natgillin/translations-raw huggingface dataset:
The slides can be stained using various methods, such as Giemsa or fluorescent DNA specific dyes.
Nõrk reaktsioonivõime
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.
Negatiivne
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).
22,62 % < sisalduse vähenemise % keskväärtus ≤ 42,47 %
Other methods for differentiation between clastogens and aneugens may be used if they have been shown to be effective and validated.
Mõõdukas reaktsioonivõime
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).
42,47 % < sisalduse vähenemise % keskväärtus ≤ 100 %
Morphological observations of nuclei could also give indications of possible aneuploidy.
Tugev reaktsioonivõime
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).
Võib esineda juhte, kus uuritaval kemikaalil (ainel või ühest või mitmest koostisosast koosneva aine või segu mõnel koostisosal) on lainepikkusel 220 nm oluline neelduvus ning peptiidiga ühesugune retentsiooniaeg (elueeruvad koos).
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.
Koelueerumise võib kõrvaldada, kui vähesel määral kohandada HPLC süsteemi nii, et paremini lahutada uuritava kemikaali elueerimise aega peptiidi elueerimise ajast.
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.
Kui koelueerimise vältimiseks kasutatakse alternatiivset HLPC süsteemi, tuleb tõendada selle samaväärsust valideeritud süsteemiga (nt kasutades 2. liites esitatud pädevuse kontrolli aineid).
Regardless of the automated platform is used to enumerate micronuclei, CBPI, RI, RPD, or RICC should be assessed concurrently.
Kui esineb koelueerimine, ei saa peptiidi piiki integreerida ning on võimatu arvutada peptiidi sisalduse vähenemise protsenti.
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.
Kui sellised uuritavad kemikaalid elueeruvad koos nii tsüsteiini sisaldavate kui ka lüsiini sisaldavate peptiididega, tuleb analüüsi tulemus lugeda ebaselgeks.
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.
Kui uuritav kemikaal elueerub ainult koos lüsiini sisaldava peptiidiga, siis võib kasutada tabelis 2 esitatud prognoosimudelit tsüsteiin 1:10.
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.
Prognoosimudel tsüsteiin 1:10 [16]
Care should be taken not to score binucleate cells with irregular shapes or where the two nuclei differ greatly in size.
Tsüsteiini (Cys) sisalduse vähenemise %
In addition, binucleate cells should not be confused with poorly spread multi-nucleate cells.
Reaktsioonivõime klass
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).
DPRA prognoos [17]
Scoring of mononucleate cells is acceptable if the test chemical is shown to interfere with cytoB activity.
0 % ≤ tsüsteiini sisalduse vähenemise % ≤ 13,89 %
A repeat test without CytoB might be useful in such cases.
13,89 % < tsüsteiini sisalduse vähenemise % ≤ 23,09 %
Scoring mononucleate cells in addition to binucleate cells could provide useful information (85) (86), but is not mandatory.
23,09 % < tsüsteiini sisalduse vähenemise % ≤ 98,24 %
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.
98,24 % < tsüsteiini sisalduse vähenemise % ≤ 100 %
When single cultures per concentration are used (see paragraph 28), at least 2000 cells per culture should be scored in this single culture.
Võib esineda muid juhte, kui uuritava kemikaali ja emma-kumma peptiidi retentsiooniaja kattumine ei ole täielik.
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.
Sellistel juhtudel võib peptiidi protsendilise sisalduse vähenemise väärtusi hinnata ning kasutada prognoosimudelit tsüsteiin 1:10 / lüsiin 1:50, kuid uuritava kemikaali reaktsioonivõime klassi ei saa täpselt määrata.
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.
Nii lüsiini kui ka tsüsteiini puhul peaks uuritava kemikaali kohta piisama ühest HPLC analüüsist, kui tulemus on selge.
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.
Kui tulemused on lähedal läviväärtusele, millega eristatakse positiivseid tulemusi negatiivsetest (st piiripealsed tulemused), võib siiski vaja minna täiendavat testimist.
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).
Kui esineb olukordi, milles prognoosimudeli tsüsteiin 1:10 / lüsiin 1:50 korral jääb keskmine sisalduse vähenemise protsent vahemikku 3–10 % või prognoosimudeli tsüsteiin 1:10 korral jääb tsüsteiini sisalduse vähenemise protsent vahemikku 9–17 %, kaalutakse teise katseseeria tegemist, ning juhul, kui selle tulemused erinevad esimesest seeriast, tuleb teha ka kolmas seeria.
