EURL ECVAM Recommendation on the Direct Peptide Reactivity Assay (DPRA) for Skin Sensitisation Testing

Identification of the skin sensitisation hazard of chemicals has traditionally relied on the use of animals. Progress in the development of alternative methods has been prompted by the increasing knowledge of the key biological mechanisms underlying this human health effect, as documented by the OECD's recent report summarising the key biological events leading to skin sensitisation ("Adverse Outcome Pathway" (AOP) for skin sensitisation). The molecular initiating event defined within this AOP is the covalent binding of chemicals with skin proteins. Thus peptide reactivity assays may provide valuable information in the context of integrated approaches such as Weight of Evidence (WoE) or Integrated Testing Strategies (ITS) for skin sensitisation hazard and safety assessment. Based on these considerations, EURL ECVAM coordinated a validation study on the Direct Peptide Reactivity Assay (DPRA) addressing mainly the test method’s transferability and within- and between-laboratory reproducibility. Following independent scientific peer review by the EURL ECVAM’s Scientific Advisory Committee (ESAC) and having considered the input from regulators, stakeholders, international partners and the general public, EURL ECVAM concluded that the DPRA may prove a valuable component of a WoE or ITS for skin sensitisation hazard assessment. In addition to this, the DPRA may also be able to contribute to the assessment of sensitising potency, e.g. by supporting sub-categorisation of sensitisers according to UN GHS. However it is recognised that further efforts are required to explore how DPRA data may contribute to potency assessmentJRC.I.5-Systems Toxicolog

EURL ECVAM Strategy for Replacement of Animal Testing for Skin Sensitisation Hazard Identification and Classification

In the absence of validated and regulatory accepted alternative methods, the assessment of the skin sensitisation potential of chemicals still relies on animal testing. Progress in the development of alternative methods has been prompted by the increasing knowledge on the key mechanisms of the skin sensitisation pathway, as recently documented in the OECD Adverse Outcome Pathway for skin sensitisation. Based on an analysis of the regulatory requirements for this endpoint within relevant pieces of EU chemicals legislation, EURL ECVAM has decided to focus its efforts on the development of non-animal testing strategies for skin sensitisation hazard identification and classification, including the subcategorisation of sensitisers, according to the GHS classification system. This would satisfy the majority of the regulatory requirements within the EU and would have a significant impact in terms of replacing animal experiments. This report describes the EURL ECVAM strategy for achieving this goal.JRC.I.5-Systems Toxicolog

Influencia del espesor de la junta de mortero en muros de ladrillo sometidos a esfuerzos de compresión

En este trabajo se analiza la influencia del espesor de la junta de mortero en la resistencia a compresión de la fábrica de ladrillo cerámico bajo esfuerzos de compresión uniaxial. Además se estudia el progresivo agrietamiento del material durante el proceso de carga. Se han ensayado nueve muros y nueve pilares, combinando tres espesores de junta de mortero relizando medidas con ultrasonidos, además se han realizado medidas con extensómetros y esclerómetro. Los resultados obtenidos muestran que a menor espesor de junta mayor carga de rotura y mayor valor de la velocidad de pulso ultrasónico. La medida con ultrasonidos muestra una buena correlación con las medidas extensométricas y permite una eficaz detección del agrietamiento interior del material durante el proceso de rotur

Evaluación Estructural Mediante Ultrasonidos del Efecto del Espesor de la Junta de Mortero en Muros de Fábrica de Ladrillo Cerámino Sometidos a Esfuerzo de Compresión hasta su Rotura.

