natgillin/translations-raw huggingface dataset:
positive og negative kontrolkemikalier, endelige koncentrationer, behandlingsbetingelser og -varighed samt restitutionstider
Cell preparation may involve hypotonic treatment, but this step is not necessary if adequate cell spreading is otherwise achieved.
anvendte metoder til præparering af objektglas og farvningsteknik
Different techniques can be used in slide preparation provided that high-quality cell preparations for scoring are obtained.
kriterier for scoring af mikrokerneceller (valg af analyserbare celler og identifikation af mikrokerner)
Cells with intact cell membrane and intact cytoplasm should be retained to allow the detection of micronuclei and (in the cytokinesis-block method) reliable identification of binucleate cells.
antal analyserede celler
The slides can be stained using various methods, such as Giemsa or fluorescent DNA specific dyes.
kriterier for bedømmelse af undersøgelsen som positiv, negativ eller tvetydig
The use of appropriate fluorescent stains (e.g. acridine orange (78) or Hoechst 33258 plus pyronin-Y (79)) can eliminate some of the artifacts associated with using a non-DNA specific stain.
anvendte statistiske analysemetoder
Anti-kinetochore antibodies, FISH with pancentromeric DNA probes, or primed in situ labelling with pancentromere-specific primers, together with appropriate DNA counterstaining, can be used to identify the contents (whole chromosomes will be stained while acentric chromosome fragments will not) of micronuclei if mechanistic information of their formation is of interest (16) (17).
metoder som f.eks. anvendelse af anti-kinetokor-antistof eller pan-centromerspecifikke prober til bestemmelse af, om mikrokernerne indeholder hele kromosomer eller kromosomfragmenter, hvor dette er relevant
Other methods for differentiation between clastogens and aneugens may be used if they have been shown to be effective and validated.
definition af acceptable celler til analyse
For example, for certain cell lines the measurements of sub-2N nuclei as hypodiploid events using techniques such as image analysis, laser scanning cytometry or flow cytometry could also provide useful information (80) (81) (82).
hvis der ikke anvendes cyto B, antal behandlede celler og antal høstede celler for hver kultur, når der benyttes cellelinjer
Morphological observations of nuclei could also give indications of possible aneuploidy.
den anvendte måling af cytotoksicitet, f.eks. CBPI eller RI ved anvendelse af cytokineseblokeringsmetoden, RICC eller RPD, når cytokineseblokeringsmetoden ikke anvendes, eventuelle andre observationer (f.eks. cellekonfluens, apoptose, nekrose, antal metafaser, forekomst af binukleære celler)
Moreover, a test for metaphase chromosome aberrations, preferably in the same cell type and protocol with comparable sensitivity, could also be a useful way to determine whether micronuclei are due to chromosome breakage (knowing that chromosome loss would not be detected in the chromosome aberration test).
data om pH og osmolalitet i behandlingsmediet, hvis disse værdier er målt
All slides, including those of the solvent and the untreated (if used) and positive controls, should be independently coded before the microscopic analysis of micronucleus frequencies.
fordeling af mono-, bi- og multinukleære celler ved anvendelse af en cytokineseblokeringsmetode
Appropriate techniques should be used to control any bias or drift when using an automated scoring system, for instance, flow cytometry, laser scanning cytometry or image analysis.
antal celler med mikrokerner, anført særskilt for hver behandlet kultur og kontrolkultur, og bestemmelse af, om de er indsamlet fra binukleære eller mononukleære celler, hvis det er relevant
Regardless of the automated platform is used to enumerate micronuclei, CBPI, RI, RPD, or RICC should be assessed concurrently.
historisk negative (opløsningsmiddel) og positive kontroldata med angivelse af intervaller, gennemsnit og standardafvigelser og kontrolgrænser på 95 % for fordelingen, samt mængden af data
In cytoB-treated cultures, micronucleus frequencies should be analysed in at least 2000 binucleate cells per concentration and control (83), equally divided among the replicates, if replicates are used.
statistisk analyse og eventuelle P-værdier.
In the case of single cultures per dose (see paragraph 28), at least 2000 binucleate cells per culture (83) should be scored in this single culture.
Kapitel B.10 i dette bilag: In vitro-test for kromosomaberrationer i celler fra pattedyr.
If substantially fewer than 1000 binucleate cells per culture (for duplicate cultures), or 2000 (for single culture), are available for scoring at each concentration, and if a significant increase in micronuclei is not detected, the test should be repeated using more cells, or at less cytotoxic concentrations, whichever is appropriate.
