natgillin/translations-raw huggingface dataset:
Both average specific growth rate (estimated from assessments of main shoot length and three additional measurement variables) and yield (estimated from the increase in main shoot length and three additional measurement variables) of untreated and treated plants should be determined.
Függetlenül attól, hogy milyen automatizált platform használatos a mikronukleuszok számolására, a CBPI, az RI, az RPD, illetve az RICC mutatót egyidejűleg vizsgálni kell.
An analytical method, with adequate sensitivity for quantification of the test chemical in the test medium, should be available.
Mutation Research, Vol. 287/1, pp.
Information on the test chemical which may be useful in establishing the test conditions includes the structural formula, purity and impurities, water solubility, stability in water and light, acid dissociation constant (pKa), partition coefficient octanol-water (Kow), vapour pressure and biodegradability.
3-15.
Water solubility and vapour pressure can be used to calculate Henry's Law constant, which will indicate if significant losses of the test chemical during the test period are likely.
Cytobios, Vol. 43/172-173, pp.
This will help indicate whether particular steps to control such losses should be taken.
233-246.
Where information on the solubility and stability of the test chemical are uncertain, it is recommended that these be assessed under the conditions of the test, i.e. growth medium, temperature, lighting regime to be used in the test.
Environmental and Molecular Mutagenesis, Vol. 35/3, pp.
The pH control of the test medium is particularly important, e.g. when testing metals or substances which are hydrolytically unstable.
167-172.
Further guidance for testing chemicals with physical-chemical properties that make them difficult to test is provided in a OECD Guidance Document (5).
Nature Protocols, Vol. 2/5, pp.
For the test to be valid, the doubling time of main shoot length in the control must be less than 14 days.
1084-1104.
Using the media and test conditions described in this test method, this criterion can be attained using a static or semi-static test regime.
Mutation Research, Vol. 161/2, pp.
The mean coefficient of variation for yield based on measurements of shoot fresh weight (i.e. from test initiation to test termination) and the additional measurement variables (see paragraph 37) in the control cultures do not exceed 35 % between replicates.
193-198.
More than 50 % of the replicates of the control group are kept sterile over the exposure period of 14 days, which means visibly free of contamination by other organisms such as algae, fungi and bacteria (clear solution).
Environmental and Molecular Mutagenesis, Vol. 13/1, pp.
Note: Guidance on how to assess sterility is provided in the ring-test report (4).
34-43.
REFERENCE CHEMICAL
Mutagenesis, Vol. 6/4, pp.
Reference chemical(s), such as 3,5-dichlorophenol used in the ring test (4), may be tested as a mean of checking the test procedure; from the ring test data, the mean EC50-values of 3,5-DCP for the different response variables (see paragraphs 37-41 of this test method) are between 3,2 mg/l and 6,9 mg/l (see ring test report for details about confidence interval for these values).
297-302.
It is advisable to test a reference chemical at least twice a year or, where testing is carried out at a lower frequency, in parallel to the determination of the toxicity of a test chemical.
Mutation Research, Vol. 392/1-2, pp.
All equipment in contact with the test media should be made of glass or other chemically inert material.
187-208.
Glassware used for culturing and testing purposes should be cleaned of chemical contaminants that might leach into the test medium and should be sterile.
Gesellschaft fur Umwelt-Mutations-forschung, Mutation Research, Vol. 410, pp.
The test vessels should be long enough for the shoot in the control vessels to grow in the water phase without reaching the surface of the test medium at the end of the test.
81-116.
Thick-walled borosilicate glass test tubes without lip, inner diameter approximately 20 mm, length approximately 250 mm, with aluminium caps are recommended.
Chemically induced micronucleus formation in V79 cells – comparison of three different test approaches.
Since the modified Andrews' medium contains sucrose (which stimulates the growth of fungi and bacteria), the test solutions have to be prepared under sterile conditions.
Mutation Research, Vol. 607/1, pp.
All liquids as well as equipment are sterilised before use.
1-152.
Sterilisation is carried out via heated air treatment (210 °C) for 4 hours or autoclaving for 20 minutes at 121 °C.
Regulatory Toxicology and Pharmacology, Vol. 59/1, pp.
In addition, all flasks, dishes, bowls etc. and other equipment undergo flame treatment at a sterile workbench just prior to use.
28-36.
The cultures and test vessels should not be kept together.
