ASPEK MAKRO DAN MIKRO PADA MANTRA PENJAGAAN (KEBUN, BADAN DAN BISA BINATANG) MASYARAKAT MELAYU KEMUJA

The problem in this research is what are the macro and micro aspects of the guarding mantra (garden, body and animal venom) of the Kemuja Malay community. The purpose of this study was to find out and describe the macro and micro aspects of the guarding mantra (garden, body, and animal venom) of the Kemuja Malay community. Keywords : Oral Literature, Guard Spells, Macro, Micro

ANIMAL VENOM FOR TREATING BREAST CANCER

Breast cancer is one of the most common malignancies found in women and is associated with increased mortality in advanced disease. With the use of improved screening techniques and improvisation in treatment, 89% of the women diagnosed with breast cancer will survive 5 years from diagnosis. Despite significant advancement in the diagnosis and treatment of breast cancer, many patients succumb to this disease. The elucidation of aberrant signaling pathways that lead to breast cancer should help develop more effective therapeutic strategies. This review focuses on the different targets of breast cancer and how they can be acted upon by different animal venoms

Antiviral activity of animal venom peptides and related compounds

Viruses exhibit rapid mutational capacity to trick and infect host cells, sometimes assisted through virus-coded peptides that counteract host cellular immune defense. Although a large number of compounds have been identified as inhibiting various viral infections and disease progression, it is urgent to achieve the discovery of more effective agents. Furthermore, proportionally to the great variety of diseases caused by viruses, very few viral vaccines are available, and not all are efficient. Thus, new antiviral substances obtained from natural products have been prospected, including those derived from venomous animals. Venoms are complex mixtures of hundreds of molecules, mostly peptides, that present a large array of biological activities and evolved to putatively target the biochemical machinery of different pathogens or host cellular structures. In addition, non-venomous compounds, such as some body fluids of invertebrate organisms, exhibit antiviral activity. This review provides a panorama of peptides described from animal venoms that present antiviral activity, thereby reinforcing them as important tools for the development of new therapeutic drugs

Animal venoms in the production of medicines

Introduction: Toxicity of animal venom is a defense mechanism known to mankind for centuries for many species of animals. We are still learning the possibilities of its use as a substance with therapeutic potential, because on its basis several drugs known and commonly used in medicine were created. Aim of the study: To review research on the known and potential use of animal toxins as medicinal substances. Material and method: Standard criteria were used to review literature data. Search for articles in the PubMed database was carried out using the following keywords: animal venoms, medical use, snake venoms, drugs from venom, Description of the state of knowledge: Animal venoms have been used by man since ancient times. Due to many different mechanisms, they are able to modulate the tissue functions of the human body. In the form of already known drugs have an effect, among others. on the cardiovascular, nervous and circulatory systems. Research is also being carried out on their use in the treatment of other diseases such as autoimmune diseases and cancer. Summary: Due to the variety of substances in animal venom and their therapeutic properties, it is possible to use them for the production of medicines. However, many studies are needed to better understand them

Modern venomics – Current insights, novel methods and future perspectives in biological and applied animal venom research

Venoms have evolved >100 times in all major animal groups, and their components, known as toxins, have been fine-tuned over millions of years into highly effective biochemical weapons. There are many outstanding questions on the evolution of toxin arsenals, such as how venom genes originate, how venom contributes to the fitness of venomous species, and which modifications at the genomic, transcriptomic, and protein level drive their evolution. These questions have received particularly little attention outside of snakes, cone snails, spiders, and scorpions. Venom compounds have further become a source of inspiration for translational research using their diverse bioactivities for various applications. We highlight here recent advances and new strategies in modern venomics and discuss how recent technological innovations and multi-omic methods dramatically improve research on venomous animals. The study of genomes and their modifications through CRISPR and knockdown technologies will increase our understanding of how toxins evolve and which functions they have in the different ontogenetic stages during the development of venomous animals. Mass spectrometry imaging combined with spatial transcriptomics, in situ hybridization techniques, and modern computer tomography gives us further insights into the spatial distribution of toxins in the venom system and the function of the venom apparatus. All these evolutionary and biological insights contribute to more efficiently identify venom compounds, which can then be synthesized or produced in adapted expression systems to test their bioactivity. Finally, we critically discuss recent agrochemical, pharmaceutical, therapeutic, and diagnostic (so-called translational) aspects of venoms from which humans benefit

Modern venomics--Current insights, novel methods, and future perspectives in biological and applied animal venom research

