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Video – Comet assay, step by step

The Norwegian Institute for Air Research (NILU), coordinator of RiskGONE, created a short video showing the main steps in conducting a comet assay, in order to assess the toxicity of nanomaterials.

One of the aims of the RiskGONE project is the verification, optimisation and development of methods for the characterization and human and eco-toxicological hazard assessment of ENMs (engineered nanomaterials). This is done through interlaboratory round robin exercises and training on selected methods. One of these methods is the Comet assay (single cell gel electrophoresis), a simple method used for measuring DNA damage in eukaryotic cells. The method is widely used for detection of strand breaks as well as specific DNA lesions, such as oxidized purines and pyrimidines, and is considered a useful method for genotoxicity testing in vitro as well as in vivo.

NILU provided RiskGONE partners with training on this method at a course that took place in February 2020, and in March 2021, through a practical online course/video learning due to the COVID-related restrictions. The video available below is part of the training material used in this course.

How the Comet Assay works

After exposure to the compound of interest (in this case, ENM), the cells are embedded in agarose on a microscope slide. After treatment with a detergent solution, membranes, cytoplasm, and most of the soluble cell contents are dissolved, and the DNA nucleoids are freed. The nucleoids are then subjected to an electrophoretic field, which makes the negatively charged DNA migrate towards the positive electrode-anode. DNA in the nucleoids is very compact and its movement is limited, but if a break is present in the DNA strands, the DNA loop is free to extend under the electrophoretic field and move towards the anode. When DNA is stained with specific dyes and examined microscopically, images resembling comets are seen; the comet tail consists of loops of DNA that, due to the presence of damage (strand break), have moved out from the nucleoid (comet head). The amount of DNA in the tail reflects the number of breaks in the DNA.

The standard comet assay measures single- and double-strand breaks. A modified version of the assay by inclusion of lesion-specific enzymes can detect specific DNA lesions, such as oxidized purines using formamidopyrimidine DNA glycosylase (Fpg).

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Comet Assay – Practical online course/video training by NILU

Comet Assay – Practical online course/video training by NILU

Making the gel drops: cells embedded in agarose are put on slides

Dates: 15th-19th March 2021

Location: remote/virtual

The Norwegian Institute for Air Research (NILU), coordinator of RiskGONE, is providing a practical online course/video learning on the Comet assay.

This training workshop represents the second edition of a series of trainings to be organised as part of RiskGONE`s training activities. The first edition of the course was physically held at NILU`s premises in Kjeller, Norway, on 17th-20th February 2020. Then, RiskGONE project partners joined the course from Swansea University, the University of Birmingham, and the University of Bergen, and they were trained on different assays for use with engineered nanomaterials.

This course welcomes participants from H2020 projects Twinalt and VISION, besides RiskGONE and the H2020-NMBP-13 network.

Due to the restrictions now imposed by the COVID-19 pandemic, the second edition of the course was re-arranged to a remote format. The course will take place for 5 days.

On the first day, the background and principles of the assay will be introduced in a series of lectures.

On the second day, the technical aspects of the assay will be presented. Part of the day will be dedicated to the planning of a real experiment. Trainees will have the opportunity to perform themselves in their own laboratory, with the constant support of the trainers.

On the following days, a hands-on experiment will be performed. In the morning, the daily work will be presented thought videos created at NILU, in which the trainers show how the experiment is performed step-by-step. In this phase, trainees will have the opportunity to interactive live with the trainers, ask questions and discuss the technical aspects of the experiment. In the afternoon, after viewing the videos, trainees will have the possibility to physically train on the experiment in their laboratory. All along, trainers will be available online to support trainees if needed.

Training materials, including the step-by-step videos, will be shared among all project partners and might also form the basis of teaching and training material to be used beyond the project course.

The training’s agenda can be accessed here

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RiskGONE partners take part in events across the continent on European Researchers Night – check the agenda!

RiskGONE partners take part in events across the continent on European Researchers Night – check the agenda!

European Union 2020

On 27 November 2020, researchers across Europe will meet the public (mostly online) and showcase their work through hands-on experiments, science shows, games, quizzes, competitions, exhibitions, and digital activities. It’s the European Researchers’ Night!

RiskGONE partners are participating in some of these important events, aimed to shorten the gap between research and the general public. We seek to raise awareness on how different kinds of nanomaterials are used in daily applications and how we can manage the risks that some of they may pose to the environment and human health.

