natgillin/translations-raw huggingface dataset:
The purpose of this test method is to describe the procedures used to evaluate the potential ocular corrosivity or severe irritancy of a test substance as measured by its ability to induce opacity and increased permeability in an isolated bovine cornea.
V tej testni metodi so opisani postopki za oceno potencialne jedkosti in močne dražilnosti testne snovi za oči glede na njeno sposobnost, da v izolirani roženici goveda povzroča motnjavo in povečano prepustnost.
Toxic effects to the cornea are measured by: (i) decreased light transmission (opacity), and (ii) increased passage of sodium fluorescein dye (permeability).
Toksični učinki na roženico se merijo z: (i) zmanjšanim prenosom svetlobe (motnjava) in (ii) povečanim prehajanjem barvila natrijevega fluoresceina (prepustnost).
The opacity and permeability assessments of the cornea following exposure to a test substance are combined to derive an In Vitro Irritancy Score (IVIS), which is used to classify the irritancy level of the test substance.
Oceni motnjave in prepustnosti roženice po aplikaciji testne snovi skupaj pomenita dražilnost in vitro (IVIS), ki se uporablja za razvrstitev testne snovi v različne stopnje draženja.
Ocular irritants that induce lesions that resolve in less than 21 days and non-irritants have also been tested using the BCOP test method.
S testno metodo BCOP so bile testirane tudi dražilne snovi, ki povzročajo poškodbe oči, ki izginejo v manj kot 21 dnevih, in nedražilne snovi.
However, the accuracy and reliability of the BCOP test method for substances in these categories have not been formally evaluated.
Vendar natančnost in zanesljivost testne metode BCOP pri snoveh iz teh dveh kategorij nista bili ocenjeni formalno.
Definitions are provided in Appendix 1.
Opredelitve so navedene v dodatku 1.
INITIAL CONSIDERATIONS AND LIMITATIONS
ZAČETNI POMISLEKI IN OMEJITVE
This test method is based on the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) BCOP test method protocol (8), which was developed following an international validation study (4)(5)(9), with contributions from the European Centre for the Validation of Alternative Methods (ECVAM) and the Japanese Center for the Validation of Alternative Methods (JaCVAM).
Ta testna metoda temelji na protokolu testne metode BCOP Medagencijskega usklajevalnega odbora za validacijo alternativnih metod (ICCVAM)(8), ki je bil pripravljen po mednarodni študiji o validaciji (4)(5)(9), pri kateri sta sodelovala Evropski center za validacijo alternativnih metod (ECVAM) in Japonski center za validacijo alternativnih metod (JaCVAM).
The protocol is based on information obtained from the Institute for In Vitro Sciences (IIVS) and INVITTOX Protocol 124 (10), which represents the protocol used for the European Community-sponsored prevalidation study of the BCOP assay conducted in 1997-1998.
Protokol temelji na informacijah, ki jih je prispeval Inštitut za raziskave in vitro (IIVS), in protokolu INVITTOX 124 (10), ki se je uporabil pri predvalidacijski študiji o preskusu BCOP, ki je bila opravljena med leti 1997–1998 in ki jo je sponzorirala Evropska skupnost.
Both of these protocols are based on the BCOP assay methodology first reported by Gautheron et al.
Oba protokola temeljita na testni metodologiji BCOP, o kateri je prvi poročal Gautheron et al.
The identified limitations for this test method are based on the high false positive rates for alcohols and ketones and the high false negative rate for solids observed in the validation database (see paragraph 44) (5).
Opredeljene omejitve za to testno metodo temeljijo na visokem številu lažno pozitivnih rezultatov za alkohole in ketone ter visokem številu lažno negativnih rezultatov za trdne snovi, zbranih v podatkovni zbirki za validacijo (glej točko 44) (5).
When substances within these chemical and physical classes are excluded from the database, the accuracy of BCOP across the EU, EPA, and GHS classification systems is substantially improved (5).
Če se snovi iz teh kemijskih in fizikalnih razredov izključijo iz podatkovne zbirke, se natančnost BCOP v sistemih za razvrščanje snovi EU, EPA in GHS bistveno izboljša (5).
Based on the purpose of this assay (i.e., to identify ocular corrosives/severe irritants only), false negative rates are not critical since such substances would be subsequently tested in rabbits or with other adequately validated in vitro tests, depending on regulatory requirements, using a sequential testing strategy in a weight of evidence approach.
Pri tem preskusu (tj. za ugotavljanje izključno jedkih in zelo dražilnih snovi za oči) lažno negativni rezultati niso kritični, saj bi se take snovi nato preskušale na zajcih ali, odvisno od regulativnih zahtev, z drugimi ustrezno potrjenimi testi in vitro, pri katerih se uporablja sekvenčna testna strategija, ki temelji na teži dokazov.
