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Search Results (211)

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27 pages, 4952 KiB  
Review
Green Innovation and Synthesis of Honeybee Products-Mediated Nanoparticles: Potential Approaches and Wide Applications
by Shaden A. M. Khalifa, Aya A. Shetaia, Nehal Eid, Aida A. Abd El-Wahed, Tariq Z. Abolibda, Abdelfatteh El Omri, Qiang Yu, Mohamed A. Shenashen, Hidayat Hussain, Mohamed F. Salem, Zhiming Guo, Abdulaziz M. Alanazi and Hesham R. El-Seedi
Bioengineering 2024, 11(8), 829; https://doi.org/10.3390/bioengineering11080829 - 14 Aug 2024
Viewed by 84
Abstract
Bee products, abundant in bioactive ingredients, have been utilized in both traditional and contemporary medicine. Their antioxidant, antimicrobial, and anti-inflammatory properties make them valuable for food, preservation, and cosmetics applications. Honeybees are a vast reservoir of potentially beneficial products such as honey, bee [...] Read more.
Bee products, abundant in bioactive ingredients, have been utilized in both traditional and contemporary medicine. Their antioxidant, antimicrobial, and anti-inflammatory properties make them valuable for food, preservation, and cosmetics applications. Honeybees are a vast reservoir of potentially beneficial products such as honey, bee pollen, bee bread, beeswax, bee venom, and royal jelly. These products are rich in metabolites vital to human health, including proteins, amino acids, peptides, enzymes, sugars, vitamins, polyphenols, flavonoids, and minerals. The advancement of nanotechnology has led to a continuous search for new natural sources that can facilitate the easy, low-cost, and eco-friendly synthesis of nanomaterials. Nanoparticles (NPs) are actively synthesized using honeybee products, which serve dual purposes in preventive and interceptive treatment strategies due to their richness in essential metabolites. This review aims to highlight the potential role of bee products in this line and their applications as catalysts and food preservatives and to point out their anticancer, antibacterial, antifungal, and antioxidant underlying impacts. The research used several online databases, namely Google Scholar, Science Direct, and Sci Finder. The overall findings suggest that these bee-derived substances exhibit remarkable properties, making them promising candidates for the economical and eco-friendly production of NPs. Full article
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<p>Flowchart of the systematic literature review strategy. n: Number of published papers.</p>
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<p>Deciphering the role of bee product metabolites in eco-friendly nanoparticle synthesis.</p>
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<p>Honey mediated nanoparticles and their possible application (figure used with permission) [<a href="#B31-bioengineering-11-00829" class="html-bibr">31</a>].</p>
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<p>Diagrammatic illustration of the eco-friendly production of AgNPs using royal jelly (<b>A</b>), and the possible antibacterial action mechanism of AgNPs against both Gram-negative and Gram-positive bacteria (<b>B</b>) (Figure used with permission) [<a href="#B97-bioengineering-11-00829" class="html-bibr">97</a>].</p>
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<p>(<b>A</b>): Synthesis of AgNPs using <span class="html-italic">Cupressus sempervirens</span> pollen extract [<a href="#B55-bioengineering-11-00829" class="html-bibr">55</a>] and (<b>B</b>): ZnO synthesis using royal jelly (figure used with permission) [<a href="#B98-bioengineering-11-00829" class="html-bibr">98</a>].</p>
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<p>(<b>A</b>) The overall diagrammatic preparation of CuNPs derived from Honey, and (<b>B</b>) illustrates the anti-cancer properties of the synthesized CuNPs (Figure used with permission) [<a href="#B114-bioengineering-11-00829" class="html-bibr">114</a>].</p>
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<p>Highlighting the diverse biological impacts of nanoparticles derived from honeybee products.</p>
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17 pages, 2640 KiB  
Review
Bee-Inspired Healing: Apitherapy in Veterinary Medicine for Maintenance and Improvement Animal Health and Well-Being
by Jevrosima Stevanović, Uroš Glavinić, Marko Ristanić, Vladimira Erjavec, Barış Denk, Slobodan Dolašević and Zoran Stanimirović
Pharmaceuticals 2024, 17(8), 1050; https://doi.org/10.3390/ph17081050 - 9 Aug 2024
Viewed by 389
Abstract
This review aims to present current knowledge on the effects of honey bee products on animals based on in vivo studies, focusing on their application in clinical veterinary practice. Honey’s best-proven effectiveness is in treating wounds, including those infected with antibiotic-resistant microorganisms, as [...] Read more.
This review aims to present current knowledge on the effects of honey bee products on animals based on in vivo studies, focusing on their application in clinical veterinary practice. Honey’s best-proven effectiveness is in treating wounds, including those infected with antibiotic-resistant microorganisms, as evidenced in horses, cats, dogs, mice, and rats. Propolis manifested a healing effect in numerous inflammatory and painful conditions in mice, rats, dogs, and pigs and also helped in oncological cases in mice and rats. Bee venom is best known for its effectiveness in treating neuropathy and arthritis, as shown in dogs, mice, and rats. Besides, bee venom improved reproductive performance, immune response, and general health in rabbits, chickens, and pigs. Pollen was effective in stimulating growth and improving intestinal microflora in chickens. Royal jelly might be used in the management of animal reproduction due to its efficiency in improving fertility, as shown in rats, rabbits, and mice. Drone larvae are primarily valued for their androgenic effects and stimulation of reproductive function, as evidenced in sheep, chickens, pigs, and rats. Further research is warranted to determine the dose and method of application of honey bee products in animals. Full article
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<p>A part of the leg of a 1-year-old female was cat cut off by a lawnmower. The wound healed completely by the second intention with the use of medical honey. The sterile syringe was used as a scale for later wound size assessment. The leg on the day of presentation (<b>a</b>), 7 days (<b>b</b>), 11 days (<b>c</b>), 19 days (<b>d</b>), 24 days (<b>e</b>), and 30 days later (<b>f</b>). Medical grade honey was applied to the wound daily for the first 10 days and thereafter every 2–3 days (Original photo, V. Erjavec).</p>
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<p>Dehiscence after unilateral mastectomy in a 12-year-old bitch: (<b>a</b>) on the day of presentation; (<b>b</b>) two days after presentation; (<b>c</b>) four days after presentation; (<b>d</b>) 14 days after presentation; (<b>e</b>) 21 days after presentation; (<b>f</b>) two months after presentation. Medical-grade honey was applied to the wound daily by the owner (Original photo, V. Erjavec).</p>
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<p>Complete skin loss across 100% of an 8-year-old female cat’s forelimb (<b>a</b>) healed completely with minimal scarring (<b>b</b>) using medical honey [<a href="#B6-pharmaceuticals-17-01050" class="html-bibr">6</a>]. (Original photo, V. Erjavec).</p>
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<p>An extensive wound on a cow’s rare leg was successfully healed with raw honey, not medical-grade honey. An infected wound with necrotic material and surrounding erythema on the first day of treatment (<b>a</b>), wound covered with some necrotic tissue and pus 14 days later (<b>b</b>), the wound is almost clean, has contracted, is covered with granulation tissue (<b>c</b>) and is epithelializing from the margins (<b>d</b>) 1.5 and 2.5 months later (Original photos donated by Clinic of ruminants, Veterinary Faculty, University of Ljubljana).</p>
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17 pages, 2027 KiB  
Review
Genetic Deficiencies of Hyaluronan Degradation
by Stephen P. Fink and Barbara Triggs-Raine
Cells 2024, 13(14), 1203; https://doi.org/10.3390/cells13141203 - 16 Jul 2024
Viewed by 462
Abstract
Hyaluronan (HA) is a large polysaccharide that is broadly distributed and highly abundant in the soft connective tissues and embryos of vertebrates. The constitutive turnover of HA is very high, estimated at 5 g per day in an average (70 kg) adult human, [...] Read more.
