natgillin/translations-raw huggingface dataset:
The most commonly used system that is recommended by default, unless otherwise justified, is a co-factor-supplemented post-mitochondrial fraction (S9) prepared from the livers of rodents (generally rats) treated with enzyme-inducing agents such as Aroclor 1254 (21) (22) (23) or a combination of phenobarbital and β-naphthoflavone (24) (25) (26) (27) (28) (29).
kemikaali identimisandmed, näiteks IUPACi või CASi nimetus, CASi number, SMILESi või InChI kood, struktuurivalem, puhtus, võimaluse ja vajaduse korral lisandite keemiline määratlus jne;
The latter combination does not conflict with the Stockholm Convention on Persistent Organic Pollutants (30) and has been shown to be as effective as Aroclor 1254 for inducing mixed-function oxidases (24) (25) (26) (28).
mitut koostisosa sisaldav aine, UVCB või segu:
The S9 fraction typically is used at concentrations ranging from 1 to 2 % (v/v) but may be increased to 10 % (v/v) in the final test medium.
võimalikult täpne omaduste kirjeldus, nt koostisosade keemiline määratlus (vt eespool), kvantitatiivne sisaldus ja asjakohased füüsikalis-keemilised omadused.
The use of products that reduce the mitotic index, especially calcium complexing products (31) should be avoided during treatment.
Lahusti:
The choice of type and concentration of exogenous metabolic activation system or metabolic inducer employed may be influenced by the class of chemicals being tested.
lahusti valiku põhjendus;
Test chemical preparation
samuti tuleb esitada lahusti protsendiline sisaldus lõppsöötmes.
Solid test chemicals should be prepared in appropriate solvents and diluted, if appropriate, prior to treatment of the cells (see paragraph 23).
Rakud:
Liquid test chemicals may be added directly to the test system and/or diluted prior to treatment of the test system.
rakkude tüüp ja päritolu;
Gaseous or volatile test chemicals should be tested by appropriate modifications to the standard protocols, such as treatment in sealed culture vessels (32) (33) (34).
karüotüübi omadused ja kasutatava rakutüübi sobivus;
Preparations of the test chemical should be made just prior to treatment unless stability data demonstrate the acceptability of storage.
rakuliinide puhul: mükoplasma puudumine;
Solvents
rakuliinide puhul: teave rakutsükli kestuse, rakupopulatsiooni kahekordistumise aja või proliferatsiooniindeksi kohta;
The solvent should be chosen to optimize the solubility of the test chemicals without adversely impacting the conduct of the assay, e.g. changing cell growth, affecting the integrity of the test chemical, reacting with culture vessels, impairing the metabolic activation system.
veredoonorite sugu, vanus ja muu asjakohane teave doonori, täisvere või eraldatud lümfotsüütide kohta, kasutatud mitogeen;
It is recommended that, wherever possible, the use of an aqueous solvent (or culture medium) should be considered first.
rakuliinide puhul: passaažide arv, kui olemas;
Well established solvents are for example water or dimethyl sulfoxide.
rakuliinide puhul: rakukultuuride säilitamise meetodid;
Generally organic solvents should not exceed 1 % (v/v) and aqueous solvents (saline or water) should not exceed 10 % (v/v) in the final treatment medium.
rakuliinide puhul: modaalne kromosoomiarv.
If not well-established solvents are used (e.g. ethanol or acetone), their use should be supported by data indicating their compatibility with the test chemicals, the test system and their lack of genetic toxicity at the concentration used.