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.
Katseprotokoll peab sisaldama järgmist teavet:
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).
ühest koostisosast koosnev aine:
When first acquiring data for an historical negative control distribution, concurrent negative controls should be consistent with published negative control data where they exist.
Kemikaali identifitseerimisandmed, nagu IUPACi või CASi nimetus(ed) ja CASi number/numbrid, SMILESi või InChI kood, struktuurivalem ja/või muud identifikaatorid;
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).
füüsiline välimus, lahustuvus vees, molekulmass ja muud asjakohased füüsikalis-keemilised omadused, kättesaadavas ulatuses;
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.
puhtus, lisandite keemiline määratlus, kui see on asjakohane ja praktiliselt teostatav jne;
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).
kontrollkemikaali töötlemine enne katset, kui asjakohane (nt soojendamine, peenestamine);
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).
uuritud kontsentratsioon(id);
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.
säilitamise tingimused ja stabiilsus säilitamisel, kättesaadavas ulatuses;
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.
Mitme koostisosaga aine, tundmatu või muutuva koostisega ained, komplekssed reaktsioonisaadused või bioloogilist päritolu materjalid ja segud:
Concurrent negative controls should ideally be within the 95 % control limits of the distribution of the laboratory's historical negative control database (87) (88).
kirjeldatakse nii palju kui võimalik, lähtudes koostisosade keemilistest identifitseerimisandmetest (vt eespool), puhtusest, kvantitatiivsest sisaldusest ja asjakohastest füüsikalis-keemilistest omadustest;
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.
välimus, lahustuvus vees ja muud asjakohased füüsikalis-keemilised omadused, kättesaadavas ulatuses;
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.
molekulmass või näiline molekulmass teadaoleva koostisega segude/polümeeride korral või muu uuringu tegemiseks asjakohane teave;
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.
säilitamise tingimused ja stabiilsus säilitamisel, kättesaadavas ulatuses;
Concurrent measures of cytotoxicity and/or cytostasis for all treated, negative and positive control cultures should be determined (16).
viide varasematele positiivse kontrolli tulemustele, mis tõendavad sobivaid analüüsitsükli nõuetekohasuse kriteeriume, kui asjakohane.
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.
Kasutatud lahusti/vehiikul ja selle koostisosade suhe, kui asjakohane;
In the absence of cytoB, the RPD or the RICC should be used (see Appendix 2).
kemikaali identifitseerimisandmed, nagu IUPACi või CASi nimetus(ed) ja CASi number/numbrid ja/või muud identifikaatorid;
The concurrent negative control is considered acceptable for addition to the laboratory historical negative control database as described in paragraph 50.
füüsiline välimus, molekulmass ja muud asjakohased füüsikalis-keemilised omadused juhul, kui kasutatakse muid lahusteid/vehiikuleid kui need, mida on katsemeetodis nimetatud, kättesaadavas ulatuses;
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.
iga uuritava kemikaali puhul lahusti valimise põhjendus;
Cell proliferation criteria in the solvent control should be fulfilled (paragraph 25-27).
atsetonitriili puhul uuring, kuidas see mõjub peptiidi stabiilsusele.
All experimental conditions were tested unless one resulted in positive results (paragraphs 36-40).
Peptiidide, positiivsete kontrollide ja uuritava kemikaali preparaatide valmistamine
Adequate number of cells and concentrations are analysable (paragraphs 28 and 44-46).
Peptiidilahuste kirjeldus (tarnija, partii, peptiidi täpne mass, põhilahusele lisatud ruumala);
The criteria for the selection of top concentration are consistent with those described in paragraphs 24-31.
positiivse kontrolli lahuse kirjeldus (positiivse kontrolli aine täpne mass, katselahusele lisatud ruumala);
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):
uuritava kemikaali lahuste kirjeldus (uuritava kemikaali täpne mass, katselahusele lisatud ruumala);
at least one of the test concentrations exhibits a statistically significant increase compared with the concurrent negative control (89)
HPLC seadmed ja analüüs
the increase is dose-related in at least one experimental condition when evaluated with an appropriate trend test (see paragraph 28)
HPLC seadme tüüp, HPLC analüütiline kolonn ja eelkolonn, detektor, proovi automaatsisestaja;
any of the results are outside the distribution of the historical negative control data (e.g. Poisson-based 95 % control limits; see paragraph 52).
