natgillin/translations-raw huggingface dataset:
Care should be taken not to score binucleate cells with irregular shapes or where the two nuclei differ greatly in size.
Na určenie toho, či je mechanizmus indukcie mikrojadier dôsledkom klastogénneho a/alebo aneugénneho pôsobenia možno použiť ďalšiu analýzu s použitím protilátok kinetochoru, sond in situ so špecifickou väzbou na centroméry alebo iných metód.
In addition, binucleate cells should not be confused with poorly spread multi-nucleate cells.
reaktivita testovanej chemikálie s rozpúšťadlom/nosičom alebo médiom na kultiváciu buniek,
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).
rozpustnosť a stabilita testovanej chemikálie v rozpúšťadle, ak je známa,
Scoring of mononucleate cells is acceptable if the test chemical is shown to interfere with cytoB activity.
prípadné meranie pH, osmolality a zrazeniny v rastovom médiu, do ktorého bola pridaná testovaná chemikália.
A repeat test without CytoB might be useful in such cases.
zdôvodnenie výberu rozpúšťadla,
Scoring mononucleate cells in addition to binucleate cells could provide useful information (85) (86), but is not mandatory.
percentuálny podiel rozpúšťadla v konečnom rastovom médiu.
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.
typ a zdroj použitých buniek,
When single cultures per concentration are used (see paragraph 28), at least 2000 cells per culture should be scored in this single culture.
vhodnosť použitého typu bunky,
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.
v prípade bunkových línií neprítomnosť mykoplazmy,
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.
v prípade bunkových línií informácie o dĺžke bunkového cyklu alebo proliferačný index,
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.
ak sa použijú lymfocyty, pohlavie darcov krvi, vek a všetky relevantné informácie o darcovi, informácie o tom či bola použitá celá krv alebo separované lymfocyty, použitý mitogén,
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).
trvanie bežného bunkového cyklu (pri negatívnej kontrole),
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.
v prípade bunkových línií metódy udržiavania bunkových kultúr,
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).
v prípade bunkových línií modálny počet chromozómov.
When first acquiring data for an historical negative control distribution, concurrent negative controls should be consistent with published negative control data where they exist.
Podmienky testovania:
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).
identita látky blokujúcej cytokinézu (napr. cytoB), ak sa použije, a jej koncentrácia a trvanie vystavenia buniek,
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.
koncentrácia testovanej chemikálie vyjadrená ako konečná koncentrácia v rastovom médiu (napr. v μg alebo mg/ml alebo mM rastového média),
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).
zdôvodnenie voľby koncentrácií a počtu kultúr vrátane údajov o cytotoxicite a obmedzeniach rozpustnosti,
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).
zloženie média, prípadne koncentrácia CO2, hladina vlhkosti,
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.
koncentrácia (a/alebo objem) rozpúšťadla a testovanej chemikálie pridaných do rastového média,
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.
teplota a doba inkubácie,
Concurrent negative controls should ideally be within the 95 % control limits of the distribution of the laboratory's historical negative control database (87) (88).
prípadne hustota buniek pri naočkovaní média,
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.
typ a zloženie použitého systému metabolickej aktivácie [zdroj S9, metóda prípravy zmesi S9, koncentrácia alebo objem zmesi S9 a S9 v konečnom rastovom médiu, kontroly akosti S9, (napr. enzymatická aktivita, sterilita, metabolická schopnosť)],
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.
chemikálie na pozitívnu a negatívnu kontrolu, konečné koncentrácie, podmienky a trvanie ovplyvňovania a obdobia obnovy,
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.
metódy prípravy preparátov a použitá technika farbenia,
Concurrent measures of cytotoxicity and/or cytostasis for all treated, negative and positive control cultures should be determined (16).
kritéria hodnotenia buniek s mikrojadrami (výber analyzovateľných buniek a identifikácia mikrojadra),
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.
počty analyzovaných buniek,
In the absence of cytoB, the RPD or the RICC should be used (see Appendix 2).
kritériá hodnotenia štúdií ako pozitívne, negatívne alebo s nejasným výsledkom,
The concurrent negative control is considered acceptable for addition to the laboratory historical negative control database as described in paragraph 50.
použitá(-é) metóda (-y) štatistickej analýzy,
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.
v uplatniteľnom prípade metódy, ako je napríklad použitie protilátok proti kinetochoru alebo špecifických pancentromérových sond na určenie toho, či mikrojadrá obsahujú celé alebo fragmentované chromozómy,
Cell proliferation criteria in the solvent control should be fulfilled (paragraph 25-27).
