Implementation of reliable methodologies allowing Reduction, Refinement, and Replacement (3Rs) of animal testing is a process that takes several decades and is still not complete. Reliable methods are essential for regulatory hazard assessment of chemicals where differences in test protocol can influence the test outcomes and thus affect the confidence in the predictive value of the organisms used as an alternative for mammals. Although test guidelines are common for mammalian studies, they are scarce for non-vertebrate organisms that would allow for the 3Rs of animal testing. Here, we present a set of 30 reporting criteria as the basis for such a guideline for Developmental and Reproductive Toxicology (DART) testing in the nematode Caenorhabditis elegans. Small organisms like C. elegans are upcoming in new approach methodologies for hazard assessment; thus, reliable and robust test protocols are urgently needed. A literature assessment of the fulfilment of the reporting criteria demonstrates that although studies describe methodological details, essential information such as compound purity and lot/batch number or type of container is often not reported. The formulated set of reporting criteria for C. elegans testing can be used by (i) researchers to describe essential experimental details (ii) data scientists that aggregate information to assess data quality and include data in aggregated databases (iii) regulators to assess study data for inclusion in regulatory hazard assessment of chemicals.
Background: In the Netherlands, general practitioners (GPs) play a key role in provider-initiated HIV testing, but opportunities for timely diagnosis are regularly missed. We implemented an educational intervention to improve HIV testing by GPs from 2015 to 2020, and observed a 7% increase in testing in an evaluation using laboratory data. The objective for the current study was to gain a deeper understanding of whether and how practices and perceptions of GPs’ HIV/sexually transmitted infection (STI) testing behaviour changed following the intervention. Methods: We performed a mixed-methods study using questionnaires and semi-structured interviews to assess self-reported changes in HIV/STI testing by participating GPs. Questionnaires were completed by participants at the end of the final educational sessions from 2017 through 2020, and participating GPs were interviewed from January through March 2020. Questionnaire data were analysed descriptively, and open question responses were categorised thematically. Interview data were analysed following thematic analysis methods. Results: In total, 101/103 participants completed questionnaires. Of 65 participants that were included in analyses on the self-reported effect of the programme, forty-seven (72%) reported it had changed their HIV/STI testing, including improved STI consultations, adherence to the STI consultation guideline, more proactive HIV testing, and more extragenital STI testing. Patients’ risk factors, patients’ requests and costs were most important in selecting STI tests ordered. Eight participants were interviewed and 15 themes on improved testing were identified, including improved HIV risk-assessment, more proactive testing for HIV/STI, more focus on HIV indicator conditions and extragenital STI testing, and tools to address HIV during consultations. However, several persistent barriers for optimal HIV/STI testing by GPs were identified, including HIV-related stigma and low perceived risk. Conclusions: Most GPs reported improved HIV/STI knowledge, attitude and testing, but there was a discrepancy between reported changes in HIV testing and observed increases using laboratory data. Our findings highlight challenges in implementation of effective interventions, and in their evaluation. Lessons learned from this intervention may inform follow-up initiatives to keep GPs actively engaged in HIV testing and care, on our way to zero new HIV infections.
While the original definition of replacement focuses on the replacement of the use of animals in science, a more contemporary definition focuses on accelerating the development and use of predictive and robust models, based on the latest science and technologies, to address scientific questions without the use of animals. The transition to animal free innovation is on the political agenda in and outside the European Union. The Beyond Animal Testing Index (BATI) is a benchmarking instrument designed to provide insight into the activities and contributions of research institutes to the transition to animal free innovation. The BATI allows participating organizations to learn from each other and stimulates continuous improvement. The BATI was modelled after the Access to Medicine Index, which benchmarks pharmaceutical companies on their efforts to make medicines widely available in developing countries. A prototype of the BATI was field-tested with three Dutch academic medical centers and two universities in 2020-2021. The field test demonstrated the usability and effectiveness of the BATI as a benchmarking tool. Analyses were performed across five different domains. The participating institutes concluded that the BATI served as an internal as well as an external stimulus to share, learn, and improve institutional strategies towards the transition to animal free innovation. The BATI also identified gaps in the development and implementation of 3R technologies. Hence, the BATI might be a suitable instrument for monitoring the effectiveness of policies. BATI version 1.0 is ready to be used for benchmarking at a larger scale.