Katsetingimused:
In the absence of that supporting data, it is important to include untreated controls (see Appendix 1) to demonstrate that no deleterious or clastogenic effects are induced by the chosen solvent.
metafaasi peatava kemikaali nimetus, kontsentratsioon ja rakkudega kokkupuute kestus;
Measuring cell proliferation and cytotoxicity and choosing treatment concentrations
uuritava kemikaali kontsentratsioon, väljendatuna lõppkontsentratsioonina söötmes (nt μg või mg/ml või mM söötmes);
When determining the highest test chemical concentration, concentrations that have the capability of producing artifactual positive responses, such as those producing excessive cytotoxicity (see paragraph 22), precipitation in the culture medium (see paragraph 23), or marked changes in pH or osmolality (see paragraph 5), should be avoided.
kontsentratsioonide ja rakukultuuride arvu valimise põhjendus, sealhulgas nt tsütotoksilisuse andmed ja lahustuvuse piirangud;
If the test chemical causes a marked change in the pH of the medium at the time of addition, the pH might be adjusted by buffering the final treatment medium so as to avoid artifactual positive results and to maintain appropriate culture conditions.
söötme koostis; CO2 kontsentratsioon, kui asjakohane, niiskusesisaldus;
Measurements of cell proliferation are made to assure that a sufficient number of treated cells have reached mitosis during the test and that the treatments are conducted at appropriate levels of cytotoxicity (see paragraphs 18 and 22).
söötmele lisatud lahusti ja uuritava kemikaali kontsentratsioon (ja/või ruumala);
Cytotoxicity should be determined with and without metabolic activation in the main experiment using an appropriate indication of cell death and growth.
inkubatsioonitemperatuur;
While the evaluation of cytotoxicity in an initial test may be useful to better define the concentrations to be used in the main experiment, an initial test is not mandatory.
inkubatsiooniaeg;
If performed, it should not replace the measurement of cytotoxicity in the main experiment.
töötluse kestus;
Relative Population Doubling (RPD) or Relative Increase in Cell Count (RICC) are appropriate methods for the assessment of cytotoxicity in cytogenetic tests (13) (15) (35) (36) (55) (see Appendix 2 for formulas).
pärast töötlemist rakkude kogumise aeg;
In case of long-term treatment and sampling times after the beginning of treatment longer than 1,5 normal cell cycle lengths (i.e. longer than 3 cell cycle lengths in total), RPD might underestimate cytotoxicity (37).
rakkude külvitihedus, vajaduse korral;
Under these circumstances RICC might be a better measure or the evaluation of cytotoxicity after 1,5 normal cell cycle lengths would be a helpful estimate using RPD.
metaboolse aktivatsiooni süsteemi tüüp ja koostis (S9-fraktsiooni allikas, S9-fraktsiooni segu valmistamise meetod, S9-fraktsiooni segu ja S9-fraktsiooni kontsentratsioon ja ruumala lõppsöötmes, S9-fraktsiooni kvaliteedikontrollid);
For lymphocytes in primary cultures, while the mitotic index (MI) is a measure of cytotoxic/cytostatic effects, it is influenced by the time after treatment it is measured, the mitogen used and possible cell cycle disruption.
positiivseks ja negatiivseks kontrolliks kasutatud kemikaalid, nende lõppkontsentratsioonid iga töötluse tingimuste puhul;
However, the MI is acceptable because other cytotoxicity measurements may be cumbersome and impractical and may not apply to the target population of lymphocytes growing in response to PHA stimulation.
mikroskoobipreparaatide valmistamise meetodid ja kasutatud värvimismeetodid;
While RICC and RPD for cell lines and MI for primary culture of lymphocytes are the recommended cytotoxicity parameters, other indicators (e.g. cell integrity, apoptosis, necrosis, cell cycle) could provide useful additional information.
katsete nõuetekohasuse kriteeriumid;
At least three test concentrations (not including the solvent and positive controls) that meet the acceptability criteria (appropriate cytotoxicity, number of cells, etc) should be evaluated.
aberratsioonide hindamise kriteeriumid;
Whatever the types of cells (cell lines or primary cultures of lymphocytes), either replicate or single treated cultures may be used at each concentration tested.
