ALTEX最新文献

{"title":"Third Virtual Summer School 3Rs for ONE Science: Alternative methods: From complexity to predictivity.","authors":"Francesca Caloni, Olga Introzzi, Alessandra Caccianiga, Leonora Buzanska, Arno C Gutleb, Helena Kándárova, Laura Ceriotti, Giulia Ranaldi, Hassan Rashidi, Doris Wilflingseder","doi":"10.14573/altex.2207112","DOIUrl":"https://doi.org/10.14573/altex.2207112","url":null,"abstract":"710 a multiple time exposure. This is the first test method submitted for regulatory acceptance to OECD for discriminating on its own the three UN GHS categories; it was adopted as OECD TG 492B in June 2022. The potential to directly identify eye irritants is important for the cosmetic industry as well as for the pharmaceutical, chemical, and pesticide sectors and will improve and speed up their safety assessment. Arno C. Gutleb, Environmental Research and Innovation (ERIN) Department, Luxembourg Institute of Science and Technology, presented “Necessary or unnecessary complexity of in vitro lung models?” The classical approach for cell culture is that in vitro models are based on single cell types cultured submerged in medium. In recent years, complex models using multiple cell types cultured at the air-liquid interphase have added a new dimension of complexity to in vitro models. Such complex models need careful characterization of the properties of the cell types, especially when culture conditions change such as in co-culture. The similarities and discrepancies among 3D-in vitro models, human tissue and animal models used in the past need to be understood. Overall, models should be as simple as possible and as complex as necessary to mimic physiological responses. Hassan Rashidi, NIHR Great Ormond Street Hospital Biomedical Research Centre, UCL Great Ormond Street Institute of Child Health, University College London, presented a lecture entitled “Human pluripotent stem cell-derived hepatocyte-like cells as a tool to predict drug-induced liver toxicity”. The use of primary human hepatocytes (PHHs) is considered the gold standard in vitro model for drug toxicity testing to predict drug-induced liver toxicity (DILI) (Gomez-Lechon et al., 2014). However, their scarcity and transient ex vivo phenotype limit their use (Lauschke et al., 2016). A variety of alternative 2-dimensional (2D) models have been developed, but none has been able to predict DILI more accurately. Recent advances in the generation of 3D liver organoids and the development of microfluidic platforms, including liver-on-chip and human-on-chip, have opened new avenues to develop more sophisticated in vitro platforms to predict DILI more accurately. Recently developed tools were discussed. Giulia Ranaldi, Food and Nutrition Research Centre, Council for Agricultural Research and Economics, CREA-AN, Rome gave a presentation entitled “Application of Confocal Laser ScanThe international Third Virtual Summer School1 was focused on the application and predictivity of complex methods and approaches in different disciplines of science. The event, held on June 8-9, 2022 and chaired by Francesca Caloni, Università degli Studi di Milano, was attended by young researchers from all over the world. Francesca Caloni, opened the summer school, emphasizing that new approach methodologies (NAMs) are useful predictive and investigative tools for a multifaceted science with a global perspective. Helena ","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":" ","pages":"710-711"},"PeriodicalIF":5.6,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40659550","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"No more secrets: Enhancing transparency in animal research - Animalfree Research Forum 2021.","authors":"Miriam A Zemanova, Silvia Frey","doi":"10.14573/altex.2111291","DOIUrl":"https://doi.org/10.14573/altex.2111291","url":null,"abstract":"","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":" ","pages":"154"},"PeriodicalIF":5.6,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39823747","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Alternative methods and citizen science.","authors":"Francesca Caloni, Paola Fossati, Thomas Hartung, Piera Anna Martino, Gianfranco Mormino, Augusto Vitale, Isabella De Angelis","doi":"10.14573/altex.2112201","DOIUrl":"https://doi.org/10.14573/altex.2112201","url":null,"abstract":"","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":" ","pages":"159-160"},"PeriodicalIF":5.6,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39824171","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A novel method for combining outcomes with different severities or gene-level