Find the agenda below! 

 

27 November 2020

Dechema LIVE Session 1 (13:00-14:00 CET):

Deutsch: Sie möchten mehr über Nanomaterialien erfahren, wie sie in Produkten genutzt werden und  wie der Stand der Sicherheitsforschung zu Nanomaterialien ist? Dann besuchen Sie unsere Veranstaltung, hören sie einen Fachvortrag von Dr. Christoph Steinbach (Dechema), nehmen Sie an einem Quiz teil und diskutieren Sie live mit Wissenschaftlerinnen und Wissenschaftlern über die Themen, die Ihnen am Herzen liegen.

Zugangslink (Plattform Zoom):  https://us02web.zoom.us/j/86964499249?pwd=WFRER0phcmVGMlU5Ry8vTTEwS2puZz09

English: Would you like to learn more about nanomaterials, how they are used in products and what the state of safety research on nanomaterials is about? Then visit our event, listen to a lecture by Dr. Christoph Steinbach (Dechema), take part in a quiz and discuss live with scientists about the topics you are interested in.

Access link (Zoom platform): https://us02web.zoom.us/j/86964499249?pwd=WFRER0phcmVGMlU5Ry8vTTEwS2puZz09

 

Dechema LIVE Session 2 (18:00-19:00 CET):

Deutsch: Sie möchten mehr über Nanomaterialien erfahren, wie sie in Produkten genutzt werden und  wie der Stand der Sicherheitsforschung zu Nanomaterialien ist? Dann besuchen Sie unsere Veranstaltung, hören sie einen Fachvortrag von Dr. Christoph Steinbach (Dechema), nehmen Sie an einem Quiz teil und diskutieren Sie live mit Wissenschaftlerinnen und Wissenschaftlern über die Themen, die Ihnen am Herzen liegen.

Zugangslink (Plattform Zoom):  https://us02web.zoom.us/j/88063302206?pwd=TGI4ZVdIU2ZxYmZNaUhvS1FIRWtZZz09

English: Would you like to learn more about nanomaterials, how they are used in products and what the state of safety research on nanomaterials is about? Then visit our event, listen to a lecture by Dr. Christoph Steinbach (Dechema), take part in a quiz and discuss live with scientists about the topics you are interested in.

Access link (Zoom platform): https://us02web.zoom.us/j/88063302206?pwd=TGI4ZVdIU2ZxYmZNaUhvS1FIRWtZZz09

 

CSIC/Universidad de Zaragoza (physical event in Zaragoza, Spain)

La noche investigación 2021

Register here: https://www.eventbrite.es/e/entradas-investiga-la-nanociencia-130232032515

 

More events by NANORIGO, RiskGONE partner project, at ERN

27 November 

At 18.30 (CET), Scotland’s ERN Explorathon is featuring  Making Nano Work for Us – a NANORIGO video and family quiz hosted by  OPTIMAT with Mark Morrison.

https://tinyurl.com/nanorigo-explorathon

 

  • The NANORIGO video is also featuring on the Scottish Research Showcase at 14.00 (CET) [@EU_NANORIGO]

 

Discussions will go about nanoscience and health:

–        What makes SARS-CoV-2 successful?

–        Viruses are biologic nanoparticles

–        What is nano and where do we find it?

–        Can nano fight Covid?

–        How safe is nano and who cares?

 

More events by Gov4Nano, RiskGONE partner project, at ERN

1.‘Life is Science‘ event: Nanotechnologie – Wie sicher sind die Produkte und wer entscheidet das? – Life is Science

  1. https://www.wissenschafftzukunft-kiel.de/nacht-der-wissenschaft/programm-2020/veranstaltung2020.html?tx_ncwzkndwevents_events%5Bevent%5D=1101&tx_ncwzkndwevents_events%5Baction%5D=show&tx_ncwzkndwevents_events%5Bcontroller%5D=Event&cHash=11c42c9359bdf366e1298a295eb886b2

 

And find here some of the materials produced for this special day!

 

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Join us in the Nanosafety Training School: From Basic Science To Risk Governance

Join us in the Nanosafety Training School: From Basic Science To Risk Governance

The training school will take place on 20-25 June 2021 in San Servolo island, in Venice, Italy.

The School week will include a variety of hands-on sessions aimed to transfer state-of-the-art knowledge on a variety of topics from key experts to the new generation of nano-environmental, health and safety, and biomedicine professionals, using interprofessional education.

