Native Venom Research Articles

КЛИНИЧЕСКИЕ ПОКАЗАТЕЛИ КРОВИ КРЫС С ХРОНИЧЕСКИМ ВОСПАЛЕНИЕМ МЯГКИХ ТКАНЕЙ СПИНЫ ПРИ КУРСОВОМ ВНУТРИМЫШЕЧНОМ ВВЕДЕНИИ РАСТВОРОВ ЯДА ГЮРЗЫ ЗАКАВКАЗСКОЙ (MACROVIPERA LEBETINA OBTUSA)

An relevant task of modern pharmacology is the search for drugs for the treatment of chronic inflammatory processes. The use of nonsteroidal anti-inflammatory drugs and drugs of the glucocorticoid group has a number of limitations due to pronounced negative effects. The use of herbal anti-inflammatory drugs often has a delayed effect. In this case, the anti-inflammatory effect is manifested in the later stages of the pathological process. For this reason, it seems interesting to us to use a solution of poison obtained from gyurza (Macrovipera lebetina obtusa) as an alternative therapy for chronic inflammatory processes. Earlier in our work, the ability of solutions of these poisons to reduce inflammation was shown. This effect was found both in native venom solutions and in those heated to 55оС. In this work, the effect of venom solutions was studied after a course of intramuscular administration. The criteria for anti-inflammatory action were blood counts in experimental chronic inflammation caused by acetic acid. Inflammation was modeled by injecting 0.5 ml of 9% acetic acid solution under the skin of the back and intraperitoneal administration of 1 ml of 10% Rheopolyglukin solution to white outbred male rats. Diclofenac in injections was chosen as a comparison drug. The assessment of clinical blood parameters was carried out on an automatic hematology analyzer BC 2800vet (Mindray). The positive dynamics of the course of the chronic aseptic inflammatory process has been revealed. A more positive dynamics of the course of the chronic aseptic inflammatory process was revealed in groups of animals receiving solutions of native venom.

Analytical and quantitative assessment of the structural components of the adrenal glands of rats under the conditions of exposure to the venom of vipers Vipera berus berus and Vipera berus nikolskii

Snakebite envenoming is a common but neglected public health problem worldwide, especially in tropical countries. Annual mortality as a result of snakebites exceeds 138,000. It is believed that this problem is underestimated, and in many countries, individual cases of bites are not subject to proper fixation. The purpose of the study is the analytical and quantitative assessment of the structural components of the rats' adrenal glands under exposure to the venom of Vipers Vipera berus berus and Vipera berus nikolskii. Experimental studies were carried out on white, non-linear male rats. Vipera berus berus and Vipera berus nikolskii viper venom were obtained from V. N. Karazin Kharkiv National University. The freeze-dried native venom was stored at -20 °C and dissolved in saline immediately before the experiment. The animals were divided into three groups (control and 2 experimental groups) of 10 individuals each. Experimental rats were injected intraperitoneally in a physiological solution with a semi-lethal dose (LD50) (1.576 mg/g-1) of Vipera berus berus and Vipera berus nikolskii venoms. Animals of the control group were injected intraperitoneally with only a physiological solution. Rats were removed from the experiment 24 hours after exposure to the poison and anesthetised by cervical dislocation. Statistical analysis of the area of the microcirculatory channel and the nuclear-cytoplasmic index was performed using Fiji: ImageJ program and processed in Excel. Administration of the venom of the vipers Vipera berus berus and Vipera berus nikolskii to rats was accompanied by a significant increase in the area of the microcirculatory bed relative to the control group (2.9 times for Vipera berus berus and 6.5 times for Vipera berus nikolskii). Exposure to Vipera berus berus viper venom was associated with a significant decrease in the nuclear-cytoplasmic index in rats of the experimental group compared to the control group (13 % and 42 %, respectively), which is evidence of a decrease in the area of the nuclei of endocrinocytes of the adrenal cortex. This indicator in rats under the administration of Vipera berus nikolskii venom was even lower and amounted to 12 %. According to the statistical analysis of the quantitative assessment of the state of the cortical substance of the adrenal glands, it is worth noting the similar effect of the poisons of both types of snakes at the cellular level. At the same time, at the tissue level, the effect of Vipera berus nikolskii venom is more pronounced than that of Vipera berus berus - this is evidenced by the higher degree of disruption of the structure of the hemomicrocirculatory channel in the adrenal cortex of animals from the group that was affected by this venom. It led to an increase in the area of vessels due to their expansion and ruptures of their walls and haemorrhages into the surrounding parenchyma and stroma.