Mutation Research Letters, Vol. 347/3-4, pp.
This is best achieved using separate environmental growth chambers, incubators, or rooms.
105-115.
Illumination and temperature should be controllable and maintained at a constant level.
Mutagenesis, Vol 23/4, pp.
Myriophyllum spicatum — a submersed aquatic dicotyledon — is a species of the water milfoils family.
Environmental Mutagenesis, Vol. 8/6, pp.
Between June and August, inconspicuous pink-white flowers protrude above the water surface.
789-886.
The plants are rooted in the ground by a system of robust rhizomes and can be found in the entire northern hemisphere in eutrophic, however non-polluted and more calciferous still waters with muddy substrate.
Environmental and Molecular Mutation., Vol. 49, pp.
Myriophyllum spicatum prefers fresh water, but is found in brackish water as well.
439-452.
For the sediment-free toxicity test, sterile plants are required.
Mutation Research, Vol. 392, pp.
If the testing laboratory does not have regular cultures of Myriophyllum spicatum, sterile plant material may be obtained from another laboratory or (unsterile) plant material might be taken from the field or provided by a commercial supplier; if plants come from the field a taxonomic verification of the species should be envisaged.
11-18.
Field sites used for collecting starting cultures have to be free of obvious sources of contamination.
825-840.
Great care should be taken to ensure that the correct species is obtained when collecting Myriophyllum spicatum from the field, especially in regions where it can hybridise with other Myriophyllum species.
Toxicology, Vol. 198/1-3, pp.
If obtained from another laboratory they should be similarly maintained for a minimum of three weeks.
315-328.
The source of plant material and the species used for testing should always be reported.
Mutation Research, Vol. 113/3-4, pp.
The quality and uniformity of the plants used for the test will have a significant influence on the outcome of the test and should therefore be selected with care.
173-215.
Young, rapidly growing plants without visible lesions or discoloration (chlorosis) should be used.
Journal of Environmental Pathology and Toxicology, Vol. 4/1, pp.
Details about preparation of the test organism are given in Appendix 4.
55-65.
Cultivation
Mutagenesis, Vol. 7, pp.
To reduce the frequency of culture maintenance (e.g. when no Myriophyllum tests are planned for a period), cultures can be held under reduced illumination and temperature (50 μE m–2 s–1, 20 ± 2 °C).
175-177.
Details of culturing are given in Appendix 3.
Environmental and Molecular Mutagenesis, Vol. 28, pp.
At least 14 to 21 days before testing, sufficient test organisms are transferred aseptically into fresh sterile medium and cultured for 14 to 21 days under the conditions of the test as a pre culture.
51-59.
Details for preparation of a pre-culture are given in Appendix 4.
Mutation Research, Vol.190/3, pp.
Only one nutrient medium is recommended for Myriophyllum spicatum in a sediment-free test system, as described in Appendix 2.
225-8.
A modification of the Andrews' medium is recommended for culturing and testing with Myriophyllum spicatum as described in (1).
Mutation Research, Vol.746/1, pp.
From five separately prepared nutrient stock solutions with addition of 3 % sucrose the modified Andrews' medium will be arranged.
29-34.
Details about preparation of the medium are given in Appendix 2.
Environmental and Molecular Mutagenesis, Vol. 52/4, pp.
A tenfold concentrated, modified Andrews' medium is needed for obtaining the test solutions (by dilution as appropriate).
280–286.
The composition of this medium is given in Appendix 2.
Mutagenesis, Vol. 25/1, pp.
Test solutions are usually prepared by dilution of a stock solution.
33-40.
Stock solutions of the test chemical are normally prepared by dissolving the chemical in demineralised (i.e. distilled or deionised) water.
Archives of Toxicology, Vol. 85/8, pp.
The addition of the nutrients will be achieved by using the tenfold concentrated, modified Andrews' medium.
873-99.
The stock solutions of the test chemical can be sterilised by autoclave at 121 °C for 20 minutes or by sterile filtration, provided that the sterilisation technique used does not denaturise the test chemical.
Mutation Research/Genetic Toxicology and Environmental Mutagenesis, Vol.723/2, pp.
Test solutions can also be prepared in sterile demineralised water or medium, under sterile conditions.
87-90.