In vitro micronucleus assay with Chinese hamster V79 cells — results of a collaborative study with in situ exposure to 26 chemical substances.
Care should be taken not to score binucleate cells with irregular shapes or where the two nuclei differ greatly in size.
A protocol for the in vitro micronucleus test, I. Contributions to the development of a protocol suitable for regulatory submissions from an examination of 16 chemicals with different mechanisms of action and different levels of activity.
In addition, binucleate cells should not be confused with poorly spread multi-nucleate cells.
Evaluation of different cytotoxic and cytostatic measures for the in vitromicronucleus test (MNVit): Introduction to the collaborative trial.
Cells containing more than two main nuclei should not be analysed for micronuclei, as the baseline micronucleus frequency may be higher in these cells (84).
Comparison of four different treatment conditions of extended exposure in the in vitro micronucleus assay using TK6 lymphoblastoid cells.
Scoring of mononucleate cells is acceptable if the test chemical is shown to interfere with cytoB activity.
Statement by the European Centre for the Validation of Alternative Methods (ECVAM) Scientific Advisory Committee (ESAC) on the scientific validity of the in vitro micronucleus test as an alternative to the in vitro chromosome aberration assay for genotoxicity testing.
A repeat test without CytoB might be useful in such cases.
ESAC's 25. møde, 16.-17. november 2006, tilgængelig på: http://ecvam.jrc.it/index.htm.
Scoring mononucleate cells in addition to binucleate cells could provide useful information (85) (86), but is not mandatory.
Tilgængelig på: http://ecvam.jrc.it/index.htm.
In cell lines tested without cytoB treatment, micronuclei should be scored in at least 2000 cells per test concentration and control (83), equally divided among the replicates, if replicates are used.
“A safe substitute for Polychlorinated Biphenyls as an Inducer of Metabolic Activation Systems”, in In Vitro Metabolic Activation in Mutagenesis Testing. de Serres, F.J. et al. (eds), Elsevier, North-Holland, pp.
When single cultures per concentration are used (see paragraph 28), at least 2000 cells per culture should be scored in this single culture.
85-88.
If substantially fewer than 1000 cells per culture (for duplicate cultures), or 2000 (for single culture), are available for scoring at each concentration, and if a significant increase in micronuclei is not detected, the test should be repeated using more cells, or at less cytotoxic concentrations, whichever is appropriate.
Tilgængelig på: http://www.pops.int/
When cytoB is used, a CBPI or an RI should be determined to assess cell proliferation (see Appendix 2) using at least 500 cells per culture.
“CHO/HGPRT Mutation Assay: Evaluation of Gases and Volatile Liquids”, in Genotoxic Effects of Airborne Agents.
When treatments are performed in the absence of cytoB, it is essential to provide evidence that the cells in culture have divided, as discussed in paragraphs 24-28.
Tice, R.R., D.L.
In order to establish sufficient experience with the assay prior to using it for routine testing, the laboratory should have performed a series of experiments with reference positive chemicals acting via different mechanisms (at least one with and one without metabolic activation, and one acting via an aneugenic mechanism, and selected from the chemicals listed in Table 1) and various negative controls (including untreated cultures and various solvents/vehicle).
Costa, K.M.
This is not applicable to laboratories that have experience, i.e. that have an historical data base available as defined in paragraphs 49 to 52.
OECD Environment, Health and Safety Publications (EHS), Series on testing and assessment, No.198, OECD Publishing, Paris.
A selection of positive control chemicals (see Table 1) should be investigated with short and long treatments in the absence of metabolic activation, and also with short treatment in the presence of metabolic activation, in order to demonstrate proficiency to detect clastogenic and aneugenic chemicals, determine the effectiveness of the metabolic activation system and demonstrate the appropriateness of the scoring procedures (microscopic visual analysis, flow cytometry, laser scanning cytometry or image analysis).
“In vitro micronucleus test”, in Encyclopedia of Biopharmaceutical Statistics, 2nd ed.
When first acquiring data for an historical negative control distribution, concurrent negative controls should be consistent with published negative control data where they exist.
Chow, S. (ed.), Marcel Dekker, Inc.
As more experimental data are added to the control distribution, concurrent negative controls should ideally be within the 95 % control limits of that distribution (87) (88).
New York, pp.
The laboratory's historical negative control database, should initially be built with a minimum of 10 experiments but would preferably consist of at least 20 experiments conducted under comparable experimental conditions.
