natgillin/translations-raw huggingface dataset:
Hvis der anvendes objektglas, kan dette ske fra to eller tre scorede objektglas pr. prøve, når der anvendes fem dyr pr. gruppe.
However, other species can be used if ethically and scientifically justified.
Flere områder af objektglasset bør observeres ved en tæthed, der sikrer, at der ikke forekommer overlapning af halerne.
For rodents, the temperature in the experimental animal room ideally should be 22 °C (± 3 °C).
Det bør undgås at foretage scoring ved kanten af objektglassene.
The relative humidity ideally should be 50-60 %, being at least 30 % and preferably not exceeding 70 % other than during room cleaning.
DNA-strengbrud i kometassayet kan måles ved hjælp af uafhængige endepunkter såsom den procentvise DNA-intensitet i komethalen, halelængde og halemoment.
Rodents should be housed in small groups (usually no more than five) of the same sex if no aggressive behaviour is expected.
Alle tre målinger kan foretages, hvis man anvender hensigtsmæssige softwaresystemer til billedanalyse.
Solid floors should be used wherever possible as mesh floors can cause serious injury (49).
Den procentvise DNA-intensitet i komethalen (også kaldet procentvis haleintensitet) anbefales dog til evaluering og fortolkning af resultater (12) (40) (41) (42) og bestemmes af intensiteten af DNA-fragmenter i halen udtrykt som en procentdel af cellens samlede intensitet (13).
Appropriate environmental enrichment must be provided.
Vævsbeskadigelse og cytotoksicitet
Animals are randomly assigned to the control and treatment groups.
Positive resultater i kometassayet skyldes muligvis ikke udelukkende genotoksicitet, idet toksicitet i målvævet også kan føre til stigninger i DNA-vandringen (12) (41).
The animals are identified uniquely and acclimated to the laboratory conditions for at least five days before the start of treatment.
Omvendt kan der ofte observeres lav eller moderat cytotoksicitet med kendte genotoksiner (12), hvilket viser, at det ikke er muligt at skelne mellem DNA-migration forårsaget af genotoksicitet og cytotoksicitet, der udelukkende forekommer i kometassayet.
The least invasive method of uniquely identifying animals must be used.
Når der imidlertid observeres stigninger i DNA-migrationen, anbefales det, at en eller flere indikatorer for cytotoksicitet undersøges, eftersom dette kan bidrage til fortolkningen af resultaterne.
Appropriate methods include ringing, tagging, micro-chipping and biometric identification.
Stigninger i DNA-migration samtidig med tilstedeværelsen af klare beviser for cytotoksicitet bør fortolkes med forsigtighed.
Toe and ear clipping are not scientifically justified in these tests.
Der har været foreslået mange målinger af cytotoxicitet, og histopatologiske forandringer anses for en relevant måling af vævstoksicitet.
Cages should be arranged in such a way that possible effects due to cage placement are minimized.
Observationer såsom inflammation, celleinfiltration og apoptotiske eller nekrotiske forandringer er blevet associeret med øget DNA-migration, men som påvist i JaCVAM-valideringstesten (12) findes der ikke en endelig liste over histopatologiske forandringer, som altid er forbundet med øget DNA-migration.
At the commencement of the study, the weight variation of animals should be minimal and not exceed ± 20 %.
Ændrede foranstaltninger inden for klinisk kemi (f.eks. AST, ALT) kan også give nyttige oplysninger om vævsskader, og man kan også overveje supplerende indikatorer såsom caspaseaktivering, TUNNEL-farvning, Annexin V-farvning osv.
Solid test chemicals should be dissolved or suspended in appropriate vehicles or admixed in diet or drinking water prior to dosing of the animals.
Der findes dog kun begrænsede offentliggjorte data, som har været anvendt til in vivo-undersøgelser, og nogle kan være mindre pålidelige end andre.
For inhalation exposures, test chemicals can be administered as gas, vapour, or a solid/liquid aerosol, depending on their physicochemical properties (50) (51).
Pindsvin (eller skyer, spøgelsesceller) er celler, der udviser et mikroskopbillede, der består af et lille eller ikke-eksisterende hoved og store, diffuse haler, og anses for at være stærkt beskadigede celler, selv om pindsvinenes ætiologi er usikker (se tillæg 3).
Vehicle
På grund af deres udseende er målinger af den procentvise DNA-intensitet i komethalen ved billedanalyse upålidelige, og derfor bør pindsvin vurderes separat.
The vehicle should not produce toxic effects at the dose volumes used, and should not be suspected of chemical reaction with the test chemicals.