HPLC analüüsi jaoks asjakohased parameetrid, nagu kolonni temperatuur, süstimisruumala, voolukiirus ja gradient.
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.
Süsteemi sobivus
Recommendations for the most appropriate statistical methods can also be found in the literature (90) (91) (92).
Iga standardproovi ja võrdluskontrolli A paralleelproovi korral peptiidipiigi alune pindala lainepikkusel 220 nm;
Providing that all acceptability criteria are fulfilled, a test chemical is considered clearly negative if, in all experimental conditions examined (see paragraphs 36-39):
lineaarse kalibreerimiskõvera graafik ja r2 väärtus;
all results are inside the distribution of the historical negative control data (e.g. Poisson-based 95 % control limits; see paragraph 52).
peptiidi kontsentratsioon iga võrdluskontrolli A paralleelproovi korral;
The test chemical is then considered unable to induce chromosome breaks and/or gain or loss in this test system.
kolme võrdluskontrolli A korral peptiidi kontsentratsiooni keskväärtus (mM), SD ja CV;
There is no requirement for verification of a clear positive or negative response.
peptiidi kontsentratsioon võrdluskontrollide A ja C korral.
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.
Analüüsiseeria
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.
Võrdluskontrolli kohta:
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.
iga võrdluskontrolli B ja C paralleelproovi peptiidipiigi alune pindala lainepikkusel 220 nm;
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.
atsetonitriilis lahustatud üheksa võrdluskontrolli B ja C korral peptiidipiigi aluse pindala keskväärtus lainepikkusel 220 nm, SD ja CV (selleks, et tõendada võrdluskontrollide stabiilsust analüüsi jooksul);
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.
iga kasutatud lahusti kohta kolme asjakohase võrdluskontrolli C korral peptiidipiigi aluse pindala keskväärtus lainepikkusel 220 nm (peptiidi sisalduse vähenemise protsendi arvutamiseks);
reactivity of the test chemicals with the solvent/vehicle or cell culture media;
iga kasutatud lahusti kohta kolme asjakohase võrdluskontrolli C korral peptiidi kontsentratsioon (mM);
solubility and stability of the test chemical in solvent, if known;
iga kasutatud lahusti kohta kolme asjakohase võrdluskontrolli C korral peptiidi kontsentratsiooni keskväärtus (mM), SD ja CV.
measurement of pH, osmolality, and precipitate in the culture medium to which the test chemical was added, as appropriate.
Positiivse kontrolli kohta:
justification for choice of solvent;
iga paralleelproovi peptiidipiigi alune pindala lainepikkusel 220 nm;
percentage of solvent in the final culture medium
iga paralleelproovi peptiidi sisalduse vähenemise protsent;
type and source of cells used;
kolme paralleelproovi peptiidi sisalduse vähenemise protsendi keskväärtus, SD ja CV.
suitability of the cell type used;
Iga uuritava kemikaali kohta:
absence of mycoplasma, in case of cell lines;
koelueerimise esinemine;
for cell lines, information on cell cycle length or proliferation index;
muude asjakohaste vaatlusandmete kirjeldus, kui asjakohane;
where lymphocytes are used, sex of blood donors, age and any relevant information on the donor, whole blood or separated lymphocytes, mitogen used;
kolme paralleelproovi peptiidi sisalduse vähenemise protsendi keskväärtus, SD ja CV;
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);
DPRA katsega saadud tulemuste arutelu;
rationale for the selection of concentrations and the number of cultures, including cytotoxicity data and solubility limitations;
katsemeetodiga saadud tulemuste arutelu ühtse lähenemisviisi kontekstis, kui on olemas muud asjakohast teavet.
concentration (and/or volume) of the solvent and test chemical added in the culture medium;
168, OECD, Paris.
incubation temperature and time;
Käesoleva lisa peatükk B.6, „Naha sensibiliseerimine“.
