natgillin/translations-raw huggingface dataset:
This assessment may be performed graphically or by a formal test (15).
kemikaal, mis pärast silma eesmisele pinnale kandmist põhjustab silmakoe kahjustuse, mis ei parane 21 päeva jooksul pärast kemikaali silmale kandmist, või põhjustab tugeva füüsilise nägemislanguse; b) kemikaal, mis klassifitseeritakse ÜRO GHSi 1. kategooriasse; ELi CLP-määruse 1. kategooriasse; või US EPA silmi ärritava toimega kemikaalide I kategooriasse.
Suitable tests are Levene's or Bartlett's.
Silmi ärritava toimega kemikaal
Failure to meet the assumption of homogeneity of variances can sometimes be corrected by logarithmic transformation of the data.
Muutused kliinilise keemia näitajates (nt AST, ALT) võivad samuti anda kasulikku teavet koekahjustuste kohta ning kaaluda võib ka täiendavaid näitajaid, nagu kaspaasi aktiveerumine, värvumine TUNELiga, värvumine anneksiin V-ga jne.
If heterogeneity of variance is extreme and cannot be corrected by transformation, analysis by methods such as step-down Jonkheere trend tests should be considered.
Vähe andmeid on avaldatud siiski viimaste kasutamise kohta in vivo uuringutes ja mõned neist võivad olla vähem usaldusväärsed kui teised.
Additional guidance on determining the NOEC can be found in (10).
Kirkland, D., G. Speit (2008), „Evaluation of the ability of a battery of three in vitro genotoxicity tests to discriminate rodent carcinogens and non-carcinogens III.
Recent scientific developments have led to a recommendation of abandoning the concept of NOEC and replacing it with regression based point estimates ECx.
Appropriate follow-up testing in vivo“, Mutation Research, Vol.
An appropriate value for x has not been established for this Myriophyllum test.
654/2, pp.
However, a range of 10 to 20 % appears to be appropriate (depending on the response variable chosen), and preferably both the EC10 and EC20 and their confidence limits should be reported.
114–32.
Reporting
Brendler-Schwaab, S. et al.
Multi-constituent substance, UVCBs or mixture:
(2005), „The in vivo Comet assay: use and status in genotoxicity testing“, Mutagenesis, Vol.
Test species
20/4, pp.
Scientific name and source.
245–54.
Test procedure used (static or semi-static).
Burlinson, B. et al.
Date of start of the test and its duration.
(2007), „Fourth International Workgroup on Genotoxicity Testing: result of the in vivo Comet assay workgroup“, Mutation Research, Vol.627/1, pp.
Test medium.
31–5.
Description of the experimental design: test vessels and covers, solution volumes, main shoot length per test vessel at the beginning of the test.
Burlinson, B. (2012), „The in vitro and in vivo Comet assays“, Methods in Molecular Biology, Vol.
Test concentrations (nominal and measured as appropriate) and number of replicates per concentration.
817, pp. 143–63.
Methods of preparation of stock and test solutions including the use of any solvents or dispersants.
Smith, C.C. et al.
Temperature during the test.
(2008), „Recommendations for design of the rat Comet assay“, Mutagenesis,Vol.
Light source, light intensity and homogeneity.
23/3, pp. 233–40.
pH values of the test and control media.
Hartmann, A. et al.
The method of analysis of test chemical with appropriate quality assessment data (validation studies, standard deviations or confidence limits of analyses).
(2003), „Recommendations for conducting the in vivo alkaline Comet assay“, Mutagenesis, Vol.18/1, pp.
Methods for determination of main shoot length and other measurement variables, e.g. total lateral branches length, total shoot length, total root length, fresh weight, dry weight or number of whorls.
45–51.
State of the culture (sterile or non-sterile) of each test and control vessel at each observation.
McKelvey-Martin, V.J. et al.
All deviations from this test method.
(1993), „The single cell gel electrophoresis assay (Comet assay): a European review“, Mutation Research, Vol.
Raw data: main shoot length and other measurement variables in each test and control vessel at each observation and occasion of analysis.
288/1, pp.
Means and standard deviations for each measurement variable.
47–63.
Growth curves for each measurement variable.
206–21.
