natgillin/translations-raw huggingface dataset:
Treatments should not be shorter than the time required for the complete induction of all the relevant metabolising enzymes, and shorter treatments may necessitate the use of multiple sampling times that are suitable for organs with different proliferation rates.
Podatki za posamezne živali naj bodo prikazani v tabelah.
In any case, all available information (e.g. on general toxicity or metabolism and pharmacokinetics) should be used when justifying a protocol, especially when deviating from the above standard recommendations.
Preskusna enota je žival.
While it may increase sensitivity, treatment times longer than 8 weeks should be explained clearly and justified, since long treatment times may produce an apparent increase in mutant frequency through clonal expansion (29).
Poročilo naj vsebuje skupno število enot, ki delajo plake ali kolonije, število mutantov in pogostnost mutantov za vsako tkivo vsake živali.
Dose Levels
Če izvedemo več reakcij pakiranja/sproščanja fagov, je treba sporočiti število reakcij na posamezni vzorec DNA.
Dose levels should be based on the results of a dose range-finding study measuring general toxicity that was conducted by the same route of exposure, or on the results of pre-existing sub-acute toxicity studies.
Sicer je treba shraniti podatke za vsako posamezno reakcijo, vendar je treba sporočiti samo skupno število enot, ki delajo plake ali kolonije.
Non-transgenic animals of the same rodent strain may be used for determining dose ranges.
Sporočiti je treba podatke o strupenosti in kliničnih znakih v skladu z odstavkom 34.
In the main test, in order to obtain dose response information, a complete study should include a negative control group (see paragraph 22) and a minimum of three, appropriately-spaced dose levels, except where the limit dose has been used (see paragraph 28).
Morebitne rezultate določanja zaporedja je treba predstaviti za vsak analizirani mutant in izračune pogostnosti mutantov prikazati za vsako žival in tkivo.
The top dose should be the Maximum Tolerated Dose (MTD).
Statistično vrednotenje in razlaga rezultatov
The MTD is defined as the dose producing signs of toxicity such that higher dose levels, based on the same dosing regimen, would be expected to produce lethality.
Za določitev pozitivnega rezultata obstaja več meril, na primer povečanje pogostnosti mutantov, povezano z odmerkom, ali očitno povečanje pogostnosti mutantov v skupini z enim odmerkom v primerjavi s kontrolno skupino s topilom/nosilcem.
Chemicals with specific biological activities at low non-toxic doses (such as hormones and mitogens), and chemicals which exhibit saturation of toxicokinetic properties may be exceptions to the dose-setting criteria and should be evaluated on a case-by-case basis.
Da bi dobili dovolj podatkov za analizo odziva na odmerek, je treba analizirati vsaj tri tretirane skupine.
The dose levels used should cover a range from the maximum to little or no toxicity.
Najprej naj se prouči biološka pomembnost rezultatov, pri vrednotenju rezultatov preskusa pa se kot pomoč lahko uporabijo ustrezne statistične metode (4) (14) (15) (25) (26).
Limit Test
V uporabljenih statističnih preskusih naj bo preskusna enota žival.
If dose range-finding experiments, or existing data from related rodent strains, indicate that a treatment regime of at least the limit dose (see below) produces no observable toxic effects,and if genotoxicity would not be expected based upon data from structurally related chemicals, then a full study using three dose levels may not be considered necessary.
Preskusno kemikalijo, pri kateri rezultati ne izpolnjujejo navedenih meril v katerem od tkiv, je treba v tem preskusu šteti za nemutageno.
For an administration period of 28 days (i.e. 28 daily treatments), the limit dose is 1000 mg/kg body weight/day.
Za biološko pomembnost negativnega rezultata je treba potrditi izpostavljenost tkiva.
For administration periods of 14 days or less, the limit dose is 2000 mg/kg/body weight/day (dosing schedules differing from 28 daily treatments should be scientifically justified in the protocol; see paragraph 26).
Za analize določanja zaporedja DNA so za pomoč pri razlagi rezultatov na voljo številni statistični pristopi (1) (5) (9) (19).
Administration of Doses
Upoštevanje tega, ali so dobljene vrednosti v razponu preteklih kontrol ali zunaj njega, lahko zagotovi smernice pri ocenjevanju biološke pomembnosti odziva (32).