vymedzenie buniek prijateľných na analýzu,
All experimental conditions were tested unless one resulted in positive results (paragraphs 36-40).
v prípade bunkových línií pri neprítomnosti cytoB počet ovplyvnených buniek a počet odobratých buniek v prípade každej kultúry,
Adequate number of cells and concentrations are analysable (paragraphs 28 and 44-46).
použitý spôsob merania cytotoxicity, napr. CBPI alebo RI v prípade metódy blokovania cytokinézy, RICC alebo RPD v prípade, že sa metódy blokovania cytokinézy nepoužijú, prípadne pozorovania, ak existujú (napr. konfluencia buniek, apoptóza, nekróza, počítanie metafáz, frekvencia dvojjadrových buniek),
The criteria for the selection of top concentration are consistent with those described in paragraphs 24-31.
údaje o hodnotách pH a osmolality testovacieho média, ak sú stanovené,
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):
distribúcia jednojadrových, dvojjadrových a viacjadrových buniek, ak sa používa metóda blokovania cytokinézy,
at least one of the test concentrations exhibits a statistically significant increase compared with the concurrent negative control (89)
prípadne počet buniek s mikrojadrami uvedený osobitne pre každú ovplyvnenú kultúru a kontrolnú kultúru a určenie, či ide o dvojjadrové alebo jednojadrové bunky,
the increase is dose-related in at least one experimental condition when evaluated with an appropriate trend test (see paragraph 28)
historické údaje o negatívnych (rozpúšťadlo) a pozitívnych kontrolách s rozsahmi, priemernými hodnotami a štandardnou odchýlkou a 95 % regulačnými medzami pre rozdelenie, ako aj počet údajov,
any of the results are outside the distribution of the historical negative control data (e.g. Poisson-based 95 % control limits; see paragraph 52).
štatistické analýzy, hodnoty p, ak existujú.
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.
Chapter B.10 of this Annex: In Vitro Mammalian Chromosome Aberration Test.
Recommendations for the most appropriate statistical methods can also be found in the literature (90) (91) (92).
In vitro micronucleus assay with Chinese hamster V79 cells – results of a collaborative study with in situ exposure to 26 chemical substances.
Providing that all acceptability criteria are fulfilled, a test chemical is considered clearly negative if, in all experimental conditions examined (see paragraphs 36-39):
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.
all results are inside the distribution of the historical negative control data (e.g. Poisson-based 95 % control limits; see paragraph 52).
Comparison of four different treatment conditions of extended exposure in the in vitro micronucleus assay using TK6 lymphoblastoid cells.
The test chemical is then considered unable to induce chromosome breaks and/or gain or loss in this test system.
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.
There is no requirement for verification of a clear positive or negative response.
ESAC 25th meeting, 16-17 November 2006, k dispozícii na webovej lokalite: http://ecvam.jrc.it/index.htm.
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.
K dispozícii na webovej lokalite: http://ecvam.jrc.it/index.htm.
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.
“A safe substitute for Polychlorinated Biphenyls as an Inducer of Metabolic Activation Systems”, in In Vitro Metabolic Activation in Mutagenesis Testing. de Serres, F.J. et al. (eds), Elsevier, North-Holland, pp.
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.
85-88.
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.
K dispozícii na webovej lokalite: http://www.pops.int/
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.
“CHO/HGPRT Mutation Assay: Evaluation of Gases and Volatile Liquids”, in Genotoxic Effects of Airborne Agents.
reactivity of the test chemicals with the solvent/vehicle or cell culture media;
Tice, R.R., D.L.
solubility and stability of the test chemical in solvent, if known;
Costa, K.M.
measurement of pH, osmolality, and precipitate in the culture medium to which the test chemical was added, as appropriate.
Corrigendum to “Report from the in vitro micronucleus assay working group”.
justification for choice of solvent;
OECD Environment, Health and Safety Publications (EHS), Series on testing and assessment, No.198, OECD Publishing, Paris.
percentage of solvent in the final culture medium
“In vitro micronucleus test”, in Encyclopedia of Biopharmaceutical Statistics, 2nd ed.
type and source of cells used;
Chow, S. (ed.), Marcel Dekker, Inc.
suitability of the cell type used;
New York, pp.
absence of mycoplasma, in case of cell lines;
463-467.
for cell lines, information on cell cycle length or proliferation index;
Analysis of Data from in vitro Cytogenetic Assays. in Statistical Evaluation of Mutagenicity Test Data, Kirkland, D.J. (ed), Cambridge University Press, Cambridge, pp.
where lymphocytes are used, sex of blood donors, age and any relevant information on the donor, whole blood or separated lymphocytes, mitogen used;
141-154.
normal (negative control) cell cycle time;
Aneugén
methods for the maintenance of cell cultures, for cell lines;
akákoľvek chemikália alebo proces, ktoré pri interakcii so zložkami mitotického alebo meiotického aparátu bunkového cyklu vedie k aneuploidii v bunkách alebo organizmoch.
