natgillin/translations-raw huggingface dataset:
It is based on the increase in uterine weight or uterotrophic response (for review, see 29).
Ne glede na morebiten izvor odziva naj bi bili na podlagi pozitivnega rezultata uterotropnega biološkega preskusa običajno začeti postopki za nadaljnja pojasnila.
It evaluates the ability of a chemical to elicit biological activities consistent with agonists or antagonists of natural oestrogens (e.g. 17ß-estradiol), however, its use for antagonist detection is much less common than for agonists.
Dodatni dokazi o estrogenosti bi bili lahko pridobljeni na podlagi preskusov in vitro, kot so preskusi vezave na estrogenske receptorje in preskusi aktivacije prepisovanja, ali na podlagi drugih preskusov in vivo, kot je preskus na podgani ženskega spola v puberteti.
The uterus responds to oestrogens in two ways.
Ob upoštevanju, da uterotropni biološki preskus služi kot presejalna študija in vivo, je bil pristop validacije koristen tako za vprašanja, povezana z dobrobitjo živali, kot za večstopenjsko strategijo preskušanja.
An initial response is an increase in weight due to water imbibition.
Zato je bilo prizadevanje usmerjeno v strogo validacijo ponovljivosti in občutljivosti za estrogenost, kar je najpomembnejše vprašanje pri številnih kemikalijah, medtem ko v razsežnost antiestrogenosti v preskusu ni bilo vloženega veliko truda.
This response is followed by a weight gain due to tissue growth (30).
Preskušen je bil samo en antiestrogen z močnim delovanjem, saj je število kemikalij z jasnim antiestrogenim profilom (ki ga ne prikrije kakšno estrogeno delovanje) zelo majhno.
The uterus responses in rats and mice qualitatively are comparable.
Zato je ta preskusna metoda namenjena estrogenemu protokolu, protokol, ki v preskusu opisuje antagonistično delovanje, pa je vključen v smernici (37).
This bioassay serves as an in vivo screening assay and its application should be seen in the context of the “OECD Conceptual Framework for the Testing and Assessment of Endocrine Disrupting Chemicals” (Appendix 2).
Ponovljivost in občutljivost preskusa za kemikalije z izključno antiestrogenim delovanjem bo jasneje opredeljena v prihodnosti, ko bo preskusni postopek že nekaj časa v uporabi in bo opredeljenih več kemikalij s tem načinom delovanja.
In this Conceptual Framework the Uterotrophic Bioassay is contained in Level 3 as an in vivo assay providing data about a single endocrine mechanism, i.e. oestrogenicity.
Priznava se, da morajo biti vsi postopki, pri katerih se uporabljajo živali, v skladu z lokalnimi standardi za varstvo živali;
The Uterotrophic Bioassay is intended to be included in a battery of in vitro and in vivo tests to identify chemicals with potential to interact with the endocrine system, ultimately leading to risk assessments for human health or the environment.
opisi oskrbe in ravnanja z živalmi v nadaljevanju so minimalni standardi, nadomestili pa jih bodo lokalni predpisi, kot je Direktiva 2010/63/EU Evropskega parlamenta in Sveta z dne 22. septembra 2010 o zaščiti živali, ki se uporabljajo v znanstvene namene (38).
The OECD validation programme used both strong and weak oestrogen agonists to evaluate the performance of the assay to identify oestrogenic chemicals (4)(5)(6)(7)(8).
Nadaljnje smernice o humanem ravnanju z živalmi je predložila OECD (25).
Thereby the sensitivity of the test procedure for oestrogen agonists was well demonstrated besides a good intra- and interlaboratory reproducibility.
Kot pri vseh preskusih, pri katerih se uporabljajo žive živali, se je treba pred pričetkom izvajanja preskusa prepričati, da so podatki resnično potrebni.
With regard to negative chemicals, only one “negative” reference chemical already reported negative by uterotrophic assay as well as in vitro receptor binding and receptor assays was included in the validation programme, but additional test data, not related to the OECD validation programme, have been evaluated, giving further support to the specificity of the Uterotrophic Bioassay for the screening of oestrogen agonists (16).