Hyaluronan (HA) is a large polysaccharide that is broadly distributed and highly abundant in the soft connective tissues and embryos of vertebrates. The constitutive turnover of HA is very high, estimated at 5 g per day in an average (70 kg) adult human, but HA turnover must also be tightly regulated in some processes. Six genes encoding homologues to bee venom hyaluronidase (HYAL1, HYAL2, HYAL3, HYAL4, HYAL6P/HYALP1, SPAM1/PH20), as well as genes encoding two unrelated G8-domain-containing proteins demonstrated to be involved in HA degradation (CEMIP/KIAA1199, CEMIP2/TMEM2), have been identified in humans. Of these, only deficiencies in HYAL1, HYAL2, HYAL3 and CEMIP have been identified as the cause or putative cause of human genetic disorders. The phenotypes of these disorders have been vital in determining the biological roles of these enzymes but there is much that is still not understood. Deficiencies in these HA-degrading proteins have been created in mice and/or other model organisms where phenotypes could be analyzed and probed to expand our understanding of HA degradation and function. This review will describe what has been found in human and animal models of hyaluronidase deficiency and discuss how this has advanced our understanding of HA’s role in health and disease. Full article
(This article belongs to the Special Issue Role of Hyaluronan in Human Health and Disease)
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<p>Organization of genes encoding HA degrading proteins in humans and mice. The organization and sizes of the human and mouse genes are based on assemblies NC_000003.12 and NC_000007.14 (human) or NC_00075.7 and NC_000072.7 (mouse). The sizes of the genes are shown above the gene in kilobase pairs. The scaling is unique for each map to allow all hyaluronidase genes in a chromosomal region to be displayed.</p>
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<p>Expression of genes encoding HA-degrading proteins in human tissues. Estimates of gene expression were determined by RNA-Seq in a previous study [<a href="#B51-cells-13-01203" class="html-bibr">51</a>]. Mean ± standard deviation is shown for the major tissues where RNA-Seq data were available from at least 3 different biological replicates. Results are graphed for <span class="html-italic">HYAL1</span>, <span class="html-italic">HYAL2</span>, <span class="html-italic">HYAL3</span>, <span class="html-italic">HYAL4</span>, <span class="html-italic">SPAM1</span>, <span class="html-italic">CEMIP1</span> and <span class="html-italic">CEMIP2</span> using GraphPad Software Version 10 (Boston, MA, USA). To facilitate viewing, some error bars are not fully graphed and are indicated by * (SD = 9.81), ** (SD = 1.84), and *** (SD = 21.8).</p>
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43 pages, 1100 KiB  
Review
Hive Products: Composition, Pharmacological Properties, and Therapeutic Applications
by Roberto Bava, Fabio Castagna, Carmine Lupia, Giusi Poerio, Giovanna Liguori, Renato Lombardi, Maria Diana Naturale, Rosa Maria Bulotta, Vito Biondi, Annamaria Passantino, Domenico Britti, Giancarlo Statti and Ernesto Palma
Pharmaceuticals 2024, 17(5), 646; https://doi.org/10.3390/ph17050646 - 16 May 2024
Cited by 1 | Viewed by 2362
Abstract
Beekeeping provides products with nutraceutical and pharmaceutical characteristics. These products are characterized by abundance of bioactive compounds. For different reasons, honey, royal jelly, propolis, venom, and pollen are beneficial to humans and animals and could be used as therapeutics. The pharmacological action of [...] Read more.
Beekeeping provides products with nutraceutical and pharmaceutical characteristics. These products are characterized by abundance of bioactive compounds. For different reasons, honey, royal jelly, propolis, venom, and pollen are beneficial to humans and animals and could be used as therapeutics. The pharmacological action of these products is related to many of their constituents. The main bioactive components of honey include oligosaccharides, methylglyoxal, royal jelly proteins (MRJPs), and phenolics compounds. Royal jelly contains jelleins, royalisin peptides, MRJPs, and derivatives of hydroxy-decenoic acid, particularly 10-hydroxy-2-decenoic acid (10-HDA), which possess antibacterial, anti-inflammatory, immunomodulatory, neuromodulatory, metabolic syndrome-preventing, and anti-aging properties. Propolis has a plethora of activities that are referable to compounds such as caffeic acid phenethyl ester. Peptides found in bee venom include phospholipase A2, apamin, and melittin. In addition to being vitamin-rich, bee pollen also includes unsaturated fatty acids, sterols, and phenolics compounds that express antiatherosclerotic, antidiabetic, and anti-inflammatory properties. Therefore, the constituents of hive products are particular and different. All of these constituents have been investigated for their properties in numerous research studies. This review aims to provide a thorough screening of the bioactive chemicals found in honeybee products and their beneficial biological effects. The manuscript may provide impetus to the branch of unconventional medicine that goes by the name of apitherapy. Full article
(This article belongs to the Special Issue Therapeutic Effects of Natural Products and Their Clinical Research)
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<p>Some general actions of hive products and anatomical sites where they exert their effects.</p>
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25 pages, 4195 KiB  
Article
Bee Venom-Loaded Niosomes as Innovative Platforms for Cancer Treatment: Development and Therapeutical Efficacy and Safety Evaluation
by Maria Beatriz Pinto, Patrícia C. Pires, Ricardo C. Calhelha, Ana Rita Silva, Maria João Sousa, Miguel Vilas-Boas, Soraia I. Falcão, Francisco Veiga, Pooyan Makvandi and Ana Cláudia Paiva-Santos
Pharmaceuticals 2024, 17(5), 572; https://doi.org/10.3390/ph17050572 - 29 Apr 2024
Viewed by 1262
Abstract
Despite past efforts towards therapeutical innovation, cancer remains a highly incident and lethal disease, with current treatments lacking efficiency and leading to severe side effects. Hence, it is imperative to develop new, more efficient, and safer therapies. Bee venom has proven to have [...] Read more.
Despite past efforts towards therapeutical innovation, cancer remains a highly incident and lethal disease, with current treatments lacking efficiency and leading to severe side effects. Hence, it is imperative to develop new, more efficient, and safer therapies. Bee venom has proven to have multiple and synergistic bioactivities, including antitumor effects. Nevertheless, some toxic effects have been associated with its administration. To tackle these issues, in this work, bee venom-loaded niosomes were developed, for cancer treatment. The vesicles had a small (150 nm) and homogeneous (polydispersity index of 0.162) particle size, and revealed good therapeutic efficacy in in vitro gastric, colorectal, breast, lung, and cervical cancer models (inhibitory concentrations between 12.37 ng/mL and 14.72 ng/mL). Additionally, they also revealed substantial anti-inflammatory activity (inhibitory concentration of 28.98 ng/mL), effects complementary to direct antitumor activity. Niosome safety was also assessed, both in vitro (skin, liver, and kidney cells) and ex vivo (hen’s egg chorioallantoic membrane), and results showed that compound encapsulation increased its safety. Hence, small, and homogeneous bee venom-loaded niosomes were successfully developed, with substantial anticancer and anti-inflammatory effects, making them potentially promising primary or adjuvant cancer therapies. Future research should focus on evaluating the potential of the developed platform in in vivo models. Full article
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<p>Schematic representation of beehive derived compounds, including a photograph of the beehive at Polytechnic Institute of Bragança’s apiary, Bragança, Portugal (produced with Biorender, melittin, phospholipase and apamin molecular structures originated from PubChem).</p>
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<p>Example chromatogram after UHPLC analysis of the bee venom sample, at 220 nm. IS—internal standard (cytochrome C, 25 µg/mL); UHPLC—ultra-high-performance liquid chromatography.</p>
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<p>Particle size and PDI values of the developed niosomes (formulation vehicle), at all tested temperatures, and with a varying number of performed extrusion cycles; data is presented as mean ± standard deviation; **** <span class="html-italic">p</span> &lt; 0.0001 and corresponds to the comparison of no extrusion with all extrusion cycles (R<sup>2</sup> 0.9991, one-way ANOVA with Tukey’s multiple comparisons test); PDI—polydispersity index.</p>
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<p>Cytotoxic potential of the developed bee venom-loaded niosomes, compared to the empty niosomes (formulation vehicle) and bee venom solution, in several different cancer cell lines; GI<sub>50</sub> corresponds to the formulation concentration required to inhibit cell growth by 50%; data is represented as mean ± standard deviation; **** <span class="html-italic">p</span> &lt; 0.0001 (R<sup>2</sup> 0.9991, one-way ANOVA with Tukey’s multiple comparisons test); AGS—gastric adenocarcinoma cell line; Caco-2—colorectal adenocarcinoma cell line; HeLa—cervical carcinoma cell line; MCF-7—breast adenocarcinoma cell line; NCI-H460—lung carcinoma cell line; PLP2—primary pig liver culture cell line; and Vero—African green monkey kidney cell line.</p>
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<p>Anti-inflammatory potential of the developed bee venom-loaded niosomes (BVN), compared to the free compound (bee venom solution, BVS), and the formulation vehicle (empty niosomes, EN), evaluated in a mouse macrophage cell line (RAW 264.7); IC<sub>50</sub> values are depicted, and correspond to formulation concentrations providing 50% of inhibition of nitric oxide production; data is represented as mean ± standard deviation; **** <span class="html-italic">p</span> &lt; 0.0001 (schematic representation produced with Biorender).</p>
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<p>Cell viability percentage (%) variation with increasing applied formulation concentrations (µg/mL), on HFF-1 cell line (left bar graphs, in blue), and HaCaT cell line (right bar graphs, in pink), including the developed bee venom loaded niosomes (BVN), formulation vehicle (empty niosomes, EN), and compound solution (bee venom solution, BVS); data is represented as mean ± standard deviation; HaCaT—skin keratinocytes cell line; HFF-1—skin fibroblasts cell line.</p>
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<p>HET-CAM assay schematic representation, and photographs of the hen’s egg chorioallantoic membranes after formulation application, either the developed bee venom-encapsulated niosomes (<b>a</b>), or the formulation vehicle (empty niosomes) (<b>b</b>); HET-CAM—hen’s egg chorioallantoic membrane.</p>
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<p>Schematic illustration of niosome composition and production, using the thin-film hydration method, followed by extrusion through a nanometric pore membrane (produced with Biorender).</p>
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13 pages, 2671 KiB  
Article
Antioxidant Activity and Mechanism of Action of Amwaprin: A Protein in Honeybee (Apis mellifera) Venom
by Bo-Yeon Kim, Kwang-Sik Lee and Byung-Rae Jin
Antioxidants 2024, 13(4), 469; https://doi.org/10.3390/antiox13040469 - 17 Apr 2024
Viewed by 911
Abstract
Bee venom contains several bioactive components, including enzymatic and non-enzymatic proteins. There is increasing interest in the bioactive components of bee venom since they have exhibited various pharmacological effects. Recently, Apis mellifera waprin (Amwaprin) was identified as a novel protein in Apis mellifera [...] Read more.