Venoms have evolved >100 times in all major animal groups, and their components, known as toxins, have been fine-tuned over millions of years into highly effective biochemical weapons. There are many outstanding questions on the evolution of toxin arsenals, such as how venom genes originate, how venom contributes to the fitness of venomous species, and which modifications at the genomic, transcriptomic, and protein level drive their evolution. These questions have received particularly little attention outside of snakes, cone snails, spiders, and scorpions. Venom compounds have further become a source of inspiration for translational research using their diverse bioactivities for various applications. We highlight here recent advances and new strategies in modern venomics and discuss how recent technological innovations and multi-omic methods dramatically improve research on venomous animals. The study of genomes and their modifications through CRISPR and knockdown technologies will increase our understanding of how toxins evolve and which functions they have in the different ontogenetic stages during the development of venomous animals. Mass spectrometry imaging combined with spatial transcriptomics, in situ hybridization techniques, and modern computer tomography gives us further insights into the spatial distribution of toxins in the venom system and the function of the venom apparatus. All these evolutionary and biological insights contribute to more efficiently identify venom compounds, which can then be synthesized or produced in adapted expression systems to test their bioactivity. Finally, we critically discuss recent agrochemical, pharmaceutical, therapeutic, and diagnostic (so-called translational) aspects of venoms from which humans benefit.This work is funded by the European Cooperation in Science and Technology (COST, www.cost.eu) and based upon work from the COST Action CA19144 – European Venom Network (EUVEN, see https://euven-network.eu/). This review is an outcome of EUVEN Working Group 2 (“Best practices and innovative tools in venomics”) led by B.M.v.R. As coordinator of the group Animal Venomics until end 2021 at the Institute for Insectbiotechnology, JLU Giessen, B.M.v.R. acknowledges the Centre for Translational Biodiversity Genomics (LOEWE-TBG) in the programme “LOEWE – Landes-Offensive zur Entwicklung Wissenschaftlich-ökonomischer Exzellenz” of Hesse's Ministry of Higher Education, Research, and the Arts. B.M.v.R. and I.K. further acknowledge funding on venom research by the German Science Foundation to B.M.v.R. (DFG RE3454/6-1). A.C., A.V., and G.Z. were supported by the European Union's Horizon 2020 Research and Innovation program through Marie Sklodowska-Curie Individual Fellowships (grant agreements No. A.C.: 896849, A.V.: 841576, and G.Z.: 845674). M.P.I. is supported by the TALENTO Program by the Regional Madrid Government (2018-T1/BIO-11262). T.H.'s venom research is funded by the DFG projects 271522021 and 413120531. L.E. was supported by grant No. 7017-00288 from the Danish Council for Independent Research (Technology and Production Sciences). N.I. acknowledges funding on venom research by the Research Fund of Nevsehir Haci Bektas Veli University (project Nos. ABAP20F28, BAP18F26). M.I.K. and A.P. acknowledge support from GSRT National Research Infrastructure structural funding project INSPIRED (MIS 5002550). G.A. acknowledges support from the Slovenian Research Agency grants P1-0391, J4-8225, and J4-2547. G.G. acknowledges support from the Institute for Medical Research and Occupational Health, Zagreb, Croatia. E.A.B.U. is supported by a Norwegian Research Council FRIPRO-YRT Fellowship No. 287462

TOXIFY: a deep learning approach to classify animal venom proteins

In the era of Next-Generation Sequencing and shotgun proteomics, the sequences of animal toxigenic proteins are being generated at rates exceeding the pace of traditional means for empirical toxicity verification. To facilitate the automation of toxin identification from protein sequences, we trained Recurrent Neural Networks with Gated Recurrent Units on publicly available datasets. The resulting models are available via the novel software package TOXIFY, allowing users to infer the probability of a given protein sequence being a venom protein. TOXIFY is more than 20X faster and uses over an order of magnitude less memory than previously published methods. Additionally, TOXIFY is more accurate, precise, and sensitive at classifying venom proteins

Development of a novel platform for high-throughput gene design and artificial gene synthesis to produce large libraries of recombinant venom peptides for drug discovery