The test chemical is usually administered by gavage using a stomach tube or a suitable intubation cannula.
identifikacijske podatke in št.
In general, the anticipated route of human exposure should be considered when designing an assay.
CAS, če je znana,
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.
izvor, številko serije, če je na voljo,
Intraperitoneal injection is not recommended since it is not a physiologically relevant route of human exposure.
agregatno stanje in čistoto,
The maximum volume of liquid that can be administered by gavage or injection at one time depends on the size of the test animal.
fizikalno-kemijske lastnosti, pomembne za izvedbo študije,
The volume should not exceed 2 ml/100 g body weight.
stabilnost preskusne kemikalije, če je znana.
The use of volumes greater than this should be justified.
Topilo/nosilec:
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.
utemeljitev izbire uporabljenega nosilca,
Sampling Time
topnost in stabilnost preskusne kemikalije v topilu/nosilcu, če sta znani,
Somatic Cells
pripravo pripravkov hrane in pitne vode ali pripravkov za vdihavanje,
The sampling time is a critical variable because it is determined by the period needed for mutations to be fixed.
analitične ugotovitve o pripravkih (npr. stabilnost, homogenost, nominalne koncentracije).
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.
uporabljeno vrsto/sev in utemeljitev izbire,
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;
število, starost in spol živali,
although the maximum mutant frequency may not manifest itself in slowly proliferating tissues under these conditions.
izvor, nastanitvene razmere, hrano itd.,
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).
težo posameznih živali na začetku preizkusa, skupaj z območjem, srednjo vrednostjo in standardnim odklonom telesne teže za vsako skupino.
In such cases, the later sampling time would replace the 3 day sampling time, and would require scientific justification.
podatke o pozitivnih in negativnih (nosilec/topilo) kontrolnih vzorcih,
Germ Cells
podatke iz študije za ugotavljanje območja,
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).
podrobnosti o pripravi preskusne kemikalije,
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).
utemeljitev načina dajanja,
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.
metode za merjenje strupenosti za živali, vključno s histopatološko ali hematološko analizo, če sta bili uporabljeni, ter pogostost opazovanja živali in tehtanja,
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).
v primeru negativnih rezultatov metode za preverjanje, ali je preskusna kemikalija dosegla tarčno tkivo ali krvni obtok,
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.
dejanski odmerek (mg/kg telesne teže/dan), preračunan na podlagi koncentracije preskusne kemikalije v hrani/pitni vodi (ppm) in porabe, če je primerno,
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.
podrobne podatke o kakovosti hrane in vode,
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).
podroben opis razporedov tretiranj in vzorčenj ter utemeljitev izbir,
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).
metode usmrtitve,
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.
postopke za izolacijo in hrambo tkiv,
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.
metode za izolacijo genomske DNA pri glodavcih, reševanje transgena iz genomske DNA in prenos transgene DNA v bakterijskega gostitelja,
The health condition of the animals should be recorded.
izvor in številko serij za vse celice, komplete in reagente (kjer je primerno),
At least twice daily, all animals should be observed for morbidity and mortality.
metode za štetje mutantov,
All animals should be weighed at least once a week, and at sacrifice.
metode za molekularno analizo mutantov in uporabo pri popravi za klonskost in/ali izračun pogostnosti mutantov, če je primerno.
Measurements of food consumption should be made at least weekly.
Rezultati:
If the test chemical is administered via the drinking water, water consumption should be measured at each change of water and at least weekly.
stanje živali pred preskusnim obdobjem in med njim, vključno z znaki strupenosti,
Animals exhibiting non-lethal indicators of excess toxicity should be euthanatised prior to completion of the test period (23).
telesno težo in težo organov ob usmrtitvi,
Tissue Collection
za vsako tkivo/žival število mutantov, število ovrednotenih plakov ali kolonij, pogostnost mutantov,
The rationale for tissue collection should be defined clearly.
za vsako skupino tkiv/živali število reakcij v obliki pakiranj na posamezni vzorec DNA, skupno število mutantov, povprečno pogostnost mutantov, standardni odklon,
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.
odvisnost med odmerkom in odzivom, kadar je to mogoče,
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.