Bee venom contains several bioactive components, including enzymatic and non-enzymatic proteins. There is increasing interest in the bioactive components of bee venom since they have exhibited various pharmacological effects. Recently, Apis mellifera waprin (Amwaprin) was identified as a novel protein in Apis mellifera (honeybee) venom and characterized as an antimicrobial agent. Herein, the novel biological function of Amwaprin as an antioxidant is described. In addition, the antioxidant effects of Amwaprin in mammalian cells were investigated. Amwaprin inhibited the growth of, oxidative stress-induced cytotoxicity, and inflammatory response in mammalian NIH-3T3 cells. Amwaprin decreased caspase-3 activity during oxidative stress and exhibited protective activity against oxidative stress-induced cell apoptosis in NIH-3T3 and insect Sf9 cells. The mechanism underlying the cell protective effect of Amwaprin against oxidative stress is due to its direct binding to the cell membrane. Furthermore, Amwaprin demonstrated radical-scavenging activity and protected against oxidative DNA damage. These results suggest that the antioxidant capacity of Amwaprin is attributed to the synergistic effects of its radical-scavenging action and cell shielding, indicating its novel role as an antioxidant agent. Full article
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<p>Schematic alignment of three waprins from the venom of lower vertebrates and invertebrates. The predicted signal sequence is boxed in white. The relative positions of the WAP domain (yellow) in the three waprins are shown. The conserved cysteine residues within the WAP domain are indicated by arrowheads. The aligned waprins include Amwaprin [<a href="#B18-antioxidants-13-00469" class="html-bibr">18</a>], <span class="html-italic">Oxyuranus microlepidotus</span> waprin [<a href="#B22-antioxidants-13-00469" class="html-bibr">22</a>], and <span class="html-italic">Ceratophrys calcarata</span> waprin [<a href="#B24-antioxidants-13-00469" class="html-bibr">24</a>].</p>
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<p>Effect of Amwaprin on the growth of mammalian cells. NIH-3T3 cells were treated with Triton X-100 (negative control) or various concentrations of Amwaprin for 24 or 48 h. Cellular growth was determined as a percentage relative to untreated cells (control). Data are represented as the mean ± SD (<span class="html-italic">n</span> = 3). Different letters indicate significant differences among the treatments (<span class="html-italic">p</span> = 0.0001).</p>
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<p>Cellular proliferation and ROS production in NIH-3T3 cells treated with Amwaprin. NIH-3T3 cells were treated with or without Amwaprin for five days, and cellular proliferation and ROS production were determined. Data are represented as the mean ± SD (<span class="html-italic">n</span> = 3). (<b>A</b>) Cells were counted for five days. **, <span class="html-italic">p</span> &lt; 0.01 and ***, <span class="html-italic">p</span> &lt; 0.001. (<b>B</b>) ROS production was measured for five days. Data are presented as ROS concentrations. **, <span class="html-italic">p</span> &lt; 0.01 and ***, <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Effect of Amwaprin on cell viability and caspase-3 activity in NIH-3T3 cells. NIH-3T3 cells were treated with (+) or without (−) Amwaprin or H<sub>2</sub>O<sub>2</sub> for 24 or 48 h. Cell viability (<b>A</b>) and caspase-3 activity (<b>B</b>) were determined as a percentage relative to the untreated cells (control). Data are represented as the mean ± SD (<span class="html-italic">n</span> = 3). Different letters indicate significant differences among the treatments (<span class="html-italic">p</span> = 0.0001).</p>
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<p>Effect of Amwaprin on the secretion of proinflammatory mediators and cytokines in NIH-3T3 cells. NIH-3T3 cells were treated with (+) or without (−) Amwaprin or H<sub>2</sub>O<sub>2</sub> for 24 or 48 h. The concentrations of IL-1β (<b>A</b>), IL-6 (<b>B</b>), and TNF-α (<b>C</b>) were determined. Data are represented as the mean ± SD (<span class="html-italic">n</span> = 3). Different letters indicate significant differences among treatments (<span class="html-italic">p</span> = 0.0001).</p>
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<p>Cell protective effect of Amwaprin against H<sub>2</sub>O<sub>2</sub>-induced cell apoptosis. NIH-3T3 (<b>A</b>) and Sf9 (<b>B</b>) cells were treated with (+) or without (−) Amwaprin or H<sub>2</sub>O<sub>2</sub>. Immunofluorescence staining was performed for the detection of apoptosis (green) and Amwaprin (red) in the cells. Representative confocal images are displayed. Merged confocal images are also included (merge). Scale bar: 10 μm.</p>
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<p>Cell protective effect of Amwaprin against H<sub>2</sub>O<sub>2</sub>-induced cell apoptosis. NIH-3T3 (<b>A</b>) and Sf9 (<b>B</b>) cells were treated with (+) or without (−) Amwaprin or H<sub>2</sub>O<sub>2</sub>. Immunofluorescence staining was performed for the detection of apoptosis (green) and Amwaprin (red) in the cells. Representative confocal images are displayed. Merged confocal images are also included (merge). Scale bar: 10 μm.</p>
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<p>Radical-scavenging activity of Amwaprin. (<b>A</b>) DPPH radical-scavenging activity of Amwaprin. Data are represented as the mean ± SD (<span class="html-italic">n</span> = 3). Different letters indicate significant differences among treatments (<span class="html-italic">p</span> = 0.0001). (<b>B</b>) Protection of DNA cleavage by Amwaprin. Lane 1, pUC18 only; lane 2, pUC18 with DTT only; lane 3, pUC18 with FeCl<sub>3</sub> only; lane 4, pUC18 with the MCO system; lanes 5–7, pUC18 with recombinant Amwaprin (50, 250, or 500 ng) in the MCO system. The nicked form (NF) and super-coiled form (SF) of the plasmid DNA are indicated by arrows.</p>
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26 pages, 2886 KiB  
Review
Natural Sources of Therapeutic Agents Used in Skin Conditions
by Monica Dinu, Alin Laurențiu Tatu, Dorin Ioan Cocoș, Lawrence Chukwudi Nwabudike, Ana Maria Chirilov, Claudia Simona Stefan, Kamel Earar and Olimpia Dumitriu Buzia
Life 2024, 14(4), 492; https://doi.org/10.3390/life14040492 - 10 Apr 2024
Cited by 3 | Viewed by 1702
Abstract
Skin conditions are numerous and often have a major impact on patients’ quality of life, and effective and safe treatment is very important. The conventional drugs used for skin diseases are usually corticosteroids and antimicrobial products that can induce various side effects, especially [...] Read more.