Furthermore, the current validation database did not allow for an adequate evaluation of some chemical or product classes (e.g., mixtures).
Poleg tega nekaterih kemijskih ali proizvodnih razredov (npr. zmesi) s trenutno validacijsko podatkovno zbirko ni bilo mogoče ustrezno oceniti.
However, investigators could consider using this test method for all types of test material (including mixtures), whereby a positive result could be accepted as indicative of an ocular corrosive or severe irritant response.
Vendar pa bi lahko preskuševalci to testno metodo uporabili pri vseh vrstah testnega materiala (tudi pri zmeseh), pri čemer bi bil pozitiven rezultat sprejemljiv kot indikator za jedko ali zelo dražilno snov za oči.
However, positive results obtained with alcohols or ketones should be interpreted cautiously due to risk of over-prediction.
Pozitivne rezultate, pridobljene z uporabo alkoholov ali ketonov, je treba razlagati previdno, saj obstaja tveganje, da so vanje vključeni tudi lažno pozitivni rezultati.
All procedures with bovine eyes and bovine corneas should follow the testing facility’s applicable regulations and procedures for handling animal-derived materials, which include, but are not limited to, tissues and tissue fluids.
Pri vseh postopkih na očesih in roženicah goveda je treba upoštevati veljavna pravila in postopke testnega urada o ravnanju z živalskim materialom, kamor spadajo med drugim tudi tkiva in tkivne tekočine.
Universal laboratory precautions are recommended (12).
Priporočajo se splošni previdnostni ukrepi za laboratorije (12).
A limitation of the test method is that, although it takes into account some of the ocular effects evaluated in the rabbit ocular irritancy test method and to some degree their severity, it does not consider conjunctival and iridal injuries.
Čeprav testna metoda upošteva nekatere učinke na očesa, ocenjene s testno metodo za ugotavljanje dražilnih snovi na očeh zajcev, in do določene mere tudi njihovo resnost, je kljub temu omejena, saj ne upošteva poškodb veznice in šarenice.
Also, although the reversibility of corneal lesions cannot be evaluated per se in the BCOP assay, it has been proposed, based on rabbit eye studies, that an assessment of the initial depth of corneal injury can be used to distinguish between irreversible and reversible effects (13).
Čeprav tudi same reverzibilnosti poškodb roženice s preskusom BCOP ni mogoče oceniti, je bilo na podlagi študij na očeh zajcev kljub temu predlagano, da se oceni prvotna globina poškodbe roženice ter tako razlikuje med ireverzibilnimi in reverzibilnimi učinki (13).
Finally, the BCOP does not allow for an assessment of the potential for systemic toxicity associated with ocular exposure.
Treba je tudi opozoriti, da s testno metodo BCOP ni mogoče oceniti potencialne sistemske toksičnosti, povezane z izpostavljenostjo oči.
Efforts are ongoing to further characterize the usefulness and limitations of the BCOP assay for identifying non-severe irritants and non-irritants (see also paragraph 45).
Trenutno potekajo prizadevanja za nadaljnjo karakterizacijo uporabnosti in omejitev preskusa BCOP za ugotavljanje lažje dražilnih in nedražilnih snovi (glej tudi točko 45).
Users are also encouraged to provide specimens and/or data to validation organizations for a formal evaluation of possible future uses of the BCOP test method, including for the identification of non-severe irritants and non-irritants.
Uporabnikom se priporoča, da organizacijam za validacijo zagotovijo vzorce in/ali podatke za uradno oceno morebitnih prihodnjih uporab testne metode BCOP, vključno za ugotavljanje lažje dražilnih in nedražilnih snovi.
For any laboratory initially establishing this assay, the proficiency chemicals provided in Appendix 2 should be used.
Laboratoriji, ki ta preskus opravljajo prvič, morajo uporabiti kemikalije za preverjanje usposobljenosti iz dodatka 2.
A laboratory can use these chemicals to demonstrate their technical competence in performing the BCOP test method prior to submitting BCOP assay data for regulatory hazard classification purposes.
Te kemikalije uporabijo, da dokažejo, da so tehnično sposobni izvajati testno metodo BCOP, preden podatke o testu BCOP predložijo za regulativno razvrstitev nevarnosti.
PRINCIPLE OF THE TEST
NAČELO TESTA
The BCOP test method is an organotypic model that provides short-term maintenance of normal physiological and biochemical function of the bovine cornea in vitro.
Testna metoda BCOP je organotipski model, ki zagotavlja kratkoročno ohranjanje običajne fiziološke in biokemijske funkcije roženice goveda in vitro.
In this test method, damage by the test substance is assessed by quantitative measurements of changes in corneal opacity and permeability with an opacitometer and a visible light spectrophotometer, respectively.