Test Conditions:
zvýšenie počtu buniek v dôsledku mitotického delenia buniek.
identity of the cytokinesis blocking substance (e.g. cytoB), if used, and its concentration and duration of cell exposure;
oblasť DNA chromozómu, v ktorej sa spájajú obe chromatídy a na ktorú sú vedľa seba pripojené oba kinetochory.
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);
sú konečné koncentrácie testovanej chemikálie v rastovom médiu.
rationale for the selection of concentrations and the number of cultures, including cytotoxicity data and solubility limitations;
akákoľvek chemikália alebo jav, ktorý spôsobuje štruktúrne chromozómové aberácie v populáciách buniek alebo eukaryotických organizmoch.
composition of media, CO2 concentration, if applicable, humidity level;
Cytokinéza
concentration (and/or volume) of the solvent and test chemical added in the culture medium;
proces delenia buniek, ktorý nasleduje ihneď po mitóze a v ktorom vznikajú dve dcérske bunky – každá s jedným jadrom.
incubation temperature and time;
Proliferačný index blokovania cytokinézy (CBPI)
cell density at seeding, if applicable;
pomer druhého delenia buniek v ovplyvnenej populácii vo vzťahu k čistej kontrole (pozri vzorec v dodatku 2).
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);
Cytostáza
positive and negative control chemicals, final concentrations, conditions and durations of treatment and recovery periods;
inhibícia rastu buniek (pozri vzorec v dodatku 2).
methods of slide preparation and the staining technique used;
v prípade testov, na ktoré sa vzťahuje táto testovacia metóda a ktoré sa vykonávajú za prítomnosti cytochalasínu B, sa cytotoxicita označuje ako zníženie proliferačného indexu blokovania cytokinézy (CBPI) alebo indexu replikácie (RI) ovplyvnených buniek v porovnaní s negatívnou kontrolou (pozri bod 26 a dodatok 2).
criteria for scoring micronucleate cells (selection of analysable cells and identification of micronucleus);
Pri skúškach, na ktoré sa vzťahuje táto testovacia metóda a ktoré sa vykonávajú za neprítomnosti cytochalasínu B, sa cytotoxicita označuje ako zníženie relatívneho zdvojenia populácie (RPD) alebo relatívneho nárastu počtu buniek (RICC) ovplyvnených buniek v porovnaní s negatívnou kontrolou (pozri bod 27 a dodatok 2).
numbers of cells analysed;
Bunky v interfáze
criteria for considering studies as positive, negative, or equivocal;
bunky, ktoré nie sú v štádiu mitotického delenia.
method(s) of statistical analysis used;
štruktúra obsahujúca proteíny, ktorá sa vytvára na centromére chromozómu a ku ktorej sa vlákna deliaceho vretienka pripájajú počas delenia buniek a umožňujú riadny pohyb dcérskych chromozómov k pólom dcérskych buniek.
methods, such as use of anti-kinetochore antibody or pan-centromeric specific probes, to characterise whether micronuclei contain whole or fragmented chromosomes, if applicable;
malé jadrá doplňujúce hlavné jadrá buniek a existujúce oddelene od nich, ktoré sa vytvárajú počas telofázy mitózy alebo meiózy z oneskorených chromozómových fragmentov alebo celých chromozómov.
definition of acceptable cells for analysis;
pomer buniek v metafáze vydelený celkovým počtom buniek pozorovaných v populácii buniek; udáva stupeň proliferácie buniek tejto populácie.
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;
Nondisjunkcia
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);
dej, pri ktorom sa spárované chromatídy riadne neoddelia a nerozdelia do vznikajúcich dcérskych buniek, čo vedie vzniku dcérskych buniek s abnormálnym počtom chromozómov.
data on pH and osmolality of the treatment medium, if determined;
numerické chromozómové aberácie v bunkách alebo organizmoch, ktoré zahŕňajú celý(-é) súbor(-y) chromozómov, na rozdiel od jedného chromozómu alebo jednotlivých chromozómov (aneuploidia).
distribution of mono-, bi-, and multi-nucleate cells if a cytokinesis block method is used;
Proliferačný index (PI)
number of cells with micronuclei given separately for each treated and control culture, and defining whether from binucleate or mononucleate cells, where appropriate;
metóda merania cytotoxicity bez použitia cytoB (pozri vzorec v dodatku 2).