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);
Käesoleva lisa peatükk B.50: „Naha sensibiliseerimine: lokaalsete lümfisõlmede katse: DA“.
criteria for scoring micronucleate cells (selection of analysable cells and identification of micronucleus);
Kättesaadav aadressil: http://ihcp.jrc.ec.europa.eu/our_labs/eurl-ecvam/eurl-ecvam-recommendations/eurl-ecvam-recommendation-on-the-direct-peptide-reactivity-assay-dpra
numbers of cells analysed;
Kättesaadav aadressil: http://ecvam-dbalm.jrc.ec.europa.eu/
criteria for considering studies as positive, negative, or equivocal;
Kättesaadav aadressil: www.fda.gov/downloads/drugs/guidancecomplianceregulatoryinformation/guidance/ucm070107.pdf – 138
method(s) of statistical analysis used;
Asjakohasusega seoses võetakse arvesse ka katsemeetodi täpsust (kordustäpsust) (21).
methods, such as use of anti-kinetochore antibody or pan-centromeric specific probes, to characterise whether micronuclei contain whole or fragmented chromosomes, if applicable;
Kahjuliku toime rada (Adverse Outcome Pathway, AOP) –
definition of acceptable cells for analysis;
sündmuste ahel alates sihtkemikaali või sarnaste kemikaalide rühma keemilisest struktuurist molekulaarse initsieerimisetapi kaudu vaadeldava toime kliinilise avaldumiseni in vivo (2).
measurement of cytotoxicity used, e.g. CBPI or RI in the case of cytokinesis-block method; RICC or RPD when cytokinesis-block methods are not used; other observations if any (e.g. cell confluency, apoptosis, necrosis, metaphase counting, frequency of binucleated cells);
Kemikaal – aine või segu.
number of cells with micronuclei given separately for each treated and control culture, and defining whether from binucleate or mononucleate cells, where appropriate;
Seda kasutatakse, et määrata lähtetaseme reaktsioon samas lahustis lahustatud uuritava kemikaaliga töödeldud proovides.
historical negative (solvent) and positive control data, with ranges, means and standard deviation and 95 % control limits for the distribution, as well as the number of data;
Mitme koostisosaga aine
statistical analysis; p-values if any.
aine, mis on määratletud oma kvantitatiivse koostisega, milles on rohkem kui üks põhikoostisosa, mille kontsentratsioon on vahemikus 10 %–80 % (massiprotsent).
Chapter B.10 of this Annex: In Vitro Mammalian Chromosome Aberration Test,.
Mitme koostisosaga aine saadakse tootmisprotsessi käigus.
In vitro micronucleus assay with Chinese hamster V79 cells — results of a collaborative study with in situ exposure to 26 chemical substances.
Erinevus segu ja mitme koostisosaga aine vahel seisneb selles, et segu saadakse kahe või enama aine kokku segamisel, ilma et toimuks keemilist reaktsiooni.
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.
Mitme koostisosaga aine saadakse keemilise reaktsiooni tulemusel.
Comparison of four different treatment conditions of extended exposure in the in vitro micronucleus assay using TK6 lymphoblastoid cells.
Ohtlikkus –
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.
mõjuri või olukorra iseloomulik omadus, mis võib põhjustada kahjulikku toimet, kui organism, süsteem või (alam-)populatsioon selle mõjuriga kokku puutub.
Available at: http://ecvam.jrc.it/index.htm.
paralleelproov, mis sisaldab kõiki katsesüsteemi komponente ja mida töödeldakse ainega, mis teadaolevalt kutsub esile positiivse reaktsiooni.
(eds), Elsevier, North-Holland, pp. 85-88.
Segu
Available at: http://www.pops.int/
ainete segu või lahus, mis koosneb kahest või enamast ainest, mis omavahel ei reageeri (1).
Corrigendum to “Report from the in vitro micronucleus assay working group”.
etalonainega enne katseseeria analüüside tegemist kindlaks määratud seadme toimivus (nt tundlikkus) (22).
“In vitro micronucleus test”, in Encyclopedia of Biopharmaceutical Statistics, 2nd ed.
Tundlikkusega mõõdetakse katsemeetodi täpsust, kui saadud tulemusi saab kategoriseerida, ning see on oluline aspekt katsemeetodi asjakohasuse hindamisel (21).
Chow, S. (ed.), Marcel Dekker, Inc.
näitab katsemeetodi tulemuste lähedust kokkulangemisele kinnitatud võrdlusväärtustega (mõõtetäpsus).
New York, pp. 463-467.
Täpsusega mõõdetakse katsemeetodi toimivust ja see on üks asjakohasuse aspektidest.
Analysis of Data from in vitro Cytogenetic Assays. in Statistical Evaluation of Mutagenicity Test Data, Kirkland, D.J.
Terminit kasutatakse ka kordustäpsuse tähenduses, mis näitab, kui sageli annab katsemeetod õige tulemuse (21).