Pri tej testni metodi se poškodba, ki jo povzroči testna snov, oceni tako, da se spremembe motnjave in prepustnosti roženice merijo kvantitativno z opacitometrom oziroma spektrofotometrom vidne svetlobe.
Both measurements are used to calculate an IVIS, which is used to assign an in vitro irritancy hazard classification category for prediction of the in vivo ocular irritation potential of a test substance (see Decision criteria).
Obe meritvi se uporabita za izračun IVIS, na podlagi katerega se dodeli kategorija nevarnosti dražilnosti in vitro, s pomočjo katere se predvidi potencialni dražilni učinek testne snovi in vivo (glej merila za odločanje).
The BCOP test method uses isolated corneas from the eyes of freshly slaughtered cattle.
Pri testni metodi BCOP se uporabljajo izolirane roženice oči sveže zaklanega goveda.
Corneal opacity is measured quantitatively as the amount of light transmission through the cornea.
Motnjava roženice se meri kvantitativno in ustreza količini svetlobe, ki prodre skozi roženico.
Permeability is measured quantitatively as the amount of sodium fluorescein dye that passes across the full thickness of the cornea, as detected in the medium in the posterior chamber.
Prepustnost se meri kvantitativno in ustreza količini barvila natrijevega fluoresceina, ki prodre skozi celotno debelino roženice in ga zazna medij v zadnjem delu držala.
Test substances are applied to the epithelial surface of the cornea by addition to the anterior chamber of the corneal holder.
Testne snovi se dodajo v sprednji del držala za roženico in se tako aplicirajo na epitelno površino roženice.
Appendix 3 provides a description and a diagram of a corneal holder used in the BCOP.
Dodatek 3 vsebuje opis in sliko držala, ki se uporablja za BCOP.
Corneal holders can be obtained commercially from different sources or can be constructed.
Držala za roženico ponujajo na trgu različna podjetja, laboratoriji pa jih lahko izdelajo tudi sami.
Source and Age of Bovine Eyes and Selection of Animal Species
Izvor in starost oči goveda ter izbor živalskih vrst
Cattle sent to slaughterhouses are typically killed either for human consumption or for other commercial uses.
Govedo, ki se zakolje v klavnicah, je običajno namenjeno za človeško prehrano ali druge komercialne uporabe.
Only healthy animals considered suitable for entry into the human food chain are used as a source of corneas for use in the BCOP.
Pri testih BCOP se kot vir roženic uporabljajo samo zdrave živali, primerne za vstop v prehransko verigo.
Because cattle have a wide range of weights, depending on breed, age, and sex, there is no recommended weight for the animal at the time of slaughter.
Ker so goveda glede na pasmo, starost in spol različno težka, ni priporočene teže za zakol.
Variations in corneal dimensions can result when using eyes from animals of different ages.
Velikost roženice se lahko pri očeh različno starih živali razlikuje.
Corneas with a horizontal diameter > 30,5 mm and central corneal thickness (CCT) values ≥ 1100 μm are generally obtained from cattle older than eight years, while those with a horizontal diameter < 28,5 mm and CCT < 900 μm are generally obtained from cattle less than five years old (14).
Roženice z vodoravnim premerom > 30,5 mm in centralno debelino roženice (CCT) ≥ 1100 μm ima običajno govedo, starejše od osem let, roženice z navpičnim premerom < 28,5 mm in CCT < 900 μm pa običajno govedo, mlajše od pet let (14).
For this reason, eyes from cattle greater than 60 months old are not typically used.
Zato se oči goveda, starejšega od 60 mesecev, običajno ne uporabljajo.
Eyes from cattle less than 12 months of age have not traditionally been used since the eyes are still developing and the corneal thickness and corneal diameter are considerably smaller than that reported for eyes from adult cattle.
Oči goveda, mlajšega od 12 mesecev, se običajno prav tako ne uporabljajo, saj še niso popolnoma razvite, debelina in premer roženice pa sta bistveno manjša kot pri očeh starejšega goveda.
However, the use of corneas from young animals (i.e., 6 to 12 months old) is permissible since there are some advantages, such as increased availability, a narrow age range, and decreased hazards related to potential worker exposure to Bovine Spongiform Encephalopathy (15).
Kljub temu se lahko uporabljajo roženice mladih živali (tj. starih od šest do 12 mesecev), saj ima njihova uporaba nekatere prednosti, npr. večja razpoložljivost, manjši starostni razpon in manjša nevarnost izpostavljenosti goveji spongiformni encefalopatiji za osebje klavnice (15).
As further evaluation of the effect of corneal size or thickness on responsiveness to corrosive and irritant substances would be useful, users are encouraged to report the estimated age and/or weight of the animals providing the corneas used in a study.
Ker bi bilo nadaljnje ocenjevanje učinka, ki ga imata velikost in debelina roženice na odzivnost jedkih in zelo dražilnih snovi za oči, koristno, se uporabnike spodbuja, da sporočijo približno starost in/ali težo živali, katerih roženice so uporabili v študiji.