Forekomsten af pindsvin noteres og rapporteres, og enhver relevant stigning, der menes at skyldes testkemikaliet, bør undersøges og fortolkes med forsigtighed.
If other than well-known vehicles are used, their inclusion should be supported with reference data indicating their compatibility in terms of test animals, route of administration and endpoint.
Kendskab til testkemikaliernes potentielle virkemåde kan hjælpe med ved sådanne overvejelser.
It should be noted that some vehicles (particularly viscous vehicles) can induce inflammation and increase background levels of DNA strand breaks at the site of contact, particularly with multiple administrations.
Dyret er forsøgsenheden, og derfor bør både data for de enkelte dyr og de opsummerede resultater præsenteres i tabelform.
At this time, a group of a minimum of 3 analysable animals of one sex, or of each sex if both are used (see paragraph 32), treated with a positive control substance should normally be included with each test.
På grund af den hierarkiske datastruktur anbefales det at bestemme middelværdien af den procentvise DNA-intensitet i komethalen for hvert objektglas og beregne gennemsnittet af middelværdierne for hvert dyr (12).
In future, it may be possible to demonstrate adequate proficiency to reduce the need for positive controls.
Gennemsnittet af middelværdierne for de enkelte dyr bestemmes derefter for at opnå et gennemsnit for gruppen.
If multiple sampling times are used (e.g. with a single administration protocol) it is only necessary to include positive controls at one of the sampling times, but a balanced design should be ensured (see paragraph 48).
Alle disse værdier bør medtages i rapporten.
It is not necessary to administer concurrent positive control substances by the same route as the test chemical, although it is important that the same route should be used when measuring site-of-contact effects.
Alternative metoder (jf. punkt 53) kan anvendes, hvis dette er videnskabeligt og statistisk begrundet.
The positive control substances should be shown to induce DNA strand breaks in all of the tissues of interest for the test chemical, and EMS is likely to be the positive control of choice since it has produced DNA strand breaks in all tissues that have been studied.
Der kan foretages en statistisk analyse ved hjælp af forskellige metoder (58) (59) (60) (61).
The doses of the positive control substances should be selected so as to produce moderate effects that critically assess the performance and sensitivity of the assay and could be based on dose-response curves established by the laboratory during the demonstration of proficiency.
Ved valget af de statistiske metoder bør der tages hensyn til behovet for at transformere dataene (f.eks. logaritme eller kvadratrod) og/eller for at tilføje et mindre tal (f.eks. 0,001) til alle (selv ikke-nul-) værdier for at mindske effekten af nul-celleværdier, jf. diskussionen i ovennævnte referencer.
The % tail DNA in concurrent positive control animals should be consistent with the pre-established laboratory range for each individual tissue and sampling time for that species (see paragraph 16).
Nærmere oplysninger om analyse af forholdet mellem behandling og køn, når begge køn anvendes, og efterfølgende analyse af data, hvor der enten konstateres eller ikke konstateres forskelle, findes i tillæg 2.
Examples of positive control substances and some of their target tissues (in rodents) are included in Table 1.
Data om toksicitet og kliniske tegn bør også anføres.
Substances other than those given in Table 1 can be selected if scientifically justified.
a. Den sideløbende negative kontrol anses for at kunne føjes til laboratoriets historiske negative kontroldatabase som beskrevet i punkt 16.
Examples of positive control substances and some of their target tissues
b. Sideløbende positive kontroller (jf. punkt 29) bør fremkalde responser, der er forenelige med responserne i den historiske positive kontroldatabase, og medføre en statistisk set signifikant stigning sammenlignet med den sideløbende negative kontrol.
Substances and CAS RN No.
c. Et tilstrækkeligt antal celler og doser er blevet analyseret (punkt 52 og 36-38).
Ethyl methanesulfonate (CAS RN 62-50-0) for any tissue
d. Kriterierne for valg af højeste dosis er i overensstemmelse med kriterierne i punkt 36.
Ethyl nitrosourea (CAS RN 759-73-9) for liver and stomach, duodenum or jejunum
Såfremt alle acceptkriterier er opfyldt, anses et testkemikalie for at være klart positivt, hvis:
Methyl methanesulfonate (CAS RN 66-27-3) for liver, stomach, duodenum or jejunum, lung and bronchoalveolar lavage (BAL) cells, kidney, bladder, lung, testis and bone marrow/blood
a. mindst en af testdoserne udviser en statistisk signifikant stigning sammenlignet med den sideløbende negative kontrol
N-Methyl-N′-nitro-N-nitrosoguanidine (CAS RN: 70-25-7) for stomach, duodenum or jejunum
c. et eller flere af resultaterne ligger uden for fordelingen af de historiske negative kontroldata for bestemte arter, bærestoffer, indgivelsesveje, væv og antal indgifter.
