natgillin/translations-raw huggingface dataset:
Therefore, other routes of exposure (such as drinking water, subcutaneous, intravenous, topical, inhalation, intratracheal, dietary, or implantation) may be acceptable where they can be justified.
Kudede kogumise põhjendus peaks olema selgelt määratletud.
Intraperitoneal injection is not recommended since it is not a physiologically relevant route of human exposure.
Kuna mutatsioonide tekkimist on võimalik uurida peaaegu igas koes, peaks kogutavate kudede valimine põhinema uuringu tegemise eesmärgil ja kõikidel uuritava kemikaali kohta saadud mutageensuse, kantserogeensuse ja mürgisuse andmetel.
The maximum volume of liquid that can be administered by gavage or injection at one time depends on the size of the test animal.
Olulised tegurid, mida tuleks kaaluda, peaksid hõlmama vähemalt järgmist: manustamisviis (põhineb inimeste tõenäolisel kokkupuuteviisil), jaotumine kudedes ja võimalik toimemehhanism.
The volume should not exceed 2 ml/100 g body weight.
Taustteabe puudumise korral tuleks koguda paljusid somaatilisi kudesid, kuna need võivad pakkuda huvi.
The use of volumes greater than this should be justified.
Nende hulgas peaks olema kiiresti kasvavaid, aeglaselt kasvavaid ja kokkupuutekoha kudesid.
Except for irritating or corrosive chemicals, which will normally reveal exacerbated effects at higher concentrations, variability in test volume should be minimised by adjusting the concentration to ensure a constant volume at all dose levels.
Lisaks tuleks koguda spermatosoide seemnejuhast ja munandimanuse sabast ning arenevaid sugurakke seemnekanalikestest (nagu on kirjeldatud punktides 32 ja 33) ja säilitada hilisema analüüsi jaoks, kui on vaja määrata mutageensust sugurakkude suhtes.
Sampling Time
Tuleks määrata elundite massid ja suuremate elundite puhul tuleks kõikidelt loomadelt koguda üks ja sama piirkond.
Somatic Cells
Kudede ja DNA säilitamine
The sampling time is a critical variable because it is determined by the period needed for mutations to be fixed.
Kudesid või nende homogenaate tuleb säilitada temperatuuril alla —70 °C ja kasutada DNA eraldamiseks viie aasta jooksul.
This period is tissue-specific and appears to be related to the turnover time of the cell population, with bone marrow and intestine being rapid responders and the liver being much slower.
Eraldatud DNA, mida säilitatakse temperatuuril 4 °C sobivas puhverlahuses, tuleks kasutada mutatsioonide analüüsiks 1 aasta jooksul.
A suitable compromise for the measurement of mutant frequencies in both rapidly and slowly proliferating tissues is 28 consecutive daily treatments (as indicated in paragraph 26) and sampling three days after the final treatment;
Kudede valimine mutantide analüüsiks
although the maximum mutant frequency may not manifest itself in slowly proliferating tissues under these conditions.
Kudede valimisel tuleks arvestada järgmist:
If slowly proliferating tissues are of particular importance, then a later sampling time of 28 days following the 28 day administration period may be more appropriate (16) (29).
Kui uuring tehakse kantserogeensusuuringu jätkuna, tuleks arvestada kantserogeensuse sihtkudesid.
In such cases, the later sampling time would replace the 3 day sampling time, and would require scientific justification.
Analüüsi jaoks kudede valimisel tuleks seada eesmärgiks selliste kemikaalide tuvastamine, mis on otse toimivad in vitro mutageenid, mis kiiresti metaboliseeritakse, on väga reaktsioonivõimelised või neelduvad halvasti või mille sihtkude on määratud manustamisteega (6).
Germ Cells
Kui taustteavet ei ole ning võttes arvesse kokkupuute kohta, mis on määratud manustamisviisiga, tuleks mutageensust hinnata maksa ja vähemalt ühe kiiresti jaguneva koe (nt näärmemagu, luuüdi) järgi.
TGR assays are well-suited for the study of gene mutation induction in male germ cells (7) (8) (27), in which the timing and kinetics of spermatogenesis have been well-defined (27).