Calculated response variables for each treatment replicate, with mean values and coefficient of variation for replicates.
Singh, N.P. et al.
Graphical representation of the concentration/effect relationship.
(1988), „A simple technique for quantitation of low levels of DNA damage in individual cells“, Experimental Cell Research, Vol.
Estimates of toxic endpoints for response variables e.g. EC50, EC10, EC20, and associated confidence intervals.
175/1, pp.
If calculated, LOEC and/or NOEC and the statistical methods used for their determination.
184–91.
If ANOVA has been used, the size of the effect which can be detected (e.g. the least significant difference).
40–69.
Any stimulation of growth found in any treatment.
OECD (2014), „Reports of the JaCVAM initiative international pre-validation and validation studies of the in vivo rodent alkaline comet assay for the detection of genotoxic carcinogens“, Series on Testing and Assessment, Nos. 195 and 196, OECD Publishing, Paris.
Any visual signs of phytotoxicity as well as observations of test solutions.
Strauss (1995), „The single cell gel electrophoresis/Comet assay: a potential tool for detecting radiation-induced DNA damage in humans“, Stem Cells, Vol.
Discussion of the results, including any influence on the outcome of the test resulting from deviations from this test method.
13/1, pp. 207–14.
ASTM Designation E 1913-04, Standard Guide for Conducting Static, Axenic, 14-Day Phytotoxicity Tests in Test Tubes with the Submersed Aquatic Macrophyte, Myriophyllum sibiricum Komarov.
Collins, A.R (2004), „The Comet assay for DNA damage and repair: principles, applications, and limitations“, Molecular Biotechnology, Vol.
Maletzki, D. et al.
26/3, pp. 249–61.
, Myriophyllum spicatum als ökotoxikologischer Testorganismus: Methodenentwicklung eines sedimentfreien Testsystems und erste Ergebnisse mit 3,5-Dichlorphenol, Umweltwiss Schadst Forsch, No. 22, pp. 702–710.
108–20.
Chapter C.26 of this Annex: Lemna sp.
Kushwaha, S. et al.
Growth Inhibition Test,
(2010), „Evaluation of multi-organ DNA damage by Comet assay from 28 days repeated dose oral toxicity test in mice: A practical approach for test integration in regulatory toxicity testing“, Regulatory Toxicology and Pharmacology, Vol.
OECD (2014), “Myriophyllum spicatum Toxicity Test: Results of an inter-laboratory ring test using a sediment-free test system”, OECD Environment, Health and Safety Publications (EHS), Series Testing and Assessment, No. 205, OECD Publishing, Paris.
58/1, pp.
OECD (2000), “Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures”, OECD Environment, Health and Safety Publications (EHS), Series on Testing and Assessment, No. 23, OECD Publishing, Paris.
145–54.
Chapter C.51 of this Annex: Water-Sediment Myriophyllum spicatum Toxicity Test
, „Combining the in vivo Comet and micronucleus assays: a practical approach to genotoxicity testing and data interpretation“, Mutagenesis, Vol.
Christensen, E.R., N. Nyholm (1984), Ecotoxicological Assays with Algae: Weibull Dose-Response Curves, Environmental Science & Technology, Vol.
25/2, pp. 187–99.
18/9, 713-718.
, „Evaluation of a multi-endpoint assay in rats, combining the bone-marrow micronucleus test, the Comet assay and the flow-cytometric peripheral blood micronucleus test“, Mutation Research, Vol.
Nyholm, N. et al.
722/1, pp. 7–19.
Bruce, R.D., D.J.
Recio, L. et al.
OECD (2006), “Current Approaches in the Statistical Analysis of Ecotoxicity Data: A Guidance to Application”, OECD Environment, Health and Safety Publications (EHS), Series on Testing and Assessment, No. 54, OECD Publishing, Paris.
(2012), „Comparison of Comet assay dose-response for ethyl methanesulfonate using freshly prepared versus cryopreserved tissues“, Environmental and Molecular Mutagenesis, Vol.
Norberg-King, T.J.
53/2, pp.
Dunnett, C.W.
149–62.
, A multiple comparisons procedure for comparing several treatments with a control, Journal of the American Statistical Association, Vol.
