Design of a Modern Liposome and Bee Venom Formulation for the Traditional VIT-Venom Immunotherapy

Traditional venom immunotherapy uses injections of whole bee venom in buffer or adsorbed in Al (OH)₃ in an expensive, time-consuming way. New strategies to improve the safety and efficacy of this treatment with a reduction of injections would, therefore, be of general interest. It would improve patient compliance and provide socio-economic benefits. Liposomes have a long tradition in drug delivery because they increase the therapeutic index and avoid drug degradation and secondary effects. However, bee venom melittin (Mel) and phospholipase (PLA₂) destroy the phospholipid membranes. Our central idea was to inhibit the PLA₂ and Mel activities through histidine alkylation and or tryptophan oxidation (with pbb, para-bromo-phenacyl bromide, and/or NBS- N-bromosuccinimide, respectively) to make their encapsulations possible within stabilized liposomes. We strongly believe that this formulation will be nontoxic but immunogenic. In this paper, we present the whole bee venom conformation characterization during and after chemical modification and after interaction with liposome by ultraviolet, circular dichroism, and fluorescence spectroscopies. The PLA₂ and Mel activities were measured indirectly by changes in turbidity at 400_{n m}, rhodamine leak-out, and hemolysis. The native whole bee venom (BV) presented 78.06%?of α-helical content. The alkylation (A-BV) and succynilation (S-BV) of BV increased 0.44 and 0.20%?of its α-helical content. The double-modified venom (S-A-BV) had a 0.74%?increase of α-helical content. The BV chemical modification induced another change on protein conformations observed by Trp that became buried with respect to the native whole BV. It was demonstrated that the liposomal membranes must contain pbb (SPC:Cho:pbb, 26:7:1) as a component to protect them from aggregation and/or fusion. The membranes containing pbb maintained the same turbidity (100%) after incubation with modified venom, in contrast with pbb-free membranes that showed a 15%?size decrease. This size decrease was interpreted as membrane degradation and was corroborated by a 50%?rhodamine leak-out. Another fact that confirmed our interpretation was the observed 100%?inhibition of the hemolytic activity after venom modification with pbb and NBS (S-A-BV). When S-A-BV interacted with liposomes, other protein conformational changes were observed and characterized by the increase of 1.93%?on S-A-BV α-helical content and the presence of tryptophan residues in a more hydrophobic environment. In other words, the S-A-BV interacted with liposomal membranes, but this interaction was not effective to cause aggregation, leak-out, or fusion. A stable formulation composed by S-A-BV encapsulated within liposomes composed by SPC:Cho:pbb, at a ratio of 26:7:1, was devised. Large unilamellar vesicles of 202.5 nm with a negative surface charge (–24.29 mV) encapsulated 95%?of S-A-BV. This formulation can, now, be assayed on VIT.     

蜂毒过敏原磷脂酶A2

蜂毒 (ｂｅｅｖｅｎｏｍ)是由工蜂毒腺和副腺分泌的、具有芳香气味的一种透明液体 ,贮藏在毒囊中 ,在蜜蜂蛰刺时由蛰针排出[1] 。蜂毒具有抗菌、消炎、镇痛、降血压、抗辐射、预防癌症等药理作用 ,可用于治疗风湿性关节炎、类风湿性关节炎、哮喘、神经痛等多种疑难杂症。目前世界上许多国家都已开展蜂针疗法 ,并有各种类型的蜂毒软膏和针剂生产。但由于蜂毒易使人产生过敏反应 ,致使蜂针疗法不能得到广泛推广。鉴于这一点 ,国内外许多学者对主要引起人类过敏的蜂毒组分———磷脂酶Ａ2 (ｐｈｏｓｐｈｏｌｉｐａｓｅＡ2 )进行了研究 ,并且取…     

Melittin, a Component of Bee Venom, Activates Unfertilized Sea Urchin Eggs

Melittin, which is known to stimulate phospholipase A , in many cells, caused as much elevation of fertilization membranes and increase in respiration of unfertilized eggs of the sea urchins Anthocidaris crassispina and Hemicentrotus pulcherrimus as normal fertilization.
In melittin-activated eggs, amino acid transport was decreased to less than that of unfertilized eggs, nucleoside transport was only slightly, activated, protein synthesis was rather inhibited and neither DNA synthesis nor cleavage was observed. It is concluded that although melittin induces the cortical reaction and activation of respiration in unfertilized eggs, its cytotoxicity prevents any "late changes".     

