{"title":"20 Years of Providing Custom Non-Animal-Derived Antibodies - A Review.","authors":"John Cardone, Achim Knappik","doi":"10.1177/02611929251333981","DOIUrl":"https://doi.org/10.1177/02611929251333981","url":null,"abstract":"<p><p>As innovators who made non-animal-derived antibodies available to researchers starting 20 years ago, we have generated more than 60,000 antibodies during this time span. Here, we review and highlight the key challenges that hampered the adoption of research antibodies generated by phage display. We describe the advantages of this method over traditional immunisation-based methods, and provide examples of specific projects. As active participants in attesting the scientific validity of non-animal-derived antibodies, we have contributed and supported key advancements in phage display for the generation of research antibodies. Such advancements have paved the way for the scientific community to switch to animal-free antibodies at a faster pace, as a goal for the near future.</p>","PeriodicalId":55577,"journal":{"name":"Atla-Alternatives To Laboratory Animals","volume":"53 3","pages":"168-174"},"PeriodicalIF":2.4,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144042727","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Holger P Behrsing, Khalid Amin, David Allen, Joseph Hughes, McKenzie Obermok, Vivek Patel
{"title":"The Development of an Animal Product-free, Precision-cut Lung Slice Cryopreservation and Post-thaw Culture Method.","authors":"Holger P Behrsing, Khalid Amin, David Allen, Joseph Hughes, McKenzie Obermok, Vivek Patel","doi":"10.1177/02611929251336446","DOIUrl":"https://doi.org/10.1177/02611929251336446","url":null,"abstract":"<p><p>As new approach methodologies (NAMs) are increasingly explored to identify dependable and accurate non-animal alternatives to predict human toxicities, several 3-D test systems have emerged as excellent models of the human respiratory tract. Among these, human precision-cut lung slices (hPCLS) are considered highly relevant, as they contain many cell types (including key immune cells), feature small airway structures and boast native respiratory parenchymal architecture. However, a lack of long-term preservation methods has hampered the use of the hPCLS model for repeat and mainstream testing. In the current study, a range of potential methods were progressively evaluated for the optimum recovery of hPCLS after thawing and multi-week culturing. These methods featured: five different cryopreservation buffer (CB) recipes; freezing either before or after culture initiation; two culture media (based on E-199 and DMEM/F12); and two culture maintenance methods (submerged and air-liquid interface (ALI)). Endpoints used for the assessment of hPCLS culture health included the WST-8 viability assay, protein content and H&E histology of slice sections. Two of the CBs and immediate cryopreservation after slicing produced hPCLS with higher post-thaw viability. While both media recipes and culture methods maintained high slice viability for approximately 2 weeks, the use of DMEM-F12-based medium in ALI culture was superior for the 3-week cultures. Applying due diligence to hPCLS cryopreservation and post-thaw method development provides researchers with an underutilised human respiratory model. Studies making use of cryopreserved banks of normal or diseased tissues (from a diverse demographic pool of donors) can now be initiated as desired, repeated, or expanded upon to interrogate numerous aspects of physiology, toxicology and drug efficacy. These can be applied as routine screening applications or complex evaluations, including those benefitting a regulatory setting.</p>","PeriodicalId":55577,"journal":{"name":"Atla-Alternatives To Laboratory Animals","volume":"53 3","pages":"138-153"},"PeriodicalIF":2.4,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144059359","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Permutation Tests Are a Useful Alternative Approach for Statistical Hypothesis Testing in Small Sample Sizes.","authors":"Theresa Unseld, Lisa Ruckerbauer, Benjamin Mayer","doi":"10.1177/02611929251326882","DOIUrl":"10.1177/02611929251326882","url":null,"abstract":"<p><p>Results interpretation and statistical analysis of animal study data is challenging, since the sample sizes involved are usually very small. The application of frequently used approaches to statistical hypothesis testing, e.g. <i>t</i>-tests or ANOVA methods, rely on specific distributional assumptions being satisfied. It can be hard to reliably assess these assumptions in animal studies with group sizes of usually less than ten animals. Non-parametric analysis methods might be considered as an alternative, but it is well-known that these approaches have lower statistical power in some situations. Following the Three Rs principles, it would be desirable to apply a class of statistical tests that is able to deal with a small number of observations, without the need for specific distributional assumptions. Thus, in this paper, we assess the application of permutation tests which seem to be able to meet both the above requirements. The performance of these permutation tests was compared with standard statistical tests by means of four real-world data examples from animal studies. The results demonstrated that permutation tests have good computational properties, leading to the conclusion that they could be a useful alternative approach when analysing small sample size animal study data for which distributional assumptions may not hold.</p>","PeriodicalId":55577,"journal":{"name":"Atla-Alternatives To Laboratory Animals","volume":" ","pages":"130-137"},"PeriodicalIF":2.4,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143674964","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"The Prevalence of Reported Gavage Incidents in Repeated Dose Toxicity Studies Involving Rats Conducted Between 2004-2023.","authors":"Katy Taylor, Laura Rego Alvarez, Emma Grange","doi":"10.1177/02611929251334721","DOIUrl":"https://doi.org/10.1177/02611929251334721","url":null,"abstract":"<p><p>In animal-based toxicology studies, oral gavage is the predominant technique for administering compounds directly into the stomach. Recognised as a stressful procedure for animals, gavage is associated with accidents and 'gavage-related reflux' (GRR), which may lead to respiratory distress and/or death. This paper estimates the prevalence of incidents (deaths and/or non-fatal clinical signs) attributed to the gavage procedure in a representative sample of repeated dose toxicity test reports from the last 20 years, sourced from the ECHA CHEM database of REACH-registered chemicals. In 21% of the 300 reviewed studies, at least one death or clinical sign in rats was attributed to the gavage procedure in the study report. An additional 16% of these studies reported clinical signs associated with GRR in the animals, with no other adverse effects, but without attribution to the gavage procedure. Among those studies with incidents attributed to the gavage procedure, incidents were three times more common in 90-day studies than they were in 28-day studies. Incidents were also more likely to have occurred with the test substance rather than with the control, with highly viscous substances, and in more recently conducted studies. There was no association between the number of studies with a gavage-attributed incident and the strain, size or age of the rats, the dose volume or the testing of irritant substances. The current study suggests that incidents attributed to gavage are more prevalent than previously thought, and that a review of the gavage procedure is urgently needed.</p>","PeriodicalId":55577,"journal":{"name":"Atla-Alternatives To Laboratory Animals","volume":"53 3","pages":"154-167"},"PeriodicalIF":2.4,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143997083","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Expression of Concern: \"Fenugreek Seed Extract Regulates Human Umbilical Vein Endothelial Cell Angiogenesis and Proliferation via the PI3K/Akt/Cyclin D1 Pathway\".","authors":"","doi":"10.1177/02611929251334753","DOIUrl":"https://doi.org/10.1177/02611929251334753","url":null,"abstract":"","PeriodicalId":55577,"journal":{"name":"Atla-Alternatives To Laboratory Animals","volume":" ","pages":"2611929251334753"},"PeriodicalIF":2.4,"publicationDate":"2025-04-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144045716","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Spotlight on Three Rs Progress.","authors":"","doi":"10.1177/02611929251323970","DOIUrl":"10.1177/02611929251323970","url":null,"abstract":"","PeriodicalId":55577,"journal":{"name":"Atla-Alternatives To Laboratory Animals","volume":" ","pages":"67-69"},"PeriodicalIF":2.4,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143558652","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}