Skin conditions are numerous and often have a major impact on patients’ quality of life, and effective and safe treatment is very important. The conventional drugs used for skin diseases are usually corticosteroids and antimicrobial products that can induce various side effects, especially with long-term use, which is why researchers are studying alternatives, especially biologically active natural products. Three products caught our attention: bee venom (BV), due to reported experimental results showing anti-inflammatory, antibacterial, antiviral, antioxidant, antimycotic, and anticancer effects, Ficus carica (FC) due to its demonstrated antioxidant, antibacterial, and anti-inflammatory action, and finally Geranium essential oil (GEO), with proven antifungal, antibacterial, anti-inflammatory, and antioxidant effects. Following a review of the literature, we produced this paper, which presents a review of the potential therapeutic applications of the three products in combating various skin conditions and for skin care, because BV, FC, and GEO have common pharmacological actions (anti-inflammatory, antibacterial, and antioxidant). We also focused on studying the safety of the topical use of BV, FC, and GEO, and new approaches to this. This paper presents the use of these natural therapeutic agents to treat patients with conditions such as vitiligo, melasma, and melanoma, as well as their use in treating dermatological conditions in patients with diabetes. Full article
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<p>Skin diseases in which the therapeutic application of BV has been studied.</p>
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<p>Skin diseases in which the therapeutic application of FC has been studied.</p>
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<p>Skin diseases in which the therapeutic application of GEO has been studied.</p>
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<p>The chemical structure of the most important constituents of GEO (<b>a</b>–<b>d</b>) and FC (<b>b</b>–<b>f</b>). Common constituents (<b>b</b>–<b>d</b>).</p>
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20 pages, 2055 KiB  
Review
Bee Venom: Composition and Anticancer Properties
by Goran Gajski, Elina Leonova and Nikolajs Sjakste
Toxins 2024, 16(3), 117; https://doi.org/10.3390/toxins16030117 - 29 Feb 2024
Cited by 2 | Viewed by 4993
Abstract
Among the various natural compounds used in alternative and Oriental medicine, toxins isolated from different organisms have had their application for many years, and Apis mellifera venom has been studied the most extensively. Numerous studies dealing with the positive assets of bee venom [...] Read more.
Among the various natural compounds used in alternative and Oriental medicine, toxins isolated from different organisms have had their application for many years, and Apis mellifera venom has been studied the most extensively. Numerous studies dealing with the positive assets of bee venom (BV) indicated its beneficial properties. The usage of bee products to prevent the occurrence of diseases and for their treatment is often referred to as apitherapy and is based mainly on the experience of the traditional system of medical practice in diverse ethnic communities. Today, a large number of studies are focused on the antitumor effects of BV, which are mainly attributed to its basic polypeptide melittin (MEL). Previous studies have indicated that BV and its major constituent MEL cause a strong toxic effect on different cancer cells, such as liver, lung, bladder, kidney, prostate, breast, and leukemia cells, while a less pronounced effect was observed in normal non-target cells. Their proposed mechanisms of action, such as the effect on proliferation and growth inhibition, cell cycle alterations, and induction of cell death through several cancer cell death mechanisms, are associated with the activation of phospholipase A2 (PLA2), caspases, and matrix metalloproteinases that destroy cancer cells. Numerous cellular effects of BV and MEL need to be elucidated on the molecular level, while the key issue has to do with the trigger of the apoptotic cascade. Apoptosis could be either a consequence of the plasmatic membrane fenestration or the result of the direct interaction of the BV components with pro-apoptotic and anti-apoptotic factors. The interaction of BV peptides and enzymes with the plasma membrane is a crucial step in the whole process. However, before its possible application as a remedy, it is crucial to identify the correct route of exposure and dosage of BV and MEL for potential therapeutic use as well as potential side effects on normal cells and tissues to avoid any possible adverse event. Full article
(This article belongs to the Section Animal Venoms)
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<p>European honey bee <span class="html-italic">Apis mellifera</span> (<b>A</b>) and lyophilized bee venom (<b>B</b>).</p>
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<p>Composition of dry bee venom.</p>
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<p>Structure of melittin (Gly-Ile-Gly-Ala-Val-Leu-Lys-Val-Leu-Thr-Thr-Gly-Leu-Pro-Ala-Leu-Ile-Ser-Trp-Ile-Lys-Arg-Lys-Arg-Gln-Gln). National Center for Biotechnology Information. PubChem Compound Summary for CID 16129627, Melitten. <a href="https://pubchem.ncbi.nlm.nih.gov/compound/Melitten" target="_blank">https://pubchem.ncbi.nlm.nih.gov/compound/Melitten</a> (Accessed on 15 January 2024).</p>
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<p>Structure of apamin (Cys-Tyr-Cys-Lys-Ala-Pro-Glu-Thr-Ala-Leu-Cys-Ala-Arg-Arg-Cys-Gln-Gln-His). National Center for Biotechnology Information. PubChem Compound Summary for CID 44134548, Apamine. <a href="https://pubchem.ncbi.nlm.nih.gov/compound/Apamine" target="_blank">https://pubchem.ncbi.nlm.nih.gov/compound/Apamine</a> (Accessed on 15 January 2024).</p>
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<p>Structure of mast cell degranulating (MCD) peptide (Ile-Lys-Cys-Asn-Cys-Lys-Arg-His-Val-Ile-Lys-Pro-His-Ile-Cys-Arg-Lys-Ile-Cys-Gly-Lys-Asn). National Center for Biotechnology Information. PubChem Compound Summary for CID 16132290, Mast cell degranulating peptide. <a href="https://pubchem.ncbi.nlm.nih.gov/compound/Mast-cell-degranulating-peptide" target="_blank">https://pubchem.ncbi.nlm.nih.gov/compound/Mast-cell-degranulating-peptide</a> (Accessed on 15 January 2024).</p>
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<p>Simplified presentation of the mechanisms of melittin action. Melittin is presented as small helices. The peptide makes pores in the plasmatic membrane and destroys some of the membrane receptors and enzymes. Inside cells, melittin damages mitochondria and binds calmodulin, resulting in apoptosis and impairment of signaling pathways.</p>
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13 pages, 3878 KiB  
Article
Bee Venom Stimulates Growth Factor Release from Adipose-Derived Stem Cells to Promote Hair Growth
by Jung Hyun Kim, Tae Yoon Kim, Bonhyuk Goo and Yeoncheol Park
Toxins 2024, 16(2), 84; https://doi.org/10.3390/toxins16020084 - 4 Feb 2024
Cited by 2 | Viewed by 2437
Abstract
Limited evidence suggests that stimulating adipose-derived stem cells (ASCs) indirectly promotes hair growth. We examined whether bee venom (BV) activated ASCs and whether BV-induced hair growth was facilitated by enhanced growth factor release by ASCs. The induction of the telogen-to-anagen phase was studied [...] Read more.