Na primer, dva pogoja, pri katerih so podatki potrebni, sta:
INITIAL CONSIDERATIONS AND LIMITATIONS
visok potencial za izpostavljenost (raven 1 temeljnega okvira, Dodatek 2) ali pokazatelji estrogenosti (raven 2), zaradi katerih je potrebno preveriti, ali se lahko takšni učinki pojavijo in vivo,
Oestrogen agonists and antagonists act as ligands for oestrogen receptors a and b and may activate or inhibit, respectively, the transcriptional action of the receptors.
učinki, ki kažejo na estrogenost v preskusih in vivo na ravni 4 ali 5, da bi ugotovili, ali so bili učinki povezani z estrogenimi mehanizmi, ki jih ni mogoče pojasniti s preskusom in vitro.
This may have the potential to lead to adverse health hazards, including reproductive and developmental effects.
Opredelitve, uporabljene v tej preskusni metodi, so navedene v Dodatku 1.
Therefore, the need exists to rapidly assess and evaluate a chemical as a possible oestrogen agonist or antagonist.
Občutljivost uterotropnega biološkega preskusa je odvisna od sistema preskušanja na živalih, pri katerem os hipotalamus – hipofiza – jajčniki ne deluje, zaradi česar so prisotne nizke endogene koncentracije cirkulirajočega estrogena.
While informative, the affinity of a ligand for an oestrogen receptor or transcriptional activation of reporter genes in vitro is only one of several determinants of possible hazard.
Tako se zagotovi nizka osnovna teža maternice in največji možni razpon odziva na prejete estrogene.
Other determinants can include metabolic activation and deactivation upon entering the body, distribution to target tissues, and clearance from the body, depending at least in part on the route of administration and the chemical being tested.
To zahtevo izpolnjujeta dve na estrogen občutljivi stanji pri glodavcih ženskega spola:
This leads to the need to screen the possible activity of a chemical in vivo under relevant conditions, unless the chemical's characteristics regarding Absorption — Distribution — Metabolism — Elimination (ADME) already provide appropriate information.
nezrele samice po odstavitvi in pred puberteto ter
Uterine tissues respond with rapid and vigorous growth to stimulation by oestrogens, particularly in laboratory rodents, where the oestrous cycle lasts approximately 4 days.
mlade odrasle samice po sterilizaciji, ko je dovolj časa, da se maternična tkiva zmanjšajo.
Rodent species, particularly the rat, are also widely used in toxicity studies for hazard characterisation.
Preskusna kemikalija se vnese dnevno, in sicer oralno z gavažo ali s podkožnim injiciranjem.
Therefore, the rodent uterus is an appropriate target organ for the in vivo screening of oestrogen agonists and antagonists.
Stopnjevani odmerki preskusne kemikalije se dajejo vsaj dvema tretiranima skupinama (za navodila glej odstavek 33) preskusnih živali, pri čemer se za vsako skupino uporabi ena velikost odmerka, in sicer v obdobju treh zaporednih dni pri metodi z nesterilizirano nezrelo samico in obdobju najmanj treh zaporednih dni pri metodi s sterilizirano odraslo samico.
This test method is based on those protocols employed in the OECD validation study which have been shown to be reliable and repeatable in intra- and interlaboratory studies (5)(7).
Približno 24 ur po prejetju zadnjega odmerka se na živalih opravi obdukcija.
Currently two methods, namely the ovariectomised adult female method (ovx-adult method) and the immature non-ovariectomised method (immature method) are available.
Pri agonistih estrogenov se na podlagi srednje teže maternice živali v tretiranih skupinah glede na nosilec oceni, ali je prišlo do statistično značilnega povečanja.
It was shown in the OECD validation test programme that both methods have comparable sensitivity and reproducibility.
Statistično značilno povečanje srednje teže maternice živali v preskusni skupini v tem biološkem preskusu pomeni pozitiven odziv.