O'Donovan, M., B. Burlinson (2013), „Maximum dose levels for the rodent comet assay to examine damage at the site of contact or to the gastrointestinal tract“, Mutagenesis, Vol.
50/272, pp. 1096-1121.
28/6, pp. 621–3.
, New tables for multiple comparisons with a control, Biometrics, Vol.
Hartmann, A. (2004), „Use of the alkaline in vivo Comet assay for mechanistic genotoxicity investigations“, Mutagenesis, Vol.
20/3, pp. 482-491.
19/1, pp. 51–9.
Williams, D.A.
Nesslany, F. (2007), „In vivo Comet assay on isolated kidney cells to distinguish genotoxic carcinogens from epigenetic carcinogens or cytotoxic compounds“, Mutation Research/Genetic Toxicology and Environmental Mutagenesis, Vol.
, A test for differences between treatment means when several dose levels are compared with a zero dose control, Biometrics, Vol.
630/1, pp. 28–41.
27/1, pp. 103-117.
Brendler-Schwaab, S.Y., B.A. Herbold (1997), „A new method for the enrichment of single renal proximal tubular cells and their first use in the Comet assay“, Mutation Research/Genetic Toxicology and Environmental Mutagenesis, Vol.
, The comparison of several dose levels with a zero dose control, Biometrics, Vol.
393/1-2, pp.
28/2, pp. 519-531.
175–8.
Brain, P., R. Cousens (1989), An equation to describe dose-responses where there is stimulation of growth at low doses, Weed Research, Vol.
Toyoizumi, T. et al.
29/2, pp. 93-96.
(2011), „Use of the in vivo skin Comet assay to evaluate the DNA-damaging potential of chemicals applied to the skin“, Mutation Research/Genetic Toxicology and Environmental Mutagenesis, Vol.
Biomass is the fresh and/or dry weight of living matter present in a population.
726/2, pp.
In this test the biomass is the sum of main shoot, all lateral branches and all roots.
175–80.
Chlorosis is the change of the color from green to yellowing of test organism especially of the whorls.
Struwe, M. et al.
ECx is the concentration of the test chemical dissolved in test medium that results in a x % (e.g. 50 %) reduction in growth of Myriophyllum spicatum within a stated exposure period (to be mentioned explicitly if deviating from full or normal test duration).
(2008), „Detection of photogenotoxicity in skin and eye in rat with the photo Comet assay“, Photochemical and Photobiological Sciences, Vol.
To unambiguously denote an EC value deriving from growth rate or yield the symbol “ErC” is used for growth rate and “EyC” is used for yield, followed by the measurement variable used, e.g. ErC (main shoot length).
7/2, pp.
Growth is an increase in the measurement variable, e.g. main shoot length, total lateral branches length, total shoot length, total root length, fresh weight, dry weight or number of whorls, over the test period.
240–9.
Growth rate (average specific growth rate) is the logarithmic increase in the measurement variable during the exposure period.
Wada, K. et al.
Note: Growth rate related response variables are independent of the duration of the test as long as the growth pattern of unexposed control organisms is exponential.
(2012), „A comparison of cell-collecting methods for the Comet assay in urinary bladders of rats“, Mutation Research/Genetic Toxicology and Environmental Mutagenesis, Vol.
Lowest Observed Effect Concentration (LOEC) is the lowest tested concentration at which the chemical is observed to have a statistically significant reducing effect on growth (at p < 0,05) when compared with the control, within a given exposure time.
742/1-2, pp.
Measurement variables are any type of variables which are measured to express the test endpoint using one or more different response variables.
26–30.
In this test method main shoot length, total lateral branches length; total shoot length, total root length, fresh weight, dry weight and number of whorls are measurement variables.
Wang, A. et al.
Monoculture is a culture with one plant species.
(2007), „Measurement of DNA damage in rat urinary bladder transitional cells: improved selective harvest of transitional cells and detailed Comet assay protocols“, Mutation Research/Genetic Toxicology and Environmental Mutagenesis, Vol.
Necrosis is dead (i.e. white or dark brown) tissue of the test organism.
634/ 1-2, pp.
No Observed Effect Concentration (NOEC) is the test concentration immediately below the LOEC.