En este trabajo se analiza la influencia del espesor de la junta de mortero en el comportamiento estructural de probetas de fábrica de ladrillo cerámico bajo esfuerzos de compresión uniaxial. El ensayo de compresión reproduce la forma habitual de trabajo de estas estructuras. Se han ensayado nueve muros y nueve pilares, combinando tres espesores de junta de mortero, sometidos a carga de compresión hasta rotura. Con el fin de evaluar el progresivo agrietamiento del material durante el proceso de carga, se han realizado medidas con ultrasonidos. Además se han realizado medidas con extensómetros y esclerómetro para contrastar la validez de las medidas de ultrasonidos. Los resultados obtenidos muestran que existe una correlación entre los valores de la fuerza de compresión que soportan las probetas y el tamaño del espesor de la junta: a menor espesor de junta mayor carga de rotura. Del estudio también se desprende que esta relación es más clara en los muros que en los pilares, ya que la esbeltez de los muros es mayor y aparecen efectos de pandeo. La medida con ultrasonidos muestra una buena correlación con las medidas extenso métricas y permite una eficaz detección del agrietamiento interior del material durante el proceso de rotura

Ability of non-animal methods for skin sensitisation to detect pre- and pro-haptens: Report and recommendations of an EURL ECVAM expert meeting

Significant progress has been made in the development, validation and regulatory acceptance of in chemico and in vitro test methods for skin sensitisation. Although these methods have been shown to perform relatively well (about 80% accuracy in predicting Local Lymph Node Assay (LLNA) classifications) a concern was raised on the regulatory acceptability of negative results since it was questioned whether these methods are able to predict chemicals that need to be activated to act as sensitisers. In order to inform ongoing discussions at the regulatory level in the EU, EURL ECVAM held an expert meeting on 10-11 November 2015 to analyse the extent to which in chemical and in vitro methods are able to correctly identify chemicals that need to be activated either through abiotic activation (pre-haptens) and/or through biotic (enzyme-mediated) mechanisms (pro-haptens) to acquire skin sensitisation potential. The expert group analysed a list of 127 chemicals, with available LLNA and in vitro data, 22% of which were considered to be pre- and/or pro-haptens. The pre-haptens, constituting the vast majority of chemicals requiring activation, where mostly correctly identified by both the in chemico and in vitro assays whereas the pro-haptens which represent a small subset of sensitising chemicals, were generally identified correctly by one of the cell-based assays. As a result, the expert group recommended that negative in vitro data should be accepted unless there is a compelling scientific argument that a substance is likely to be an exclusively metabolically activated pro-hapten.JRC.I.5-Systems Toxicolog

Анализ методов вибродиагностики металлорежущих станков

Цель работы - выработка рекомендаций по применению методов вибродиагностики металлорежущих станков в конкретной задаче. Объект исследования - методы и комплексы вибродиагностики металлорежущих станков. Предмет исследования – систематизация и обобщение методов вибродиагностики металлорежущих станков. Актуальность - отсутствие простой для реализации методики виброиспытаний. В процессе работы были рассмотрены различные методы вибродиагностики металлорежущих станков, сделаны предложения по применению методов вибродиагностики металлорежущих станков в каждой конкретной задаче, создана универсальная методика проведения вибродиагностики металлорежущих станков диагностическим комплексом "Виброрегистратор-М2".The aim of the work is to develop recommendations on the application of vibration diagnostics methods for metal-cutting machine tools in a specific task. The object of research is methods and complexes of vibration diagnostics of metal cutting machines. The subject of the study is the systematization and generalization of methods of vibration diagnostics of metal-cutting machines. Actuality is the absence of a simple vibration testing technique. In the course of the work various methods of vibration diagnostics of metal cutting machines were considered, suggestions were made on the application of vibration diagnostics methods for metal cutting machines in each specific task, a universal technique for performing vibration diagnostics of metal cutting machines with the Vibroregistrator-M2

ECVAM Technical Report on the Status of Alternative Methods for Cosmetics Testing (2008-2009)

The ECVAM technical report presents the progress made in the development and validation of alternative methods for the human health effects relevant to the Cosmetics Directive. It provides an update on the activities described by ECVAM in 2005 , 2006 and 2007 . The report intends to present the latest scientific and technical developments in the field during 2008-2009. As required by Directive 2003/15/EC, the seventh amendment to Directive 76/768/EEC, developments in refinement and reduction methods are also described (EU, 2003). Most successes in the development of alternative methods are in acute local toxicity and short-term testing, such as e.g. skin and eye irritation/corrosion, phototoxicity and skin penetration The test methods consuming a high number of animals, however, are in long-term testing and systemic toxicity, such as e.g. reproductive toxicity and repeated dose toxicity. In these complex fields, several research initiatives are ongoing. However full replacement approaches are still lacking.JRC.DG.I.3-In-vitro method

Excess of NPM-ALK oncogenic signaling promotes cellular apoptosis and drug dependency.