analüüsitud metafaaside arv;
While the use of duplicate cultures is advisable, single cultures are also acceptable provided that the same total number of cells are scored for either single or duplicate cultures.
tsütotoksilisuse mõõtmise meetodid;
The use of single cultures is particularly relevant when more than 3 concentrations are assessed (see paragraph 31).
muu täiendav asjakohane teave tsütotoksilisuse ja kasutatud meetodi kohta;
The results obtained in the independent replicate cultures at a given concentration can be pooled for the data analysis (38).
kriteeriumid, mille alusel uuringutulemust käsitatakse positiivse, negatiivse või ebaselgena;
For test chemicals demonstrating little or no cytotoxicity, concentration intervals of approximately 2 to 3 fold will usually be appropriate.
pH, osmolaalsuse ja sademe määramiseks kasutatud meetodid.
Where cytotoxicity occurs, the test concentrations selected should cover a range from that producing cytotoxicity as described in paragraph 22 and including concentrations at which there is moderate and little or no cytotoxicity.
töödeldud rakkude arv ja kogutud rakkude arv iga rakukultuuri kohta, kui kasutatakse rakuliine;
Many test chemicals exhibit steep concentration response curves and in order to obtain data at low and moderate cytotoxicity or to study the dose response relationship in detail, it will be necessary to use more closely spaced concentrations and/or more than three concentrations (single cultures or replicates), in particular in situations where a repeat experiment is required (see paragraph 47).
tsütotoksilisuse mõõtmise tulemused, nt RPD, RICC, MI või muud vaatlusandmed, kui on olemas;
If the maximum concentration is based on cytotoxicity, the highest concentration should aim to achieve 55 ± 5 % cytotoxicity using the recommended cytotoxicity parameters (i.e. reduction in RICC and RPD for cell lines and reduction in MI for primary cultures of lymphocytes to 45 ± 5 % of the concurrent negative control).
rakutsükli kestuse, rakupopulatsiooni kahekordistumise aja või proliferatsiooniindeksi andmed rakuliinide puhul;
Care should be taken in interpreting positive results only to be found in the higher end of this 55 ± 5 % cytotoxicity range (13).
sademe tekke tunnused ja määramise aeg;
For poorly soluble test chemicals that are not cytotoxic at concentrations lower than the lowest insoluble concentration, the highest concentration analysed should produce turbidity or a precipitate visible by eye or with the aid of an inverted microscope at the end of the treatment with the test chemical.
aberratsioonide, sh tühikute määratlemine;
Even if cytotoxicity occurs above the lowest insoluble concentration, it is advisable to test at only one concentration producing turbidity or with a visible precipitate because artifactual effects may result from the precipitate.
loendatud rakkude arv, kromosoomaberratsioonidega rakkude arv ning kromosoomaberratsioonide arv tüübiti eraldi iga töödeldud või kontrolli kultuuri kohta, tühikuid arvestades või välja jättes;
At the concentration producing a precipitate, care should be taken to assure that the precipitate does not interfere with the conduct of the test (e.g. staining or scoring).
ploidsuse muutused (polüploidsed rakud ja endoreduplitseeritud kromosoomidega rakud esitada eraldi), kui neid täheldati;
The determination of solubility in the culture medium prior to the experiment may be useful.
kontsentratsiooni-toime vaheline seos, kui võimalik;
If no precipitate or limiting cytotoxicity is observed, the highest test concentration should correspond to 10 mM, 2 mg/ml or 2 μl/ml, whichever is the lowest (39) (40) (41).
paralleelse negatiivse (lahusti) ja positiivse kontrolli andmed (kontsentratsioonid ja lahustid);
When the test chemical is not of defined composition, e.g. a substance of unknown or variable composition, complex reaction products or biological material (UVCB) (42), environmental extract etc., the top concentration may need to be higher (e.g. 5 mg/ml), in the absence of sufficient cytotoxicity, to increase the concentration of each of the components.
varasemad negatiivse ja positiivse kontrolli andmed, sh vahemik, keskväärtused, standardhälbed ja jaotuse usaldusvahemik usaldatavustasemel 95 %, samuti selliste andmete arv;
It should be noted however that these requirements may differ for human pharmaceuticals (43).
statistiline analüüs; p-väärtused, kui olemas.