The thermo-stability and the adsorption on different surfaces should the taken into account in the selection of the sterilisation procedure of the stock solutions of the test chemical.
valamint:
Because of that, it is recommended that the stock solutions be prepared under sterile conditions, i.e. using sterile material for dissolving the test chemical under sterile conditions (e.g. flame sterilisation, laminar-flow hoods, etc.) into sterile water.
A vizsgálati módszertől várható előrejelzések reprodukálhatóságának mértéke a laboratóriumokon belül 85 %, a laboratóriumok között pedig 80 % (10).
This technique of preparation of sterile stock solutions is valid for both substances and mixtures.
A validálási vizsgálatban (18) és a közzétett tanulmányokban (19) kapott eredmények összességében azt jelzik, hogy a DPRA a szenzibilizáló (azaz az ENSZ-GHS/a CLP-rendelet szerinti 1. kategóriába tartozó) anyagokat az LLNA vizsgálati módszer eredményeihez képest 80 %-os (N=157) pontossággal különbözteti meg a nem szenzibilizáló anyagoktól, 80 % az érzékenysége (88/109), a specifikussága pedig 77 % (37/48).
The highest tested concentration of the test chemical should normally not exceed its water solubility under the test conditions.
A vizsgálati vegyi anyagnak 100 mM végső koncentrációjú, megfelelő oldószerben oldhatónak kell lennie (lásd a 18. pontot).
For test chemicals of low water solubility it may be necessary to prepare a concentrated stock solution or dispersion of the chemical using an organic solvent or dispersant in order to facilitate the addition of accurate quantities of the test chemical to the test medium and aid in its dispersion and dissolution.
Az említett koncentrációnál nem oldható vizsgálati vegyi anyagok azonban alacsonyabb oldható koncentrációkban még vizsgálhatók.
Every effort should be made to avoid the use of such materials.
Ilyen esetben a pozitív eredmény továbbra is felhasználható a vizsgálati vegyi anyag bőrszenzibilizáló hatásúként történő azonosítására, a negatív eredmény alapján azonban nem szabad a reakcióképesség hiányára következtetni.
There should be no phytotoxicity resulting from the use of auxiliary solvents or dispersants.
kicsapódás megjelenése a reakciókeverékben az inkubációs idő végén, ha megfigyelték; a kicsapódást újra feloldották-e vagy centrifugálták-e;
For example, commonly used solvents which do not cause phytotoxicity at concentrations up to 100 μl/l, include acetone and dimethylformamide.
Egyesült Nemzetek Szervezete (ENSZ) (2013).
If a solvent or dispersant is used, its final concentration should be reported and kept to a minimum (≤ 100 μl/l), and all treatments and controls should contain the same concentration of solvent or dispersant.
90–100
Further guidance on the use of dispersants is given in (5).
0–24
Prior knowledge of the toxicity of the test chemical to Myriophyllum spicatum from a range-finding test will help in selecting suitable test concentrations.
0–5,5
In the definitive toxicity test, there should normally be five (like in the Lemna growth inhibition test, Chapter C.26 of this Annex) to seven test concentrations arranged in a geometric series; they should be chosen in order that the NOEC and EC50 values are bracketed by the concentration range (see below).
Technical Working Group on Pesticides – (Quantitative) Structure Activity Relationship ((Q)SAR) Guidance Document.
Preferably the separation factor between test concentrations should not exceed 3,2; however, a larger value may be used where the concentration-response curve is flat.
A klinikai kémiai változások (például AST, ALT) szintén szolgálhatnak a szövetkárosodásra vonatkozó hasznos információkkal, és figyelembe vehetők további mutatók is, mint a kaszpáz-aktiválás, a TUNEL-festés, az Anexin V-festés stb.
Justification should be provided when fewer than five concentrations are used.
Mindemellett kevés közzétett adat van e mutatók in vivo vizsgálatokhoz történő felhasználására vonatkozóan, és egyes mutatók nem annyira megbízhatóak, mint mások.
At least five replicates should be used at each test concentration.
Appropriate follow-up testing in vivo, Mutation Research, Vol. 654/2, pp.
In setting the range of test concentrations (for range-finding and/or for the definitive toxicity test), the following should be considered:
114-32.
To determine an ECx, test concentrations should bracket the ECx value to ensure an appropriate level of confidence.
Burlinson, B. (2012), The in vitro and in vivo Comet assays, Methods in Molecular Biology, Vol. 817, pp.