za vsako tkivo/žival število neodvisnih mutantov in povprečno pogostnost mutantov, če je bila izvedena molekularna analiza mutantov,
In the absence of any background information, several somatic tissues as may be of interest should be collected.
podatke o hkratni in pretekli negativni kontroli z območji, povprečji in standardnimi odkloni,
These should represent rapidly proliferating, slowly proliferating and site of contact tissues.
podatke o hkratni pozitivni kontroli (ali nehkratni pozitivni kontroli za DNA),
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.
analitične ugotovitve, če so na voljo (npr. koncentracija DNA pri pakiranju, podatki o določanju zaporedja DNA),
Organ weights should be obtained, and for larger organs, the same area should be collected from all animals.
uporabljene statistične analize in metode.
Storage of Tissues and DNA
Sklepna ugotovitev
Tissues (or tissue homogenates) should be stored at or below – 70 °C and be used for DNA isolation within 5 years.
Skopek (1987), ‚Statistical Test for the Comparison of Samples from Mutational Spectra‘, J. Mol.
Isolated DNA, stored refrigerated at 4 °C in appropriate buffer, should be used optimally for mutation analysis within 1 year.
, ‚A more Efficient Big Blue® Protocol Improves Transgene Rescue and Accuracy in an Adduct and Mutation Measurement‘, Mutation Res., 518:
Selection of Tissues for Mutant Analysis
Gossen and J. Vijg (1995), ‚Plasmid-based Transgenic Mouse Model for Studying in vivo Mutations‘Nature, 377(6550):
The choice of tissues should be based on considerations such as:
Gorelick (1995), ‚Statistical Design and Analysis of Mutation Studies in Transgenic Mice‘, Environ.
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);
Gorelick (1996), ‚Mutational Spectra in Transgenic Animal Research: Data Analysis and Study Design Based upon the Mutant or Mutation Frequency‘, Environ.
and 2) pharmacokinetic parameters observed in general toxicity studies, which indicate tissue disposition, retention or accumulation, or target organs for toxicity.
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):
If studies are conducted to follow up carcinogenicity studies, target tissues for carcinogenicity should be considered.
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.
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).
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):
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.
Tindall (2000), ‚Bayesian Analysis of Mutational Spectra‘, Genetics, 156:
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.
Knook and J. Vijg (1989), ‚Efficient Rescue of Integrated Shuttle Vectors from Transgenic Mice: a Model for Studying Mutations in vivo‘, Proc.
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.
Gossen, J.A. and J. Vijg (1993), ‚A Selective System for lacZ-Phage using a Galactose-sensitive E. coli Host‘, Biotechniques, 14(3):
Methods of Measurement
, ‚Somatic Gene Mutation and Human Disease other than Cancer‘, Mutation Res., 543:
Standard laboratory or published methods for the detection of mutants are available for the recommended transgenic models:
, ‚Somatic Gene Mutation and Human Disease other than Cancer: an Update‘, Mutation Res., 705:
lacZ lambda bacteriophage and plasmid (30);
Douglas and D. Krewski (1998), ‚Statistical Analysis of lacZ Mutant Frequency Data from Muta™Mouse Mutagenicity Assays‘, Mutagenesis, 13(3):
lacI mouse (2) (18);
Skopek, V. Thybaud, K.R.Tindall and N. Yajima (2000), ‚In vivo Transgenic Mutation Assays‘, Environ.
gpt delta mouse (22);
Douglas (2003), ‚Treatment and Sampling Protocols for Transgenic Mutation Assays‘, Environ.
gpt delta rat (28);
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.
Modifications should be justified and properly documented.
Short (1990), ‚The Use of Transgenic Mice for Short-term, in vivo Mutagenicity Testing‘, Genet.
Data from multiple packagings can be aggregated and used to reach an adequate number of plaques or colonies.
Burns (2008), ‚Comparison of Induced and Cancer-associated Mutational Spectra using Multivariate Data Analysis‘, Carcinogenesis, 29(4):
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.
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.
In such cases, data should be considered cautiously because they may be unreliable.
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.
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.
Masumura (2000), 'Recent Advances in the Protocols of Transgenic Mouse Mutation Assays', Mutation Res., 455(1–2):
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).
Tindall (1995), 'Study Design and Sample Sizes for a lacI Transgenic Mouse Mutation Assay', Environ.