51–9.
Response variable is a variable for the estimation of toxicity derived from any measured variable describing biomass by different methods of calculation.
31–5.
For this test method growth rate and yield are response variables derived from measurement variables like main shoot length, total shoot length, fresh weight, dry weight, or number of whorls.
Jackson, P. et al.
Semi-static (renewal) test is a test in which the test solution is periodically replaced at specific intervals during the test.
(2013), „Validation of use of frozen tissues in high-throughput comet assay with fully-automatic scoring“, Mutagenesis, Vol.
Static test is a test method without renewal of the test solution during the test.
28/6, pp.
Test endpoint describes the general factor that will be changed relative to control by the test chemical as aim of the test.
486–500.
In this test method the test endpoint is inhibition of growth which may be expressed by different response variables which are based on one or more measurement variables.
629–799.
Test medium is the complete synthetic growth medium on which test plants grow when exposed to the test chemical.
Sekihashi, K. et al.
The test chemical will normally be dissolved in the test medium.
(2002), „Comparative investigations of multiple organs of mice and rats in the Comet assay“, Mutation Research, Vol.
UVCB is a substance of unknown or variable composition, complex reaction product or biological material
517/1-2, pp.
Yield is value of a measurement variable to express biomass at the end of the exposure period minus the measurement variable at the start of the exposure period.
53–74.
Note: When the growth pattern of unexposed organisms is exponential, yield-based response variables will decrease with the test duration.
Speit, G, M. Vasquez, A. Hartmann (2009), „The comet assay as an indicator test for germ cell genotoxicity“, Mutation Research, Vol.
MODIFIED ANDREWS' MEDIUM FOR STOCK CULTURE AND PRE-CULTURE
681/1, pp. 3–12.
From five separately prepared nutrient stock solutions the modified Andrews' medium required for stock culture and pre culture will be prepared, with addition of 3 % sucrose.
Olive (1997), „Influence of oxygen on radiation-induced DNA damage in testicular cells of C3H mice“, International Journal of Radiation Biology, Vol.
Table 1
71/3, pp. 275–282.
Composition of Andrews' nutrient solution: (ASTM Designation E 1913-04)
Cordelli, E. et al.
Production of nutrient stock solutions
(2003), „Evaluation of DNA damage in different stages of mouse spermatogenesis after testicular X irradiation“, Journal of Radiation Research, Vol. 160/4, pp.
Production of nutrient solution
443–451.
Stock solution
Merk, O., G. Speit (1999), „Detection of crosslinks with the Comet assay in relationship to genotoxicity and cytotoxicity“, Environmental and Molecular Mutagenesis, Vol.
Initial weight per 1000 ml
33/2, pp. 167–72.
ml per 5 l nutrient solution
Wolf (1996), „Detection of DNA-crosslinking agents with the alkaline Comet assay“, Environmental and Molecular Mutagenesis, Vol.
74,6 mg
27/3, pp. 196–201.
8,08 g
Jones (2012), „Assessment of DNA interstrand crosslinks using the modified alkaline Comet assay“, Methods in Molecular Biology, Vol.
18,88 g
817, pp. 165–81.
9,86 g
Hartley (2010), „Measurement of DNA interstrand crosslinking in individual cells using the Single Cell Gel Electrophoresis (Comet) assay“, Methods in Molecular Biology, Vol.
See below stock solution 3.1
613, pp. 267–282.
2,72 g
Jha (2006), „Reliable Comet assay measurements for detecting DNA damage induced by ionizing radiation and chemicals“, Mutation Research/Genetic Toxicology and Environmental Mutagenesis, Vol.
0,278 g
605(1-2), pp. 7–16.
0,372 g
31–5.
Stock solutions can be kept in a refrigerator for 6 months (at 5-10 °C).
Kumaravel, T.S. et al.
Only stock solution No. 5 has a reduced shelf life (two months).
(2009), „Comet Assay measurements: a perspective“, Cell Biology and Toxicology, Vol.
Production of stock solution 3.1 for preparing stock solution 3
25/1, pp.
Initial weight g/100 ml
53–64.