However, the immature, as it has an intact hypothalamic-pituitary-gonadal (HPG) axis, is somewhat less specific but covers a larger scope of investigation than the ovariectomised animal because it can respond to chemicals that interact with the HPG axis rather than just the oestrogen receptor.
OPIS PRESKUSNE METODE
The HGP axis of the rat is functional at about 15 days of age.
Lahko se uporabijo laboratorijski sevi glodavcev, ki se običajno uporabljajo.
Prior to that, puberty cannot be accelerated with treatments like GnRH.
Med validacijo so bili na primer uporabljeni sevi podgan Sprague-Dawley in Wistar.
As the females begin to reach puberty, prior to vaginal opening, the female will have several silent cycles that do not result in vaginal opening or ovulation, but there are some hormonal fluctuations.
Sevi z maternicami, za katere je znano ali pa se sumi, da so manj odzivne, se ne smejo uporabiti.
If a chemical stimulates the HPG axis directly or indirectly, precocious puberty, early ovulation and accelerated vaginal opening result.
Laboratorij bi moral dokazati občutljivost uporabljenega seva v skladu z odstavkoma 26 in 27.
Not only chemicals that act on the HPG axis do this but some diets with higher metabolisable energy levels than others will stimulate growth and accelerate vaginal opening without being oestrogenic.
V uterotropnem biološkem preskusu se od 30-ih let prejšnjega stoletja običajno uporabljajo podgane in miši.
Such chemicals would not induce an uterotrophic response in OVX adult animals as their HPG axis does not work.
Validacijske študije OECD so bile izvedene samo s podganami na podlagi prepričanja, da sta obe vrsti enakovredni, zato bi morala biti ena vrsta dovolj za globalno validacijo, da bi prihranili sredstva in živali.
For animal welfare reasons preference should be given to the method using immature rats, avoiding surgical pre-treatment of the animals and avoiding also a possible non-use of those animals which indicate any evidence entering oestrous (see paragraph 30).
Podgane so prva izbira pri večini študij strupenosti za razmnoževanje in razvoj.
The uterotrophic response is not entirely of oestrogenic origin, i.e. chemicals other than agonists or antagonists of oestrogens may also provide a response.
Ker za miši obstaja obsežna zbirka preteklih podatkov, in bi tako razširili področje uporabe uterotropnega biološkega preskusa na glodavcih na uporabo miši kot preskusne vrste, je bila na miših opravljena omejena nadaljevalna validacijska študija (16).
For example, relatively high doses of progesterone, testosterone, or various synthetic progestins may all lead to a stimulative response (30).
V skladu s prvotnim namenom ohranjanja sredstev in živali je bil izbran premostitveni pristop z omejenim številom preskusnih kemikalij in sodelujočih laboratorijev ter brez preskušanja kodiranih vzorcev.
Any response may be analysed histologically for keratinisation and cornification of the vagina (30).
Ta premostitvena validacijska študija za uterotropni biološki preskus na steriliziranih mladih odraslih miših dokazuje, da se podatki, pridobljeni s podganami in mišmi, kvalitativno in kvantitativno dobro dopolnjujejo.
Irrespective of the possible origin of the response, a positive outcome of an Uterotrophic Bioassay should normally initiate actions for further clarification.
Če bo po uterotropnem biološkem preskusu morda izvedena dolgotrajna študija, bo tako mogoče v obeh študijah uporabiti živali iste vrste, seva in izvora.
Additional evidence of oestrogenicity could come from in vitro assays, such as the ER binding assays and transcriptional activation assays, or from other in vivo assays such as the female pubertal assay.
Premostitveni pristop je bil uporabljen samo za sterilizirane odrasle miši, poročilo pa ne zagotavlja zanesljivih podatkov za validacijo modela z nezrelimi samicami, zato model z nezrelimi mišmi ne sodi na področje uporabe obstoječe preskusne metode.
Taking into account that the Uterotrophic Bioassay serves as an in vivo screening assay, the validation approach taken served both animal welfare considerations and a tiered testing strategy.
