natgillin/translations-raw huggingface dataset:
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.
vsi rezultati nahajajo znotraj porazdelitve podatkov o negativnih kontrolah iz preteklih preskusov (npr. 95-odstotna kontrolna meja, ki temelji na Poissonovi porazdelitvi; glej odstavek 39).
Chromosome preparation
Nato se šteje, da preskusna kemikalija ne more povzročiti nastanka kromosomskih aberacij v kultiviranih celicah sesalcev v tem preskusnem sistemu.
Cell cultures are treated with colcemid or colchicine usually for one to three hours prior to harvesting.
Preverjanje jasno pozitivnega ali jasno negativnega odziva ni potrebno.
Each cell culture is harvested and processed separately for the preparation of chromosomes.
Če odziv ni niti jasno negativen niti jasno pozitiven, kot je opisano zgoraj, ali za podporo pri ugotavljanju biološke pomembnosti rezultatov, je treba podatke oceniti s strokovno presojo in/ali nadaljnjimi preiskavami.
Chromosome preparation involves hypotonic treatment of the cells, fixation and staining.
Štetje dodatnih celic (po potrebi) ali izvedba ponovitve poskusa je lahko koristna, po možnosti z uporabo spremenjenih preskusnih pogojev (npr. razmik med koncentracijami, drugi pogoji za presnovno aktivacijo (tj. koncentracija ali izvor S9)).
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.
Zbirka podatkov v redkih primerih celo po izvedbi nadaljnjih preiskav onemogoča sprejetje sklepa o pozitivnih ali negativnih rezultatih, zato se zaključi, da je odziv na preskusno kemikalijo dvoumen.
Because these mitotic cells are easily detached, they can be lost when the medium containing the test chemical is removed.
Povečanje števila poliploidnih celic kaže na to, da ima preskusna kemikalija potencial za zaviranje mitotičnih procesov in povzročanje numeričnih kromosomskih aberacij (52).
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.
Povečanje števila celic z endoredupliciranimi kromosomi lahko pomeni, da lahko preskusne kemikalije zavirajo potek celičnega cikla (53) (54) (glej odstavek 2).
Analysis
Pojav poliploidnih celic in celic z endoredupliciranimi kromosomi je zato treba zabeležiti ločeno.
All slides, including those of the positive and negative controls, should be independently coded before microscopic analysis for chromosomal aberrations.
vir, številka serije, rok uporabe, če je na voljo;
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.
stabilnost preskusne kemikalije, če je znana;
At least 300 well-spread metaphases should be scored per concentration and control to conclude a test chemical as clearly negative (see paragraph 45).
topnost in stabilnost preskusne kemikalije v topilu/nosilcu, če sta znani;
The 300 cells should be equally divided among the replicates, when replicate cultures are used.
meritve vrednosti pH, osmolarnosti in oborine v gojišču, ki mu je bila dodana preskusna kemikalija, če je ustrezno.
When single cultures are used per concentration (see paragraph 21), at least 300 well spread metaphases should be scored in this single culture.
Snov iz ene sestavine:
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).
fizični videz, topnost v vodi in dodatne ustrezne fizikalno-kemijske lastnosti;
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.
kemijski identifikacijski podatki, kot so ime po nomenklaturi IUPAC ali ime CAS, številka CAS, koda po sistemu SMILES ali identifikatorju InChI, strukturna formula, čistost, kemijska identiteta nečistoč, kot je ustrezno in praktično izvedljivo, itd.
Cells with structural chromosomal aberration(s) including and excluding gaps should be scored.
Snov z več sestavinami, UVCB in zmesi:
Breaks and gaps are defined in Appendix 1 according to (45) (46).
čim obsežnejša opredelitev lastnosti s kemijsko identiteto (glej zgoraj), kvantitativnim pojavljanjem in ustreznimi fizikalno-kemijskimi lastnostmi sestavin.
Chromatid- and chromosome-type aberrations should be recorded separately and classified by sub-types (breaks, exchanges).
