OBJECTIVE: Melittin, a cell-penetrating peptide, improves the efficiency of many non-viral gene delivery vectors, yet its application in viral vectors has not been well studied. The non-pathogenic recombinant adeno-associated virus (rAAV) vector is an ideal in vivo gene delivery vector. However, its full potential will only be achieved after improvement of its transduction efficiency. To improve the transduction efficiency of rAAV2 vectors, we attempted to develop a melittin-based rAAV2 vector delivery strategy. METHODS: The melittin peptide was inserted into the rAAV2 capsid either in the loop VIII of all viral proteins (VPs) or at the N terminus of VP2. Various rAAV2-gfp or -fluc vectors were subjected to quantitative real-time polymerase chain reaction and Western blot assays to determine their titers and integrity of capsid proteins, respectively. Alternatively, the vectors based on wild-type capsid were pre-incubated with melittin, followed by transduction of cultured cells or tail vein administration of the mixture to C57BL/6 and BALB/c nude mice. In vivo bioluminescence imaging was performed to evaluate the transgene expression. RESULTS: rAAV2 vectors with melittin peptide inserted in the loop VIII of VPs had low transduction efficiency, probably due to dramatically reduced ability to bind to the target cells. Fusing the melittin peptide at the N-terminus of VP2 produced vectors without the VP2 subunit. Interestingly, among the commonly used rAAV vectors, pre-incubation of rAAV2 and rAAV6 vectors with melittin significantly enhanced their transduction efficiency in HEK293 and Huh7 cells in vitro. Melittin also had the ability to increase the rAAV2-mediated transgene expression in mouse liver in vivo. Mechanistically, melittin did not change the vector-receptor interaction. Moreover, cell counting kit-8 assays of cultured cells and serum transaminase levels indicated melittin had little cytotoxicity. CONCLUSION Pre-incubation with melittin, but not insertion of melittin into the rAAV2 capsid, significantly enhanced rAAV2-mediated transgene expression. Although further in vivo evaluations are required, this research not only expands the pharmacological potential of melittin, but also provides a new strategy to improve gene therapy mediated by rAAV vectors.
Mice ; Animals ; Humans ; Melitten/genetics* ; Dependovirus/genetics* ; Serogroup ; HEK293 Cells ; Mice, Nude ; Mice, Inbred C57BL ; Transgenes ; Genetic Vectors/genetics*

Acupuncture ; Acupuncture Therapy/adverse effects* ; Bee Venoms/adverse effects* ; Humans

PURPOSE: To report a patient stung by a bee, who was diagnosed with sterile endopthalmitis and another patient diagnosed with optic neuritis, with decreasing visual acuity, after refined bee venom injection around the orbital tissue. CASE SUMMARY: A 82-year-old female visited our hospital for decreased visual acuity in the right eye and ocular pain due to a bee sting. The bee sting penetrated the sclera into the vitreous. In the anterior segment, severe cornea edema and anterior chamber cells were seen. Using ultrasonography, inflammation was seen around the intravitreal area. After 3 months, intravitreal inflammation regressed but the patient's visual acuity was light perception negative, and corneal opacity, neovascularization, and phthisis bulbi were detected. A 55-year-old male visited our hospital for ocular pain in the right eye and decreasing visual acuity after refined bee venom injection around the orbital tissue. The best-corrected visual acuity in the right eye was 15/100, there was moderate injection on the conjunctiva. A relative afferent pupillary defect, abnormal color vision test results, and a defect in the visual field test were observed. There was no pain during external ocular movement, and other general blood tests, and a brain MRI were normal. Based on these symptoms, methylprednisolone megatherapy was started for treatment of optic neuritis. After treatment, visual acuity of the right eye was 9/10 and all other clinical optic neuritis symptoms regressed. CONCLUSIONS: Based on these two cases, ocular toxicity from bee venom could result from both direct and indirect courses. Treatment using refined bee venom might be harmful, and caution is recommended in its use.
Aged, 80 and over ; Anterior Chamber ; Bee Venoms ; Bees ; Bites and Stings ; Brain ; Color Vision ; Conjunctiva ; Cornea ; Corneal Opacity ; Edema ; Female ; Hematologic Tests ; Humans ; Inflammation ; Magnetic Resonance Imaging ; Male ; Methylprednisolone ; Middle Aged ; Optic Neuritis ; Orbit ; Pupil Disorders ; Sclera ; Ultrasonography ; Visual Acuity ; Visual Field Tests