MnSO4 × 4 H2O
Ersson, C., L. Möller (2011), „The effects on DNA migration of altering parameters in the Comet assay protocol such as agarose density, electrophoresis conditions and durations of the enzyme or the alkaline treatments“, Mutagenesis, Vol.
ZnSO4 × 7 H2O
26/6, pp. 689–95.
CuSO4 × 5 H2O
Møller, P. et al.
6Mo7O24 × 4 H2O
(2010), „Assessment and reduction of Comet assay variation in relation to DNA damage: studies from the European Comet Assay Validation Group“, Mutagenesis, Vol.
After having produced stock solution 3.1 (Table 2), deep-freeze this solution in approximately 11 ml-aliquots (at – 18 °C at least).
25/2, pp.
The deep-frozen portions have a shelf life of five years.
109–11.
To produce stock solution 3, defrost stock solution 3.1, fill 10 ml of it into a 1 l volumetric flask and add ultra-pure water up to the flask's mark.
Forchhammer, L. et al.
To obtain modified Andrews' medium, fill approximately 2500 ml ultra-pure water into a 5 l volumetric flask.
(2010), „Variation in the measurement of DNA damage by Comet assay measured by the ECVAG inter-laboratory validation trial“, Mutagenesis, Vol.
After adding 50 ml of each stock solution, fill 90 % of the volumetric flask with ultra-pure water and set pH to 5,8.
25/2, pp.
After this, add 150 g dissolved sucrose (3 % per 5 l); then, fill the volumetric flask with ultra-pure water up to the mark.
113–23.
Finally, the nutrient solution is filled into 1 l Schott flasks and autoclaved at 121 °C for 20 minutes.
Azqueta, A. et al.
The nutrient solution thus yielded can be kept sterile in a refrigerator (at 5-10 °C) for three months.
(2011), „Towards a more reliable comet assay: Optimising agarose concentration, unwinding time and electrophoresis conditions“, Mutation Research, Vol.
Modified Andrews' medium for Sediment-free toxicity test
724/1-2, pp.
From the five nutrient stock solutions already mentioned in Tables 1 and 2, a tenfold concentrated, modified Andrews' medium required for obtaining the test solutions will be prepared, with addition of 30 % sucrose.
41–45.
To do so, fill approximately 100 ml ultra-pure water into a 1 l volumetric flask.
87–90.
After adding 100 ml of each of the stock solutions, set pH to 5,8.
Douglas (1984), „Transient DNA lesions induced by benzo[a]pyrene in Chinese hamster ovary cells“, Mutation Research, Vol.
After this, add 30 % dissolved sucrose (300 g per 1000 ml); then, fill the volumetric flask with ultra-pure water up to the mark.
140/2-3, pp. 141–45.
Finally, the nutrient solution is filled into 0,5 l Schott flasks and autoclaved at 121 °C for 20 minutes.
Douglas (1990), „The role of excision repair in the removal of transient benzo[a]pyrene-induced DNA lesions in Chinese hamster ovary cells“, Mutation Research, Vol.
The tenfold concentrated modified nutrient solution thus yielded can be kept sterile in a refrigerator (at 5-10 °C) for three months.
236/1, pp. 35–41.
MAINTENANCE OF STOCK CULTURE
Nakajima, M. (2012), „Tissue sample preparation for in vivo rodent alkaline Comet assay“, Genes and Environment, Vol.
In this Appendix 3 the stock culture of Myriophyllum spicatum L [1], a submersed aquatic dicotyledon, a species of the water milfoils family is described.
34/1, pp. 50–4.
For sediment-free stock culture under laboratory conditions, sterile plants are required.
45–51.
Sterile plants are available from the ecotoxicology laboratory of the German Umweltbundesamt (Federal Environment Agency of Germany).
Lovell, D.P., G. Thomas, R. Dubow (1999), „Issues related to the experimental design and subsequent statistical analysis of in vivo and in vitro Comet studies“, Teratogenesis Carcinogenesis Mutagenesis, Vol.
Alternatively, test organisms can be prepared from non-sterile plants in accordance with ASTM designation E 1913-04.
19/2, pp. 109–19.