Topilo:
Procedures in use in the laboratory should ensure that analysis of chromosomal aberrations is performed by well-trained scorers and peer-reviewed if appropriate.
utemeljitev izbire topila;
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.
navesti je treba tudi odstotek topila v končnem gojišču.
(See paragraph 2).
Celice:
Proficiency of the laboratory
vrsta in izvor celic;
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).
lastnosti kariotipa in ustreznost uporabljene vrste celic;
These positive and negative control responses should be consistent with the literature.
za celične linije – odsotnost mikoplazme;
This is not applicable to laboratories that have experience, i.e. that have an historical data base available as defined in paragraph 37.
za celične linije – informacije o trajanju celičnega cikla, podvojitvenem času ali indeksu proliferacije;
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.
spol donorjev krvi, starost in vse pomembne informacije o donorju, uporabljeni krvi v celoti ali ločenih limfocitih, mitogenih;
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.
za celične linije – število pasaž, če je na voljo;
Historical control data
za celične linije – metode vzdrževanja celičnih kultur;
The laboratory should establish:
za celične linije – modalno število kromosomov.
A historical positive control range and distribution,
Preskusni pogoji:
A historical negative (untreated, solvent) control range and distribution.
identiteta kemikalije, ki zavira metafazo, njena koncentracija in čas izpostavljenosti celic;
When first acquiring data for an historical negative control distribution, concurrent negative controls should be consistent with published control data, where they exist.
koncentracija preskusne kemikalije, izražena kot končna koncentracija v gojišču (npr. v μg ali mg/ml ali mM gojišča);
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).
razlogi za izbiro koncentracij in števila kultur, vključno z npr. podatki o citotoksičnosti in mejah topnosti;
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.
sestava gojišča, koncentracija CO2, če je na voljo, stopnja vlažnosti;
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).
koncentracija (in/ali volumen) topila in preskusne kemikalije, ki se dodata v gojišče;
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 (47).
temperatura inkubacije;
Any changes to the experimental protocol should be considered in terms of their consistency with the laboratory's existing historical control databases.
čas inkubacije;
Any major inconsistencies should result in the establishment of a new historical control database.
trajanje tretiranja;
Negative control data should consist of the incidence of cells with chromosome aberrations from a single culture or the sum of replicate cultures as described in paragraph 21.
odvzem celic po tretiranju;
Concurrent negative controls should ideally be within the 95 % control limits of the distribution of the laboratory's historical negative control database (44) (47).
gostota celic ob nasaditvi, če je ustrezno;
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 37) and evidence of absence of technical or human failure.
vrsta in sestava sistema presnovne aktivacije (vir S9, način priprave zmesi S9, koncentracija ali volumen zmesi S9 in S9 v končnem gojišču, nadzor kakovosti S9);
Presentation of the results
kemikalije pozitivne in negativne kontrole, končne koncentracije za vsak pogoj tretiranja;
The percentage of cells with structural chromosomal aberration(s) should be evaluated.
metode priprave preparatov in uporabljene tehnike barvanja;
Chromatid- and chromosome-type aberrations classified by sub-types (breaks, exchanges) should be listed separately with their numbers and frequencies for experimental and control cultures.
merila za sprejemljivost poskusov;
Gaps are recorded and reported separately but not included in the total aberration frequency.
merila za štetje aberacij;
Percentage of polyploidy and/or endoreduplicated cells are reported when seen.
število analiziranih metafaz;
Concurrent measures of cytotoxicity for all treated, negative and positive control cultures in the main aberration experiment(s) should be recorded.
metode merjenja citotoksičnosti;
Individual culture data should be provided.
vsi dodatni podatki, pomembni za citotoksičnost in uporabljeno metodo;
Additionally, all data should be summarised in tabular form.
merila za obravnavanje študij kot pozitivnih, negativnih ali dvoumnih;
Acceptability Criteria
uporabljene metode za določitev vrednosti pH, osmolarnosti in obarjanja.