Controls
Tulemuste arutelu.
Concurrent negative controls (see paragraph 15), consisting of solvent alone in the treatment medium and treated in the same way as the treatment cultures, should be included for every harvest time.
Järeldused.
Concurrent positive controls are needed to demonstrate the ability of the laboratory to identify clastogens under the conditions of the test protocol used and the effectiveness of the exogenous metabolic activation system, when applicable.
Series on Testing and Assessment No.
Examples of positive controls are given in the table 1 below.
234, OECD, Paris.
Alternative positive control chemicals can be used, if justified.
, „Cytological Methods for Detecting Chemical Mutagens“, in Chemical Mutagens, Principles and Methods for their Detection, Vol.
Because in vitro mammalian cell tests for genetic toxicity are sufficiently standardized, the use of positive controls may be confined to a clastogen requiring metabolic activation.
4, Hollaender, A. (ed.), Plenum Press, New York ja London, pp. 1–29.
Provided it is done concurrently with the non-activated test using the same treatment duration, this single positive control response will demonstrate both the activity of the metabolic activation system and the responsiveness of the test system.
Ishidate, M. Jr., T. Sofuni (1985), „The in vitro Chromosomal Aberration Test Using Chinese Hamster Lung (CHL) Fibroblast Cells in Culture“, in Progress in Mutation Research, Vol.
Long term treatment (without S9) should however have its own positive control as the treatment duration will differ from the test using metabolic activation.
5, Ashby, J. et al. (eds.), Elsevier Science Publishers, Amsterdam-NewYork-Oxford, pp.
Each positive control should be used at one or more concentrations expected to give reproducible and detectable increases over background in order to demonstrate the sensitivity of the test system (i.e. the effects are clear but do not immediately reveal the identity of the coded slides to the reader), and the response should not be compromised by cytotoxicity exceeding the limits specified in the test method.
427–432.
Reference chemicals recommended for assessing laboratory proficiency and for selection of positive controls
318–327.
Category
Käesoleva lisa peatükk B.49, „Mikrotuumade tekke in vitro katse imetajarakkudega“.
Clastogens active without metabolic activation
85–88.
Methyl methanesulphonate
Kättesaadav aadressil: http://www.pops.int/.
Mitomycin C
McCooey (1982), „CHO/HGPRT Mutation Assay: Evaluation of Gases and Volatile Liquids“, väljaandes Genotoxic Effects of Airborne Agents, Tice, R.R., D.L. Costa, K.M.
itroquinoline-N-Oxide
Schaich (eds.), Plenum, New York, pp.
Cytosine arabinoside
91–103.
Clastogens requiring metabolic activation
(ed.)
Benzo(a)pyrene
Cambridge University Press, Cambridge, pp.
Cyclophosphamide
141–154.
PROCEDURE
Nõude esitamisel kättesaadav.
Treatment with test chemical
Kättesaadav aadressil: https://federalregister.gov/a/2012-13774.
Proliferating cells are treated with the test chemical in the presence and absence of a metabolic activation system.
OECD (2014), „Statistical analysis supporting the revision of the genotoxicity Test Guidelines“, OECD Environment, Health and Safety Publications (EHS), Series on Testing and Assessment No. 198, OECD Publishing, Paris.
Culture harvest time
(ed.), Cambridge University Press, Cambridge, pp.
For thorough evaluation, which would be needed to conclude a negative outcome, all three of the following experimental conditions should be conducted using a short term treatment with and without metabolic activation and long term treatment without metabolic activation (see paragraphs 43, 44 and 45):
62–86.