Enamikul juhtudel võib eespool nimetatud nõuded täita kahe hoolikalt valitud koe analüüsiga, kuid mõnel juhul oleks vaja analüüsida kolme või enamat kude.
The low numbers of ova available for analysis, even after super-ovulation, and the fact that there is no DNA synthesis in the oocyte, preclude the determination of mutation in female germ cells using transgenic assays (31).
Kui on põhjust eriti muretseda mõju pärast sugurakkudele, sealhulgas positiivse tulemuse pärast somaatilistes rakkudes, tuleks mutatsioonide suhtes uurida sugurakkude kudesid.
The sampling times for male germ cells should be selected so that the range of exposed cell types throughout germ cell development is sampled, and so that the stage targeted in the sampling has received sufficient exposure.
Mõõtmismeetodid
The time for the progression of developing germ cells from spermatogonial stem cells to mature sperm reaching the vas deferens/cauda epididymisis ~ 49 days for the mouse (36) and ~70 days for the rat (34) (35).
Standardsed avaldatud laborimeetodid mutantide avastamiseks on kättesaadavad järgmiste soovitatavate transgeenimudelite jaoks:
Following a 28-day exposure with a subsequent three day sampling period, accumulated sperm collected from the vas deferens/cauda epididymis (7)(8) will represent a population of cells exposed during approximately the latter half of spermatogenesis, which includes the meiotic and postmeiotic period, but not the spermatogonial or stem cell period.
lacZ λ-bakteriofaag ja -plasmiid (30);
In order to adequately sample cells in the vas deferens/cauda epididymis that were spermatogonial stem cells during the exposure period, an additional sampling time at a minimum of 7 weeks (mice) or 10 weeks (rat), after the end of treatment is required.
lacI hiir (2, 18);
Cells extruded from seminiferous tubules after a 28 + 3 day regimen comprise a mixed population enriched for all stages of developing germ cells (7) (8).
gpt delta hiir (22);
Sampling these cells for gene mutation detection does not provide as precise an assessment of the stages at which germ cell mutations are induced as can be obtained from sampling spermatozoa from the vas deferens/cauda epididymis (since there is a range of germ cell types sampled from the tubules, and there will be some somatic cells contaminating this cell population).
gpt delta rott (28);
However, sampling cells from seminiferous tubules in addition to spermatozoa from the vas deferens/cauda epididymis following only a 28 + 3 day sampling regimen would provide some coverage of cells exposed across the majority of phases of germ cell development, and may be useful for detecting some germ cell mutagens.
Muudatused peavad olema nõuetekohaselt põhjendatud ja dokumenteeritud.
General clinical observations should be made at least once a day, preferably at the same time(s) each day and considering the peak period of anticipated effects after dosing.
Mitme pakendi andmed saab koondada ja kasutada selleks, et saavutada piisav arv lüüsilaike või kolooniaid.
The health condition of the animals should be recorded.
Kui vajaliku arvu lüüsilaikude saamiseks on vaja suurt arvu pakendamisreaktsioone, võib see näidata DNA halba kvaliteeti.
Measurements of food consumption should be made at least weekly.
Üldiselt on vaja vähemalt 125000 — 300000 lüüsilaiku, kui spontaanne mutatsioonisagedus on suurusjärgus 3 × 10–5 (15).
If the test chemical is administered via the drinking water, water consumption should be measured at each change of water and at least weekly.
Big Blue® lacI katse puhul on oluline näidata, et kogu mutantide värvusfenotüüpide vahemikku saab avastada; selleks lisatakse sobivad värvuse kontrollid paralleelselt igale plaadile.
Animals exhibiting non-lethal indicators of excess toxicity should be euthanatised prior to completion of the test period (23).
Koed ja saadavad proovid (näidised) tuleks töödelda ja analüüsida plokkskeemi alusel, milles lahusti/kandeaine kontrollrühma, positiivse kontrolli rühma (kui seda kasutatakse) või positiivse kontrolli DNA (kui see on asjakohane) ja iga doosirühma näidised töödeldakse koos.
Tissue Collection
Andmete töötlemine
The rationale for tissue collection should be defined clearly.