Acceptance of a test is based on the following criteria:
število tretiranih celic in število odvzetih celic za vsako kulturo, kadar se uporabljajo celične linije;
The concurrent negative control is considered acceptable for addition to the laboratory historical negative control database as described in paragraph 39.
meritve citotoksičnosti, npr. relativna podvojitev populacije, relativno povečanje števila celic in mitotični indeks, morebitna druga opažanja;
Concurrent positive controls (see paragraph 26) should induce responses that are compatible with those generated in the historical positive control data base and produce a statistically significant increase compared with the concurrent negative control.
informacije o trajanju celičnega cikla, času podvojitve ali indeksu proliferacije v primeru celičnih linij;
Cell proliferation criteria in the solvent control should be fulfilled (paragraphs 17 and 18).
znaki obarjanja in čas odkritja;
All three experimental conditions were tested unless one resulted in positive results (see paragraph 28).
opredelitev aberacij, vključno z vrzelmi;
Adequate number of cells and concentrations are analysable (paragraphs 31 and 21).
število štetih celic, število celic s kromosomskimi aberacijami in vrsta kromosomskih aberacij, ki se navedejo ločeno za vsako tretirano in kontrolno kulturo, vključno z vrzelmi in brez njih;
The criteria for the selection of top concentration are consistent with those described in paragraphs 22, 23 and 24.
spremembe v ploidnosti (poliploidne celice in celice z endoredupliciranimi kromosomi, navedene ločeno), če so opažene;
Evaluation and interpretation of results
razmerje med koncentracijo in odzivom, kadar je mogoče;
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 paragraph 28):
podatki o sočasnih negativnih (s topilom) in pozitivnih kontrolah (koncentracije in topila);
at least one of the test concentrations exhibits a statistically significant increase compared with the concurrent negative control,
podatki o negativnih (s topilom) in pozitivnih kontrolah iz preteklih preskusov, skupaj z razponi, srednjimi vrednostmi in standardnimi odkloni ter 95-odstotnimi kontrolnimi mejami za porazdelitev, pa tudi število podatkov;
the increase is dose-related when evaluated with an appropriate trend test,
statistične analize, p-vrednosti, če obstajajo.
any of the results are outside the distribution of the historical negative control data (e.g. Poisson-based 95 % control limits; see paragraph 39).
Razprava o rezultatih
When all of these criteria are met, the test chemical is then considered able to induce chromosomal aberrations in cultured mammalian cells in this test system.
Sklepne ugotovitve
Recommendations for the most appropriate statistical methods can be found in the literature (49) (50) (51).
Overview of the set of OECD Genetic Toxicology Test Guidelines and updates performed in 2014–2015.
Providing that all acceptability criteria are fulfilled, a test chemical is considered clearly negative if, in all experimental conditions examined (see paragraph 28):
Series on Testing and Assessment, št. 234, OECD, Pariz.
none of the test concentrations exhibits a statistically significant increase compared with the concurrent negative control,
4, Hollaender, A. (ed.), Plenum Press, New York in London, str. 1–29.
there is no concentration-related increase when evaluated with an appropriate trend test,
Ishidate, M. Jr. in Sofuni, T. (1985). „The in vitro Chromosomal Aberration Test Using Chinese Hamster Lung (CHL) Fibroblast Cells in Culture“ in Progress in Mutation Research, zv.
all results are inside the distribution of the historical negative control data (e.g. Poisson-based 95 % control limits; see paragraph 39).
5, Ashby, J. idr.
The test chemical is then considered unable to induce chromosomal aberrations in cultured mammalian cells in this test system.
(eds.), Elsevier Science Publishers, Amsterdam – New York – Oxford, str. 427–432.
There is no requirement for verification of a clearly positive or negative response.
„Improving dose selection and identification of aneugens in the in vitro chromosome aberration test by integration of flow cytometry-based methods“, Environmental and Molecular Mutagenesis, zv.