Function and Mechanism of Antiviral Wasp Venom Peptide Protopolybia-MP III and Its Derivatives against HSV-1.

Viruses are one of the leading causes of human disease, and many highly pathogenic viruses still have no specific treatment drugs. Therefore, producing new antiviral drugs is an urgent matter. In our study, we first found that the natural wasp venom peptide Protopolybia-MP III had a significant inhibitory effect on herpes simplex virus type 1 (HSV-1) replication in vitro by using quantitative real-time PCR (qPCR), Western blotting, and plaque-forming assays. Immunofluorescence analysis showed Protopolybia-MP III could enter cells, and it inhibited multiple stages of the HSV-1 life cycle, including the attachment, entry/fusion, and post-entry stages. Furthermore, ultracentrifugation and electron microscopy detected that Protopolybia-MP III significantly suppressed HSV-1 virion infectivity at different temperatures by destroying the integrity of the HSV-1 virion. Finally, by comparing the antiviral activity of Protopolybia-MP III and its mutants, a series of peptides with better anti-HSV-1 activity were identified. Overall, this work found the function and mechanism of the antiviral wasp venom peptide Protopolybia-MP III and its derivatives against HSV-1 and laid the foundation for the research and development of wasp venom-derived antiviral candidate peptide drugs.

Application of a new inhibitor for the corrosion of iron in acidic solution: Electrochemical effect of a scorpion venom

In this study, the venom of the species Leiurus abdullahbayrami, known as the yellow scorpion from the Buthidae family, was applied for the first time as a natural corrosion inhibitor for iron in hydrochloric acid solution. The effectiveness of scorpion venom as an eco-friendly and natural inhibitor was determined by electrochemical methods such as electrochemical impedance spectroscopy (EIS), potentiodynamic polarization (Tafel extrapolation method) and linear polarization resistance (LPR) after an hour of immersion. Four different concentrations were determined for the green and natural inhibitor scorpion venom in 1.0 M HCl, and it was observed that the corrosion of iron in these solutions was significantly inhibited. In general, the inhibition efficiency was above 80%. According to the potentiodynamic polarization data, it has been determined that the Leiurus abdullahbayrami venom acts as a cathodic-type inhibitor on the Fe surface. Finally, the surface images of the iron electrodes in 1.0 M HCl solutions without and with Leiurus abdullahbayrami venom after 1 h immersion were examined by field emission scanning electron microscope (FE-SEM), it was concluded that the surface containing scorpion venom had a flatter compared to the uninhibited surface.

The inhibitory activity of a new scorpion venom-derived antimicrobial peptide Hp1470 against Gram-positive bacteria

Antimicrobial peptides (AMPs) are a new type of antibiotic and target a variety of microbes, including antibiotic-resistant strains; thus, AMPs have attracted widespread interest. Scorpion venoms contain many bioactive peptides, including AMPs, and have become an important natural resource of peptide-based drugs. Here, the antibacterial peptide gene Hp1470 from the venom of the scorpion Heterometrus petersii was characterized, and its antibacterial activity was determined. The cDNA sequence of Hp1470 is 300 nt in length and contains an open reading frame (ORF) of 207 nt. The ORF was shown to encode 68 amino acid residues, including a signal peptide (23 aa), a mature peptide (13 aa), a C-terminal posttranslational processing signal (3 aa), and a propeptide (29 aa). Multiple sequence alignment results indicated that Hp1470 is an antibacterial peptide. The mature peptide Hp1470, which has a molecular mass of 1564.09 Da, was further chemically synthesized with a purity of greater than 95%. Antimicrobial assays showed that the synthesized Hp1470 exerted an inhibitory effect on Gram-positive bacteria and clinical drug-resistant strains, including PRSA and MRSA, but not Gram-negative bacteria. Hp1470 was further found to protect mice from MRSA infection, suggesting its potential application as an in vivo antimicrobial agent. Interestingly, Hp1470 only inhibited bacterial growth but did not kill bacteria, which was consistent with scanning electron microscopy results showing that Hp1470 did not lyse the cell membrane of Staphylococcus aureus. Our work provides a new direction for developing antibacterial agents with different modes of action from natural scorpion venoms.