Cells should be exposed to the test chemical without metabolic activation for 3-6 hours, and sampled at a time equivalent to about 1,5 normal cell cycle lengths after the beginning of treatment (18),
(ed.), Cambridge University Press, Cambridge, pp.
Cells should be exposed to the test chemical with metabolic activation for 3-6 hours, and sampled at a time equivalent to about 1,5 normal cell cycle lengths after the beginning of treatment (18),
141–154.
Cells should be continuously exposed without metabolic activation until sampling at a time equivalent to about 1,5 normal cell cycle lengths.
liide
Certain chemicals (e.g. nucleoside analogues) may be more readily detected by treatment/sampling times longer than 1,5 normal cell cycle lengths (24).
MÕISTED
In the event that any of the above experimental conditions lead to a positive response, it may not be necessary to investigate any of the other treatment regimens.
Aneuploidsus
Chromosome preparation
iga kõrvalekalle normaalsest diploidsest (või haploidsest) kromosoomiarvust ühe või enama kromosoomi võrra, kuid mitte terve(te) kromosoomikomplekti(de) võrra (polüploidsus).
Cell cultures are treated with colcemid or colchicine usually for one to three hours prior to harvesting.
Apoptoos
Each cell culture is harvested and processed separately for the preparation of chromosomes.
aine või segu.
Chromosome preparation involves hypotonic treatment of the cells, fixation and staining.
Interfaasirakud
In monolayers, mitotic cells (identifiable as being round and detaching from the surface) may be present at the end of the 3-6 hour treatment.
Endoreduplikatsioon
Because these mitotic cells are easily detached, they can be lost when the medium containing the test chemical is removed.
Selle tulemusena tekivad kromosoomid, milles on 4, 8, 16, … kromatiidi.
If there is evidence for a substantial increase in the number of mitotic cells compared with controls, indicating likely mitotic arrest, then the cells should be collected by centrifugation and added back to cultures, to avoid losing cells that are in mitosis, and at risk for chromosome aberration, at the time of harvest.
Genoommutatsioon
Analysis
Lahusti kontroll
All slides, including those of the positive and negative controls, should be independently coded before microscopic analysis for chromosomal aberrations.
üldtermin, millega tähistatakse kontrollikultuure, kuhu lisatakse ainult uuritava kemikaali lahustamiseks kasutatud lahusti.
Since fixation procedures often result in a proportion of metaphase cells which have lost chromosomes, the cells scored should, therefore, contain a number of centromeres equal to the modal number +/- 2.
Maksa S9-fraktsioon
At least 300 well-spread metaphases should be scored per concentration and control to conclude a test chemical as clearly negative (see paragraph 45).
üldtermin, mis hõlmab kõiki DNA või kromosoomi kahjustuse tüüpe, sealhulgas murde, deletsioone, adukte, nukleotiidide modifikatsioone ja sidemete teket, ümberpaigutusi, geenimutatsioone, kromosoomaberratsioone ja aneuploidsust.
The 300 cells should be equally divided among the replicates, when replicate cultures are used.
Mitoosiindeks
When single cultures are used per concentration (see paragraph 21), at least 300 well spread metaphases should be scored in this single culture.
suhtarv, mis saadakse, kui vaatlusaluse rakupopulatsiooni metafaasis olevate rakkude arv jagatakse rakkude koguarvuga; näitab rakkude proliferatsioonitaset kõnealuses populatsioonis.
Scoring 300 cells has the advantage of increasing the statistical power of the test and in addition, zero values will be rarely observed (expected to be only 5 %) (44).
p53 staatus
The number of metaphases scored can be reduced when high numbers of cells with chromosome aberrations are observed and the test chemical considered as clearly positive.
p53 on valk, mis on seotud rakutsükli, apoptoosi ja DNA reparatsiooni reguleerimisega.
Cells with structural chromosomal aberration(s) including and excluding gaps should be scored.