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.
Gorelick (1997), ‚Sources of Variability in Data from a Positive Selection lacZ Transgenic Mouse Mutation Assay: an Interlaboratory Study‘, Mutation.
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.
Yauk (2006), ‚Detection of Induced Male Germline Mutation: Correlations and Comparisons between Traditional Germline Mutation Assays, Transgenic Rodent Assays and Expanded Simple Tandem Repeat Instability Assays‘, Mutation.
Treatment of Results
Kurata, M. Takamune, M. Yamada, H. Sanada, T. Umemura, A. Nishikawa and T. Nohmi (2010), 'Integration of in vivo Genotoxicity and Short-term Carcinogenicity Assays using F344 gpt delta Transgenic Rats: in vivo Mutagenicity of 2,4-diaminotoluene and 2,6-diaminotoluene Structural Isomers', Toxicol.
Individual animal data should be presented in tabular form.
Martus, J.C. Mirsalis, T. Suzuki and N. Yajima (2003), ‚In vivo Transgenic Mutation Assays‘, Mutation Res., 540:
The experimental unit is the animal.
Douglas (1996), 'Bacteriophage λ and Plasmid lacZ Transgenic Mice for studying Mutations in vivo' in:
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.
Douglas (2005), ‚A lacZ Transgenic Mouse Assay for the Detection of Mutations in Follicular Granulosa Cells‘, Mutation Res., 578(1–2):
If there are multiple packaging/rescue reactions, the number of reactions per DNA sample should be reported.
Martus, V. Thybaud (2011), ‚Compilation and Use of Genetic Toxicity Historical Control Data‘, Mutation Res., doi:10.1016/j.mrgentox.2010.09.007.
While data for each individual reaction should be retained, only the total pfu or cfu need be reported.
, ‚Kinetics of spermatogenesis in mammals seminiferous epithelium cycle and spermatogonial renewal‘.
Data on toxicity and clinical signs as per paragraph 34 should be reported.
, ‚The Epididymis‘, in Neil, J.D., Pfaff, D.W., Chalis, J.R.G., de Kretser, D.M., Richards, J.S., and P. M, Wassarman (eds.), Physiology of Reproduction, Elsevier, the Netherlands, str. 1071–1148.
Any sequencing results should be presented for each mutant analysed, and resulting mutation frequency calculations for each animal and tissue should be shown.
, ‚Effects of male germ-cell stage on the frequency, nature, and spectrum of induced specific-locus mutations in the mouse‘, Genetica, 122:
Statistical Evaluation and Interpretation of Results
Obdobje odmerjanja
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.
celotno obdobje, v katerem žival prejema odmerek.
At least three treated dose groups should be analysed in order to provide sufficient data for dose-response analysis.
Substitucija baznih parov
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).
vrsta mutacije, ki povzroči zamenjavo enega samega nukleotidnega baznega para v DNA z drugim nukleotidnim baznim parom DNA.
Statistical tests used should consider the animal as the experimental unit.
Kapsida
A test chemical for which the results do not meet the above criteria in any tissue is considered non-mutagenic in this assay.
proteinski plašč, ki ovija virusni delec.
For biological relevance of a negative result, tissue exposure should be confirmed.
snov ali zmes.
For DNA sequencing analyses, a number of statistical approaches are available to assist in interpreting the results (1) (5) (9) (19).
Klonalna ekspanzija
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).
nastanek številnih celic iz ene same (mutirane) celice.
identification data and CAS no, if known;
Enote, ki delajo kolonije
source, lot number if available;
merilo števila bakterij, sposobnih za razmnoževanje.
physical nature and purity;
Konkatemer
physiochemical properties relevant to the conduct of the study;
dolga neprekinjena biomolekula, ki jo sestavlja več enakih kopij, povezanih v serijo.
stability of the test chemical, if known;
Kohezivni konec
Solvent/vehicle:
odsek enoverižne DNA z 12 nukleotidi, ki je na obeh koncih dvovijačnega genoma bakteriofaga lambda.
justification for choice of vehicle;
Delecija
solubility and stability of the test chemical in the solvent/vehicle, if known;
mutacija, pri kateri genom izgubi enega ali več (zaporednih) nukleotidov.