Limited evidence suggests that stimulating adipose-derived stem cells (ASCs) indirectly promotes hair growth. We examined whether bee venom (BV) activated ASCs and whether BV-induced hair growth was facilitated by enhanced growth factor release by ASCs. The induction of the telogen-to-anagen phase was studied in mice. The underlying mechanism was investigated using organ cultures of mouse vibrissa hair follicles. When BV-treated ASCs were injected subcutaneously into mice, the telogen-to-anagen transition was accelerated and, by day 14, the hair weight increased. Quantitative polymerase chain reaction (qPCR) revealed that BV influenced the expression of several molecules, including growth factors, chemokines, channels, transcription factors, and enzymes. Western blot analysis was employed to verify the protein expression levels of extracellular-signal-regulated kinase (ERK) and phospho-ERK. Both the Boyden chamber experiment and scratch assay confirmed the upregulation of cell migration by BV. Additionally, ASCs secreted higher levels of growth factors after exposure to BV. Following BV therapy, the gene expression levels of alkaline phosphatase (ALP), fibroblast growth factor (FGF)-1 and 6, endothelial cell growth factor, and platelet-derived growth factor (PDGF)-C were upregulated. The findings of this study suggest that bee venom can potentially be utilized as an ASC-preconditioning agent for hair regeneration. Full article
(This article belongs to the Section Animal Venoms)
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<p>Results (mean + SEM) of MTT (tetrazolium dye) assay to assess toxicity of bee venom to adipose stem cells (ASCs). N = 1 × 10<sup>5</sup> cells/well for each mean. MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide.</p>
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<p>(<b>A</b>) Changes in mouse hair at 14 days with adipose-derived stem cells (ASCs) and bee venom (BV). (<b>B</b>) Hair weight changes (mean + SEM) after ASC and ASC + BV pretreatment, N = 32 for each mean, ** <span class="html-italic">p</span> &lt; 0.01 for ASC only relative to control and ASC + BV relative to ASC only. (<b>C</b>) The effect of ASC and ASC + BV pretreatment demonstrated by HE staining. (<b>D</b>) The effect of ASC and ASC + BV pretreatment demonstrated by DAPI and Ki67+ staining. (<b>E</b>) The number of Ki67+ cells (mean + SEM) after pretreatment with ASC and ASC + BV, N = 32 for each mean, *** <span class="html-italic">p</span> &lt; 0.001 for ASC only relative to control and ASC + BV relative to ASC only.</p>
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<p>(<b>A</b>) Results (mean + SEM) of scratch assay with bee venom (BV), N = 31 for each mean, *** <span class="html-italic">p</span> &lt; 0.001 for BV 1 µg relative to control and BV 2 µg relative to BV 1 µg. (<b>B</b>) Results of Boyden chamber assay with BV. (<b>C</b>) The effect (mean + SEM) of BV on in vitro angiogenesis proliferation, N = 31 for each mean, *** <span class="html-italic">p</span> &lt; 0.001 for BV 1 µg relative to control and BV 2 µg relative to BV 1 µg.</p>
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<p>Results (mean + SEM) of qPCR in vivo, N = 31 for each mean; * <span class="html-italic">p</span> = 0.027, approaching significance after Bonferroni correction; *** <span class="html-italic">p</span> &lt; 0.001; ASC only is compared to control and ASC + BV is compared to ASC only. The adipose stem cell + bee venom (ASC + BV) group is statistically relevant to the ASC-only group.</p>
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<p>(<b>A</b>) Vibrissal hair follicle morphological changes after 1 and 14 days of bee venom (BV) treatment. (<b>B</b>) Follicle length changes (mean + SEM) after 1 and 14 days of BV treatment, N = 29 for each mean, *** <span class="html-italic">p</span> &lt; 0.001 relative to control (CON).</p>
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<p>(<b>A</b>) Western blot results of apical stem cell (ASC) and ASC + bee venom (ASC + BV), N = 32 for each mean. (<b>B</b>) Protein expression levels (mean + SEM) demonstrated with extracellular-signal-regulated kinase (ERK) and phosphorylated ERK (p-ERK) pathways, N = 29 for each mean, * <span class="html-italic">p</span> = 0.031, which approached significance after Bonferroni correction.</p>
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16 pages, 2194 KiB  
Article
Anti-Herpes Simplex Virus and Anti-Inflammatory Activities of the Melittin Peptides Derived from Apis mellifera and Apis florea Venom
by Pichet Praphawilai, Thida Kaewkod, Sureeporn Suriyaprom, Aussara Panya, Terd Disayathanoowat and Yingmanee Tragoolpua
Insects 2024, 15(2), 109; https://doi.org/10.3390/insects15020109 - 4 Feb 2024
Viewed by 1832
Abstract
Herpes simplex virus (HSV) is known to cause cold sores and various diseases in humans. Importantly, HSV infection can develop latent and recurrent infections, and it is also known to cause inflammation. These infections are difficult to control, and effective treatment of the [...] Read more.
Herpes simplex virus (HSV) is known to cause cold sores and various diseases in humans. Importantly, HSV infection can develop latent and recurrent infections, and it is also known to cause inflammation. These infections are difficult to control, and effective treatment of the disease remains a challenge. Thus, the search for new antiviral and anti-inflammatory agents is a necessity. Melittin is a major peptide that is present in the venom of the honeybee. It possesses a number of pharmacological properties. In this study, the effects of the melittin peptides from A. mellifera (MEL-AM) and A. florea (MEL-AF) against HSV-1 and HSV-2 were evaluated at different stages during the viral multiplication cycle in an attempt to define the mode of antiviral action using plaque reduction and virucidal assays. The results revealed a new finding that melittin at 5 µg/mL demonstrated the highest inhibitory effect on HSV through the direct inactivation of viral particles, and MEL-AF displayed a greater virucidal activity. Moreover, melittin was also observed to interfere with the process of HSV attachment to the host cells. MEL-AM exhibited anti-HSV-1 and anti-HSV-2 effects with EC50 values of 4.90 ± 0.15 and 4.39 ± 0.20 µg/mL, while MEL-AF demonstrated EC50 values of 4.47 ± 0.21 and 3.95 ± 0.61 µg/mL against HSV-1 and HSV-2, respectively. However, non-cytotoxic concentrations of both types of melittin produced only slight degrees of HSV-1 and HSV-2 inhibition after viral attachment, but melittin at 5 µg/mL was able to reduce the plaque size of HSV-2 when compared to the untreated group. In addition, MEL-AM and MEL-AF also exhibited anti-inflammatory activity via the inhibition of nitric oxide production in LPS-stimulated RAW 264.7 macrophage cells, and they were also found to down-regulate the expressions of the iNOS, COX-2 and IL-6 genes. The highest inhibition of IL-6 mRNA expression was found after treatment with 10 µg/mL of MEL-AM and MEL-AF. Therefore, melittin peptides have displayed strong potential to be used as an alternative treatment for HSV infection and inflammatory diseases in the future. Full article
(This article belongs to the Special Issue Bee Products: Status, Properties, Opportunities, and Challenges)
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<p>Effect of melittin on cell viability. The percentage of cell viability was measured using MTT assay. The percentage of Vero cell viability was measured after treatment with melittin at various concentrations for 72 h (<b>a</b>), as was the effect of melittin on RAW 264.7 cell viability after treatment for 24 h (<b>b</b>). Data are presented as the mean percentage of cell viability ± SD from three independent experiments. * Values were significantly different from the untreated group (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effect of melittin on HSVs during viral infection. Cells were infected with HSVs and treated with non-cytotoxic concentrations of melittin simultaneously. Relative amounts of plaque formation of HSV-1 (<b>a</b>) and HSV-2 (<b>b</b>) in the treated groups were compared to those of the untreated group. Data are presented as mean values of relative plaque formation (%) ± SD from three independent experiments. * Values were significantly different from the untreated group (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effect of melittin on HSVs after viral infection. Cells were infected with HSVs followed by being treated with non-cytotoxic concentrations of melittin. The relative amounts of plaque formation of HSV-1 (<b>a</b>) and HSV-2 (<b>b</b>) in the treated groups were compared with those of the untreated group and ACV positive drug control. The plaque size of HSV-2 was reduced after treatment with melittin at 5 µg/mL (<b>c</b>). Data are presented as mean values of relative plaque formation (%) ± SD from three independent experiments. * Values were significantly different from the untreated group (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Virucidal effects of melittin on infectivity of HSV-1 and HSV-2. The viral suspension was incubated with melittin at indicated concentrations at 4 °C for 30 min. The infectivity of HSVs was measured using plaque titration assay. HSV-1 and HSV-2 suspensions of 300 to 400 PFU/mL (<b>a</b>,<b>b</b>) and 3 × 10<sup>3</sup> to 4 × 10<sup>3</sup> PFU/mL (<b>c</b>,<b>d</b>) were treated with non-cytotoxic concentrations of melittin. The titers of HSV-1 and HSV-2 in the treated group were compared to those in the untreated group. Data are presented as mean values of vital titer ± SD from three independent experiments. * Values were significantly different from the untreated group (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Inhibitory effect of MEL-AM (<b>a</b>) and MEL-AF (<b>b</b>) at indicated concentrations (1.25–10 µg/mL) on nitric oxide (NO) production in LPS-stimulated RAW 264.7 cells. The cells with or without LPS stimulation were used as the positive control and negative control, respectively. Data are presented as mean ± SD values from three independent experiments. * Values were significantly different from the untreated group (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effect of melittin on the level of pro-inflammatory gene expression after cells were stimulated with LPS (1 µg/mL) and treated with non-cytotoxic concentrations of melittin for 3 h. Expressions of the inflammatory gene were measured using reverse transcription PCR. The relative mRNA expressions of inducible nitric oxide synthase (<span class="html-italic">iNOS</span>) (<b>a</b>) cyclooxygenase-2 (<span class="html-italic">COX-2</span>) (<b>b</b>) and interleukin 6 (<span class="html-italic">IL-6</span>) (<b>c</b>) were normalized to the <span class="html-italic">β-actin</span> internal control. The results are representative of those obtained from three independent experiments. * Values were significantly different from the untreated group (<span class="html-italic">p</span> &lt; 0.05).</p>
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19 pages, 5130 KiB  
Article
Discovery of Melittin as Triple-Action Agent: Broad-Spectrum Antibacterial, Anti-Biofilm, and Potential Anti-Quorum Sensing Activities
by Hongyan Yang, Rong Ma, Jiarou Chen, Qian Xie, Wenhui Luo, Pinghua Sun, Zheng Liu and Jialiang Guo
Molecules 2024, 29(3), 558; https://doi.org/10.3390/molecules29030558 - 23 Jan 2024
Cited by 2 | Viewed by 1587
Abstract
The development of antibiotic-resistant microorganisms is a major global health concern. Recently, there has been an increasing interest in antimicrobial peptides as a therapeutic option. This study aimed to evaluate the triple-action (broad-spectrum antibacterial, anti-biofilm, and anti-quorum sensing activities) of melittin, a membrane-active [...] Read more.