Polüploidsus
Breaks and gaps are defined in Appendix 1 according to (45) (46).
tähendavad uuritava kemikaali lõppkontsentratsioone söötmes.
Chromatid- and chromosome-type aberrations should be recorded separately and classified by sub-types (breaks, exchanges).
tsütotoksilisuse mõõtmise meetod, kui cytoB-d ei kasutata (vt valem, 2. liide).
Procedures in use in the laboratory should ensure that analysis of chromosomal aberrations is performed by well-trained scorers and peer-reviewed if appropriate.
struktuurne kromosoomikahjustus, mis väljendub üksikkromatiidide katkemisena või kromatiidide katkemise ja nendevahelise taasühinemisena.
Although the purpose of the test is to detect structural chromosomal aberrations, it is important to record polyploidy and endoreduplication frequencies when these events are seen.
Polüploidsus
(See paragraph 2).
rakkude või organismi genoommutatsioon, mis hõlmab terveid kromosoomistikke, erinevalt sellisest genoommutatsioonist, mis hõlmab üksikute kromosoomide arvu muutust (aneuploidsus).
Proficiency of the laboratory
Proliferatsiooniindeks (Proliferation Index, PI)
In order to establish sufficient experience with the test prior to using it for routine testing, the laboratory should have performed a series of experiments with reference positive chemicals acting via different mechanisms and various negative controls (using various solvents/vehicle).
Kromosoomaberratsioon
These positive and negative control responses should be consistent with the literature.
struktuurne kromosoomikahjustus, mis väljendub mõlema kromatiidi katkemisena või katkemise ja taasühinemisena samas kohas.
This is not applicable to laboratories that have experience, i.e. that have an historical data base available as defined in paragraph 37.
rakkude arvu suurenemine mitootilise rakujagunemise tagajärjel.
A selection of positive control chemicals (see Table 1 in paragraph 26) 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 chemicals and determine the effectiveness of the metabolic activation system.
üldtermin, millega tähistatakse kontrollikultuure, millele lisatakse ainult uuritava kemikaali lahustamiseks kasutatud lahusti.
A range of concentrations of the selected chemicals should be chosen so as to give reproducible and concentration-related increases above the background in order to demonstrate the sensitivity and dynamic range of the test system.
S9-fraktsiooni segu
Historical control data
S9-fraktsiooni ja metaboolsete ensüümide aktiivsuse jaoks vajalike kofaktorite segu.
The laboratory should establish:
Tsentromeer
A historical positive control range and distribution,
Mitoosiindeks (MI) suhtarv, mis saadakse, kui vaatlusaluse rakupopulatsiooni metafaasis olevate rakkude arv jagatakse rakkude koguarvuga; on selle rakupopulatsiooni proliferatsioonitaseme näitaja.
A historical negative (untreated, solvent) control range and distribution.
Mutageenne tekitab päriliku muutuse geenide DNA aluspaaride järjestuses või kromosoomide struktuuris (kromosoomaberratsioonid). p53 staatus
When first acquiring data for an historical negative control distribution, concurrent negative controls should be consistent with published control data, where they exist.
Tsütokinees
As more experimental data are added to the control distribution, concurrent negative controls should ideally be within the 95 % control limits of that distribution (44) (47).
mitoosile vahetult järgnev rakujagunemise protsess, mille tulemusel tekivad kaks tütarrakku, milles mõlemas on üksainus tuum.
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.
kemikaaliga kokku puutunud rakukultuuris rakkude arvu suurenemine, võrreldes töötlemata kultuuriga; suhtarv väljendatakse protsentides.
Laboratories should use quality control methods, such as control charts (e.g. C-charts or X-bar charts (48)), to identify how variable their positive and negative control data are, and to show that the methodology is ‘under control’ in their laboratory (44).
Rakkude paljunemine rakkude arvu suurenemine mitootilise rakujagunemise tagajärjel.