natgillin/translations-raw huggingface dataset:
Other methods for differentiation between clastogens and aneugens may be used if they have been shown to be effective and validated.
Za sve tretirane i kontrolne kulture potrebno je izračunati CBPI ili RI kako bi se izmjerilo kašnjenje staničnog ciklusa ako se primjenjuje metoda blokirane citokineze.
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).
U odsutnosti citohalazina B mjeri se RPD ili RICC (vidjeti Dodatak 2.).
Morphological observations of nuclei could also give indications of possible aneuploidy.
istodobna negativna kontrola smatra se prihvatljivom za dodavanje u bazu podataka laboratorija o prijašnjim negativnim kontrolama kako je opisano u stavku 50.,
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).
istodobne pozitivne kontrole (vidjeti stavak 50.) trebale bi inducirati odgovore koji su u skladu s onima koji su dobiveni u bazi podataka laboratorija o prijašnjim pozitivnim kontrolama te uzrokovati statistički značajno povećanje u usporedbi s istodobnom negativnom kontrolom,
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.
trebali bi biti ispunjeni kriteriji proliferacije stanica u kontroli s otapalom (stavci 25. – 27.),
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.
ispitani su svi pokusni uvjeti, osim ako su u jednom od njih dobiveni pozitivni rezultati (stavci 36. – 40.),
Regardless of the automated platform is used to enumerate micronuclei, CBPI, RI, RPD, or RICC should be assessed concurrently.
moguće je analizirati odgovarajući broj stanica i koncentracija (stavak 28. i stavci 44. – 46.),
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.
kriteriji za odabir najveće koncentracije u skladu su s kriterijima opisanima u stavcima 24. – 31.
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.
Pod uvjetom da su svi kriteriji prihvatljivosti ispunjeni, ispitivana kemikalija smatra se jasno pozitivnom ako u bilo kojem ispitanom pokusnom uvjetu (vidjeti stavke 36. – 39.):
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.
najmanje jedna od ispitnih koncentracija pokazuje statistički značajno povećanje u usporedbi s istodobnom negativnom kontrolom (89.),
Care should be taken not to score binucleate cells with irregular shapes or where the two nuclei differ greatly in size.
odgovarajući test trenda pokaže da je u najmanje jednom pokusnom uvjetu povećanje povezano s dozom (vidjeti stavak 28.),
In addition, binucleate cells should not be confused with poorly spread multi-nucleate cells.
bilo koji od rezultata nalazi se izvan raspona distribucije prijašnjih podataka o negativnim kontrolama (npr. kontrolne granice od 95 % prema Poissonovoj distribuciji; vidjeti stavak 52.).
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).
Ako su svi navedeni kriteriji ispunjeni, smatra se da ispitivana kemikalija može uzrokovati lomove i/ili povećanje ili gubitak kromosoma u tom ispitnom sustavu.
Scoring of mononucleate cells is acceptable if the test chemical is shown to interfere with cytoB activity.
Preporuke o najprikladnijim statističkim metodama dostupne su i u literaturi (90., 91., 92.).
A repeat test without CytoB might be useful in such cases.
Pod uvjetom da su svi kriteriji prihvatljivosti ispunjeni, ispitivana kemikalija smatra se jasno negativnom ako u svim ispitanim pokusnim uvjetima (vidjeti stavke 36. – 39.):
Scoring mononucleate cells in addition to binucleate cells could provide useful information (85) (86), but is not mandatory.
svi se rezultati nalaze unutar raspona distribucije prijašnjih podataka o negativnim kontrolama (npr. kontrolne granice od 95 % prema Poissonovoj distribuciji; vidjeti stavak 52.).
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.
Tada se smatra da ispitivana kemikalija ne može uzrokovati lomove i/ili povećanje ili gubitak kromosoma u tom ispitnom sustavu.
When single cultures per concentration are used (see paragraph 28), at least 2000 cells per culture should be scored in this single culture.
Jasno pozitivan ili negativan odgovor nije potrebno provjeravati.
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.
U slučaju kada reakcija nije ni jasno negativna niti jasno pozitivna kako je prethodno opisano i/ili kako bi se utvrdila biološka relevantnost rezultata, podatke bi trebalo ocijeniti stručnom prosudbom i/ili dodatnim istraživanjima.