For example, if estimating the EC50, the highest test concentration should be greater than the EC50 value.
143-63.
If the EC50 value lies outside of the range of test concentrations, associated confidence intervals will be large and a proper assessment of the statistical fit of the model may not be possible.
, Recommendations for design of the rat Comet assay, Mutagenesis, Vol. 23/3, pp.
In addition, the highest test concentration should be high enough so that growth is significantly lower than that in the control.
, Recommendations for conducting the in vivo alkaline Comet assay, Mutagenesis, Vol. 18/1, pp.
If this is not the case, the test will have to be repeated using a different concentration range (unless the highest concentration is at the limit of solubility or the maximum required limit concentration, e.g. 100 mg/l).
45-51.
Every test should include controls consisting of the same nutrient medium, test organism (choosing plant material as homogeneous as possible, fresh lateral branches from pre-cultures, shortened to 2,5 cm from base), environmental conditions and procedures as the test vessels but without the test chemical.
, A simple technique for quantitation of low levels of DNA damage in individual cells, Experimental Cell Research, Vol. 175/1, pp.
If an auxiliary solvent or dispersant is used, an additional control treatment with the solvent/dispersant present at the same concentration as that in the vessels with the test chemical should be included.
184-91.
If determination of NOEC is not required, the test design may be altered to increase the number of concentrations and reduce the number of replicates per concentration.
40-69.
However, in any case the number of control replicates should be at least ten.
, 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. 722/1, pp.
Exposure
7-19.
Fresh lateral branches from pre-culture shortened to 2,5 cm from base are assigned randomly to the test vessels under aseptic conditions; each test vessel should contain one 2,5 cm lateral branch that should have an apical meristem on one end.
, Dose-response assessment of four genotoxic chemicals in a combined mouse and rat micronucleus (MN) and Comet assay protocol, The Journal of Toxicological Science, Vol. 35/2, pp.
The chosen plant material should be the same quality in each test vessel.
149-62.
A randomised design for location of the test vessels in the incubator is required to minimise the influence of spatial differences in light intensity or temperature.
Hartmann, A. (2004), Use of the alkaline in vivo Comet assay for mechanistic genotoxicity investigations, Mutagenesis, Vol. 19/1, pp.
If a preliminary stability test shows that the test chemical concentration cannot be maintained (i.e. the measured concentration falls below 80 % of the measured initial concentration) over the test duration (14 days), a semi-static test regime is recommended.
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. 630/1, pp.
In this case, the plants should be exposed to freshly prepared test and control solutions on at least one occasion during the test (e.g. day 7).
28-41.
The frequency of exposure to fresh medium will depend on the stability of the test chemical; a higher frequency may be needed to maintain near-constant concentrations of highly unstable or volatile chemicals.
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. 393/1-2, pp.
The method of light detection and measurement, in particular the type of sensor, will affect the measured value.
175-80.
Spherical sensors (which respond to light from all angles above and below the plane of measurement) and “cosine” sensors (which respond to light from all angles above the plane of measurement) are preferred to unidirectional sensors, and will give higher readings for a multi-point light source of the type described here.
, Detection of photogenotoxicity in skin and eye in rat with the photo Comet assay, Photochemical and Photobiological Sciences, Vol. 7/2, pp.
The temperature in the test vessels should be 23 ± 2 °C.
240-9.
Additional care is needed on pH drift in special cases such as when testing unstable chemicals or metals; the pH should remain in a range of 6-9.
, A comparison of cell-collecting methods for the Comet assay in urinary bladders of rats, Mutation Research/Genetic Toxicology and Environmental Mutagenesis, Vol. 742/1-2, pp.
See (5) for further guidance.
26-30.
The main shoot length of test organism appearing normal or abnormal needs to be determined at the beginning of the test, at least once during the 14-day exposure period and at test termination.
53-74.
Significant features of the test medium (e.g. presence of undissolved material, growth of algae, fungi and bacteria in the test vessel) should also be noted.
, Evaluation of DNA damage in different stages of mouse spermatogenesis after testicular X irradiation, Journal of Radiation Research, Vol. 160/4, pp.
In addition to determinations of main shoot length during the test, effects of the test chemical on three (or more) of the following measurement variables should be also assessed:
443-451.
Total lateral branches length
Wolf (1996), Detection of DNA-crosslinking agents with the alkaline Comet assay, Environmental and Molecular Mutagenesis, Vol. 27/3, pp.