See below — extracted from the ASTM Standard Guide — the procedure for culturing Myriophyllum sibiricum collected from field:
Wiklund, S.J., E. Agurell (2003), „Aspects of design and statistical analysis in the Comet assay“, Mutagenesis, Vol.
Since the modified Andrews' medium contains sucrose (which stimulates the growth of fungi and bacteria), all material, solutions and culturing be conducted under sterile conditions.
18/2, pp. 167–75.
In addition, all flasks, dishes, bowls etc and other equipment undergo flame treatment at the sterile workbench just prior to use.
Bright, J. et al.
Stock cultures can be maintained under reduced illumination and temperature (50 μE m– 2 s– 1, 20 ± 2 °C) for longer times without needing to be re-established.
(2011), „Recommendations on the statistical analysis of the Comet assay“, Pharmaceutical Statistics, Vol.
The Myriophyllum growth medium should be the same as that used for testing but other nutrient rich media can be used for stock cultures.
10/6, pp.
The plant segments are distributed axenically over several 500 ml Erlenmeyer or/and 2000 ml Fernbach flasks, each filled with approximately 450 respectively 1000 ml modified Andrews' medium.
485–93.
Then, the flasks are axenically cellulose plug stoppered.
Lovell, D.P., T. Omori (2008), „Statistical issues in the use of the Comet assay“, Mutagenesis, Vol.
In addition, thorough flame treatment of equipment at the sterile workbench just prior to use is absolutely necessary.
23/3, pp. 171–82.
Dependent on number and size, the plants are to be transferred into fresh nutrient solution approximately every three weeks.
Box, G.E.P, Hunter, W.G. ja Hunter, J.S.
Apices as well as segments of the stem middle part for this renewed culture can be used.
Mead, R. (1990), The Design of Experiments.
Number and size of transferred plants (or segments of plants) are dependent on how many plants are needed.
Wu, C.F.J ja Hamada, M.S.
For example, you can transfer five shoot segments into one Fernbach flask and three shoot segments into one Erlenmeyer flask, each with a length of 5 cm.
Guerard, M., C. Marchand, U. Plappert-Helbig (2014), „Influence of Experimental Conditions on Data Variability in the Liver Comet Assay“, Environmental and Molecular Mutagenesis, Vol.
Discard any rooted, flowering, dead or otherwise conspicuous parts.
55/2, pp. 114–21.
Cutting of plants for the stock and pre culture after 3 weeks of cultivation
699–707.
waste
427–32.
pre culture
Recio L, Hobbs C, Caspary W, Witt KL, (2010), „Dose-response assessment of four genotoxic chemicals in a combined mouse and rat micronucleus (MN) and Comet assay protocol“, J. Toxicol.
stock culture
35:149–62.
Culturing of plants is to be performed in 500 ml Erlenmeyer and 2000 ml Fernbach flasks in a cooling incubator at 20 ± 2 °C with continuously light at approximately 100-150 μE m– 2 s– 1 or 6000-9000 Lux (emitted by chamber illumination with colour temperature “warm white light”).
101–13.
Culturing of plants in a cooling incubator with chamber illumination
tabeli kujul esitatud andmed igal proovivõtmisel määratud aine sisalduse kohta kalades (iga üksikut kala iseloomustav Cf) ja vees (Cw) ning vajaduse korral asjaomane keskväärtus, standardhälve ja vahemik kontrollrühmas ja katserühma(de)s (Cf-i väljendatakse milligrammides terve kala või konkreetsete kudede, näiteks rasvkoe märgmassi kilogrammi kohta ja Cw-d milligrammides liitri kohta); Cw väärtus kontrollrühmas (tuleks esitada ka taustsisaldus);
Chemically clean (acid-washed) and sterile glass culture vessels should be used and aseptic handling techniques employed.
2004. aasta alguses esitati püsivate, bioakumuleeruvate ja mürgiste kemikaalide ELi töörühmale sellise meetodi kavand (38), mis oli ette nähtud võimaliku bioakumuleerumise tuvastamiseks selliste vees raskesti lahustuvate orgaaniliste ainete puhul, mida ei saa uurida veekaudsel kokkupuutel toimuva biokontsentreerumise hindamise standardmeetodiga; nimetatud kavandiga koos esitati ka seda toetav taustdokument (39).