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.
Moglo bi biti korisno pregledati dodatne stanice (prema potrebi) ili ponoviti pokus u eventualno izmijenjenim pokusnim uvjetima (npr. razmak između koncentracija, drugi uvjeti metaboličke aktivacije (tj. koncentracija S9 ili podrijetlo S9)).
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.
U rijetkim slučajevima, čak i nakon dodatnih istraživanja, skupom podataka neće se omogućiti donošenje zaključka o tome je li rezultat pozitivan ili negativan te će se stoga zaključiti da je dvosmislen.
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).
U in vitro mikronukleus-testu ispitivane kemikalije mogu inducirati mikronukleuse zato što uzrokuju lom kromosoma, gubitak kromosoma ili kombinaciju tih dviju pojava.
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.
Kako bi se odredilo je li mehanizam indukcije mikronukleusa posljedica klastogenog i/ili aneugenog djelovanja, mogu se provesti dodatne analize primjenom antikinetohornih protutijela, in situ sondi specifičnih za centromere ili drugih metoda.
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).
reaktivnost ispitivanih kemikalija s otapalom/nosačem ili hranjivom podlogom za uzgoj stanica,
When first acquiring data for an historical negative control distribution, concurrent negative controls should be consistent with published negative control data where they exist.
topljivost i stabilnost ispitivane kemikalije u otapalu, ako je poznata,
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).
izmjerene vrijednosti za pH, osmolalnost i taloženje u hranjivoj podlozi za uzgoj kulture u koju je dodana ispitivana kemikalija, prema potrebi.
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.
obrazloženje odabira nosača,
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).
postotak nosača u konačnoj hranjivoj podlozi za uzgoj kulture.
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).
vrsta i podrijetlo upotrijebljenih stanica,
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.
prikladnost upotrijebljene vrste stanice,
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.
odsutnost mikoplazme, u slučaju staničnih linija,
Concurrent negative controls should ideally be within the 95 % control limits of the distribution of the laboratory's historical negative control database (87) (88).
za stanične linije, informacije o duljini staničnog ciklusa ili indeksu proliferacije,
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.
ako su upotrijebljeni limfociti, spol darivatelja krvi, dob i sve bitne informacije o darivatelju, punoj krvi ili izoliranim limfocitima, upotrijebljenom mitogenu;
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.
normalno trajanje staničnog ciklusa (negativna kontrola),
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.
metode održavanja staničnih kultura, za stanične linije,
Concurrent measures of cytotoxicity and/or cytostasis for all treated, negative and positive control cultures should be determined (16).
modalni broj kromosoma, za stanične linije.
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.
Uvjeti ispitivanja:
In the absence of cytoB, the RPD or the RICC should be used (see Appendix 2).
identitet tvari koja blokira citokinezu (npr. citohalazin B), ako je upotrijebljena, njezina koncentracija i trajanje izloženosti stanice,
The concurrent negative control is considered acceptable for addition to the laboratory historical negative control database as described in paragraph 50.
koncentracija ispitivane kemikalije izražena kao konačna koncentracija u hranjivoj podlozi za uzgoj kulture (npr. μg ili mg/mL, ili mM hranjive podloge),
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.
obrazloženje za odabir koncentracija i broja kultura, uključujući podatke o citotoksičnosti i ograničenja topljivosti,
Cell proliferation criteria in the solvent control should be fulfilled (paragraph 25-27).
sastav hranjivih podloga, koncentracija CO2, ako je primjenjivo, stupanj vlažnosti,
All experimental conditions were tested unless one resulted in positive results (paragraphs 36-40).
koncentracija (i/ili volumen) otapala i ispitivane kemijske tvari dodane u hranjivu podlogu za uzgoj kulture,
Adequate number of cells and concentrations are analysable (paragraphs 28 and 44-46).
temperatura i vrijeme inkubacije,
The criteria for the selection of top concentration are consistent with those described in paragraphs 24-31.