Discovery of a novel jellyfish venom metalloproteinase inhibitor from secondary metabolites isolated from jellyfish-derived fungus Aspergillus versicolor SmT07

The major jellyfish stings that occur in China are caused by scyphozoan Nemopilema nomurai, whose venom exhibits significant metalloproteinase activity that contributes to the toxic effects of jellyfish envenomation. Researching effective inhibitors suppressing the metalloproteinase activity of jellyfish venom represents a new attempt to cure jellyfish envenomations. In the present study, secondary metabolites produced by the jellyfish-associated fungus Aspergillus versicolor SmT07 were isolated and evaluated for their anti-proteolytic activities. Two xanthones, sterigmatocystin (JC-01) and oxisterigmatocystin C (JC-06), and four alkaloids, cottoquinazoline A (JC-02), phenazine-1-carboxylic acid (JC-03), viridicatin (JC-04) and viridicatol (JC-05), were isolated and identified. Only phenazine-1-carboxylic acid (PCA) showed significant anti-proteolytic activity of jellyfish venom assayed on azocasein, and the IC50 value was 2.16 mM. PCA also significantly inhibited fibrinogenolytic activity, protecting the Bβ chain of fibrinogen from degradation when preincubated with jellyfish venom at a ratio of >1:0.6 (PCA:venom, w/w). Molecular docking with several well-characterized snake venom metalloproteinases suggested the venom metalloproteinases inhibitory property of PCA by forming complex interactions with the active site via hydrogen bonds, π-π stacking and salt bridges, which was distinct from the binding mode of batimastat. The present study represents the first study identifying natural jellyfish venom metalloproteinase inhibitors from marine natural products, which may provide an alternative to develop therapeutic agents for treating jellyfish envenomations.

Long-term antibody response and protective effect induced by attenuated scorpion toxins: Involvement of memory plasma cells

In Algeria, Androctonus australis hector scorpion envenomation remains a major problem of public health because of non-efficient therapy. The development of safe vaccine against scorpion venom could be one key strategy for the envenomation prevention. The irradiation of venom by γ-rays develops suitable immunogens which produced effective antivenom and safe vaccine. In this study, we investigated the ability of the irradiated toxic fraction (γ-FtoxG50) to induce long-term memory humoral response in immunized animals (mice and rabbits), by involving the long-lived plasma cells to prevent efficiently the lethality of scorpion envenomation. For this purpose, an appropriate immunization schedule was established in mice and rabbits using three (3) similar doses of γ-FtoxG50 associated with Alum adjuvant.Obtained results indicate that the long-term immunogenicity of γ-FtoxG50 is able to induce the long-term memory humoral response with a high level of specific antibodies. The long-term persistence of antibody levels could depend on bone marrow memory plasma cells. These cells produce continuously antibodies without antigen stimulus. Furthermore, an enhanced memory response was obtained post-repeated envenomation with toxic native venom that leads to improved protection of animals. Together, pre-existing protective antibodies and the activation of memory B-cells could induce a rapid neutralization of scorpion toxins and long-term protection against scorpion envenomation.