Zato se lahko v nekaterih primerih namesto podgan uporabijo miši.
To this end, effort was directed at rigorously validating reproducibility and sensitivity for oestrogenicity — the main concern for many chemicals-, while little effort was directed at the antioestrogenicity component of the assay.
Uporabo te vrste je treba utemeljiti na podlagi toksikoloških, farmakokinetičnih in/ali drugih meril.
Only one antioestrogen with strong activity was tested since the number of chemicals with a clear antioestrogenic profile (not obscured by some oestrogenic activity) is very limited.
Za miši bodo morda potrebne spremembe protokola.
Thus this test method is dedicated to the oestrogenic protocol, while the protocol describing the antagonist mode of the assay is included in a Guidance Document (37).
Na primer, poraba hrane pri miših glede na telesno težo je večja kot pri podganah, zato mora biti vsebnost fitoestrogena v hrani za miši manjša kot v hrani za podgane (9) (20) (22).
The reproducibility and sensitivity of the assay for chemicals with purely anti-oestrogenic activity will be more clearly defined later on, after the test procedure has been in routine use for some time and more chemicals with this modality of action are identified.
Vsi postopki morajo biti skladni z vsemi lokalnimi standardi za varstvo živali v laboratorijih.
It is acknowledged that all animal based procedures will conform to local standards of animal care;
Ti opisi oskrbe in ravnanja z živalmi so minimalni standardi, ki jih bodo nadomestili lokalni predpisi, kot je Direktiva 2010/63/EU Evropskega parlamenta in Sveta z dne 22. septembra 2010 o zaščiti živali, ki se uporabljajo v znanstvene namene (38).
the descriptions of care and treatment set forth below are minimal performance standards, and will be superseded by local regulations such as Directive 2010/63/EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes (38).
Temperatura v prostoru s preskusnimi živalmi naj bo 22 °C (z odstopanjem približno ± 3 °C).
Further guidance of the humane treatment of animals is given by the OECD (25).
Relativna vlažnost naj bo najmanj 30 % in po možnosti naj ne presega 70 %, razen med čiščenjem prostora.
As with all assays using live animals, it is essential to ensure that the data are truly necessary prior to the start of the assay.
Cilj je 50–60-odstotna relativna vlažnost.
For example, two conditions where the data may be required are:
Osvetlitev naj bo umetna.
high exposure potential (Level 1 of the Conceptual Framework, Appendix 2) or indications for oestrogenicity (Level 2) to investigate whether such effects may occur in vivo;
Dnevno zaporedje osvetlitve naj bo 12 ur svetlobe, 12 ur teme.
effects indicating oestrogenicity in Level 4 or 5 in vivo tests to substantiate that the effects were related to an oestrogenic mechanism that cannot be elucidated using an in vitro test.
Laboratorijsko prehrano in pitno vodo je treba priskrbeti ad libitum.
Definitions used in this test method are given in Appendix 1.
Mlade odrasle živali so lahko nastanjene posamično ali pa so v kletke razvrščene v skupine po največ tri živali.
The Uterotrophic Bioassay relies for its sensitivity on an animal test system in which the hypothalamic-pituitary-ovarian axis is not functional, leading to low endogenous levels of circulating oestrogen.
Zaradi mladosti nezrelih živali je priporočljiva družabna skupinska nastanitev.
This will ensure a low baseline uterine weight and a maximum range of response to administered oestrogens.
Znano je, da visoka vsebnost fitoestrogenov v laboratorijski prehrani povzroči tolikšno povečanje teže maternice pri glodavcih, da to vpliva na uterotropni biološki preskus (13) (14) (15).
Two oestrogen sensitive states in the female rodent meet this requirement:
Pri nezrelih živalih lahko visoka vsebnost fitoestrogenov in presnovljiva energija v laboratorijski prehrani vplivata tudi na pojav zgodnje pubertete.
immature females after weaning and prior to puberty; and
Fitoestrogeni so prisotni v glavnem zaradi vsebnosti izdelkov iz soje in lucerne v laboratorijski prehrani, koncentracija fitoestrogenov v standardni laboratorijski prehrani pa se razlikuje od serije do serije (23).
young adult females after ovariectomy with adequate time for uterine tissues to regress.