gustoća stanica kod nasađivanja, ako je primjenjivo,
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):
vrsta i sastav sustava metaboličke aktivacije, izvor S9, metoda pripreme smjese S9, koncentracija ili volumen mješavina S9 i samog S9 u konačnoj hranjivoj podlozi za uzgoj kulture, kontrole kvalitete S9 (npr. enzimsko djelovanje, sterilnost, metabolička sposobnost),
at least one of the test concentrations exhibits a statistically significant increase compared with the concurrent negative control (89)
kemikalije pozitivne i negativne kontrole, konačne koncentracije, uvjeti i trajanja razdoblja tretiranja i obnavljanja,
the increase is dose-related in at least one experimental condition when evaluated with an appropriate trend test (see paragraph 28)
metode pripreme predmetnih stakalaca i primijenjene tehnike bojenja,
any of the results are outside the distribution of the historical negative control data (e.g. Poisson-based 95 % control limits; see paragraph 52).
kriteriji za pregledavanje stanica s mikronukleusima (odabir stanica koje se mogu analizirati te identifikacija mikronukleusa),
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.
broj analiziranih stanica,
Recommendations for the most appropriate statistical methods can also be found in the literature (90) (91) (92).
kriteriji u skladu s kojima se studiju smatra pozitivnom, negativnom ili dvosmislenom,
Providing that all acceptability criteria are fulfilled, a test chemical is considered clearly negative if, in all experimental conditions examined (see paragraphs 36-39):
upotrijebljena metoda (upotrijebljene metode) statističke analize,
all results are inside the distribution of the historical negative control data (e.g. Poisson-based 95 % control limits; see paragraph 52).
metode, kao što su uporaba antikinetohornih antitijela ili sonda specifičnih za centromere, kojima se određuje sadržavaju li mikronukleusi čitave ili fragmentirane kromosome, ako je to primjenjivo,
The test chemical is then considered unable to induce chromosome breaks and/or gain or loss in this test system.
definicija stanica prihvatljivih za analizu,
There is no requirement for verification of a clear positive or negative response.
u odsutnosti citohalazina B, broj tretiranih stanica i broj izdvojenih stanica za svaku kulturu u slučaju staničnih linija,
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.
upotrijebljena metoda mjerenja citotoksičnosti, npr.
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.
CBPI ili RI u slučaju metode blokirane citokineze; RICC ili RPD ako nisu primijenjene metode blokirane citokineze; druga zapažanja, ako postoje (npr. konfluencija stanica, aptoza, nekroza, brojanje stanica u metafazi, učestalost dvojezgrenih stanica),
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.
podaci o pH i osmolalnosti podloge za tretiranje, ako su utvrđeni,
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.
raspodjela jednojezgrenih, dvojezgrenih i višejezgrenih stanica ako je primijenjena metoda blokirane citokineze,
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.
broj stanica s mikronukleusima, koji se navodi odvojeno za svaku tretiranu i kontrolnu kulturu, i određivanje jesu li iz dvojezgrenih ili jednojezgrenih stanica, prema potrebi,
reactivity of the test chemicals with the solvent/vehicle or cell culture media;
podaci o prijašnjim negativnim kontrolama (s otapalom) i prijašnjim pozitivnim kontrolama, zajedno s rasponima, prosjecima i standardnom devijacijom te kontrolnim granicama od 95 % za distribuciju, kao i broj podataka,
solubility and stability of the test chemical in solvent, if known;
statistička analiza, vrijednosti p, ako postoje.
measurement of pH, osmolality, and precipitate in the culture medium to which the test chemical was added, as appropriate.
Poglavlje B.10. ovog Priloga: In vitro test kromosomskih aberacija kod sisavaca.
justification for choice of solvent;
In vitro micronucleus assay with Chinese hamster V79 cells – results of a collaborative study with in situ exposure to 26 chemical substances.
percentage of solvent in the final culture medium
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.
type and source of cells used;
Evaluation of different cytotoxic and cytostatic measures for the in vitromicronucleus test (MNVit): Introduction to the collaborative trial.
suitability of the cell type used;
Comparison of four different treatment conditions of extended exposure in the in vitro micronucleus assay using TK6 lymphoblastoid cells.
absence of mycoplasma, in case of cell lines;
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.
for cell lines, information on cell cycle length or proliferation index;
sastanak ESAC-a, 16. – 17. studenoga 2006., dostupno na: http://ecvam.jrc.it/index.htm.