Applications of venom biodiversity in agriculture

Agriculture can benefit from the biotechnological use of animal venom biodiversity. We explored the patent database to find the prevailing applications of scorpion, snake, spider, bee, and wasp venoms for agricultural purposes. We searched for patents registered worldwide using the keywords “animal species” AND “venom” associated with the Cooperative Patent Classification (CPC) subclasses, based on Google Patents by November 2020. The results indicate an increasing frequency in patents related to animal venoms. Scorpion, snake, and wasp appeared most frequently, besides other species. The inventors concentrate on two subclasses, namely new plant varieties development (A01H) and biopesticides (A01N). Monsanto and other giant agricultural and biotechnology companies lead the list in the number of patents. New varieties of crops, like soybean, corn, cotton, rice, and wheat, are the most frequent inventions. The benefits of using natural venoms in insect-resistant plant varieties and biopesticides are pertinent to agriculture, the environment, and human health.

Description of a serpin toxin in Loxosceles (Brown spider) venoms: Cloning, expression in baculovirus-infected insect cells and functional characterization

Several classes of toxins are present in the venom of Brown spiders (Loxosceles genus), some of them are highly expressed and others are less expressed. In this work, we aimed to clone the sequence of a little expressed novel toxin from Loxosceles venom identified as a serine protease inhibitor (serpin), as well as to express and characterize its biochemical and biological properties. It was named LSPILT, derived from Loxoscelesserine protease inhibitor-like toxin. Multiple alignment analysis revealed high identity between LSPILT and other serpin molecules from spiders and crab. LSPILT was produced in baculovirus-infected insect cells, resulting in a 46-kDa protein fused to a His-tag. Immunological assays showed epitopes in LSPILT that resemble native venom toxins of Loxosceles spiders. The inhibitory activity of LSPILT on trypsin was found both by reverse zymography and fluorescent gelatin-degradation assay. Additionally, LSPILT inhibited the complement-dependent lysis of Trypanosoma cruzi epimastigotes, reduced thrombin-dependent clotting and suppressed B16-F10 melanoma cells migration. Results described herein prove the existence of conserved serpin-like toxins in Loxosceles venoms. The availability of a recombinant serpin enabled the determination of its biological and biochemical properties and indicates potential applications in future studies regarding the pathophysiology of the envenoming or for biotechnological purposes.

Mass Spectrometry-based Sequencing of Venom Peptides (Conotoxins) from Vermivorous Cone Snail, Conus Loroisii: Toxicity of its Natural Venom

Conus loroisii is a marine vermivorous snail found profusely in the southern seas of India. They harbor several toxic peptide components commonly called as ‘conotoxins’. In this study, we have identified and sequenced five conotoxins using proteome based tandem mass spectrometry analysis through Data analysis 4.1 software. Among them, we found Lo959 as contryphan which is previously described. All other conotoxins Lo1702, Lo1410, Lo1385 and Lo1686 belong to M-Superfamily conotoxins and novel to C. loroisii. Lo1410 is completely novel to conotoxin research with 3 disulfides and the amino acid sequence is derived as CCSTNCAVCIPCCP. All the identified M-Superfamily conotoxins are sub categorised to mini M2 superfamily conotoxins. Lo1702 and Lo1686 possess C- terminal amidation which is the key feature in conotoxins. Moreover, we have screened the natural venom for the occurrence of toxicity in the zebrafish model and brine shrimp.

Mass spectrometric identification and denovo sequencing of novel conotoxins from vermivorous cone snail (Conus inscriptus), and preliminary screening of its venom for biological activities in vitro and in vivo.

Venom of Conus inscriptus, a vermivorous cone snail found abundantly in the southern coastal waters was studied to yield conotoxins through proteomic analysis. A total of 37 conotoxins (4 with single disulfide bonds, 20 with two disulfide bonds and 11 three disulfide-bonded peptides) were identified using mass spectrometric analysis. Among them, amino acid sequences of 11 novel conopeptides with one, two and three disulfides belonging to different classes were derived through manual de novo sequencing. Based on the established primary sequence, they were pharmacologically classified into α conotoxins, µ conotoxins and contryphans. Except In1696 all other conopeptides have undergone C-terminal amidation. The natural venom exhibited 50% lethality at 304.82 µg/mL against zebrafish embryo and 130.31 µg/mL against brine shrimp nauplii. The anticonvulsant study of natural venom effectively reduced the locomotor activity against pentylenetetrazole (PTZ) treated zebrafish. This concludes that the venom peptides from Conus inscriptus exhibit potential anticonvulsant function, which leads to the discovery of lead molecules against seizures.