In case the response is neither clearly negative nor clearly positive as described above 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.
49/4, str. 318–327.
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.
Poglavje B.49 te priloge: Preskus mikronukleusov in vitro pri celicah sesalcev.
In rare cases, even after further investigations, the data set will preclude making a conclusion of positive or negative results, and therefore the test chemical response will be concluded to be equivocal.
„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. idr.
An increase in the number of polyploid cells may indicate that the test chemicals have the potential to inhibit mitotic processes and to induce numerical chromosomal aberrations (52).
(eds.), Elsevier, North-Holland, str 85–88.
An increase in the number of cells with endoreduplicated chromosomes may indicate that the test chemicals have the potential to inhibit cell cycle progress (53) (54) (see paragraph 2).
Na voljo na: http://www.pops.int/.
Therefore, incidence of polyploid cells and cells with endoreduplicated chromosomes should be recorded separately.
Tice, R. R., Costa, D. L. in Schaich, K. M. (eds.), Plenum, New York, str. 91–103.
source, lot number, limit date for use, if available
V: Statistical Evaluation of Mutagenicity Test Data, Kirkland, D.J.
stability of the test chemical itself, if known;
(ed.)
solubility and stability of the test chemical in solvent, if known.
Cambridge University Press, Cambridge, str. 141–154.
measurement of pH, osmolality and precipitate in the culture medium to which the test chemical was added, as appropriate.
Na voljo na zahtevo.
Mono-constituent substance:
Na voljo na: https://federalregister.gov/a/2012-13774.
physical appearance, water solubility, and additional relevant physicochemical properties;
OECD (2014).
chemical identification, such as IUPAC or CAS name, CAS number, SMILES or InChI code, structural formula, purity, chemical identity of impurities as appropriate and practically feasible, etc.
Statistical analysis supporting the revision of the genotoxicity Test Guidelines, OECD Environment, Health and Safety Publications (EHS), Series on Testing and Assessment, št. 198, OECD Publishing, Pariz.
Multi-constituent substance, UVCBs and mixtures:
Dodatek 1
characterised as far as possible by chemical identity (see above), quantitative occurrence and relevant physicochemical properties of the constituents.
OPREDELITEV POJMOV
Solvent:
Anevplodija
justification for choice of solvent.
vsako odstopanje od normalnega diploidnega (ali haploidnega) števila kromosomov za en ali več kot en kromosom, vendar ne za celotni set kromosomov (poliploidija).
percentage of solvent in the final culture medium should also be indicated.
Apoptoza
Cells:
programirana celična smrt, za katero je značilna vrsta korakov, ki vodijo v razgradnjo celic na delce, obdane z membrano, ki se nato odstranijo s fagocitozo ali osipanjem.
type and source of cells
Celična proliferacija
karyotype features and suitability of the cell type used;
povečanje števila celic kot posledica mitotične delitve celic.
absence of mycoplasma, for cell lines;
snov ali zmes.
for cell lines, information on cell cycle length, doubling time or proliferation index;
Kromatidni prelom
sex of blood donors, age and any relevant information on the donor, whole blood or separated lymphocytes, mitogen used;
prekinitev ene kromatide, pri čemer je jasno vidna neporavnanost ene od kromatid.
number of passages, if available, for cell lines;
Kromatidna vrzel
methods for maintenance of cell cultures, for cell lines;
neobarvano območje (akromatska lezija) posamezne kromatide, pri čemer je neporavnanost kromatide minimalna.
modal number of chromosomes, for cell lines.