Most of the anaplastic large-cell lymphoma (ALCL) cases carry the t(2;5; p23;q35) that produces the fusion protein NPM-ALK (nucleophosmin-anaplastic lymphoma kinase). NPM-ALK-deregulated kinase activity drives several pathways that support malignant transformation of lymphoma cells. We found that in ALK-rearranged ALCL cell lines, NPM-ALK was distributed in equal amounts between the cytoplasm and the nucleus. Only the cytoplasmic portion was catalytically active in both cell lines and primary ALCL, whereas the nuclear portion was inactive because of heterodimerization with NPM1. Thus, about 50% of the NPM-ALK is not active and sequestered as NPM-ALK/NPM1 heterodimers in the nucleus. Overexpression or relocalization of NPM-ALK to the cytoplasm by NPM genetic knockout or knockdown caused ERK1/2 (extracellular signal-regulated protein kinases 1 and 2) increased phosphorylation and cell death through the engagement of an ATM/Chk2- and γH2AX (phosphorylated H2A histone family member X)-mediated DNA-damage response. Remarkably, human NPM-ALK-amplified cell lines resistant to ALK tyrosine kinase inhibitors (TKIs) underwent apoptosis upon drug withdrawal as a consequence of ERK1/2 hyperactivation. Altogether, these findings indicate that an excess of NPM-ALK activation and signaling induces apoptosis via oncogenic stress responses. A 'drug holiday' where the ALK TKI treatment is suspended could represent a therapeutic option in cells that become resistant by NPM-ALK amplification.We thank Maria Stella Scalzo for technical support, Dr Emanuela Colombo for kindly providing MEFs that lack NPM1 (MEF NPM−/−p53−/−) and control fibroblasts (MEF p53−/−), Dr Guido Serini for the use of his confocal microscopy unit at the Candiolo Cancer Institute—IRCCS, Torino, Italy. We also thank Ariad Pharmaceutical, Pfizer, Astellas and Novartis that kindly provided all drugs used in this study. This work was supported by the Regione Lombardia (ID14546A) and Fondazione Berlucchi Onlus Grant 2014 (to CGP), and by grants FP7 ERC-2009-StG (Proposal No. 242965—‘Lunely’); Associazione Italiana per la Ricerca sul Cancro (AIRC) Grant IG-12023; Koch Institute/DFCC Bridge Project Fund; Ellison Foundation Boston; Worldwide Cancer Research Association (former AICR) grant 12-0216; the Grant for Oncology Innovation by Merck-Serono and R01 CA196703-01 (to RC).This is the author accepted manuscript. The final version is available from NPG via http://dx.doi.org/10.1038/onc.2015.45

EURL ECVAM Status Report on the Development, Validation and Regulatory Acceptance of Alternative Methods and Approaches (2013-April 2014)

The EURL ECVAM status report provides an update on the progress made in the development, validation and regulatory acceptance of alternative methods and approaches since the last report published in April 2013. It is informing on ongoing research and development activities, validation studies, peer reviews, recommendations, strategies and international acceptance of alternative methods and approaches. R&D activities are ongoing for the complex endpoints where the toxicological processes and the mechanistic understanding have not been sufficiently elucidated yet and for which 3Rs solutions are more difficult to find. On the other hand, good progress In the validation and regulatory acceptance is made in areas where non-animal alternative methods have been developed and validated and where the focus lies in an intelligent combination/ integration of the various non-animal approaches.JRC.I.5-Systems Toxicolog