1999

• Journal of Oriental Neuropsychiatry
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• v.19 no.3
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• pp.117-127
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• 2008

Objective: Bee venom (BV) has been used for the treatment of inflammatory diseases such as rheumatoid arthritis and relief of pain in Oriental medicine. The two main components of BV are melittin and phospholipase A2 (PLA2). Of these, melittin, the major active ingredient of BV, has been reported to induce apoptosis and to possess anti tumor effects. Several studies have established that the agents inducing apoptosis in target organs suppress tumorigenesis. As the other component, PLA2 has been reported to induce neurite outgrowth in PC12 cells. However, there was no report about proliferative effect of BV in neuronal cells. In order to examine the effect of BV on glioma cell, human glioma cell line, U87 was used. Methods: Analysis of proliferation was confirmed by MTT assay. BV increased cell number through dose and duration dependent manner and these effects are apparent at a concentration of 10 ug/ml. To observe which signaling molecules will be activated by BV, phosphorylation of Akt, MAPK, PYK2 or CREB were examined by Western blot analysis. To study the long term effect of BV in U87 cells, the image of cells treated with BV for 4 days were obtained. Results: The phosphorylation levels of PYK2 and Akt were increased at 5 min after addition of 10 ug/ml of BV and sustained to 2 hours. On the other hand, phosphorylation of MAPK and CREB were increased at 5 min, maximum at 10 min, and returned to 30 min. These imply that BV may activate two different signaling pathways, PYK2/Akt and MAPK/CREB. BV treated cells showed increased neurite number and length. Conclusion: These results propose that BV may induce differentiation as well as proliferation of U87 cells through the activation of PYK2/ Akt and MAPK/ CREB.