Andmed üksikute loomade kohta esitatakse tabeli kujul.
Since it is possible to study mutation induction in virtually any tissue, the selection of tissues to be collected should be based upon the reason for conducting the study and any existing mutagenicity, carcinogenicity or toxicity data for the chemical under investigation.
Katseühikuks on loom.
Important factors for consideration should include the route of administration (based on likely human exposure route(s)), the predicted tissue distribution, and the possible mechanism of action.
Protokollis tuleks esitada lüüsilaike moodustavate ühikute (plaque-forming units, PFU) või kolooniaid moodustavate ühikute (colony-forming units, CFU) koguarv, mutantide arv ja mutatsioonide sagedus iga looma iga koe puhul.
In the absence of any background information, several somatic tissues as may be of interest should be collected.
Kui tegemist on paljude pakendamis- või vabastamisreaktsioonidega, tuleks esitada reaktsioonide arv iga DNA-näidise kohta.
These should represent rapidly proliferating, slowly proliferating and site of contact tissues.
Andmed iga üksiku reaktsiooni kohta tuleks säilitada, kuid protokollis on vaja esitada ainult vaid PFU-de või CFU-de üldarv.
In addition, spermatozoa from the vas deferens/cauda epididymis and developing germ cells from the seminiferous tubules (as described in paragraphs 32 and 33) should be collected and stored in case future analysis of germ cell mutagenicity is required.
Tuleks esitada andmed mürgisuse kohta ja kliinilised tunnused, vt punkt 34.
Organ weights should be obtained, and for larger organs, the same area should be collected from all animals.
Iga analüüsitud mutandi puhul tuleb esitada kõik sekveneerimisandmed, samuti nendest tulenevad mutatsioonisageduse andmed iga looma ja iga koe kohta.
Storage of Tissues and DNA
Tulemuste statistiline hindamine ja tõlgendamine
Tissues (or tissue homogenates) should be stored at or below – 70 °C and be used for DNA isolation within 5 years.
On mitmeid kriteeriume positiivse tulemuse määramiseks, nagu muteerumissageduse suurenemine sõltuvalt kemikaali doosist või mutatsioonide sageduse selge suurenemine ühes doosirühmas, võrreldes lahusti/kandeaine kontrollrühmaga.
Isolated DNA, stored refrigerated at 4 °C in appropriate buffer, should be used optimally for mutation analysis within 1 year.
Doosi-toime sõltuvuse jaoks piisavate andmete saamiseks on vaja uurida vähemalt kolme doosirühma.
Selection of Tissues for Mutant Analysis
Peamine arvesse võetav tegur on tulemuste bioloogiline asjakohasus, kuid katsetulemuste hindamiseks võib kasutada sobivaid statistilisi meetodeid (4, 14, 15, 25, 26).
The choice of tissues should be based on considerations such as:
Kasutatavate statistiliste testide korral tuleks eksperimendiühikuks pidada üksiklooma.
the route of administration or site of first contact (e.g. glandular stomach if administration is oral, lung if administration is through inhalation, or skin if topical application has been used);
Uuritavat kemikaali, millega saadavad tulemused ühegi koe puhul ei vasta eespool esitatud kriteeriumidele, peetakse käesoleva katsemeetodi kohaselt negatiivseks.
and 2) pharmacokinetic parameters observed in general toxicity studies, which indicate tissue disposition, retention or accumulation, or target organs for toxicity.
Et negatiivset tulemust saaks pidada bioloogiliselt asjakohaseks, tuleks koe kokkupuudet kemikaaliga tõendada.
If studies are conducted to follow up carcinogenicity studies, target tissues for carcinogenicity should be considered.
DNA sekveneerimisanalüüside puhul võib tulemuste tõlgendamisel kasutada mitmeid statistilisi lähenemisviise (1, 5, 9, 19).
The choice of tissues for analysis should maximise the detection of chemicals that are direct-acting in vitro mutagens, rapidly metabolised, highly reactive or poorly absorbed, or those for which the target tissue is determined by route of administration (6).
Toime bioloogilise olulisuse hindamisel võib juhinduda sellest, kas leitud väärtused on varasemate kontrollkatsete vahemikus või väljaspool seda vahemikku (32).