The development of antibiotic-resistant microorganisms is a major global health concern. Recently, there has been an increasing interest in antimicrobial peptides as a therapeutic option. This study aimed to evaluate the triple-action (broad-spectrum antibacterial, anti-biofilm, and anti-quorum sensing activities) of melittin, a membrane-active peptide present in bee venom. The minimum inhibitory concentration and minimum bactericidal concentration of the melittin were determined using the microdilution method and agar plate counting. Growth curve analysis revealed that melittin showed a concentration-dependent antibacterial activity. Scanning electron microscope analysis revealed that melittin treatment altered the morphology. Confocal laser scanning microscope revealed that melittin increased the membrane permeability and intracellular ROS generation in bacteria, all of which contribute to bacterial cell death. In addition, the crystal violet (CV) assay was used to test the anti-biofilm activity. The CV assay demonstrated that melittin inhibited biofilm formation and eradicated mature biofilms. Biofilm formation mediated by quorum sensing (QS) plays a major role in this regard, so molecular docking and molecular dynamics analysis confirmed that melittin interacts with LasR receptors through hydrogen bonds, and further evaluates the anti-QS activity of melittin through the production of virulence factors (pyocyanin, elastase, and rhamnolipid), exopolysaccharides secretion, and bacterial motility, that may be the key to inhibiting the biofilm formation mechanism. The present findings highlight the promising role of melittin as a broad-spectrum antibacterial, anti-biofilm agent, and potential QS inhibitor, providing a new perspective and theoretical basis for the development of alternative antibiotics. Full article
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<p>Bacterial growth curves at different melittin concentrations. Different concentrations of melittin inhibited the growth of <span class="html-italic">E. coli</span> ATCC 25922, <span class="html-italic">S. aureus</span> 1, <span class="html-italic">P. aeruginosa</span> PAO1, and <span class="html-italic">A. baumannii</span> 34230 at 1/4 MIC, 1/2 MIC, MIC, and MBC, respectively. Data are presented as mean ± standard deviation, <span class="html-italic">n</span> = 3. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001 compared with control (0-melittin) group.</p>
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<p>Effects of melittin on cell morphology at MIC and MBC. <span class="html-italic">E. coli</span> ATCC 25922, <span class="html-italic">S. aureus</span> 1, <span class="html-italic">P. aeruginosa</span> PAO1, <span class="html-italic">A. baumannii</span> 34230. PBS was used as the control. The red arrows indicate the site of the cell damage. The scale bars represent 1 µm.</p>
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<p>Effect of melittin at MIC and MBC on bacterial cell membrane permeability. <span class="html-italic">E. coli</span> ATCC 25922, <span class="html-italic">S. aureus</span> 1, <span class="html-italic">P. aeruginosa</span> PAO1, <span class="html-italic">A. baumannii</span> 34230, and PBS-treated bacteria were incubated for 10 h and stained with Propidium iodide (PI) and SYTO-9. The cells were observed through Confocal laser scanning microscope (CLSM) for red fluorescence (PI) and green fluorescence (SYTO-9) at excitation and emission. The scale bars represent 10 µm.</p>
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<p>Effects of melittin at MIC and MBC on reactive oxygen species (ROS) production in bacterial cells. (<b>A</b>) Fluorescence of ROS of four bacteria under laser confocal observation (<span class="html-italic">E. coli</span> ATCC 25922, <span class="html-italic">S</span>. <span class="html-italic">aureus</span> 1, <span class="html-italic">P</span>. <span class="html-italic">aeruginosa</span> PAO1, and <span class="html-italic">A. baumannii</span> 34230). The scale bars represent 10 µm. (<b>B</b>) Quantitative analysis of the intracellular ROS fluorescence intensity. PBS was used as the control. ** <span class="html-italic">p</span> &lt; 0.01, and *** <span class="html-italic">p</span> &lt; 0.001 compared with PBS-treated group.</p>
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<p>Biofilm inhibition of melittin against <span class="html-italic">E. coli</span> ATCC 25922, <span class="html-italic">S. aureus</span> 1, <span class="html-italic">P. aeruginosa</span> PAO1, and <span class="html-italic">A. baumannii</span> 34230. Compared to the positive control group (PC = positive control, azithromycin), data are shown as mean ± SD, and individual biological replicates are shown (<span class="html-italic">n</span> = 3); * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001 compared with PC group.</p>
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<p>Biofilm eradication of melittin against <span class="html-italic">E. coli</span> ATCC 25922, <span class="html-italic">S. aureus</span> 1, <span class="html-italic">P. aeruginosa</span> PAO1, and <span class="html-italic">A. baumannii</span> 34230. Compared to the positive control group (PC = positive control, azithromycin), data are shown as mean ± SD, and individual biological replicates are shown (<span class="html-italic">n</span> = 3); * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001 compared with PC group.</p>
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<p>Molecular docking 3D image of melittin (<b>A</b>), LL-37 (<b>B</b>), and LasR protein model. Blue represents the receptor LasR, yellow represents the polypeptide ligand, and green represents the amino acid residues of the polypeptide-protein interaction.</p>
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<p>(<b>A</b>) RMSD profile and (<b>B</b>) RMSF profile of LasR-Melittin and LasR-LL-37 during molecular dynamics simulations at 500 ns.</p>
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<p>Effect of melittin on EPS in <span class="html-italic">P. aeruginosa</span> PAO1 biofilms. Data are shown as the mean ± SD, and individual biological replicates are shown (<span class="html-italic">n</span> = 3); * <span class="html-italic">p</span> &lt; 0.05 compared with control (0-melittin) group.</p>
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<p>Effects of different concentrations of melittin on virulence factors. (<b>A</b>) pyocyanin, (<b>B</b>) elastase, and (<b>C</b>) rhamnolipid production. <span class="html-italic">n</span> = 3. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001 compared with control (0-melittin) group.</p>
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<p>Effects of different concentrations of melittin on swarming, swimming, and twitching motility. (<b>A</b>) Swarming motility, (<b>B</b>) swimming motility, and (<b>C</b>) twitching motility. The results shown are representative of the results of three independent experiments.</p>
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<p>Schematic illustration of the multiple actions of melittin against bacteria.</p>
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19 pages, 3471 KiB  
Article
Synergistic Antibacterial Efficacy of Melittin in Combination with Oxacillin against Methicillin-Resistant Staphylococcus aureus (MRSA)
by Ana Flávia Marques Pereira, Alessandra Aguirra Sani, Tatiane Baptista Zapata, Débora Silva Marques de Sousa, Bruno César Rossini, Lucilene Delazari dos Santos, Vera Lúcia Mores Rall, Carla dos Santos Riccardi and Ary Fernandes Júnior
Microorganisms 2023, 11(12), 2868; https://doi.org/10.3390/microorganisms11122868 - 27 Nov 2023
Cited by 2 | Viewed by 1321
Abstract
Methicillin-resistant Staphylococcus aureus (MRSA) often cause infections with high mortality rates. Antimicrobial peptides are a source of molecules for developing antimicrobials; one such peptide is melittin, a fraction from the venom of the Apis mellifera bee. This study aimed to evaluate the antibacterial [...] Read more.