Total shoot length
196-201.
Fresh weight
165-81.
Dry weight
Hartley (2010), Measurement of DNA interstrand crosslinking in individual cells using the Single Cell Gel Electrophoresis (Comet) assay, Methods in Molecular Biology, Vol. 613, pp.
Note 1:
Jha (2006), Reliable Comet assay measurements for detecting DNA damage induced by ionizing radiation and chemicals, Mutation Research/Genetic Toxicology and Environmental Mutagenesis, Vol. 605(1-2), pp.
The observations made during the range-finding test could help in selecting relevant additional measurements among the six variables listed above.
7-16.
The determination of the fresh and dry weights (parameters iv and v) is highly desirable.
113-23.
Note 3:
, Compilation and use of genetic toxicity historical control data, Mutation Research, Vol. 723/2, pp.
Due to the fact that sucrose and light (exposure of roots to light during the test) may have an influence on auxin (plant growth hormone) transport carriers, and that some chemicals may have an auxin-type mode of action, the inclusion of root endpoints (parameter iii) is questionable.
87-90.
Note 4:
Douglas (1984), Transient DNA lesions induced by benzo[a]pyrene in Chinese hamster ovary cells, Mutation Research, Vol. 140/2-3, pp.
The ring test results show high coefficients of variation (> 60 %) for the total lateral branch length (parameter i).
141-45.
Total lateral branch length is in any case encompassed within the total shoot length measurement (parameter ii) which shows more acceptable coefficients of variation of < 30 %.
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. 236/1, pp.
Note 5:
35-41.
Resulting from the above considerations, the recommended main measurement endpoints are: total shoot length, fresh weight and dry weight (parameters ii, iv and v); parameter vi — number of whorls — is left to the experimenter's judgment.
Nakajima, M. (2012), Tissue sample preparation for in vivo rodent alkaline Comet assay, Genes and Environment, Vol. 34/1, pp.
Main shoot length and number of whorls have an advantage, in that they can be determined for each test and control vessel at the start, during, and at the end of the test by photography and image analysis, although a (sterile) ruler can also be used.
50-4.
Total lateral branches length, total root length (as a sum of all lateral branches or roots) and total shoot length (as a sum of main shoot length and total lateral branches length) can be measured with a ruler at the end of exposure.
, Recommendations on the statistical analysis of the Comet assay, Pharmaceutical Statistics, Vol. 10/6, pp.
The fresh and/or dry weight should be determined at the start of the test from a sample of the pre-culture representative of what is used to begin the test, and at the end of the test with the plant material from each test and control vessel.
485-93.
Total lateral branches length, total shoot length, total root length, fresh weight, dry weight and number of whorls may be determined as follows:
, Validation of use of frozen tissues in high-throughput comet assay with fully-automatic scoring, Mutagenesis, Vol. 28/6, pp.
The lateral branch length may be determined by measuring all lateral branches with a ruler at the end of exposure.
699-707.
The main shoot length may be determined by image analysis or using a ruler.
101-13.
The total shoot length is the sum of the total lateral branches length and the main shoot length of each test and control vessel at the end of exposure.
2004 elején olyan protokolltervezetet (38) nyújtottak be az EU PBT munkacsoportjához, amelynek célja a vízben csekély mértékben oldódó szerves anyagok bioakkumulációs potenciáljának mérése, amely anyagok esetében a standard, vízi expozíción alapuló biokoncentrációs módszer nem megvalósítható; e tervezetet háttérdokumentum kísérte (39).
The root length may be determined by measuring all roots with a ruler at the end of exposure.
A módszer tekintetében megadott további indok az volt, hogy a vízben csekély mértékben oldódó (azaz, ahol log KOW >5) ezen anyagoknak való potenciális környezeti expozíció jórészt az étrend útján alakulhat ki (vö. (40) (41) (42) (43) (44)).
The fresh weight may be determined by weighing the test organisms at the end of exposure.
Fel kell ismerni ugyanakkor, hogy az itt bemutatott módszerben a vizes fázisú expozíciót gondosan kerülik, így az e vizsgálati módszerből adódó BMF érték közvetlenül nem hasonlítható össze a terepvizsgálat során nyert BMF értékkel (ahol kombinálható a vízi és az étrendi expozíció).