In the absence of background information and taking into consideration the site of contact due to route of administration, the liver and at least one rapidly dividing tissue (e.g. glandular stomach, bone marrow) should be evaluated for mutagenicity.
identifitseerimisandmed ja CASi nr, kui see on teada;
In most cases, the above requirements can be achieved from analyses of two carefully selected tissues, but in some cases, three or more would be needed.
allikas, partii number, kui on olemas;
If there are reasons to be specifically concerned about germ cell effects, including positive responses in somatic cells, germ cell tissues should be evaluated for mutations.
füüsikaline olek ja puhtus;
Methods of Measurement
uuringu tegemisel olulised füüsikalis-keemilised omadused;
Standard laboratory or published methods for the detection of mutants are available for the recommended transgenic models:
uuritava kemikaali stabiilsus, kui see on teada.
lacZ lambda bacteriophage and plasmid (30);
Lahusti/kandeaine:
lacI mouse (2) (18);
kandeaine valiku põhjendus;
gpt delta mouse (22);
uuritava kemikaali lahustuvus ja püsivus lahustis/kandeaines, kui on teada;
gpt delta rat (28);
sööda, joogivee või sissehingamise kaudu manustamiseks sobiva vormi valmistamine;
Modifications should be justified and properly documented.
osutatud vormi analüüsid (nt, stabiilsus, ühtlus, nominaalsed kontsentratsioonid).
Data from multiple packagings can be aggregated and used to reach an adequate number of plaques or colonies.
kasutatud liik/liin ja selle valimise põhjendus;
However, the need for a large number of packaging reactions to reach the appropriate number of plaques may be an indication of poor DNA quality.
loomade arv, vanus ja sugu;
In such cases, data should be considered cautiously because they may be unreliable.
päritolu, pidamistingimused, sööt jne;
The optimal total number of plaques or colonies per DNA sample is governed by the statistical probability of detecting sufficient numbers of mutants at a given spontaneous mutant frequency.
iga looma mass katse alustamisel, sh kehamassi vahemik, keskväärtus ja standardhälve iga rühma kohta.
In general, a minimum of 125000 to 300000 plaques is required if the spontaneous mutant frequency is in the order of 3 × 10–5 (15).
positiivseid ja negatiivseid (lahus/kandeaine) kontrollkatseid käsitlevad andmed;
For the Big Blue® lacI assay, it is important to demonstrate that the whole range of mutant colour phenotypes can be detected by inclusion of appropriate colour controls concurrent with each plating.
vahemiku leidmise uuringu andmed;
Tissues and the resulting samples (items) should be processed and analysed using a block design, where items from the vehicle/solvent control group, the positive control group (if used) or positive control DNA (where appropriate), and each treatment group are processed together.
uuritud kemikaali preparaadi üksikasjad;
Treatment of Results
manustamistee põhjendus;
Individual animal data should be presented in tabular form.
looma suhtes avalduva mürgisuse mõõtmise meetodid, sealhulgas histopatoloogilised või hematoloogilised analüüsid, kui need on tehtud, ja loomade vaatluste ja kehamassi määramise sagedus;
The experimental unit is the animal.
meetodid, millega kontrolliti, et uuritav kemikaal jõudis sihtkoesse või üldvereringesse, kui tulemused olid negatiivsed;
The report should include the total number of plaque-forming units (pfu) or colony-forming units (cfu), the number of mutants, and the mutant frequency for each tissue from each animal.
tegelik doos (mg kehakaalu kg kohta päevas), mis arvutatakse sööda/joogivee tarbimise ja selles uuritava kemikaali sisalduse (ppm) järgi, kui see on kohaldatav;
If there are multiple packaging/rescue reactions, the number of reactions per DNA sample should be reported.
sööda ja vee kvaliteedi üksikasjad;
While data for each individual reaction should be retained, only the total pfu or cfu need be reported.
manustamis- ja proovivõtukavade detailne kirjeldus ja nende valiku põhjendused;
Data on toxicity and clinical signs as per paragraph 34 should be reported.
humaanse surmamise meetod;
Any sequencing results should be presented for each mutant analysed, and resulting mutation frequency calculations for each animal and tissue should be shown.