Methicillin-resistant Staphylococcus aureus (MRSA) often cause infections with high mortality rates. Antimicrobial peptides are a source of molecules for developing antimicrobials; one such peptide is melittin, a fraction from the venom of the Apis mellifera bee. This study aimed to evaluate the antibacterial and antibiofilm activities of melittin and its association with oxacillin (mel+oxa) against MRSA isolates, and to investigate the mechanisms of action of the treatments on MRSA. Minimum inhibitory concentrations (MICs) were determined, and synergistic effects of melittin with oxacillin and cephalothin were assessed. Antibiofilm and cytotoxic activities, as well as their impact on the cell membrane, were evaluated for melittin, oxacillin, and mel+oxa. Proteomics evaluated the effects of the treatments on MRSA. Melittin mean MICs for MRSA was 4.7 μg/mL and 12 μg/mL for oxacillin. Mel+oxa exhibited synergistic effects, reducing biofilm formation, and causing leakage of proteins, nucleic acids, potassium, and phosphate ions, indicating action on cell membrane. Melittin and mel+oxa, at MIC values, did not induce hemolysis and apoptosis in HaCaT cells. The treatments resulted in differential expression of proteins associated with protein synthesis and energy metabolism. Mel+oxa demonstrated antibacterial activity against MRSA, suggesting a potential as a candidate for the development of new antibacterial agents against MRSA. Full article
(This article belongs to the Section Antimicrobial Agents and Resistance)
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<p>Curve of the inhibitory activity (OD<sub>600</sub>) for ATCC MRSA (<b>A</b>) and isolate MRSA (<b>C</b>) and time-kill curve of the bactericidal activity, in Log CFU/mL, for ATCC MRSA (<b>B</b>) and isolate MRSA (<b>D</b>).</p>
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<p>The percentage (%) of biofilm inhibition at subinhibitory concentrations of 25%, 50%, and 75% MIC for melittin, oxacillin, and the melittin-oxacillin combination against ATCC MRSA (<b>A</b>) and MRSA isolate (<b>B</b>). Distinct letters represent significant differences in percentage of biofilm inhibition among the tested treatments when <span class="html-italic">p</span> ≤ 0.05.</p>
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<p>The protein leakage, measured in μg/mL, after a 4-h exposure of both ATCC MRSA (<b>A</b>) and MRSA isolate (<b>B</b>) to treatments with MIC and 2× MIC of melittin, oxacillin, and the melittin and oxacillin combination. Distinct letters represent significant differences in leakage of proteins among the tested treatments when <span class="html-italic">p</span> ≤ 0.05.</p>
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<p>The release of nucleic acids, measured at an optical density of 260 nm, after a 4-h exposure of both ATCC MRSA (<b>A</b>) and MRSA isolate (<b>B</b>) to treatments with MIC and 2× MIC of melittin, oxacillin, and the melittin and oxacillin combination. Distinct letters represent significant differences in release of nucleic acids among the tested treatments when <span class="html-italic">p</span> ≤ 0.05.</p>
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<p>Analysis of HaCaT cells was conducted following treatment with melittin, oxacillin, and their combination (mel+oxa) to assess the induction of early apoptosis and late apoptosis/necrosis. The evaluation employed annexin V labeling (FITC) and propidium iodide. The treatments were represented as follows: (<b>A</b>) Basal negative control (autofluorescence); (<b>B</b>) MIC Melittin; (<b>C</b>) MIC Oxacillin; (<b>D</b>) MIC Mel+Oxa; (<b>E</b>) Basal negative control; (<b>F</b>) 2× MIC Melittin; (<b>G</b>) 2× MIC Oxacillin; (<b>H</b>) 2× MIC Mel+Oxa (Quadrant UL: represents late apoptosis or necrotic cells—anexxin V negative and propidium iode positive; Quadrant UR: represents late apoptotic or necrotic cells—anexxin V positive and propidium iodide positive; Quadrant LL: represents live cells—anexxin V negative and propidium iodide negative; Quadrant LR: represents early apoptotic cells—anexxin V positive and propidium iodide negative).</p>
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<p>Venn diagram of all proteins identified in the proteomic analysis of the three treatments (melittin, oxacillin, and mel+oxa) in comparison to the proteins identified in the control group.</p>
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<p>The molecular function of subinhibitory treatments with melittin (<b>A</b>), oxacillin (<b>C</b>), and mel+oxa (<b>E</b>), as well as the biological processes influenced by the treatments with melittin (<b>B</b>), oxacillin (<b>D</b>), and mel+oxa (<b>F</b>) in the differentially expressed proteins of MRSA when exposed to the treatments, considering both upregulation and downregulation, and compared to the control group.</p>
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<p>Protein–protein interactions networks for the most relevant proteins influenced by the treatment melittin in association with oxacillin (mel+oxa) against MRSA. Strong associations between proteins are depicted by connecting lines. rsmH: S-adenosyl-methyltransferase, ftsZ: Cell division protein FtsZ, pstB: Phosphate ABC transporter ATP-binding protein, metK: S-adenosylmethionine synthetase, eno: Enolase, ABD30684.1: 6-phosphogluconate dehydrogenase, decarboxylating, ABD30150.1: Phosphoenolpyruvate-protein phosphotransferase, ABD29862.1: ABC transporter, ATP-binding protein, ychF: Ribosome-binding ATPase YchF, rplR: Ribosomal protein L18, rplK: Ribosomal protein L11, aroB: DAHP synthetase, guaB: inosine-5′-monophosphate dehydrogenase, guaC: Guanosine monophosphate reductase, argJ: N-acetyltransferase, pflB: Formate acetyltransferase.</p>
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14 pages, 1179 KiB  
Review
Bee Venom and Its Two Main Components—Melittin and Phospholipase A2—As Promising Antiviral Drug Candidates
by Carole Yaacoub, Rim Wehbe, Rabih Roufayel, Ziad Fajloun and Bruno Coutard
Pathogens 2023, 12(11), 1354; https://doi.org/10.3390/pathogens12111354 - 15 Nov 2023
Cited by 3 | Viewed by 2087
Abstract
Viruses are known to infect most types of organisms. In humans, they can cause several diseases that range from mild to severe. Although many antiviral therapies have been developed, viral infections continue to be a leading cause of morbidity and mortality worldwide. Therefore, [...] Read more.
Viruses are known to infect most types of organisms. In humans, they can cause several diseases that range from mild to severe. Although many antiviral therapies have been developed, viral infections continue to be a leading cause of morbidity and mortality worldwide. Therefore, the discovery of new and effective antiviral agents is desperately needed. Animal venoms are a rich source of bioactive molecules found in natural goods that have been used since ancient times in alternative medicine to treat a variety of human diseases. Recently, and with the onset of the COVID-19 pandemic, scientists have regained their interest in the possible use of natural products, such as bee venom (BV), as a potential antiviral agent to treat viral infections. BV is known to exert many therapeutic activities such as anti-proliferative, anti-bacterial, and anti-inflammatory effects. However, there is limited discussion of the antiviral activity of BV in the literature. Therefore, this review aims to highlight the antiviral properties of BV and its two primary constituents, melittin (MEL) and phospholipase A2 (PLA2), against a variety of enveloped and non-enveloped viruses. Finally, the innovative strategies used to reduce the toxicity of BV and its two compounds for the development of new antiviral treatments are also considered. Full article
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Graphical abstract
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<p>Figure showing BV composition, characteristics, and 3D representation of its two main components, MEL (PDB code 1BH1) and PLA2 (PDB code 1POA).</p>
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<p>Different mechanisms of action of BV against different families of viruses.</p>
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12 pages, 10442 KiB  
Review
Bee Products for Poultry and Rabbits: Current Challenges and Perspectives
by Ayman Abd El-Aziz, Mahmoud Abo Ghanima, Daniel Mota-Rojas, Anjum Sherasiya, Francesca Ciani and Karim El-Sabrout
Animals 2023, 13(22), 3517; https://doi.org/10.3390/ani13223517 - 14 Nov 2023
Cited by 6 | Viewed by 1706
Abstract
Poultry and rabbit production are important and rapidly growing agricultural subsectors, particularly in several developing countries. To ensure the sustainability of poultry and rabbit production, realistic poultry and rabbit farming practices must be improved. Apitherapy is a traditional alternative medicine that involves the [...] Read more.