classifications.","authors":"Salomon Sand","doi":"10.14573/altex.2004212","DOIUrl":"https://doi.org/10.14573/altex.2004212","url":null,"abstract":"<p><p>Chemical risk assessment is currently based on consideration of health effects individually. The present work discusses a method for combining data by characterizing the dose-related sequence of the development of lower- to higher-order toxicological effects or the range of bioactivity observed at genomic level caused by a chemical/mixture. A reference point profile (RPP) is defined as the relation between benchmark doses for considered effects (or bioactivity measures) and a standardized severity or rank score determined for these effects. For a given dose of a chemical/mixture, the probability for exceeding the RPP can be assessed. An overall toxicological response can also be derived at the same dose by integrating contributions across all effects, with a rational for severity weighing. Conversely, dose equivalents corresponding to specified responses can be estimated. The variation in RPPs across chemicals suggests that joint consideration of effects under the proposed concept differentiates the consequence of chemical exposure, both at genomic and apical levels, to a higher extent compared to using a specific effect as a basis. This may help to refine the development of points of departure or sets of such values describing a range of health concerns. Analysis and comparison of apical and genomic RPPs, as well as consideration of functional relations between gene sets within such analyses, may aid in the transition towards a new approach method-based risk assessment paradigm that to a higher degree may require methods for combination of effect data compared to relying on specific outcomes.</p>","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":" ","pages":"480–497"},"PeriodicalIF":5.6,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39288638","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluation of an in vitro three-dimensional HepaRG spheroid model for genotoxicity testing using the high-throughput CometChip platform.","authors":"Ji-Eun Seo,&nbsp;Xiaobo He,&nbsp;Levan Muskhelishvili,&nbsp;Pritpal Malhi,&nbsp;Nan Mei,&nbsp;Mugimane Manjanatha,&nbsp;Matthew Bryant,&nbsp;Tong Zhou,&nbsp;Timothy Robison,&nbsp;Xiaoqing Guo","doi":"10.14573/altex.2201121","DOIUrl":"https://doi.org/10.14573/altex.2201121","url":null,"abstract":"<p><p>Three-dimensional (3D) culture systems are increasingly being used for genotoxicity studies due to improved cell-to-cell interactions and tissue-like structures that are limited or lacking in 2D cultures. The present study optimized a 3D culture system using metabolically competent HepaRG cells for in vitro genotoxicity testing. 3D HepaRG spheroids, formed in 96- or 384-well ultra-low attachment plates, were exposed to various concentrations of 34 test articles, including 8 direct-acting and 11 indirect-acting genotoxicants/carcinogens as well as 15 compounds that show different genotoxic responses in vitro and in vivo. DNA damage was evaluated using the high-throughput CometChip assay with con-current cytotoxicity assessment by the ATP assay in both 2D and 3D cultures. 3D HepaRG spheroids maintained a stable phenotype for up to 30 days with higher levels of albumin secretion, cytochrome P450 gene expression, and enzyme activities compared to 2D cultures. 3D spheroids also demonstrated a higher sensitivity than 2D cultures for detecting both direct- and indirect-acting genotoxicants/carcinogens, indicating a better prediction of in vivo genotoxicity responses. When DNA damage dose-response data were quantified using PROAST software, 3D spheroids generally had lower or similar benchmark dose values compared to 2D HepaRG cells and were more comparable with primary human hepatocytes. These results demonstrate that 3D models can be adapted to the CometChip technology for high-throughput genotoxicity testing and that 3D HepaRG spheroids may be used as a reliable and pragmatic in vitro approach to better support the hazard identification and risk assessment of potential human genotoxic carcinogens.