463-467.
Laboratories should use quality control methods, such as control charts (e.g. C-charts or X-bar charts (88)), to identify how variable their positive and negative control data are, and to show that the methodology is ‘under control’ in their laboratory (83).
Analysis of Data from in vitro Cytogenetic Assays. in Statistical Evaluation of Mutagenicity Test Data, Kirkland, D.J. (ed), Cambridge University Press, Cambridge, pp.
Further recommendations on how to build and use the historical data (i.e. criteria for inclusion and exclusion of data in historical data and the acceptability criteria for a given experiment) can be found in the literature (87).
141-154.
Any changes to the experimental protocol should be considered in terms of the consistency of the data with the laboratory's existing historical control databases.
Alle kemikalier eller processer, der ved interaktion med komponenterne i det mitotiske og det meiotiske celledelingscyklusapparat forårsager aneuploidi i celler eller organismer.
Negative control data should consist of the incidence of micronucleated cells from a single culture or the sum of replicate cultures as described in paragraph 28.
Programmeret celledød karakteriseret ved en række trin, der forårsager opløsning af celler i membranbundne partikler, som herefter elimineres ved fagocytose eller afstødning.
Concurrent negative controls should ideally be within the 95 % control limits of the distribution of the laboratory's historical negative control database (87) (88).
Forøgelse af celleantal som følge af mitotisk celledeling.
Where concurrent negative control data fall outside the 95 % control limits, they may be acceptable for inclusion in the historical control distribution as long as these data are not extreme outliers and there is evidence that the test system is ‘under control’ (see paragraph 50) and there is evidence of absence of technical or human failure.
DNA-region af et kromosom, hvor begge kromatider holdes sammen, og hvorpå begge kinetokorer er bundet side om side.
If the cytokinesis-block technique is used, only the frequencies of binucleate cells with micronuclei (independent of the number of micronuclei per cell) are used in the evaluation of micronucleus induction.
De endelige koncentrationer af testkemikaliet i dyrkningsmediet.
The scoring of the numbers of cells with one, two, or more micronuclei can be reported separately and could provide useful information, but is not mandatory.
Alle kemikalier eller processer, der forårsager strukturelle kromosomaberrationer i populationer af celler eller organismer.
Concurrent measures of cytotoxicity and/or cytostasis for all treated, negative and positive control cultures should be determined (16).
Cytokinese
The CBPI or the RI should be calculated for all treated and control cultures as measurements of cell cycle delay when the cytokinesis-block method is used.
Celledeling umiddelbart efter mitose, hvorved der dannes to datterceller, som hver indeholder en enkelt kerne.
In the absence of cytoB, the RPD or the RICC should be used (see Appendix 2).
Proliferationsindeks for cytokineseblokerende stof (CBPI)
The concurrent negative control is considered acceptable for addition to the laboratory historical negative control database as described in paragraph 50.
Andelen af celler efter anden deling i den behandlede population i forhold til den ubehandlede kontrol (se formlen i tillæg 2).
Concurrent positive controls (see paragraph 50) should induce responses that are compatible with those generated in the laboratory's historical positive control data base and produce a statistically significant increase compared with the concurrent negative control.
Cytostase
Cell proliferation criteria in the solvent control should be fulfilled (paragraph 25-27).
Hæmning af cellevækst (se formlen i tillæg 2)
All experimental conditions were tested unless one resulted in positive results (paragraphs 36-40).
For de assays, som er omfattet af denne forsøgsmetode, der gennemføres i fravær af cytochalasin B, identificeres cytotoksicitet som en reduktion i proliferationsindekset for cytokineseblokering (CPPI) eller replikationsindekset (RI) i de behandlede celler sammenlignet med den negative kontrol (jf. punkt 26 og tillæg 2).
Adequate number of cells and concentrations are analysable (paragraphs 28 and 44-46).
For de assays, som er omfattet af denne forsøgsmetode, der gennemføres i fravær af cytochalasin B, identificeres cytotoksicitet som en reduktion i den relative populationsfordobling (RPD) eller den relative forøgelse af celletal (RICC) i de behandlede celler sammenlignet med den negative kontrol (jf. punkt 27 og tillæg 2).
The criteria for the selection of top concentration are consistent with those described in paragraphs 24-31.