Tese de Doutoramento em Ciências Veterinárias na Especialidade de Ciências Biológicas e BiomédicasAnimal venoms are complex mixtures of biologically active molecules that, while presenting low immunogenicity, target with high selectivity and efficacy a variety of membrane receptors. It is believed that animal venoms comprise a natural library of more than 40 million different natural compounds that have been continuously fine-tuned during the evolutionary process to disturb cellular function. Within animal venoms, reticulated peptides are the most attractive class of molecules for drug discovery. However, the use of animal venoms to develop novel pharmacological compounds is still hampered by difficulties in obtaining these low molecular mass cysteine-rich polypeptides in sufficient amounts. Here, a high-throughput gene synthesis platform was developed to produce synthetic genes encoding venom peptides. The final goal of this project is the production of large libraries of recombinant venom peptides that can be screened for drug discovery. A robust and efficient Polymerase Chain Reaction (PCR) methodology was refined to assemble overlapping oligonucleotides into small artificial genes (< 500 bp) with high-fidelity. In addition, two bioinformatics tools were constructed to design multiple optimized genes (ATGenium) and overlapping oligonucleotides (NZYOligo designer), in order to allow automation of the high-throughput gene synthesis platform. The platform can assemble 96 synthetic genes encoding venom peptides simultaneously, with an error rate of 1.1 mutations per kb. To decrease the error rate associated with artificial gene synthesis, an error removal step using phage T7 endonuclease I was designed and integrated into the gene synthesis methodology. T7 endonuclease I was shown to be highly effective to specifically recognize and cleave DNA mismatches allowing a dramatically reduction of error frequency in large synthetic genes, from 3.45 to 0.43 errors per kb. Combining the knowledge acquired in the initial stages of the work, a comprehensive study was performed to investigate the influence of gene design, presence of fusion tags, cellular localization of expression, and usage of Tobacco Etch Virus (TEV) protease for tag removal, on the recombinant expression of disulfide-rich venom peptides in Escherichia coli. Codon usage dramatically affected the levels of recombinant expression in E. coli. In addition, a significant pressure in the usage of the two cysteine codons suggests that both need to be present at equivalent levels in genes designed de novo to ensure high levels of expression. This study also revealed that DsbC was the best fusion tag for recombinant expression of disulfide-rich peptides, in particular when expression of the fusion peptide was directed to the bacterial periplasm. TEV protease was highly effective for efficient tag removal and its recognition sites can tolerate all residues at its C-terminal, with exception of proline, confirming that no extra residues need to be incorporated at the N-terminus of recombinant venom peptides. This study revealed that E. coli is a convenient heterologous host for the expression of soluble and potentially functional venom peptides. Thus, this novel high-throughput gene synthesis platform was used to produce ~5,000 synthetic genes with a low error rate. This genetic library supported the production of the largest library of recombinant venom peptides constructed until now. The library contains 2736 animal venom peptides and it is presently being screened for the discovery of novel drug leads related to different diseases.RESUMO - Desenvolvimento de uma nova plataforma de alta capacidade para desenhar e sintetizar genes artificiais, para a produção de péptidos venómicos recombinantes - Os venenos animais são misturas complexas de moléculas biologicamente activas que se ligam com elevada selectividade e eficácia a uma grande variedade de receptores de membrana. Embora apresentem baixa imunogenicidade, os venenos podem afectar a função celular actuando ao nível dos seus receptores. Actualmente, pensa-se que os venenos de animais constituam uma biblioteca natural de mais de 40 milhões de moléculas diferentes que têm sido continuamente aperfeiçoadas ao longo do processo evolutivo. Tendo em conta a composição dos venenos, os péptidos reticulados são a classe mais atractiva de moléculas com interesse farmacológico. No entanto, a utilização de venenos para o desenvolvimento de novos fármacos está limitada por dificuldades em obter estas moléculas em quantidades adequadas ao seu estudo. Neste trabalho desenvolveu-se uma plataforma de alta capacidade para a síntese de genes sintéticos codificadores de péptidos venómicos, com o objectivo de produzir bibliotecas de péptidos venómicos recombinantes que possam ser rastreadas para a descoberta de novos medicamentos. Com o objectivo de sintetizar genes pequenos (< 500 pares de bases) com elevada fidelidade e em simultâneo, desenvolveu-se uma metodologia de PCR (polymerase chain reaction) robusta e eficiente, que se baseia na extensão de oligonucleótidos sobrepostos. Para possibilitar a automatização da plataforma de síntese de genes, foram construídas duas ferramentas bioinformáticas para desenhar simultaneamente dezenas a milhares de genes optimizados para a expressão em Escherichia coli (ATGenium) e os respectivos oligonucleótios sobrepostos (NZYOligo designer). Esta plataforma foi optimizada para sintetizar em simultâneo 96 genes sintéticos, tendo-se obtido uma taxa de erro de 1.1 mutações por kb de DNA sintetizado. A fim de diminuir a taxa de erro associada à produção de genes sintéticos, desenvolveu-se um método para remoção de erros utilizando a enzima T7 endonuclease I. A enzima T7 endonuclease I mostrou-se muito eficaz no reconhecimento e clivagem de moléculas DNA que apresentam emparelhamentos incorrectos, reduzindo drasticamente a frequência de erros identificados em genes grandes, de 3.45 para 0.43 erros por kb de DNA sintetizado. Investigou-se também a influência do desenho dos genes, da presença de tags de fusão, da localização celular da expressão e da actividade da protease Tobacco Etch Virus (TEV) para a remoção eficiente de tags, na expressão de péptidos venómicos ricos em cisteínas em E. coli. A utilização de codões meticulosamente escolhidos afectou drasticamente os níveis de expressão em E. coli. Para além disso, os resultados mostram que existe uma pressão significativa no uso dos dois codões que codificam para o resíduo cisteína, o que sugere que ambos os codões têm de estar presentes, em níveis equivalentes, nos genes que foram desenhados e optimizados para garantir elevados níveis de expressão. Este trabalho indicou também que o tag de fusão DsbC foi o mais apropriado para a expressão eficiente de péptidos venómicos ricos em cisteínas, particularmente quando os péptidos recombinantes foram expressos no periplasma bacteriano. Confirmou-se que a protease TEV é eficaz na remoção de tags de fusão, podendo o seu local de reconhecimento conter quaisquer aminoácidos na extremidade C-terminal, com excepção da prolina. Desta forma, verificou-se não ser necessário incorporar qualquer aminoácido extra na extremidade N-terminal dos péptidos venómicos recombinantes. Reunindo todos os resultados, verificou-se que a E. coli é um hospedeiro adequado para a expressão, na forma solúvel, de péptidos venómicos potencialmente funcionais. Por último, foram produzidos, com uma taxa de erro reduzida, ~5000 genes sintéticos codificadores de péptidos venómicos utilizando a nova plataforma de elevada capacidade para a síntese de genes aqui desenvolvida. A nova biblioteca de genes sintéticos foi usada para produzir a maior biblioteca de péptidos venómicos recombinantes construída até agora, que inclui 2736 péptidos venómicos. Esta biblioteca recombinante está presentemente a ser rastreada com o objectivo de descobrir novas drogas com interesse para a saúde humana