Bee Venom and Its Component Apamin as Neuroprotective Agents in a Parkinson Disease Mouse Model

Bee venom has recently been suggested to possess beneficial effects in the treatment of Parkinson disease (PD). For instance, it has been observed that bilateral acupoint stimulation of lower hind limbs with bee venom was protective in the acute 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model of PD. In particular, a specific component of bee venom, apamin, has previously been shown to have protective effects on dopaminergic neurons in vitro. However, no information regarding a potential protective action of apamin in animal models of PD is available to date. The specific goals of the present study were to (i) establish that the protective effect of bee venom for dopaminergic neurons is not restricted to acupoint stimulation, but can also be observed using a more conventional mode of administration and to (ii) demonstrate that apamin can mimic the protective effects of a bee venom treatment on dopaminergic neurons. Using the chronic mouse model of MPTP/probenecid, we show that bee venom provides sustained protection in an animal model that mimics the chronic degenerative process of PD. Apamin, however, reproduced these protective effects only partially, suggesting that other components of bee venom enhance the protective action of the peptide.     

Wanted: entrepreneurial parasitologists

At the recent World Water '86 conference in London, the Director of Water Supply and Urban Development in the Operations Policy Department of the World Bank chastised the assembled engineers for behaving like religious fanatics, convinced that their work is good and righteous but not willing (or able) to demonstrate this. There must be an opening here for an entrepreneurial parasitologist.     

Determination of the Effects of Bee Venom on Triple Negative Breast Cancer Cells in Vitro

Honeybees provide multiple products such as bee venom (BV) which are used for various nutritional and medicinal purposes. BV has received great attention due to its wide range of bioactive components with potential anti-cancer effects on different cancers. Triple negative breast cancer (TNBC) is defined as an aggressive type of breast cancer and new therapeutic targets are required for its treatment. In the current literature information is varied about the composition and quantity of BV bioactive compounds as well as the origin of BV and its significance. In this context, the cytotoxic and apoptotic effects of BV with a higher rate of mellitin from Apis mellifera anatoliaca (Muğla ecotype) on MDA-MB-231 cells was evaluated, in vitro. The cytotoxic, apoptotic and morphological effects of BV were determined by WST-1, Annexin V, cell cycle analysis and Acridine Orange staining. The results showed that BV caused apoptotic cell death in TNBC cells at a lower dose (0.47 μg/mL, p<0.01). This study suggests that BV could be developed as a potential therapeutic agent for cancer treatment. However, the mechanism of BV-induced apoptosis death should be clarified at the molecular level.     

Bee Venom Ameliorates Cognitive Dysfunction Caused by Neuroinflammation in an Animal Model of Vascular Dementia

Vascular dementia (VaD) is caused by the reduction of blood supply by vessel occlusion and is characterized by progressive cognitive decline. VaD incidence has been growing due to the aging population, placing greater strain on social and economic resources. However, the pathological mechanisms underlying VaD remain unclear. Many studies have used the bilateral common carotid artery occlusion (BCCAO) animal model to investigate potential therapeutics for VaD. In this study, we investigated whether bee venom (BV) improves cognitive function and reduces neuroinflammation in the hippocampus of BCCAO animals. Animals were randomly divided into three groups: a sham group (n = 15), BCCAO control group (n = 15), and BV-treated BCCAO group (n = 15). BCCAO animals were treated with 0.1 μg/g BV at ST36 (“Joksamli” acupoint) four times every other day. In order to investigate the effect of BV treatment on cognitive function, we performed a Y-maze test. In order to uncover any potential relationship between these results and neuroinflammation, we also performed Western blotting in the BCCAO group. Animals that had been treated with BV showed an improved cognitive function and a reduced expression of neuroinflammatory proteins in the hippocampus, including Iba-1, TLR4, CD14, and TNF-α. Furthermore, we demonstrated that BV treatment increased pERK and BDNF in the hippocampus. The present study thus underlines the neuroprotective effect of BV treatment against BCCAO-induced cognitive impairment and neuroinflammation. Our findings suggest that BV may be an effective complementary treatment for VaD, as it may improve cognitive function and attenuate neuroinflammation associated with dementia.