Telesna teža je pomembna spremenljivka, ker je z njo povezana količina zaužite hrane.
The test chemical is administered daily by oral gavage or subcutaneous injection.
Zato se lahko dejanski odmerek fitoestrogenov, zaužitih z enako prehrano, razlikuje glede na vrsto in starost (9).
Graduated test chemical doses are administered to a minimum of two treatment groups (see paragraph 33 for guidance) of experimental animals using one dose level per group and an administration period of three consecutive days for immature method and a minimum administration period of three consecutive days for ovx-adult method.
Pri nezrelih podganah ženskega spola je lahko poraba hrane glede na telesno težo približno dvakrat večja od porabe hrane pri steriliziranih mladih odraslih samicah.
The animals are necropsied approximately 24 hours after the last dose.
Pri mladih odraslih miših je lahko poraba hrane glede na telesno težo približno štirikrat večja od porabe hrane pri steriliziranih mladih odraslih podganah ženskega spola.
For oestrogen agonists, the mean uterine weight of the treated animal groups relative to the vehicle group is assessed for a statistically significant increase.
Rezultati uterotropnega biološkega preskusa (9) (17) (18) (19) kažejo, da je določena količina fitoestrogenov v prehrani sprejemljiva in ne zmanjša občutljivosti biološkega preskusa.
A statistically significant increase in the mean uterine weight of a test group indicates a positive response in this bioassay.
Praviloma naj raven fitoestrogenov v prehrani nezrelih podganah ženskega spola sevov Sprague Dawley in Wistar (6) (9) ne bi presegla 350 μg ekvivalentov genisteina/gram laboratorijske prehrane.
DESCRIPTION OF THE METHOD
Takšna prehrana bi morala biti primerna tudi pri preskušanju na mladih odraslih steriliziranih podganah, ker je poraba hrane glede na telesno težo pri mladih odraslih živalih manjša v primerjavi s porabo hrane pri nezrelih živalih.
Commonly used laboratory rodent strains may be used.
Če se uporabijo odrasle sterilizirane miši ali podgane, ki so bolj občutljive na fitoestrogen, je treba upoštevati sorazmerno zmanjšanje ravni fitoestrogenov v prehrani (20).
As an example, Sprague-Dawley and Wistar strains of rats were used during the validation.
Poleg tega lahko zaradi razlik v razpoložljivi presnovni energiji iz različne prehrane pride do časovnega zamika začetka pubertete (21) (22).
Strains with uteri known or suspected to be less responsive should not be used.
Pred začetkom študije je treba skrbno izbrati prehrano brez povečane vrednosti fitoestrogenov (za navodila glej (6) (9)) ali presnovljive energije, ki lahko vplivajo na rezultate (15) (17) (19) (22) (36).
The laboratory should demonstrate the sensitivity of the strain used as described in paragraphs 26 and 27.
Zagotavljanje ustreznega delovanja sistema preskušanja, ki ga uporablja laboratorij in je določen v odstavkih 26 in 27, je pomembno za preverjanje obeh dejavnikov.
The rat and mouse have been routinely used in the Uterotrophic Bioassay since the 1930s.
Kot varovalo v skladu z dobro laboratorijsko prakso je treba za vsako serijo prehrane, uporabljene med študijo, opraviti reprezentativno vzorčenje zaradi morebitne analize vsebnosti fitoestrogenov (npr. v primeru visoke kontrolne teže maternice v primerjavi s preteklimi kontrolnimi vzorci ali nezadostnega odziva na referenčni estrogen, 17-alfa-etinilestradiol).
The OECD validation studies were only performed with rats based on an understanding that both species are expected to be equivalent and therefore one species should be enough for the world-wide validation in order to save resources and animals.