Interfaseceller
Providing that all acceptability criteria are fulfilled, a test chemical is considered to be clearly positive if, in any of the experimental conditions examined (see paragraphs 36-39):
Celler, der ikke er i mitosefase.
at least one of the test concentrations exhibits a statistically significant increase compared with the concurrent negative control (89)
Proteinstruktur, der samles ved et kromosoms centromer, hvortil tentrådene er påhæftet under celledeling, således at datterkromosomerne kan ordnes i forhold til dattercellernes poler.
the increase is dose-related in at least one experimental condition when evaluated with an appropriate trend test (see paragraph 28)
Små ekstrakerner, der er adskilt fra cellens hovedkerne, og som dannes under mitosens eller meiosens telofase ud fra efterladte kromosomfragmenter eller hele kromosomer.
any of the results are outside the distribution of the historical negative control data (e.g. Poisson-based 95 % control limits; see paragraph 52).
Forholdet mellem antallet af celler i metafase og det samlede antal celler i en population; viser populationens formeringsgrad.
When all of these criteria are met, the test chemical is then considered able to induce chromosome breaks and/or gain or loss in this test system.
Non-disjunction
Recommendations for the most appropriate statistical methods can also be found in the literature (90) (91) (92).
Forbundne kromatiders manglende evne til at dele sig og spalte sig korrekt til datterceller under udvikling, hvorved der dannes datterceller med anormale kromosomtal.
Providing that all acceptability criteria are fulfilled, a test chemical is considered clearly negative if, in all experimental conditions examined (see paragraphs 36-39):
Numeriske kromosomaberrationer i celler eller organismer, der involverer et eller flere komplette kromosomsæt, men ikke et enkelt kromosom eller kromosomer (aneuploidi).
all results are inside the distribution of the historical negative control data (e.g. Poisson-based 95 % control limits; see paragraph 52).
Proliferationsindeks (PI)
The test chemical is then considered unable to induce chromosome breaks and/or gain or loss in this test system.
Metode til måling af cytotoksicitet, hvis der ikke anvendes cytoB (se formlen i tillæg 2).
There is no requirement for verification of a clear positive or negative response.
Relativ forøgelse af celletal (RICC)
In case the response is neither clearly negative nor clearly positive as described above and/or in order to assist in establishing the biological relevance of a result, the data should be evaluated by expert judgement and/or further investigations.
Replikationsindeks (RI)
Scoring additional cells (where appropriate) or performing a repeat experiment possibly using modified experimental conditions (e.g. concentration spacing, other metabolic activation conditions [i.e. S9 concentration or S9 origin]) could be useful.
Andel af afsluttede celledelingscyklusser i den behandlede kultur i forhold til den ubehandlede kontrol i eksponerings- og restitutionsfasen (se formlen i tillæg 2).
In rare cases, even after further investigations, the data set will not allow a conclusion of positive or negative, and will therefore be concluded as equivocal.
Blanding af S9-leverfraktionen og andre medvirkende faktorer, som er nødvendige for at skabe metabolisk enzymaktivitet.
Test chemicals that induce micronuclei in the MNvit test may do so because they induce chromosome breakage, chromosome loss, or a combination of the two.
Ubehandlet kontrol
Further analysis using anti-kinetochore antibodies, centromere specific in situ probes, or other methods may be used to determine whether the mechanism of micronucleus induction is due to clastogenic and/or aneugenic activity.
Hvis cytoB anvendes, baseres evalueringen af cytotoksicitet på proliferationsindekset for cytokineseblokerende stof (CBPI) eller replikationsindekset (RI) (17) (69).
reactivity of the test chemicals with the solvent/vehicle or cell culture media;
CBPI angiver det gennemsnitlige antal kerner pr. celle og kan anvendes til at beregne celleproliferationen.
solubility and stability of the test chemical in solvent, if known;
RI angiver det relative antal cellecyklusser pr. celle i perioden med eksponering for cytoB i behandlede kulturer sammenholdt med kontrolkulturerne og kan anvendes til at beregne cytostase i procent.
measurement of pH, osmolality, and precipitate in the culture medium to which the test chemical was added, as appropriate.