Size measurement plays an essential role for micro-/nanoparticle characterization and property evaluation. Due to high costs, complex operation or resolution limit, conventional characterization techniques cannot satisfy the growing demand of routine size measurements in various industry sectors and research departments, e.g., pharmaceuticals, nanomaterials and food industry etc. Together with start-up SeeNano and other partners, we will develop a portable compact device to measure particle size based on particle-impact electrochemical sensing technology. The main task in this project is to extend the measurement range for particles with diameters ranging from 20 nm to 20 um and to validate this technology with realistic samples from various application areas. In this project a new electrode chip will be designed and fabricated. It will result in a workable prototype including new UMEs (ultra-micro electrode), showing that particle sizing can be achieved on a compact portable device with full measuring range. Following experimental testing with calibrated particles, a reliable calibration model will be built up for full range measurement. In a further step, samples from partners or potential customers will be tested on the device to evaluate the application feasibility. The results will be validated by high-resolution and mainstream sizing techniques such as scanning electron microscopy (SEM), dynamic light scattering (DLS) and Coulter counter.
The missing link in diagnostic testing for rheumatoid arthritis (RA) is an agglutination assay, easy to perform and dedicated to decentralized testing. Approximately 75% of RA patients produce autoantibodies to citrullinated proteins (ACPA), which can be detected using an agglutination-based diagnostic test. Such a diagnostic test will be cheaper, less laborious and faster than current tests and does not require sophisticated equipment. Novio Catalpa is developing this alternative test for ACPA in collaboration with Radboud University. To develop this test, specifically tagged and citrullinated nanobodies are needed, but the production is still challenging. Current methods for the production of ACPA diagnostics involve chemical synthesis, in which a variety of toxic chemicals are used in each step. The alternative assay involves nanobodies fused with RA-biomarker target entities, which can be completely produced by ‘green synthesis’ in the yeast Pichia pastoris using the expertise of HAN BioCentre. The yeast P. pastoris has proven to be able to produce nanobodies and is a fast and cost-effective platform that often results in high protein yields. Goal of the project is therefore to determine the feasibility and best green route to produce purified nanobodies tagged with citrullinated ACPA targets that can be used for developing an agglutination assay for RA. P. pastoris does not produce endogenous PAD enzymes which are needed for citrullination of the nanobodies in order to be able to use it in a RA agglutination test. Therefore, PAD enzymes from other sources need to be tested and applied. The project results will be directly used by Novio Catalpa to further develop the innovative test for RA. This project will contribute to and finally result in a single-step agglutination assay suitable for both point-of-care testing and automation in clinical laboratories.
Lab-on-a-Chip (LOC) devices worden op dit moment voor het overgrote deel in de wetenschap toegepast, in laboratoria met apparatuur die daar gebruikelijk is. LOC-devices hebben echter een grote potentie buiten de laboratoria, in bijvoorbeeld Point-Of-Care Testing situaties in de gezondheidszorg, en het verrichten van metingen ter plaatse t.b.v. de voedsel- en watersector. Ook in andere applicatievelden zoals forensisch recherchewerk, en environmental monitoring kunnen LOC devices een enorme impact hebben. Voorwaarde is wel dat zij hanteerbaarder, goedkoper en robuuster worden, en qua productiewijze minder gecompliceerd. De ontwikkeling van de huidige LOCs vereist een aanzienlijke investering om tot een werkend en betrouwbaar product te komen. Traditioneel wordt hierbij gebruikt gemaakt van kostbare fotolithografische technieken in combinatie met chemisch etsen in zogenoemde cleanrooms. Dit maakt de initiële drempel voor nieuwe toepassingen extreem hoog. Daarnaast is de apparatuur om deze chips aan te sluiten, aan te sturen en een meting te verrichten in veel gevallen zeer specialistisch en daardoor blijft de inzet van deze technologie beperkt tot een goed geoutilleerd lab. Met het LOC2Use project van Saxion lectoraten NanoBio en Industrial Design in samenwerking met het bedrijfsleven wordt beoogd om de LOC-technologie uit het lab te halen, door technologische toepassingen te ontwikkelen op basis van meer toegankelijke technologie. Vertrekpunt voor de toepassingen zijn de use-cases, die in samenwerking met stakeholders en MKB-partners in verschillende toepassingsgebieden worden opgesteld. Beoogd resultaat is een catalogus vol met getoetste technische oplossingen, gekoppeld aan de eisen uit de use-cases. Aan de hand van de gevonden technologische oplossingen worden een viertal illustratieve cases uitgewerkt. En worden er uit de catalogus een aantal bruikbare technologische bouwstenen geselecteerd voor rapid prototyping en kleine series microfluidic devices in een FabLab omgeving, een eerste stap in de richting van een LOC-fieldlab. Met dit project wordt LOC betaalbaar en breder toepasbaar voor het MKB.