Functional Impact of BeKm-1, a High-Affinity hERG Blocker, on Cardiomyocytes Derived from Human-Induced Pluripotent Stem Cells.

IKr current, a major component of cardiac repolarization, is mediated by human Ether-à-go-go-Related Gene (hERG, Kv11.1) potassium channels. The blockage of these channels by pharmacological compounds is associated to drug-induced long QT syndrome (LQTS), which is a life-threatening disorder characterized by ventricular arrhythmias and defects in cardiac repolarization that can be illustrated using cardiomyocytes derived from human-induced pluripotent stem cells (hiPS-CMs). This study was meant to assess the modification in hiPS-CMs excitability and contractile properties by BeKm-1, a natural scorpion venom peptide that selectively interacts with the extracellular face of hERG, by opposition to reference compounds that act onto the intracellular face. Using an automated patch-clamp system, we compared the affinity of BeKm-1 for hERG channels with some reference compounds. We fully assessed its effects on the electrophysiological, calcium handling, and beating properties of hiPS-CMs. By delaying cardiomyocyte repolarization, the peptide induces early afterdepolarizations and reduces spontaneous action potentials, calcium transients, and contraction frequencies, therefore recapitulating several of the critical phenotype features associated with arrhythmic risk in drug-induced LQTS. BeKm-1 exemplifies an interesting reference compound in the integrated hiPS-CMs cell model for all drugs that may block the hERG channel from the outer face. Being a peptide that is easily modifiable, it will serve as an ideal molecular platform for the design of new hERG modulators displaying additional functionalities.

The interaction between the natural metalloendopeptidase inhibitor BJ46a and its target toxin jararhagin analyzed by structural mass spectrometry and molecular modeling

Snakebite envenoming affects millions of people worldwide, being officially considered a neglected tropical disease by the World Health Organization. The antivenom is effective in neutralizing the systemic effects of envenomation, but local effects are poorly neutralized, often leading to permanent disability. The natural resistance of the South American pit viper Bothrops jararaca to its venom is partly attributed to BJ46a, a natural snake venom metalloendopeptidase inhibitor. Upon complex formation, BJ46a binds non-covalently to the metalloendopeptidase, rendering it unable to exert its proteolytic activity. However, the structural features that govern this interaction are largely unknown. In this work, we applied structural mass spectrometry techniques (cross-linking-MS and hydrogen-deuterium exchange MS) and in silico analyses (molecular modeling, docking, and dynamics simulations) to understand the interaction between BJ46a and jararhagin, a metalloendopeptidase from B. jararaca venom. We explored the distance restraints generated from XL-MS experiments to guide the modeling of BJ46a and jararhagin, as well as the protein-protein docking simulations. HDX-MS data pinpointed regions of protection/deprotection at the interface of the BJ46a–jararhagin complex which, in addition to the molecular dynamics simulation data, reinforced our proposed interaction model. Ultimately, the structural understanding of snake venom metalloendopeptidases inhibition by BJ46a could lead to the rational design of drugs to improve anti-snake venom therapeutics, alleviating the high morbidity rates currently observed.

Improvement of Naja haje snake antivenom production using gamma radiation and a biotechnological technique

This study aimed to improve antivenom production using gamma radiation and calcium phosphate nanoparticles (CPN) as adjuvant. Toxicological and immunological properties of the Naja haje venom before and after exposure to radiation were estimated. The immune response of immunized rabbits was evaluated. Toxicity of irradiated venom was reduced six times as compared to native venom. Moreover, no change in antigenic reactivity between native and irradiated venoms. Serum titer produced with irradiated venom loaded on CPN showed highest titer and neutralization capacity as compared to other sera. Thus, irradiation and nanoparticles adjuvant could be a new approach for snake antivenom production.