Poultry and rabbit production are important and rapidly growing agricultural subsectors, particularly in several developing countries. To ensure the sustainability of poultry and rabbit production, realistic poultry and rabbit farming practices must be improved. Apitherapy is a traditional alternative medicine that involves the prevention and treatment of some diseases with several bee products including propolis, royal jelly, pollen, and venom. More feeding investigations on the numerous benefits of bee products for poultry and rabbits are crucial to be addressed. Poultry and rabbit production has recently experienced numerous challenges, including climate change, disease spread, and antibiotic misuse. Improving animal welfare, health, and production is a top priority for all livestock farms, as is supplying consumers with safe and healthy products. Therefore, this review aims to collect and investigate recent relevant literature on the use of bee products, as feed additives, drinking water supplements, and injections, for poultry and rabbits to improve animal health and production. From the current findings, bee products can improve the growth and immunological performance of small-livestock animals, such as poultry and rabbits, by activating digestive enzymes, maintaining microbial balance, and promoting vitamin synthesis. Therefore, bee products could be a promising natural alternative to growth promoters, reproductive stimulants, and immunological enhancers in poultry and rabbit farms to provide safe and healthy products for humans. Full article
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<p>The potential role of bee products on poultry and rabbit performance.</p>
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13 pages, 5277 KiB  
Article
Effect of CM15 on Supported Lipid Bilayer Probed by Atomic Force Microscopy
by Olivia D. Walsh, Leona Choi and Krishna P. Sigdel
Membranes 2023, 13(11), 864; https://doi.org/10.3390/membranes13110864 - 28 Oct 2023
Viewed by 1811
Abstract
Antimicrobial peptides are key components of the immune system. These peptides affect the membrane in various ways; some form nano-sized pores, while others only produce minor defects. Since these peptides are increasingly important in developing antimicrobial drugs, understanding the mechanism of their interactions [...] Read more.
Antimicrobial peptides are key components of the immune system. These peptides affect the membrane in various ways; some form nano-sized pores, while others only produce minor defects. Since these peptides are increasingly important in developing antimicrobial drugs, understanding the mechanism of their interactions with lipid bilayers is critical. Here, using atomic force microscopy (AFM), we investigated the effect of a synthetic hybrid peptide, CM15, on the membrane surface comprising E. coli polar lipid extract. Direct imaging of supported lipid bilayers exposed to various concentrations of the peptide revealed significant membrane remodeling. We found that CM15 interacts with supported lipid bilayers and forms membrane-spanning defects very quickly. It is found that CM15 is capable of remodeling both leaflets of the bilayer. For lower CM15 concentrations, punctate void-like defects were observed, some of which re-sealed themselves as a function of time. However, for CM15 concentrations higher than 5 µM, the defects on the bilayers became so widespread that they disrupted the membrane integrity completely. This work enhances the understanding of CM15 interactions with the bacterial lipid bilayer. Full article
(This article belongs to the Special Issue Membrane Interaction between Lipids, Proteins and Peptides)
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<p>Peptides at a glance. Helical wheel of CM15 peptide that demonstrates the relative positions of amino acids in the peptide along with a table of amino acid sequences of parent peptides and hybrid CM15 peptide. The helical wheel is prepared using reference [<a href="#B15-membranes-13-00864" class="html-bibr">15</a>].</p>
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<p>CM15 concentration dependence on defect formation. (<b>A</b>–<b>F</b>) Empty lipid bilayer, 0.2 <math display="inline"><semantics> <mi mathvariant="sans-serif">μ</mi> </semantics></math>M, 0.5 <math display="inline"><semantics> <mi mathvariant="sans-serif">μ</mi> </semantics></math>M, 1 <math display="inline"><semantics> <mi mathvariant="sans-serif">μ</mi> </semantics></math>M, 2.5 <math display="inline"><semantics> <mi mathvariant="sans-serif">μ</mi> </semantics></math>M, and 5 <math display="inline"><semantics> <mi mathvariant="sans-serif">μ</mi> </semantics></math>M peptide concentration, respectively. When the concentration of peptide was increased, the defects grew larger and, accordingly, the effective roughness of the surface was increased. The 1-micron lateral scale bar applies to all images.</p>
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<p>RMS roughness as a function of peptide CM15 concentration. The RMS roughness of the surface after exposure to CM15 peptides.</p>
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<p>Characterizing defects: (<b>A</b>) A representative image of a supported lipid bilayer with 0.5 <math display="inline"><semantics> <mi mathvariant="sans-serif">μ</mi> </semantics></math>M of CM15. (<b>B</b>) A depth profile that is taken along the white line shown in (<b>A</b>) and demonstrates ~4 nm depth of the defect. (<b>C</b>) A smooth histogram of the depth of defects (<span class="html-italic">N</span> = 3027) below the lipid bilayer caused by CM15 shows the three major depth populations indicated by shaded gaussians. Note that these histograms include data from peptide concentrations of 0.5, 1, and 2.5 <math display="inline"><semantics> <mi mathvariant="sans-serif">μ</mi> </semantics></math>M. 3D rendering of representative images for each peak, peak value, and their corresponding weightage are also shown. (<b>D</b>) Probability density of the areal footprint of the defects shows several populations indicated by different gaussians.</p>
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<p>Time-lapse imaging: time-lapse imaging of the defects on the supported lipid bilayer caused by CM15 peptides at 1 <math display="inline"><semantics> <mi mathvariant="sans-serif">μ</mi> </semantics></math>M concentration. The first image was completed ~20 min after the addition of the peptide on the supported lipid bilayer. Most of the defects are stable except some of them are observed to seal up (dashed circle with arrow).</p>
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<p>Time-lapse imaging for 2.5 <math display="inline"><semantics> <mi mathvariant="sans-serif">μ</mi> </semantics></math>M peptide: time-lapse imaging of the defects on the supported lipid bilayer caused by CM15 peptides at 2.5 <math display="inline"><semantics> <mi mathvariant="sans-serif">μ</mi> </semantics></math>M concentration. The defects are stable, except some of them are sealing up (please follow the circular marks with an arrow). Lower panel: a smooth histogram of the depth of the voids for an image at t = 20 min, 28 min, and 60 min, respectively. Depth profile evolution can clearly be seen as a function of time. Different populations are indicated by shaded gaussians in each profile. In addition, the number of voids per image and average depth of the voids are also decreasing.</p>
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<p>Membrane disruption: (<b>A</b>,<b>B</b>) Sample images of the defects on the supported lipid bilayer caused by CM15 peptides at 5 <math display="inline"><semantics> <mi mathvariant="sans-serif">μ</mi> </semantics></math>M concentration. CM15 affects the bilayer very quickly, and, thereafter, the defect patterns remain fairly similar. The images show nominally the same areas imaged at the time interval of about 30 min with similar defect patterns. (<b>C</b>) Depth profiles taken at the locations of red and blue dotted lines (in (<b>A</b>,<b>B</b>)) demonstrate similar defect patterns. The lateral displacement between features is due to lateral drift. The black vertical scale represents the scale of the depth profile.</p>
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<p>(<b>A</b>–<b>D</b>) Time-lapse imaging of void-like defects on the supported lipid bilayer caused by CM15 peptides. (<b>E</b>,<b>F</b>) The depth profile of the feature along the scan line is shown in (<b>A</b>). The profiles in (<b>E</b>) show the depth profile for features 1 and 2 for each image and (<b>F</b>) shows the profile for feature 3.. The times are listed next to each line as well. Two pores close at the end completely.</p>
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