% cytostase = 100 – 100(CBPIT – 1) ÷ (CBPIC – 1)
justification for choice of solvent;
T kultur til behandling af testkemikalier
percentage of solvent in the final culture medium
C kontrolkultur
type and source of cells used;
antal mononukl.celler
suitability of the cell type used;
antal binukl.celler
absence of mycoplasma, in case of cell lines;
antal multinukl.celler
for cell lines, information on cell cycle length or proliferation index;
antal celler i alt
where lymphocytes are used, sex of blood donors, age and any relevant information on the donor, whole blood or separated lymphocytes, mitogen used;
et CBPI på 1 (alle celler er mononukleære) svarer til en cytostase på 100 %.
normal (negative control) cell cycle time;
Cytostase = 100-RI
methods for the maintenance of cell cultures, for cell lines;
T behandlede kulturer
modal number of chromosomes, for cell lines;
C kontrolkulturer
Test Conditions:
Et RI på 53 % angiver således, at i forhold til antallet af celler, der delte sig til binukleære og multinukleære celler i kontrolkulturen, var der kun 53 % af disse celler, der delte sig i den behandlede kultur, dvs. 47 % cytostase.
identity of the cytokinesis blocking substance (e.g. cytoB), if used, and its concentration and duration of cell exposure;
Hvis cytoB ikke anvendes, anbefales evaluering af cytotoksicitet baseret på relativ forøgelse af celletal (RICC) eller relativ populationsfordobling (RPD) (69), da der ved begge metoder tages højde for den andel af cellepopulationen, som har delt sig.
concentration of the test chemical expressed as a final concentration in the culture medium (e.g. μg or mg/mL, or mM of culture medium);
Et RICC eller RPD på 53 % angiver således 47 % cytotoksicitet/cytostase.
rationale for the selection of concentrations and the number of cultures, including cytotoxicity data and solubility limitations;
Ved anvendelse af et proliferationsindeks (PI) kan cytotoksiciteten vurderes ved optælling af antallet af kloner med 1 celle (cl1), 2 celler (cl2), 3-4 celler (cl4) og 5-8 celler (cl8).
composition of media, CO2 concentration, if applicable, humidity level;
PI er også blevet anvendt som en værdifuldt og pålideligt cytotoksicitetsparameter for cellelinjer dyrket in vitro uden cytoB (35) (36) (37) (38) og kan betragtes som en nyttig supplerende parameter.
concentration (and/or volume) of the solvent and test chemical added in the culture medium;
Under alle omstændigheder bør antallet af celler inden behandling være det samme for behandlede og negative kontrolkulturer.
incubation temperature and time;
Selv om RCC (dvs. antal celler i behandlede kulturer/antal celler i kontrolkulturer) tidligere er blevet brugt som cytotoksicitetsparameter, anbefales dette ikke længere, da cytotoksiciteten kan undervurderes.
cell density at seeding, if applicable;
Ved hjælp automatiserede scoringssystemer, f.eks. flowcytometri, laserskanningcytometri eller billedanalyse, kan antallet af celler i formlen erstattes af antallet af kerner.
type and composition of metabolic activation system, (source of S9, method of preparation of the S9 mix, the concentration or volume of S9 mix and S9 in the final culture medium, quality controls of S9 (e.g. enzymatic activity, sterility, metabolic capability);
I del B indsættes følgende kapitler:
methods of slide preparation and the staining technique used;
In chemico-hudsensibilisering: direkte peptidreaktivitetsassay (DPRA)
criteria for scoring micronucleate cells (selection of analysable cells and identification of micronucleus);
Denne forsøgsmetode svarer til OECD's testvejledning (Test Guideline (TG)) 442C (2015).
numbers of cells analysed;
Et hudsensibiliserende stof er et stof, der fremkalder en allergisk reaktion ved hudkontakt som defineret ved De Forenede Nationers globalt harmoniserede system til klassificering og mærkning af kemikalier (UN GHS) (1) og forordning (EF) nr. 1272/2008 om klassificering, mærkning og emballering af stoffer og blandinger (CLP-forordningen) [12].
criteria for considering studies as positive, negative, or equivocal;
Den nuværende viden om kemiske og biologiske mekanismer i forbindelse med hudsensibilisering er sammenfattet i form af en Adverse Outcome Pathway (AOP) (2) fra den molekylære udløsende begivenhed gennem de mellemliggende begivenheder til den skadelige virkning, nemlig allergisk kontaktdermatitis hos mennesker eller kontaktoverfølsomhed hos gnavere.
method(s) of statistical analysis used;
Vurderingen af hudsensibilisering har typisk omfattet brug af laboratoriedyr.
definition of acceptable cells for analysis;
Med de klassiske metoder baseret på marsvin, Guinea Pig Maximisation Test (GPMT) og Buehler-testen (TM B.6 (3)), undersøger man både induktions- og udløsningsfasen ved hudsensibilisering.
in the absence of cyto B, the number of cells treated and the number of cells harvested for each culture in case of cell lines;