Snake Venom Cathelicidins as Natural Antimicrobial Peptides.

Bioactive small molecules isolated from animals, plants, fungi and bacteria, including natural antimicrobial peptides, have shown great therapeutic potential worldwide. Among these peptides, snake venom cathelicidins are being widely exploited, because the variation in the composition of the venom reflects a range of biological activities that may be of biotechnological interest. Cathelicidins are short, cationic, and amphipathic molecules. They play an important role in host defense against microbial infections. We are currently facing a strong limitation on pharmacological interventions for infection control, which has become increasingly complex due to the lack of effective therapeutic options. In this review, we will focus on natural snake venom cathelicidins as promising candidates for the development of new antibacterial agents to fight antibiotic-resistant bacteria. We will highlight their antibacterial and antibiofilm activities, mechanism of action, and modulation of the innate immune response.

Structural analysis of the natural snake venom metalloendopeptidase inhibitor BJ46a and its target toxin, jararhagin, based on XL-MS and HDX-MS

Structural analysis of the natural snake venom metalloendopeptidase inhibitor BJ46a and its target toxin, jararhagin, based on XL-MS and HDX-MS

Proteome Based de novo Sequencing of Novel Conotoxins from Marine Molluscivorous Cone Snail Conus amadis and Neurological Activities of Its Natural Venom in Zebrafish Model.

Conus amadis is a carnivorous snail found abundantly in coastal waters of India. Despite its abundance in southern coastal waters of India and the fact that most of the conotoxin act in neuronal system, research work on Conus amadis venom was not much focused. So we have made a brief study on the venom complex of Conus amadis to identify the library of novel conotoxins and to screen the natural venom for neurological function. De novo sequencing of novel conopeptides from the venom cocktail of Conus amadis and to screen its natural venom for the presence of biological activities in zebrafish model. Proteome based MALDI-TOF and LC-MS-MS analysis for identification of novel conotoxins and subsequent sequencing. Due to the complex disulfide rich nature of the venom peptides, the study also involves global chemical modification experiments of the venom extract to unambiguously determine the sequence of novel conotoxins. Biological function analysis of natural venom was tested in zebrafish model to ascertain anti-epileptic properties. In this study, we have identified 19 novel conotoxins containing 1, 2 & 3 disulfides, belonging to different classes. Among them, 2 novel contryphans, 3 T-superfamily conotoxins, 2 A-superfamily conotoxins and 2 Mini M-Superfamily conotoxins were sequenced to its amino acid level from the fragmented spectrum of singly and doubly charged parent ions using de novo sequencing strategies. ama1054, a contryphan peptide toxin, possesses post translationally modified bromo tryptophan at its seventh position. Except ama1251, all the sequenced peptide toxins possess modified C-terminal amidation. Crude venom exhibited anticonvulsant properties in pentylenetetrazole-induced seizure in zebrafish larvae, which suggested anti-epileptic property of the venom cocktail. Acetylcholinesterase activity was also identified in the venom complex. Based on the preliminary evidence, if this study is extended further through bioassay guided purification, could possibly yield peptide toxins with anticonvulsant and other neurologically active molecules.

Biochemical Modulation of Venom by Spiders is Achieved Via Compartmentalized Toxin Production and Storage

Animal venoms are complex chemical arsenals containing hundreds or even thousands of diverse small molecules, peptides, and proteins that affect a myriad of molecular targets. Because this biochemical arsenal is metabolically expensive, venom must be used sparingly and therefore one might predict that venom evolution would result in retention of only the most essential toxins for predation and/or defence. However, this presents a strong dichotomy in the functional requirements of venoms that are used for both predation and defence, which results in extensive functional redundancy. Here we show that spiders can overcome this potential metabolic burden by modulating the biochemical composition of their venom. We demonstrate that the venom peptidome of the lethal Australian funnel-web spider Hadronyche infensa changes both qualitatively and quantitatively throughout a series of defensive secretions — secretion of the most potent insecticidal toxins is preceded by secretion of non-insecticidal, presumably defensive, toxins. Using mass spectrometry imaging, we show that modulation of toxin secretion is facilitated by differential storage of toxins in the venom gland. We propose that the establishment of distinct venom-gland zipcodes for toxin production is an adaptation that reduces the metabolic expense of venom production and perhaps also serves to minimize effects from development of toxin resistance during predator-prey co-evolution. Our results highlight the importance of considering behavioral aspects of natural venom secretions in understanding toxin function and evolution.