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;
Relatívne zvýšenie počtu buniek (RICC)
statistical analysis; p-values if any.
Index replikácie (RI)
Chapter B.10 of this Annex: In Vitro Mammalian Chromosome Aberration Test,.
pomer bunkových cyklov, ktoré boli dokončené v ovplyvnenej kultúre, vo vzťahu k čistej kontrole počas doby expozície a obnovy (pozri vzorec v dodatku 2).
In vitro micronucleus assay with Chinese hamster V79 cells — results of a collaborative study with in situ exposure to 26 chemical substances.
zmes frakcie pečene S9 a kofaktorov nepotrebných pre metabolickú aktiváciu enzýmov.
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.
Čistá kontrola
Evaluation of different cytotoxic and cytostatic measures for the in vitro micronucleus test (MNVit): Introduction to the collaborative trial.
Pri použití cytoB by hodnotenie cytotoxicity malo byť založené na proliferačnom indexe blokovania cytokinézy (CBPI) alebo na indexe replikácie (RI) (17) (69).
Comparison of four different treatment conditions of extended exposure in the in vitro micronucleus assay using TK6 lymphoblastoid cells.
CBPI udáva priemerný počet jadier na bunku a môže sa použiť na výpočet proliferácie buniek.
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.
RI udáva relatívny počet bunkových cyklov na bunku počas obdobia vystavenia cytoB v ovplyvnených kultúrach v porovnaní s kontrolnými kultúrami a možno ho použiť na percentuálny výpočet cytostázy:
ESAC 25th meeting, 16-17 November 2006, Available at: http://ecvam.jrc.it/index.htm.
% cytostáza = 100 – 100(CBPIT – 1) ÷ (CBPIC – 1)
Available at: http://ecvam.jrc.it/index.htm.
T kultúra ovplyvnená testovanou chemikáliou
“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.
C kontrolná kultúra,
(eds), Elsevier, North-Holland, pp. 85-88.
počet jednojadrových buniek
Available at: http://www.pops.int/
počet dvojjadrových buniek
“CHO/HGPRT Mutation Assay: Evaluation of Gases and Volatile Liquids”, in Genotoxic Effects of Airborne Agents.
počet viacjadrových buniek
Tice, R.R., D.L.
celkový počet buniek
Corrigendum to “Report from the in vitro micronucleus assay working group”.
CBPI zodpovedajúci 1 (všetky bunky sú jednojadrové) sa teda rovná 100 % cytostáze.
OECD Environment, Health and Safety Publications (EHS), Series on testing and assessment, No. 198, OECD Publishing, Paris.
Cytostáza = 100 – RI
“In vitro micronucleus test”, in Encyclopedia of Biopharmaceutical Statistics, 2nd ed.
T ovplyvnené kultúry
Chow, S. (ed.), Marcel Dekker, Inc.
C kontrolné kultúry
New York, pp. 463-467.
Hodnota RI zodpovedajúca 53 % teda znamená, že v porovnaní s počtami buniek, ktorých delením vznikli v kontrolnej kultúre dvojjadrové a viacjadrové bunky, len 53 % z tohto počtu sa delilo v ovplyvnenej kultúre, t. j.
Analysis of Data from in vitro Cytogenetic Assays. in Statistical Evaluation of Mutagenicity Test Data, Kirkland, D.J.
47 % cytostáza.
(ed), Cambridge University Press, Cambridge, pp. 141-154.
Ak sa cytoB nepoužije, odporúča sa vykonať hodnotenie cytotoxicity na základe relatívneho nárastu počtu buniek (RICC) alebo relatívneho zdvojenia populácie (RPD) (69), keďže v oboch prípadoch sa zohľadňuje podiel populácie buniek, ktoré sa delili.
Aneugen
Preto 53 % RICC alebo RDP indikuje 47 % cytotoxicitu/cytostázu.
any chemical or process that, by interacting with the components of the mitotic and meiotic cell division cycle apparatus, leads to aneuploidy in cells or organisms.
Pri použití proliferačného indexu (PI) možno cytotoxicitu hodnotiť pomocou počítania počtu klonov obsahujúcich jednu bunku (cl1), 2 bunky (cl2), 3 až 4 bunky (cl4) a 5 až 8 buniek (cl8).