</p>","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":" ","pages":"583-604"},"PeriodicalIF":5.6,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40474586","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ready for regulatory use: NAMs and NGRA for chemical safety assurance.","authors":"Paul L Carmichael,&nbsp;Maria T Baltazar,&nbsp;Sophie Cable,&nbsp;Stella Cochrane,&nbsp;Matthew Dent,&nbsp;Hequn Li,&nbsp;Alistair Middleton,&nbsp;Iris Muller,&nbsp;Georgia Reynolds,&nbsp;Carl Westmoreland,&nbsp;Andrew White","doi":"10.14573/altex.2204281","DOIUrl":"https://doi.org/10.14573/altex.2204281","url":null,"abstract":"<p><p>New approach methodologies (NAMs) that do not use experimental animals are, in certain settings, entirely appropriate for assuring the safety of chemical ingredients, although regulatory adoption has been slow. In this opinion article we discuss how scientific advances that utilize NAMs to certify systemic safety are available now and merit broader acceptance within the framework of next generation risk assessments (NGRA).</p>","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":" ","pages":"359–366"},"PeriodicalIF":5.6,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40477651","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A cell-based alternative to the mouse potency assay for pharmaceutical type E botulinum antitoxins.","authors":"Eran Diamant,&nbsp;Amram Torgeman,&nbsp;Eyal Epstein,&nbsp;Adva Mechaly,&nbsp;Alon Ben David,&nbsp;Lilach Levin,&nbsp;Arieh Schwartz,&nbsp;Eyal Dor,&nbsp;Meni Girshengorn,&nbsp;Ada Barnea,&nbsp;Ohad Mazor,&nbsp;Ran Zichel","doi":"10.14573/altex.2105251","DOIUrl":"https://doi.org/10.14573/altex.2105251","url":null,"abstract":"<p><p>The pharmacopeia mouse neutralization assay (PMNA) is the standard method for determining the potency of phar­maceutical botulinum antitoxins. However, a PMNA requires a large number of mice, and, thus, an alternative in vitro method to replace it is needed. Herein, we developed an in vitro SiMa cell line-based neutralization assay (SBNA), compatible with a PMNA design, for therapeutic antitoxins against type E botulinum neurotoxin (BoNT/E). The SBNA measures the residual cellular activity of BoNT/E following antitoxin neutralization in the SiMa lysate using a specific quantitative sandwich ELISA for its cleaved cellular target protein SNAP-25. The potencies of different pharmaceutical antitoxin preparations were determined by applying two different quantification approaches: (1) a cutoff value, in accor­dance with the pharmacopeia concept, and (2) nonlinear regression of a standard curve generated by serial dilutions of a standard antitoxin. Both approaches achieved accurate potencies compared to the PMNA (average %RE of ~16%). Furthermore, the SBNA was able to determine in vitro, for the first time, the accurate neutralizing activity (%RE ≤ 20) of next-generation equine and rabbit therapeutic antitoxins. Collectively, a high correlation between SBNA and PMNA results was obtained for all antitoxin preparations (r = 0.99, P < 0.0001 for the standard curve approach, and r = 0.97, p < 0.0001 for the cutoff approach). In conclusion, the SBNA can potentially replace the PMNA and markedly reduce the need for laboratory animals for the approval of botulinum antitoxin preparations.</p>","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":" ","pages":"113-122"},"PeriodicalIF":5.6,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39906868","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engagement of scientists with the public and policymakers to promote alternative methods.","authors":"Sonja Von Aulock,&nbsp;Francois Busquet,&nbsp;Paul Locke,&nbsp;Kathrin Herrmann,&nbsp;Thomas Hartung","doi":"10.14573/altex.2209261","DOIUrl":"https://doi.org/10.14573/altex.2209261","url":null,"abstract":"<p><p>Scientists are usually good at teaching, sometimes even to lay audiences. But communicating with journalists, activists, or policymakers can be a different story - hesitancy to make mistakes as well as the temptation to disproportionally promote one's own case come into play. The multitude of social media and other web-based outlets has diversified and accelerated the communication of science. Real-time reactions, sharing of data, tools and results, increasing invitation of personal opinion, demand for transparency, political correctness, and loss of trust in experts are challenges to researchers in general. The field of alternatives to animal testing is more political and important to lay audiences than many others, so its scientists must be especially aware of these challenges. Public engagement offers the opportunity to form community and create wide support for non-animal research and its implementation. This requires scientists to step out of the ivory tower of higher education and engage with diverse interest groups by outreach activities, interviews, and press releases, etc. by employing tailored communication.