BACKGROUND/AIMS: Interstitial cells play important roles in gastrointestinal (GI) neuro-smooth muscle transmission. The underlying mechanisms of colonic dysmotility have not been well illustrated. We established a partial colon obstruction (PCO) mouse model to investigate the changes of interstitial cells and the correlation with colonic motility. METHODS: Western blot technique was employed to observe the protein expressions of Kit, platelet-derived growth factor receptor-α (Pdgfra), Ca²⁺-activated Cl⁻ (Ano1) channels, and small conductance Ca²⁺- activated K⁺ (SK) channels. Colonic migrating motor complexes (CMMCs) and isometric force measurements were employed in control mice and PCO mice. RESULTS: PCO mice showed distended abdomen and feces excretion was significantly reduced. Anatomically, the colon above the obstructive silicone ring was obviously dilated. Kit and Ano1 proteins in the colonic smooth muscle layer of the PCO colons were significantly decreased, while the expression of Pdgfra and SK3 proteins were significantly increased. The effects of a nitric oxide synthase inhibitor (L-NAME) and an Ano1 channel inhibitor (NPPB) on CMMC and colonic spontaneous contractions were decreased in the proximal and distal colons of PCO mice. The SK agonist, CyPPA and antagonist, apamin in PCO mice showed more effect to the CMMCs and colonic smooth muscle contractions. CONCLUSIONS: Colonic transit disorder may be due to the downregulation of the Kit and Ano1 channels and the upregulation of SK3 channels in platelet-derived growth factor receptor-α positive (PDGFRα⁺) cells. The imbalance between interstitial cells of Cajal-Ano1 and PDGFRα-SK3 distribution might be a potential reason for the colonic dysmotility.
Abdomen ; Animals ; Apamin ; Blotting, Western ; Chloride Channels ; Colon ; Down-Regulation ; Feces ; Interstitial Cells of Cajal ; Mice ; Muscle, Smooth ; Myoelectric Complex, Migrating ; Nitric Oxide Synthase ; Platelet-Derived Growth Factor ; Silicon ; Silicones ; Small-Conductance Calcium-Activated Potassium Channels ; Up-Regulation

STUDY DESIGN: Case report. OBJECTIVES: We report a case of widespread lumbosacral subdural abscess in a patient who underwent bee venom therapy. SUMMARY OF LITERATURE REVIEW: Subdural abscess is rare, but has a poor prognosis. Therefore, prompt recognition and appropriate treatment are paramount. MATERIALS AND METHODS: A 54-year-old woman was hospitalized due to severe back pain. Two days previously, she had undergone bee venom therapy. The patient then visited the emergency room because of severe back pain. However, a paraspinal infection was not detected on enhanced magnetic resonance imaging (MRI). Six days after admission, the patient showed signs of meningeal irritation and an emergency cerebrospinal fluid analysis showed typical findings of bacterial meningitis. Although adequate antibiotic treatment was administered, 20 days after admission, the patient's symptoms became aggravated. Pachymeningeal enhancement, myelomeningitis, and subdural abscess compressing the cauda equina were found on enhanced MRI. Thus, laminectomy between L3–L4 and L5–S1 was performed, as well as subdural abscess drainage. Antibiotic agents were applied for 6 weeks after the operation, and resolution of the subdural abscess was identified on follow-up MRI. RESULTS: In this patient, lumbosacral subdural abscess occurred due to bee venom therapy. It was cured by adequate surgical and antibiotic treatment. CONCLUSIONS: Bee venom therapy can cause subdural abscess of the spinal cord. Even if it is a rare case, this possibility is worth consideration in the Korean medical context.
Abscess ; Back Pain ; Bee Venoms ; Bees ; Cauda Equina ; Cerebrospinal Fluid ; Drainage ; Emergencies ; Emergency Service, Hospital ; Female ; Follow-Up Studies ; Humans ; Laminectomy ; Magnetic Resonance Imaging ; Meningitis, Bacterial ; Middle Aged ; Prognosis ; Spinal Cord ; Spine

Severe systemic responses including neurologic complications such as myasthenia gravis, myeloradiculopathy, optic neuropathy, parkinsonism, stroke and Guillain-barré syndrome can occur after bee stings. This case describes a 78-year-old female who presented with symptoms of acute progressive bilateral symmetrical weakness in both lower legs after multiple bee stings. Nerve conduction study findings were consistent with acute sensorimotor axonal neuropathy and recovered by treatment with intravenous immunoglobulin. This case highlights that bee stings can result in acute onset Guillain-barré syndrome, although the pathophysiologies of bee venoms need to be investigated accurately.
Aged ; Axons ; Bee Venoms ; Bees* ; Bites and Stings* ; Female ; Guillain-Barre Syndrome* ; Humans ; Immunoglobulins ; Leg ; Myasthenia Gravis ; Neural Conduction ; Optic Nerve Diseases ; Parkinsonian Disorders ; Polyradiculoneuropathy ; Stroke