From foe to friend: using animal toxins to investigate ion channel function

Ion channels are vital contributors to cellular communication in a wide range of organisms, a distinct feature that renders this ubiquitous family of membrane-spanning proteins a prime target for toxins found in animal venom. For many years, the unique properties of these naturally-occurring molecules have enabled researchers to probe the structural and functional features of ion channels and to define their physiological roles in normal and diseased tissues. To illustrate their considerable impact on the ion channel field, this review will highlight fundamental insights into toxin-channel interactions as well as recently developed toxin screening methods and practical applications of engineered toxins

Evaluation of the lethal potency of scorpion and snake venoms and comparison between intraperitoneal and intravenous injection routes.

International audienceScorpion stings and snake bites are major health hazards that lead to suffering of victims and high mortality. Thousands of injuries associated with such stings and bites of venomous animals occur every year worldwide. In North Africa, more than 100,000 scorpion stings and snake bites are reported annually. An appropriate determination of the 50% lethal doses (LD50) of scorpion and snake venoms appears to be an important step to assess (and compare) venom toxic activity. Such LD50 values are also commonly used to evaluate the neutralizing capacity of specific anti-venom batches. In the present work, we determined experimentally the LD50 values of reference scorpion and snake venoms in Swiss mice, and evaluated the influence of two main venom injection routes (i.e., intraperitoneal (IP) versus intravenous (IV)). The analysis of experimental LD50 values obtained with three collected scorpion venoms indicates that Androctonus mauretanicus (Am) is intrinsically more toxic than Androctonus australis hector (Aah) species, whereas the latter is more toxic than Buthus occitanus (Bo). Similar analysis of three representative snake venoms of the Viperidae family shows that Cerastes cerastes (Cc) is more toxic than either Bitis arietans (Ba) or Macrovipera lebetina (Ml) species. Interestingly, the venom of Elapidae cobra snake Naja haje (Nh) is far more toxic than viper venoms Cc, Ml and Ba, in agreement with the known severity of cobra-related envenomation. Also, our data showed that viper venoms are about three-times less toxic when injected IP as compared to IV, distinct from cobra venom Nh which exhibited a similar toxicity when injected IP or IV. Overall, this study clearly highlights the usefulness of procedure standardization, especially regarding the administration route, for evaluating the relative toxicity of individual animal venoms. It also evidenced a marked difference in lethal activity between venoms of cobra and vipers, which, apart from the nature of toxins, might be attributed to the rich composition of high molecular weight enzymes in the case of viper venoms