</p>","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":" ","pages":"543-559"},"PeriodicalIF":5.6,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40659549","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Application of high-throughput transcriptomics for mechanism-based biological read-across of short-chain carboxylic acid analogues of valproic acid.","authors":"Nanette G Vrijenhoek,&nbsp;Matthias M Wehr,&nbsp;Steven J Kunnen,&nbsp;Lukas S Wijaya,&nbsp;Giulia Callegaro,&nbsp;Martijn J Moné,&nbsp;Sylvia E Escher,&nbsp;Bob Van de Water","doi":"10.14573/altex.2107261","DOIUrl":"https://doi.org/10.14573/altex.2107261","url":null,"abstract":"<p><p>Chemical read-across is commonly evaluated without specific knowledge of the biological mechanisms leading to observed adverse outcomes in vivo. Integrating data that indicate shared modes of action in humans will strengthen read-across cases. Here we studied transcriptomic responses of primary human hepatocytes (PHH) to a large panel of carboxylic acids to include detailed mode-of-action data as a proof-of-concept for read-across in risk assessment. In rodents, some carboxylic acids, including valproic acid (VPA), are known to cause hepatic steatosis, whereas others do not. We investigated transcriptomics responses of PHHs exposed for 24 h to 18 structurally different VPA analogues in a concentration range to determine biological similarity in relation to in vivo steatotic potential. Using a targeted high-throughput screening assay, we assessed the differential expression of ~3,000 genes covering relevant biological pathways. Differentially expressed gene analysis revealed differences in potency of carboxylic acids, and expression patterns were highly similar for structurally similar compounds. Strong clustering occurred for steatosis-positive versus steatosis-negative carboxylic acids. To quantitatively define biological read-across, we combined pathway analysis and weighted gene co-expression network analysis. Active carboxylic acids displayed high similarity in gene network modulation. Importantly, free fatty acid synthesis modulation and stress pathway responses are affected by active car­boxylic acids, providing coherent mechanistic underpinning for our findings. Our work shows that transcriptomic analysis of cultured human hepatocytes can reinforce the prediction of liver injury outcome based on quantitative and mechanistic biological data and support its application in read-across.</p>","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":" ","pages":"207–220"},"PeriodicalIF":5.6,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39830262","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The nearest neighbor nuclei method to objectify analysis of pertussis toxin-induced clustering.","authors":"Marieke E Hoonakker,&nbsp;Ed Remarque,&nbsp;Jennifer Veth,&nbsp;Arjen Sloots,&nbsp;Jeffrey J Bajramovic","doi":"10.14573/altex.2012171","DOIUrl":"https://doi.org/10.14573/altex.2012171","url":null,"abstract":"<p><p>The in vivo histamine sensitization test (HIST) has historically been performed to guarantee the safety of acellular per­tussis vaccine batches. Non-compliance of batches is primarily associated with the presence of low levels of pertussis toxin (PTx). Because of ethical, standardization and scientific reasons, a variety of alternative in vitro approaches have been studied to replace the lethal HIST. A broadly applied and partially accepted method is the CHO cell clustering test, which is based on the clustered growth pattern of CHO cells when exposed to minute amounts of PTx. One of the major hurdles for global application of the CHO clustering test is the manual assessment of the clusters, which is associated with suboptimal reproducibility of test outcomes and is time-consuming. Here, various parameters of CHO cell nuclei were evaluated in search for a reliable, objective read-out parameter. We demonstrate that the distance between each nucleus and its nearest neighbor (3N method) is the most suitable parameter to assess clustered cell growth. This method detects 2.8 mIU PTx/mL and thereby complies with the requirement set for the sensitivity of the CHO clustering test based on visual reading. In commercial acellular pertussis vaccines spiked with PTx, the method detects 45 mIU/mL PTx, which is substantially lower than the 181-725 mIU/mL PTx detected by visual interpretation. The 3N method thus allows objective and sensitive assessment of CHO clustering and thereby encourages broad and global implementation of the in vitro test as an alternative to the HIST.</p>","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":" ","pages":"140-148"},"PeriodicalIF":5.6,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39525187","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}