kudede eraldamise ja säilitamise kord;
Statistical Evaluation and Interpretation of Results
näriliste genoomse DNA eraldamise, transgeeni genoomsest DNA-st vabastamise ja transgeense DNA bakteriaalsesse peremeesorganismi ülekandmise meetodid;
There are several criteria for determining a positive result, such as a dose-related increase in the mutant frequency, or a clear increase in the mutant frequency in a single dose group compared to the solvent/vehicle control group.
kõikide rakkude, katsekomplektide ja reaktiivide allikad ja partiide numbrid (vajaduse korral);
At least three treated dose groups should be analysed in order to provide sufficient data for dose-response analysis.
mutantide loendamine meetodid;
While biological relevance of the results should be the primary consideration, appropriate statistical methods may be used as an aid in evaluating the test results (4) (14) (15) (25) (26).
mutantide molekulaarse analüüsi, kloonsuse paranduse kasutamise ja mutatsioonide sageduse arvutamise meetodid, kui see on kohaldatav.
Statistical tests used should consider the animal as the experimental unit.
Tulemused:
A test chemical for which the results do not meet the above criteria in any tissue is considered non-mutagenic in this assay.
loomade seisund enne katset ja kogu katse ajal, sealhulgas mürgistuse tunnused;
For biological relevance of a negative result, tissue exposure should be confirmed.
keha ja elundite massid surmamisel;
For DNA sequencing analyses, a number of statistical approaches are available to assist in interpreting the results (1) (5) (9) (19).
iga koe ja looma kohta mutantide arv, hinnatud lüüsilaikude või kolooniate arv, mutatsioonide sagedus;
Consideration of whether the observed values are within or outside of the historical control range can provide guidance when evaluating the biological significance of the response (32).
iga koe ja loomade rühma kohta pakendamisreaktsioonide arv DNA proovi kohta, mutantide üldarv, keskmine muteerumissagedus, standardhälve;
identification data and CAS no, if known;
doosi ja toime suhe, võimaluse korral;
source, lot number if available;
iga koe ja looma kohta sõltumatute mutantide arv ja keskmine mutatsioonisagedus, kui tehti mutatsioonide molekulaarne analüüs;
physical nature and purity;
paralleelsed ja varasemad negatiivse kontrolli andmed, nende vahemikud, keskväärtused ja standardhälbed;
physiochemical properties relevant to the conduct of the study;
paralleelse positiivse kontrolli (või mitteparalleelse positiivse kontrolli DNA) andmed;
stability of the test chemical, if known;
analüütilised määramised, kui need on kättesaadavad (nt DNA kontsentratsioon, mida kasutati pakendamisel, DNA sekveneerimise andmed);
Solvent/vehicle:
kasutatud statistilised analüüsid ja meetodid.
justification for choice of vehicle;
Kokkuvõte
solubility and stability of the test chemical in the solvent/vehicle, if known;
Skopek (1987), „Statistical Test for the Comparison of Samples from Mutational Spectra”, J. Mol.
preparation of dietary, drinking water or inhalation formulations;
, „A more Efficient Big Blue® Protocol Improves Transgene Rescue and Accuracy in an Adduct and Mutation Measurement”, Mutation Res., 518:
analytical determinations on formulations (e.g. stability, homogeneity, nominal concentrations);
Gossen and J. Vijg (1995), „Plasmid-based Transgenic Mouse Model for Studying in vivo Mutations”Nature, 377(6550):
species/strain used and justification for the choice;
Gorelick (1995), „Statistical Design and Analysis of Mutation Studies in Transgenic Mice”, Environ.
number, age and sex of animals;
Gorelick (1996), „Mutational Spectra in Transgenic Animal Research: Data Analysis and Study Design Based upon the Mutant or Mutation Frequency”, Environ.