APOPTOTIC AND NECROTIC EFFECTS OF CHITOSAN NANOPARTICLES LOADED WITH THE HONEYBEE, APIS MELLIFERA VENOM ON DIFFERENT CANCER CELL LINES

Nanoparticles encapsulated with natural venoms have a considerable attention to improve illnesses therapies by overcoming toxic effects secreted by different pathogens. The present study explored conjugation of chitosan extracted from cuticle of naturally died honeybees nanoparticles (B-Cs-NPs) with the honeybee, Apis mellifera venom (BV). Detected related toxicity and anticancer potential of BV and BV loaded NPs- against Colon (CaCO-2), Larynx (HEp-2) and breast (MCF-7) cancer cell lines. The Inotropic gelation of chitosan (CS) with sodium tripolyphosphate (TPP) was the method used to prepare nanoparticles. Cytotoxic effects of BV and BV loaded NPs nanoparticles against the three cancer cell lines studied, then they were assessed using MTT stain assay. Data recorded revealed that combination of bee venom with bee chitosan nanoparticles enhanced the inhibitory effect against cancer cells studied when compared with bee venom alone. Also, it was cleared that by treating CaCO-2, HEp-2 & MCF-7 cell lines with IC50 concentrations of BV alone and BV loaded chitosan nanoparticles, this induced apoptotic features as cell shrinking, peripheral condensation of chromatin, apoptoticbodies in late stage, nucleoli fragmentation, membrane blebbing, shrunken nucleus with some necrotic features. Control cells showed normal pathway of mitotic divisions of obvious cell membrane and nucleus when stained by (H&E) and by visualization using inverted microscope.

ВПЛИВ ДЕЯКИХ АНЕСТЕТИКІВ ТА ПРИРОДНИХ ОТРУТ НА ФУНКЦІОНУВАННЯ LCC-КАНАЛІВ ЯДЕРНОЇ МЕМБРАНИ КАРДІОМІОЦИТІВ ТА НЕЙРОНІВ ПУРКІНЬЄ МОЗОЧКА

The investigation of pharmacological sensitivity of the cationic channels in nuclear membrane to the influence of anesthetics and natural venoms is relevant since it was shown that some modulators of N-cholinoreceptors (dithylinum, atracurium) affecting the large conductance cation channels (LCC-channels) functional activity are used in medicine during surgery. In addition, some injectable forms of toxins from the snake venom are used as drugs with an analgesic effect. Therefore, the aim of the study was to investigate the pharmacological sensitivity of the LCC-channels to the muscle relaxants, anaesthetics (mydocalm, diprofol) and natural venoms (neurotoxin II, α-Cobratoxin). The influence of these substances was evaluated based on changes in biophysical parameters of functioning of the LCC-channels of nuclear membrane of the cardiomyocytes and cerebellar Purkinje neurons. Ion currents through these channels were registered in the nucleus-attached or excised patch configuration and the voltage-clamp mode of the patch-clamp technique. We found that mydocalm (2 mM), diprofol (2 mM) and α-Cobratoxin (1 mM) reduced several times the probability of the channels being in the open state. Under the influence of mydocalm and α-Kobratoxin in high concentrations (1-2 mM) the effect of channels flickering was observed which indicates the channel pore blocking in its open state. At the same time, the average amplitude of the K+ current through the LCC-channels decreased by 13 % under the influence of NT II (25 μM). The results will be the basis for identification of new, more effective inhibitors of the LCC-channels that will be promising for the physiological relevance and structure of the channels investigation.