1,2-Dimethylhydrazine 2HCl (CAS RN 306-37-6) for liver and intestine
Når alle disse kriterier er opfyldt, anses testkemikaliet for at kunne fremkalde strengbrud på DNA i de vævstyper, der undersøges i dette testsystem.
N-methyl-N-nitrosourea (CAS RN 684-93-5) for liver, bone marrow, blood, kidney, stomach, jejunum, and brain.
Hvis kun et eller to af disse kriterier er opfyldt, se da afsnit 62.
A group of negative control animals, treated with vehicle alone, and otherwise treated in the same way as the treatment groups, should be included with each test for every sampling time and tissue.
Såfremt alle acceptkriterier er opfyldt, anses et testkemikalie for at være klart negativt, hvis:
The % tail DNA in negative control animals should be within the pre-established laboratory background range for each individual tissue and sampling time for that species (see paragraph 16).
b. der ikke indtræder en koncentrationsafhængig stigning ved evaluering med en passende trend test
In the absence of historical or published control data showing that no deleterious or genotoxic effects are induced by the chosen vehicle, by the number of administrations or by the route of administration, initial studies should be performed prior to conducting the full study, in order to establish acceptability of the vehicle control.
c. alle resultater ligger inden for fordelingen af de historiske negative kontroldata for bestemte arter, bærestoffer, ruter, væv og antal indgifter
Number and Sex of Animals
d. der er påvist direkte eller indirekte evidens, som støtter eksponering af eller toksicitet over for målvævet.
Although there is little data on female animals from which to make comparison between sexes in relation to the comet assay, in general, other in vivo genotoxicity responses are similar between male and female animals and therefore most studies could be performed in either sex.
Testkemikaliet anses i så fald for ikke at kunne fremkalde strengbrud på DNA i de vævstyper, der undersøges i dette testsystem.
Data demonstrating relevant differences between males and females (e.g. differences in systemic toxicity, metabolism, bioavailability, etc. including e.g. in a range-finding study) encourage the use of both sexes.
Hvis responsen hverken er klart negativ eller positiv (dvs. at ikke alle kriterier i punkt 59 eller 60 er opfyldt) og for at bidrage til at fastslå den biologiske relevans af et resultat, bør dataene evalueres af eksperter, og/eller der bør foretages yderligere undersøgelser, hvis det er videnskabeligt begrundet.
In this case, it may be appropriate to perform a study in both sexes e.g. as part of a repeated dose toxicity study.
Det kan være nyttigt at score flere celler eller foretage en ny undersøgelse, evt. med optimerede forsøgsbetingelser (f.eks. dosisintervaller, andre indgivelsesveje, andre prøveudtagningstidspunkter eller andet væv).
Group sizes at study initiation (and during establishment of proficiency) should be established with the aim of providing a minimum of 5 analysable animals of one sex, or of each sex if both are used, per group (less in the concurrent positive control group — see paragraph 29).
I sjældne tilfælde, selv efter yderligere undersøgelser, giver dataene ikke mulighed for at konkludere, om resultaterne er positive eller negative; konklusionen er derfor, at resultaterne er tvetydige.
As a guide to maximum typical animal requirements, a study conducted according the parameters established in paragraph 33 with three dose groups and concurrent negative and positive controls (each group composed of five animals of a single sex) would require between 25 and 35 animals.
For at vurdere den biologiske relevans af et positivt eller tvetydigt resultat er der behov for oplysninger om cytotoksicitet i målvævet (jf. punkt 54-55).
TREATMENT SCHEDULE
I tilfælde af at der alene forekommer positive eller usikre resultater, hvis der er klar evidens for cytotoksicitet, vil man konkludere, at undersøgelsen er tvetydig for genotoksicitet, medmindre der foreligger tilstrækkelige oplysninger til at underbygge en endelig konklusion.
Animals should be given daily treatments over a duration of 2 or more days (i.e. two or more treatments at approximately 24 hour intervals), and samples should be collected once at 2-6 h (or at the Tmax) after the last treatment (12).
I tilfælde af et negativt undersøgelsesresultat, hvor der er tegn på toksicitet ved alle testede doser, kan det være tilrådeligt at foretage yderligere undersøgelser ved ikke-toksiske doser.