Aberacija kromatidnega tipa
Test conditions:
strukturna poškodba kromosoma, izražena kot prelom posamezne kromatide ali prelom in združitev med kromatidama.
identity of the metaphase-arresting chemical, its concentration and duration of cell exposure;
Aberacija kromosomskega tipa
concentration of test chemical expressed as final concentration in the culture medium (e.g. μg or mg/mL or mM of culture medium).
strukturna poškodba kromosoma, izražena kot prelom ali prelom in združitev obeh kromatid na istem mestu.
rationale for selection of concentrations and number of cultures including, e.g. cytotoxicity data and solubility limitations;
Klastogen
composition of media, CO2 concentration if applicable, humidity level;
vsaka kemikalija, ki povzroči strukturne kromosomske aberacije v populacijah celic ali evkariotskih organizmih.
concentration (and/or volume) of solvent and test chemical added in the culture medium;
Koncentracije
incubation temperature;
se nanašajo na končne koncentracije preskusne kemikalije v gojišču.
incubation time;
Citotoksičnost
duration of treatment;
strukturna poškodba kromosoma, izražena kot prelom ali prelom in združitev obeh kromatid na istem mestu.
harvest time after treatment;
Endoreduplikacija
cell density at seeding, if appropriate;
proces, pri katerem v jedru po podvajanju DNK v fazi S ne začne potekati mitoza, temveč takoj nastopi druga faza S. Rezultat tega je, da nastanejo kromosomi s 4, 8, 16, … kromatidami.
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);
Genotoksiče
positive and negative control chemicals, final concentrations for each conditions of treatment;
splošni pojem, ki zajema vse vrste poškodb DNK ali kromosomov, vključno s prelomi, delecijami, adukti, nukleotidnimi spremembami in povezavami, prerazporeditvami, mutacijami genov, kromosomskimi aberacijami in anevploidijo.
methods of slide preparation and staining technique used;
Vsi genotoksični učinki ne vplivajo na nastanek mutacij ali trajno poškodbo kromosomov.
criteria for acceptability of assays;
Mitotični indeks (MI)
criteria for scoring aberrations;
razmerje med številom celic v metafazi in skupnim številom celic v opazovani populaciji celic; kazalnik stopnje proliferacije te populacije.
number of metaphases analysed;
Mitoza
methods for the measurements of cytotoxicity;
delitev celičnega jedra, ki je običajno razdeljena na profazo, prometafazo, metafazo, anafazo in telofazo.
any supplementary information relevant to cytotoxicity and method used;
Mutagen
criteria for considering studies as positive, negative or equivocal;
povzroča dedno spremembo zaporedij baznih parov DNK v genih ali strukturi kromosomov (kromosomske aberacije).
methods used to determine pH, osmolality and precipitation.
Numerična aberacija
the number of cells treated and the number of cells harvested for each culture when cell lines are used
sprememba števila kromosomov, ki se razlikuje od običajnega števila, ki je značilno za uporabljene celice.
cytotoxicity measurements, e.g. RPD, RICC, MI, other observations if any;
Poliploidija
information on cell cycle length, doubling time or proliferation index in case of cell lines;
numerične kromosomske aberacije v celicah ali organizmih, ki ne vplivajo na posamezen kromosom ali kromosome (anevploidija), temveč na celoten set kromosomov.
signs of precipitation and time of the determination;
Status beljakovine p53
Number of cells scored, number of cells with chromosomal aberrations and type of chromosomal aberrations given separately for each treated and control culture, including and excluding gaps;
Celice brez funkcionalne beljakovine p53, ki ne morejo zaustaviti celičnega cikla ali odstraniti poškodovanih celic z apoptozo ali drugimi mehanizmi (npr. indukcijo popravljanja DNK), povezanimi s funkcijami beljakovine p53, s katerimi se ta odzove na poškodbo DNK, so teoretično bolj nagnjene h genskim mutacijam ali kromosomskim aberacijam.
changes in ploidy (polyploid cells and cells with endoreduplicated chromosomes, given separately) if seen;
Relativno povečanje števila celic (RICC)
concentration-response relationship, where possible;
povečanje števila celic v kulturah, izpostavljenih kemikalijam, v primerjavi s povečanjem v netretiranih kulturah, pri čemer je razmerje izraženo kot odstotek.
concurrent negative (solvent) and positive control data (concentrations and solvents);
Relativna podvojitev populacije (RPD)