Alikvote je treba analizirati v okviru študije ali pa jih zamrzniti pri –20 °C oziroma tako, da se vzorec pred analizo ne razgradi.
The rat is the species of choice in most reproductive and developmental toxicity studies.
Nekatere vrste stelje lahko vsebujejo naravno prisotne estrogene ali antiestrogene kemikalije (na primer, znano je, da koruzni storž vpliva na cikličnost pri podganah in očitno deluje antiestrogeno).
Taking into consideration that a vast historical database exists for mice and thus to broaden the scope of the Uterotrophic Bioassay test method in rodents to the use of mice as test species, a limited follow-up validation study was carried out in mice (16).
Izbrana stelja naj bi vsebovala najmanjšo možno koncentracijo fitoestrogenov.
A bridging approach with a limited number of test chemicals, participating laboratories and without coded sample testing has been selected in keeping with the original intent to save resources and animals.
Priprava živali
This bridging validation study shows for the Uterotrophic Bioassay in young adult ovariectomised mice that, qualitatively and quantitatively, the data obtained in rats and mice correspond well with each other.
Preskusne živali, ki ne kažejo nobenih znakov bolezni ali fizičnih nepravilnosti, se naključno dodelijo v kontrolne in tretirane skupine.
Where the Uterotrophic Bioassay result may be preliminary to a long-term study, this allows animals from the same strain and source to be used in both studies.
Živali je treba označiti z edinstvenimi oznakami.
The bridging approach was limited to the OVX mice and the report does not provide a robust data set to validate the immature model, thus the immature model for mice is not considered under the scope of the current test method.
Najbolje je, da so nezrele živali med prilagajanjem do odstavitve v kletkah skupaj z materami ali nadomestnimi materami.
Thus, in some cases mice may be used instead of rats.
Obdobje prilagajanja pred začetkom študije naj bi pri mladih odraslih živalih in nezrelih živalih, ki so prispele skupaj z materami ali nadomestnimi materami, trajalo približno 5 dni.
A rationale should be given for this species, based on toxicological, pharmacokinetic, and/or other criteria.
Če so prispele nezrele živali, odstavljene in brez mater, bo morda potrebno krajše obdobje prilagajanja, saj naj bi se dajanje odmerkov pričelo takoj po odstavitvi (glej odstavek 29).
Modifications of the protocol may be necessary for mice.
Preverjanje strokovnosti laboratorija
For example, the food consumption of mice on a body weight basis is higher than that of rats and therefore the phyto-oestrogen content in food should be lower for mice than for rats (9)(20)(22).
Za preverjanje strokovnosti laboratorija sta na voljo dve različni možnosti:
All procedures should conform with local standards of laboratory animal care.
redno preverjanje na podlagi prvotne študije osnovne pozitivne kontrole (glej odstavek 27).
These descriptions of care and treatment are minimum standards and will be superseded by local regulations such as Directive 2010/63/EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes (38).
Vsaj vsakih 6 mesecev in vedno, ko pride do spremembe, ki bi lahko vplivala na uspešnost preskusa (npr. nova sestava prehrane, menjava osebja, ki opravlja obdukcije, menjava seva živali ali dobavitelja itd.), je treba odzivnost sistema preskušanja (živalski model) preveriti s primernim odmerkom (na podlagi študije osnovne pozitivne kontrole iz poglavja 27) referenčnega estrogena:
The temperature in the experimental animal room should be 22 °C (with an approximate range ± 3 °C).
etinilestradiol (št.
The relative humidity should be a minimum of 30 % and preferably should not exceed a maximum 70 %, other than during room cleaning.
CAS 57-63-6) (EE),
The aim should be relative humidity of 50-60 %.
uporaba sočasnih kontrolnih vzorcev z vključitvijo skupine, ki je prejela ustrezni odmerek referenčnega estrogena, v vsak preskus.
Lighting should be artificial.
Če se sistem ne odzove v skladu s pričakovanji, je treba pogoje preskusa preveriti in ustrezno spremeniti.