Samples from extended dose regimens (e.g. 28-day daily dosing) are acceptable.
kilde, evt. partinummer
Successful combination of the comet and the erythrocyte micronucleus test has been demonstrated (10) (19).
testkemikaliets stabilitet, sidste anvendelsesdato eller dato for ny analyse, hvis denne er kendt.
However careful consideration should be given to the logistics involved in tissue sampling for comet analysis alongside the requirements of tissue sampling for other types of toxicological assessments.
Opløsningsmiddel/bærestof:
Harvest 24 hours after the last dose, which is typical of a general toxicity study, is not appropriate in most cases (see paragraph 40 on sampling time).
begrundelse for valg af opløsningsmiddel/bærestof
The use of other treatment and sampling schedules should be justified (see Appendix 3).
fremstilling af dosisformuleringer
For example single treatment with multiple sampling could be used however, it should be noted that more animals will be required for a study with a single administration study because of the need for multiple sampling times, but on occasions this may be preferable, e.g. when the test chemical induces excessive toxicity following repeated administrations.
analytiske bestemmelser af formuleringer (f.eks. stabilitet, homogenitet og nominel koncentration).
Whatever way the test is performed, it is acceptable as long as the test chemical gives a positive response or, for a negative study, as long as direct or indirect evidence supportive of exposure of, or toxicity to, the target tissue(s) has been demonstrated or if the limit dose is achieved (see paragraph 36).
anvendte arter/stammer samt videnskabelige og etiske begrundelser for valget
Test chemicals also may be administered as a split dose, i.e., two treatments on the same day separated by no more than 2-3 hours, to facilitate administering a large volume.
oprindelse, miljø, føde, berigelse osv.
Under these circumstances, the sampling time should be scheduled based on the time of the last dosing (see paragraph 40).
de enkelte dyrs vægt ved testens start og afslutning, samt interval for legemsvægt samt middel- og standardafvigelse for legemsvægten for hver enkelt gruppe
Dose Levels
resultater fra (en eventuel) undersøgelse til bestemmelse af dosisinterval
If a preliminary range-finding study is performed because there are no suitable data available from other relevant studies to aid in dose selection, it should be performed in the same laboratory, using the same species, strain, sex, and treatment regimen to be used in the main study according to current approaches for conducting dose range-finding studies.
begrundelse for indgivelsesvej
The study should aim to identify the maximum tolerated dose (MTD), defined as the dose inducing slight toxic effects relative to the duration of the study period (for example, clear clinical signs such as abnormal behaviour or reactions, minor body weight depression or target tissue cytotoxicity), but not death or evidence of pain, suffering or distress necessitating euthanasia.
injektionssted (for subkutane og intravenøse undersøgelser)
For a non-toxic test chemical, with an administration period of 14 days or more, the maximum (limit) dose is 1000 mg/kg bodyweight/day.
metoder til fremstilling af prøver, om muligt histopatologiske analyser, især for et kemikalie med positiv komet-respons
For administration periods of less than 14 days the maximum (limit) dose is 2000 mg/kg bodyweight/day.
begrundelse for valg af væv
For certain types of test chemicals (e.g. human pharmaceuticals) covered by specific regulations these limits may vary.
metoder til kontrol af, at testkemikaliet er nået frem til det almindelige kredsløb eller målvævet om der er opnået negative resultater
Chemicals that exhibit saturation of toxicokinetic properties, or induce detoxification processes that may lead to a decrease in exposure after long-term administration, may be exceptions to the dose-setting criteria and should be evaluated on a case-by-case basis.
detaljeret beskrivelse af behandling og prøveudtagningsplaner samt begrundelser for valg (f.eks. eventuelle toksikokinetiske data)
However, the dose levels used should also preferably cover a range from the maximum to one producing little or no toxicity.
metode til smertelindring, analgesi
When target tissue toxicity is observed at all dose levels tested, further study at non-toxic doses is advisable (see paragraphs 54-55).
procedurer for isolering og opbevaring af væv
Studies intending to more fully investigate the shape of the dose-response curve may require additional dose group(s).
metoder til fremstilling af enkeltcelle-/enkeltkernesuspension
Administration of Doses
kilde og partinummer for alle reagenser (når det er muligt)
In any case the route should be chosen to ensure adequate exposure of the target tissue(s).
metoder til evaluering af cytotoksicitet
Intraperitoneal injection is generally not recommended since it is not a typical relevant route of human exposure, and should only be used with specific justification (e.g. some positive control substances, for investigative purposes, or for some drugs that are administered by the intraperitoneal route).
elektroforese
The volume should not exceed 1 ml/100 g body weight, except in the case of aqueous solutions where 2 ml/100g body weight may be used.
anvendte farvningsteknikker
The use of volumes greater than this (if permitted by animal welfare legislation) should be justified.
metoder til scoring og måling af kometer.