source, housing conditions, diet, etc.;
Brooks, B. Burlinson, J. Mirsalis, B. Myhr, L. Recio and V. Thybaud (1999), „Transgenic Mouse Mutation Assay Systems can Play an important Role in Regulatory Mutagenicity Testing in vivo for the Detection of Site-of-contact Mutagens”, Mutagenesis, 14(1):
individual weight of the animals at the start of the test, including body weight range, mean and standard deviation for each group;
Soper(1995), „Temporal and Molecular Characteristics of Mutations Induced by Ethylnitrosourea in Germ Cells Isolated from Seminiferous Tubules and in Spermatozoa of lacZ Transgenic Mice”, Proc.
positive and negative (vehicle/solvent) control data;
Soper and J. Jiao (1997), „Toward an Understanding of the Use of Transgenic Mice for the Detection of Gene Mutations in Germ Cells”, Mutation Res., 388(2–3):
data from the range-finding study;
Tindall (2000), „Bayesian Analysis of Mutational Spectra”, Genetics, 156:
details of test chemical preparation;
Knook and J. Vijg(1989), „Efficient Rescue of Integrated Shuttle Vectors from Transgenic Mice: a Model for Studying Mutations in vivo”, Proc.
rationale for route of administration;
Gossen, J.A. and J. Vijg (1993), „A Selective System for lacZ-Phage using a Galactose-sensitive E. coli Host”, Biotechniques, 14(3):
methods for measurement of animal toxicity, including, where available, histopathological or haematological analyses and the frequency with which animal observations and body weights were taken;
, „Somatic Gene Mutation and Human Disease other than Cancer”, Mutation Res., 543:
methods for verifying that the test chemical reached the target tissue, or general circulation, if negative results are obtained;
, „Somatic Gene Mutation and Human Disease other than Cancer: an Update”, Mutation Res., 705:
actual dose (mg/kg body weight/day) calculated from diet/drinking water test chemical concentration (ppm) and consumption, if applicable;
Douglas and D. Krewski (1998), „Statistical Analysis of lacZ Mutant Frequency Data from Muta™Mouse Mutagenicity Assays”, Mutagenesis, 13(3):
details of food and water quality;
Skopek, V. Thybaud, K.R.Tindall and N. Yajima (2000), „In vivo Transgenic Mutation Assays”, Environ.
detailed description of treatment and sampling schedules and justifications for the choices;
Douglas (2003), „Treatment and Sampling Protocols for Transgenic Mutation Assays”, Environ.
method of euthanasia;
Muller, B. Paul, S. Adhya and S. Garges (1996), „Analysis of Genetic Instability during Mammary Tumor Progression using a novel Selection-based Assay for in vivo Mutations in a Bacteriophage λ Transgene Target”, Proc.
procedures for isolating and preserving tissues;
Short (1990), „The Use of Transgenic Mice for Short-term, in vivo Mutagenicity Testing”, Genet.
methods for isolation of rodent genomic DNA, rescuing the transgene from genomic DNA, and transferring transgenic DNA to a bacterial host;
Burns (2008), „Comparison of Induced and Cancer-associated Mutational Spectra using Multivariate Data Analysis”, Carcinogenesis, 29(4):
source and lot numbers of all cells, kits and reagents (where applicable);
Nohmi, T., M. Katoh, H. Suzuki, M. Matsui, M. Yamada, M. Watanabe, M. Suzuki, N. Horiya, O. Ueda, T. Shibuya, H. Ikeda and T. Sofuni (1996), „A new Transgenic Mouse Mutagenesis Test System using Spi– and 6-thioguanine Selections”, Environ.
methods for enumeration of mutants;
Nohmi, T., M. Suzuki, K. Masumura, M. Yamada, K. Matsui, O. Ueda, H. Suzuki, M. Katoh, H. Ikeda and T. Sofuni (1999), „Spi – Selection: an Efficient Method to Detect γ-ray-induced Deletions in Transgenic Mice”, Environ.
methods for molecular analysis of mutants and use in correcting for clonality and/or calculating mutation frequencies, if applicable;
Masumura (2000), „Recent Advances in the Protocols of Transgenic Mouse Mutation Assays”, Mutation Res., 455(1–2):
Results:
OECD (2000), Guidance Document on the Recognition, Assessment and Use of Clinical Signs as Humane Endpoints for Experimental Animals Used in Safety Evaluation, Series on Testing and Assessment, No19, ENV/JM/MONO(2000)7, OECD, Paris.