About the School

This year the Venice Nano Training School celebrates its 10th anniversary.

The School will feature keynote speeches, hands-on sessions and a dedicated Young Scientist Forum Day during which early career researchers (PhD students, PhD candidates and Post-Docs) will have the opportunity to present their work.

The School week will include a variety of hands-on sessions aimed to transfer state-of-the-art knowledge on a variety of topics from key experts to the new generation of nano-environmental, health and safety, and biomedicine professionals, using interprofessional education.

A variety of networking activities (a special 10th anniversary welcome cocktail, social event and a social dinner) will enable plenty of time and opportunities for you to widen your network and foster academic exchange.

Become part of an interactive, exciting week and enrich your knowledge by developing multidisciplinary expertise!

 

Who should attend?

  • Early-stage researchers
  • PhD students and Post-Docs
  • Senior researchers
  • Industry
  • Governmental Agencies
  • Medical Personnel
  • Anyone interested in Safe Nanotechnology, Risk Assessment and Nano-Medicine

School Topics

  • Hazard to Human Health & Environment
  • Fate & Exposure Assessment
  • Nanomedicine: from the lab to the market
  • Modelling
  • Grouping & Read Across Approaches
  • Risk Governance

 

More Information about the Young Scientist Forum Day

Early career researchers will have the opportunity to present their work in the form of oral and poster presentations. You can choose between the following themes:

State of the art approaches and challenges towards nanomaterial characterisation
Strategies towards testing ENM toxicology (human and environmental)
Modelling approaches towards ENM toxicology
Risk assessment and legal regulation of ENM
Approaches towards defining human and environmental exposure to EN

 

School Location

The training school will take place in San Servolo (Venice, Italy).

 

School Certificates

Each participant will be given a Certificate of attendance upon request.

 

Further information and updates available here. 

 

Contacts

Scientific enquiries:

Danail Hristozov, GreenDecision (Italy) | danail.hristozov@greendecision.eu

Stella Stoycheva, Yordas Group (UK) | s.stoycheva@yordasgroup.com

Susanne Resch, BioNanoNet (Austria) | susanne.resch@bionanonet.at

Logistics, local support and administration:

Paola Basso, GreenDecision (Italy) | management@greendecision.eu

 

Sponsors

GreenDecision (Venice, IT)

Institute of Occupational Medicine (Edinburgh, UK)

Yordas Group (Forchheim, DE)

Warrant Hub S.p.A (Casalecchio di Reno, IT)

BioNanoNet Forschungsgesellschaft mbH (Graz, AT)

NC3RS (London, UK)

Aarhus University (Aarhus, DK)

RIVM (Utrecht, NL)

NILU (Kjeller, NO)

These projects have received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 760928 (BIORIMA), No 814401 (Gov4Nano), No 760840 (GRACIOUS), No 814426 (NanoInformaTIX), No 814530 (NANORIGO), No 760813 (PATROLS), and No 814425 (RiskGONE).

 

 

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Webinar recording: identifying health hazards with AOPs – watch it anytime!

Webinar recording: identifying health hazards with AOPs – watch it anytime!

On 5th June, RiskGONE held its second webinar, attended by more than 40 participants. This time project partners focused the discussion on how to use Adverse Outcome Pathways (AOPs), a risk-assessment method, to analyse the possible hazards nanomaterials can pose to human health.

Experts from KU Leuven (Belgium), the Institute for Medical Research and Occupational Health (Croatia) and the QSAR lab (Poland) explained how they are applying this method to tracing nanomaterials risks.

AOPs could help researchers find negative effects from nanomaterials more efficiently, and reduce the need for animal testing. So, how do AOPs work in practice? The idea behind is that researchers create pathways that link a first event observable at molecular level with subsequent effects in the human body and the final problems they can pose to human health, which can be as serious as cancer, neurodegeneration or decreased fertility.

These pathways are living documents that can be accessed online through the AOP Wiki website and be modified as fresh evidence reveals new or different effects of nanomaterials in human health. The AOPs system is a tool launched by the OECD in 2012 to systematically assess toxicological risks in chemicals. Researchers in the RiskGONE project propose using these AOPs to map possible hazards in using nanomaterials too.

Eager to know more? We have recorded the webinar for you! Simply click on the links below to watch the webinar sections of your interest:

 

Access all the presentation slides from speakers here.