Wherever possible different dose levels should be achieved by adjusting the concentration of the dosing formulation to ensure a constant volume in relation to body weight at all dose levels.
eventuelle almindelige kliniske observationer, før og i løbet af testperioden for hvert dyr
Sampling Time
dokumentation for cytotoksicitet, hvis det er relevant
The sampling time is a critical variable because it is determined by the period needed for the test chemicals to reach maximum concentration in the target tissue and for DNA strand breaks to be induced but before those breaks are removed, repaired or lead to cell death.
for undersøgelser af længere end en uges varighed: dyrenes individuelle vægt i løbet af undersøgelsen, herunder interval for kropsvægt, middel- og standardafvigelse for hver gruppe samt fødeindtag
The persistence of some of the lesions that lead to the DNA strand breaks detected by the comet assay may be very short, at least for some chemicals tested in vitro (52) (53).
dosis/respons-forhold, hvor det kan beregnes
Accordingly, if such transient DNA lesions are suspected, measures should be taken to mitigate their loss by ensuring that tissues are sampled sufficiently early, possibly earlier than the default times given below.
for hver vævstype/hvert dyr, den procentvise DNA-intensitet i komethalen (eller andre målinger, hvis dette vælges) og middelværdier pr. objektglas, gennemsnitsværdier pr. dyr og middelværdier pr. gruppe
The optimum sampling time(s) may be chemical- or route-specific resulting in, for example, rapid tissue exposure with intravenous administration or inhalation exposure.
sideløbende og historiske negative kontroldata med intervaller, medianer/gennemsnit og standardafvigelser for hver evalueret vævstype
Accordingly, where available, sampling times should be determined from kinetic data (e.g. the time (Tmax) at which the peak plasma or tissue concentration (Cmax) is achieved, or at the steady state for multiple administrations).
data fra sideløbende negative og positive kontrolprøver
In the absence of kinetic data a suitable compromise for the measurement of genotoxicity is to sample at 2-6 h after the last treatment for two or more treatments, or at both 2-6 and 16-26 h after a single administration, although care should be taken to necropsy all animals at the same time after the last (or only) dose.
for andet væv end lever, en dosis/respons-kurve, hvortil den positive kontrol anvendes.
Information on the appearance of toxic effects in target organs (if available) may also be used to select appropriate sampling times.
Dette kan være fra data, der er indsamlet i forbindelse med godtgørelse af kompetence (jf. punkt 16-17) og bør ledsages af en begrundelse med henvisninger til aktuel litteratur for at fastslå hensigtsmæssigheden af størrelsen og spredningen af responserne på kontrollerne i dette væv
General clinical observations related to the health of the animals should be made and recorded at least once a day preferably at the same time(s) each day and considering the peak period of anticipated effects after dosing (54).
anvendte statistiske analyser og metoder samt kriterier for, hvornår en respons kan anses som positiv, negativ eller tvetydig
At least twice daily, all animals should be observed for morbidity and mortality.
hyppigheden af pindsvin i hver gruppe og pr. dyr.
For longer duration studies, all animals should be weighed at least once a week, and at completion of the test period.
Kapitel B.8 til dette bilag: Subakut toksicitet (inhalation): 28-dages undersøgelse.
Food consumption should be measured at each change of food and at least weekly.
Kapitel B.29 til dette bilag: Subkronisk toksicitet (inhalation): 90-dages undersøgelse.
Animals exhibiting non-lethal indicators of excessive toxicity should be euthanized prior to completion of the test period, and are generally not used for comet analysis.
Alkalisk encellet gelelektroforese
Tissue Collection
Følsom metode til påvisning af primær DNA-skade på celle-/kerneniveau.
Since it is possible to study induction of DNA strand breaks (comets) in virtually any tissue, the rationale for selection of tissue(s) to be collected should be clearly defined and based upon the reason for conducting the study together with any existing ADME, genotoxicity, carcinogenicity or other toxicity data for the test chemicals under investigation.
Komet
Important factors for consideration should include the route of administration (based on likely human exposure route(s)), the predicted tissue distribution and absorption, the role of metabolism and the possible mechanism of action of the test chemicals.