where lymphocytes are used, sex of blood donors, age and any relevant information on the donor, whole blood or separated lymphocytes, mitogen used;
Dostupno na: http://ecvam.jrc.it/index.htm.
normal (negative control) cell cycle time;
‚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, str.
methods for the maintenance of cell cultures, for cell lines;
85.–88.
modal number of chromosomes, for cell lines;
Dostupno na: http://www.pops.int/
Test Conditions:
‚CHO/HGPRT Mutation Assay: Evaluation of Gases and Volatile Liquids’, in Genotoxic Effects of Airborne Agents.
identity of the cytokinesis blocking substance (e.g. cytoB), if used, and its concentration and duration of cell exposure;
Corrigendum to ‚Report from the in vitro micronucleus assay working group’.
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);
OECD Environment, Health and Safety Publications (EHS), Series on testing and assessment, No.198, OECD Publishing, Paris.
rationale for the selection of concentrations and the number of cultures, including cytotoxicity data and solubility limitations;
‚In vitro micronucleus test’, in Encyclopedia of Biopharmaceutical Statistics, 2nd ed.
concentration (and/or volume) of the solvent and test chemical added in the culture medium;
New York, str.
incubation temperature and time;
463.–467.
cell density at seeding, if applicable;
Analysis of Data from in vitro Cytogenetic Assays. in Statistical Evaluation of Mutagenicity Test Data, Kirkland, D.J.
positive and negative control chemicals, final concentrations, conditions and durations of treatment and recovery periods;
141.–154.
methods of slide preparation and the staining technique used;
svaka kemikalija ili postupak koji u interakciji s elementima mitotičkog i mejotičkog ciklusa diobe stanice uzrokuje aneuploidiju u stanicama ili organizmima.
criteria for scoring micronucleate cells (selection of analysable cells and identification of micronucleus);
programirana smrt stanice koju obilježava niz koraka koji vode do razgradnje stanica u čestice vezane na membranu koje se potom eliminiraju fagocitozom ili rasipanjem.
numbers of cells analysed;
povećanje broja stanica kao posljedica mitotičke diobe stanice.
criteria for considering studies as positive, negative, or equivocal;
DNK sekvenca kromosoma gdje su spojene obje kromatide i na kojoj su na obje strane pričvršćena oba kinetohora.
method(s) of statistical analysis used;
odnosi se na konačne koncentracije ispitivane kemikalije u hranjivoj podlozi za uzgoj kulture.
definition of acceptable cells for analysis;
Citokineza
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;
proces diobe stanice koji slijedi neposredno nakon mitoze i kojim nastaju dvije stanice kćeri od kojih svaka sadržava jednu jezgru.
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);
Indeks proliferacije s blokiranom citokinezom (CBPI)
data on pH and osmolality of the treatment medium, if determined;
udio stanica u fazi druge diobe u tretiranoj populaciji u odnosu na netretiranu kontrolu (vidjeti Dodatak 2. za formulu).
distribution of mono-, bi-, and multi-nucleate cells if a cytokinesis block method is used;
Citostaza
number of cells with micronuclei given separately for each treated and control culture, and defining whether from binucleate or mononucleate cells, where appropriate;
sprječavanje rasta stanica (vidjeti Dodatak 2. za formulu).
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;
za analize obuhvaćene ovom ispitnom metodom koje se provode u prisutnosti citohalazina B citotoksičnost se definira kao smanjenje indeksa proliferacije stanica s blokiranom citokinezom (CBPI) ili indeksa ponavljanja (RI) tretiranih stanica u odnosu na negativnu kontrolu (vidjeti stavak 26. i Dodatak 2.).
statistical analysis; p-values if any.
Za ispitivanja obuhvaćena ovom ispitnom metodom koja se provode u odsutnosti citohalazina B citotoksičnost se definira kao smanjenje relativnog udvostručenja populacije (RPD) ili relativnog povećanja broja stanica (RICC) kod tretiranih stanica u usporedbi s negativnom kontrolom (vidjeti stavak 27. i Dodatak 2.).
Chapter B.10 of this Annex: In Vitro Mammalian Chromosome Aberration Test,.
Interfazne stanice