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

Investigation of immunomodulatory efficacy of beta-glucan and cephalaria spp. saponin formulation

[No Abstract Available

Interleukin-33 Levels in Gingival Crevicular Fluid, Saliva, or Plasma Do Not Differentiate Chronic Periodontitis

WOS: 000301620500013PubMed ID: 21859321Background: This study investigates whether gingival crevicular fluid (GCF), saliva, and plasma levels of interleukin-33 (IL-33) can differentiate individuals with chronic periodontitis from individuals with healthy periodontium. Methods: GCF, whole saliva, and plasma samples together with full-mouth clinical periodontal recordings were obtained from 32 otherwise healthy, non-smoker chronic periodontitis individuals and 25 systemically and periodontally healthy, non-smoker individuals. IL-33 levels in the biofluid samples were determined by enzyme-linked immunosorbent assay. Data were tested statistically by Mann-Whitney U test. Results: The GCF concentrations of IL-33 were significantly lower in chronic periodontitis individuals than in healthy individuals (P 0.05). The salivary and plasma contrations of IL-33 were indifferent in the two study groups (P>0.05). Conclusions: According to the present findings, the GCF, saliva or plasma levels of IL-33 could not differentiate chronic periodontitis individuals and periodontally healthy individuals. Larger-scale intervention studies may better clarify this issue. J Periodontot 2012;83:362-368

Salivary and Plasma Levels of Toll-Like Receptor 2 and Toll-Like Receptor 4 in Chronic Periodontitis

WOS: 000291828300012PubMed ID: 21138350Background: This cross-sectional study was planned to investigate whether patients with chronic periodontitis exhibit different salivary or plasma concentrations of Toll-like receptor (TLR) 2 and TLR4 compared to subjects who are clinically healthy. Methods: Whole saliva and plasma samples were obtained and full-mouth clinical periodontal measurements were recorded from 22 otherwise healthy patients with chronic periodontitis and 21 systemically and periodontally healthy control subjects. Salivary and plasma TLR2 and TLR4 levels were determined by enzyme-linked immunoassays. Data were tested statistically using Mann-Whitney U test. Results: The healthy group exhibited significantly lower values in all clinical measurements (P 0.05). The patients with chronic periodontitis exhibited significantly higher salivary TLR4 (P <0.01) and plasma TLR2 and TLR4 levels (P<0.05). Conclusion: The present findings support a hypothesis that inflammation increases expression of TLRs which leads to an increased detection of TLRs in saliva and plasma, which could be useful as a diagnostic test for periodontal diseases. J Periodontol 2011;82:878-884

Investigation of immunomodulation potential of dental pulp stem cells on infectious bronchitis virus infected lung alveolar epithelial cells

[No Abstract Available

In Vitro Cytotoxic and Anti-inflammatory Activities of Tanacetum argenteum (Lam.) Willd. subsp argenteum Extract

WOS: 000417379400003Objectives: The objective of this study was to examine the anti-inflammatory and cytotoxic potential of n-hexane, ethyl acetate, and methanolic extracts of Tanacetum argenteum subsp. argenteum. Materials and Methods: Tanacetum L. is the third largest genus of Asteraceae family and is represented by 60 taxa in Turkish flora. Sesquiterpene lactones and pyrethrins are the main chemical groups of the genus. T. argenteum subsp. argenteum is an endemic taxa that is distributed in the Central and South Anatolia. Results: In vitro anti-inflammatory activity was assayed using iNOS and NF-kappa B inhibition tests on RAW264.7 and HeLa cells. The cytotoxic activities were tested against ten cell lines using MTT assays. Conclusion: As a result, the n-hexane extract was found more active than the positive control parthenolide in iNOS test (IC50 : 0.627 +/- 0.16 mu g/mL) and cytotoxic experiments against PC3 and MPANC-96 cell lines (IC50 : 2.85 +/- 0.51 mu g/mL and 5.35 +/- 1.24 mu g/mL, respectively)

Production of Monoclonal Antibody Against Salmonella H: g,m Flagellar Antigen and Potential Diagnostic Application

WOS: 000283852200006PubMed ID: 21050043In this study, the flagella antigen was detached at high speed by shaking vigorously with glass pearl beads, from Salmonella enteritidis in a yield of 3.9 mg/mL(-1) after being concentrated with polyethylene glycol (PEG). A monoclonal antibody (MAb), designated D7 clone, was generated from Balb/c mice immunized with Salmonella enteritidis flagella using the conventional hybridoma method. The D7 clone secreting MAb was classified as IgG2a isotype. ELISA analyses of D7 MAb to Salmonella-specific flagella demonstrated that the antibody reacted with H: m. However, Western immunoblot analyses of D7 clone appear to be secreting heavy chain of IgG2a antibody, which was eligible for the diagnosis of Salmonella enteritidis

Metabolic Activity and Monoclonal Antibody Production of Salmonella Enteritidis O and H Antigen Specific Hybridoma Cells in Static Culture

WOS: 000290030800012PubMed ID: 21529293The aim of the present work was to study the kinetics of two hybridomas that produce monoclonal antibody against Salmonella Enteritidis O (5A8) and H (D7) antigen. The hybridomas originated from the Ag8x653 (5A8) and Sp2/O (D7) myeloma cell lines. The relationship between the uptake of glucose and glutamine and the release of the lactate and ammonia and monoclonal antibody production into hybridoma growth were investigated in static culture with serum-containing DMEM/F:12 medium for the determination of pilot-production strategies of the hybridomas. Results showed that glucose and glutamine concentrations were reduced, with an increase in ammonia and lactate concentration in the culture medium. The hybridoma cell line 5A8 has shown lower metabolic activities compared with D7, whereas its monoclonal antibody productivity was found to be two-fold higher than the D7. MAb production by the hybridoma cell line 5A8 seems promising, considering the moderate level of productivity compared to that found in the literature.Ege UniversityEge University [06MUH05]This work was supported in part by a grant from the Research Fund of Ege University (06MUH05). We thank Melis Kuban for helpful discussions and a critical review of this manuscript

Production of Monoclonal Antibodies in a Mouse Model via Lipopolysaccharide Conjugates with Synthetic Polymers Specific to Salmonella Enteritidis O Antigen

WOS: 000284837300011PubMed ID: 20707736Monoclonal antibodies (Mabs) specific for lipopolysaccharide (LPS) of Salmonella Enteritidis were evaluated in a model of LPS conjugated synthetic polymer immunization of Balb/c mice by conventional hybridoma method. Nine hybridoma cell lines were determined as antibody positive against LPS. The clone 5A8 secreting the highest antibody was selected for further characterization. Evaluated results indicate that the synthetic polymer can be used as an effective adjuvant in immunization with LPS, because the 5A8 Mab were obtained using synthetic polymer as an adjuvant. 5A8 Mab was classified as IgG2a isotype by antibody capture enzyme-linked immunosorbent assay. The reactivity of the Mab against lipid A and different LPS of Salmonella were investigated using an indirect enzyme-linked immunosorbent assay. Mab presented a wide spectrum of reactivity, coupling with antigens against Salmonella. The hybridoma 5A8 determined in this study has a great potential to be used in the development of diagnostic, prophylactic, and therapeutic agents specific for Salmonella Enteritidis and Salmonella LPS.Ege UniversityEge University [2002FEN09]This work was financially supported in part by a grant from Research Fund of Ege University (2002FEN09)