Lymphedema is a common complication associated with cancer itself or with cancer treatment. Lymphedema infrequently occurs after drug therapy. Bee venom is one of the materials used in acupuncture, and it has been used in the treatment of a variety of inflammatory diseases including arthritis. We report a 74-year-old male patient with late-onset post-radiation lymphedema provoked by bee venom therapy. He was free of lymphedema for 5 years after the complete remission of prostate cancer which had been treated with transurethral resection and radiation therapy. The patient developed left leg swelling after undergoing bee venom therapy for left hip pain. Computed tomography and lymphoscintigraphy showed lymphedema without tumor recurrence or infection. The lymphatic system was suspected to be injured by bee venom therapy and lymphedema was provoked. Bee venom therapy should be used cautiously in patients prone to lymphedema.
Acupuncture ; Aged ; Arthritis ; Bee Venoms* ; Bees* ; Drug Therapy ; Hip ; Humans ; Leg ; Lymphatic System ; Lymphedema* ; Lymphoscintigraphy ; Male ; Prostatic Neoplasms ; Recurrence

BACKGROUND: Bee venom (BV) has been widely investigated for potential medical uses. Recent inadvertent uses of BV based products have shown to mitigate signs of fungal infections. However, the component mediating the antifungal effect has not been identified. OBJECTIVE: This investigation compares bee venom in its whole and partial forms to evaluate the possible component responsible for the antifungal effect. METHODS: Forty-eight plates inoculated with Trichophyton rubrum were allocated into four groups. The groups were treated with raw BV (RBV), melittin, apamin and BV based mist (BBM) respectively and each group was further allocated accordingly to three different concentrations. The areas were measured every other day for 14 days to evaluate the kinetic changes of the colonies. RESULTS: The interactions of ratio differences over interval were confirmed in groups treated with RBV and BBM. In RBV, the level of differences were achieved in groups treated with 10 mg/100 µl (p=0.026) and 40 mg/100 µl (p=0.000). The mean difference of ratio in groups treated with RBV was evident in day 3 and day 5. The groups that were treated with melittin or apamin did not show any significant interaction. In BBM groups, the significant levels of ratio differences over time intervals were achieved in groups treated with 200 µl/100 µl (p=0.000) and 300 µl/100 µl (p=0.030). CONCLUSION: The the bee venom in its whole form delivered a significant level of inhibition and we concluded that the venom in separated forms are not effective. Moreover, BV based products may exert as potential antifungal therapeutics.
Antifungal Agents ; Apamin ; Bee Venoms* ; Bees* ; Melitten ; Negotiating ; Trichophyton* ; Venoms

Bee venom immunotherapy (b-VIT) can be combined with omalizumab therapy in order to suppress systemic reactions developing due to b-VIT itself. Omalizumab acts as a premedication and gains time for the immunotherapy to develop its immunomodulatory effects. However, the combination of omalizumab and b-VIT is not always effective enough. Herein we present a patient in whom successful immunotherapy cannot be achieved with combination of omalizumab to b-VIT.
Anaphylaxis ; Bee Venoms ; Bees ; Humans ; Immunotherapy ; Omalizumab ; Premedication

BACKGROUND: Allergic reactions developing after bee sting can be severe and life-threatening. According to epidemiological data, serious systemic reactions range between 1.2%–3%, and this is 2–3 times higher (6%) in beekeeping. In different beekeepers' populations, risk factors of systemic reactions have been investigated and diverse results have been found. OBJECTIVE: The aim of this study is to evaluate the level of knowledge of beekeepers about venom allergy, epidemiological data, systemic reaction rates, risk factors for systemic reactions, and the rate of emergency admissions after bee sting. METHODS: With the collaboration of Uludağ University Beekeeping Development Research Center and Beekeepers Association, a questionnaire consisting of 19 questions was applied to 242 beekeepers in Bursa and Yalova. Two hundred twenty-one beekeepers who completed the questionnaire were involved in the study. RESULTS: The mean age of the beekeepers was 49.9 years (range, 18–75 years). The systemic reaction to bee sting in beekeepers was 37.6%. Allergic rhinitis was found to be a risk factor for systemic reaction. Although 80% of the beekeepers recognized that bee venom could be lethal, only 60% of the beekeepers were aware of immunotherapy, and only 30% were aware of the adrenaline auto-injector drug. CONCLUSION: Similar to previous studies, we found that the systemic response to the bee sting in beekeepers was higher compared to normal population. Considering the occupational exposure to bee venom and thus higher risk, the awareness of venom allergy in this high risk population was low, and they were poorly informed about the treatment options.
Bee Venoms ; Beekeeping ; Bees ; Bites and Stings ; Cooperative Behavior ; Emergencies ; Epinephrine ; Hypersensitivity ; Immunotherapy ; Occupational Exposure ; Rhinitis, Allergic ; Risk Factors ; Venoms