Collection and Transport of Eyes to the Laboratory
Odvzem in prevoz oči v laboratorij
Eyes are collected by slaughterhouse employees.
Oči odvzame osebje klavnice.
To minimize mechanical and other types of damage to the eyes, the eyes should be enucleated as soon as possible after death.
Da bi čim bolj zmanjšali mehanske in druge vrste poškodb oči, jih morajo odvzeti takoj po zakolu.
To prevent exposure of the eyes to potentially irritant substances, the slaughterhouse employees should not use detergent when rinsing the head of the animal.
Da oči ne bi bile izpostavljene potencialno dražilnim snovem, osebje klavnice pri spiranju glav živali ne sme uporabljati detergentov.
Eyes should be immersed completely in Hanks’ Balanced Salt Solution (HBSS) in a suitably sized container, and transported to the laboratory in such a manner as to minimize deterioration and/or bacterial contamination.
Oči je treba dati v dovolj veliko posodo, jih preliti s Hanksovo uravnoteženo raztopino soli (HBSS) in prepeljati v laboratorij tako, da se njihovo stanje čim manj poslabša in/ali da ne pride do bakterijske okužbe. Ker se oči odvzamejo med zakolom, lahko pridejo v stik s krvjo in drugimi biološkimi snovmi, vključno z bakterijami in drugimi mikroorganizmi.
Because the eyes are collected during the slaughter process, they might be exposed to blood and other biological substances, including bacteria and other microorganisms.
Zato mora biti tveganost okužbe čim manjša (tj. posoda, v kateri so oči, se mora hraniti na mokrem ledu, HBSS, ki se uporablja za hrambo oči med prevozom, pa je treba dodati antibiotike [tj.
Therefore, it is important to ensure that the risk of contamination is minimized (e.g., by keeping the container containing the eyes on wet ice, by adding antibiotics to the HBSS used to store the eyes during transport [e.g., penicillin at 100 IU/mL and streptomycin at 100 μg/mL]).
100 IU/mL penicilina in 100 μg/mL streptomicina]).
The time interval between collection of the eyes and use of corneas in the BCOP should be minimized (typically collected and used on the same day) and should be demonstrated to not compromise the assay results.
Čas med odvzemom oči in uporabo roženic mora biti pri BCOP čim krajši (običajno se odvzamejo in uporabijo isti dan) in ne sme vplivati na zanesljivost testnih rezultatov.
These results are based on the selection criteria for the eyes, as well as the positive and negative control responses.
Ti temeljijo na merilih za izbor oči ter pozitivnih in negativnih kontrolnih reakcijah.
All eyes used in the assay should be from the same group of eyes collected on a specific day.
Vsa očesa, uporabljena v preskusu, morajo pripadati isti skupini oči, odvzeti na določen dan.
Selection Criteria for Eyes Used in the BCOP
Merila za izbor oči za uporabo pri testu BCOP
The eyes, once they arrive at the laboratory, are carefully examined for defects including increased opacity, scratches, and neovascularisation.
Po prihodu v laboratorij se oči skrbno pregledajo, ali so poškodovane, vključno s povečano motnjavo, praskami in neovaskularizacijo.
Only corneas from eyes free of such defects are to be used.
Uporabljajo se lahko samo roženice nepoškodovanih oči.
The quality of each cornea is also evaluated at later steps in the assay.
Kakovost vsake roženice se oceni tudi v poznejši fazi preskusa.
Corneas that have an opacity greater than seven opacity units (NOTE: the opacitometer should be calibrated with opacity standards that are used to establish the opacity units, see Appendix 3) after an initial one hour equilibration period are to be discarded.
Roženice z motnjavo več kot sedem enot motnjave (OPOMBA: opacitometer je treba kalibrirati v skladu s standardi za določevanje enot motnjave, glej dodatek 3) je treba po enournem kalibriranju izločiti.
Each treatment group (test substance, concurrent negative and positive controls) consists of a minimum of three eyes.
Vsaka testna skupina (testna snov, primerjalne negativne in pozitivne kontrole) je sestavljena iz najmanj treh oči.
Three corneas should be used for the negative control corneas in the BCOP assay.
Pri preskusu BCOP je treba za negativno kontrolo uporabiti tri roženice.
Since all corneas are excised from the whole globe, and mounted in the corneal chambers, there is the potential for artefacts from handling upon individual corneal opacity and permeability values (including negative control).
Ker se roženice odstranijo s celotnega zrkla in pritrdijo v držala za roženice, se lahko zaradi artefaktov, ki nastanejo zaradi prijemanja, vrednosti motnjave in prepustnosti v posameznih primerih spremenijo (tudi pri negativni kontroli).
Furthermore, the opacity and permeability values from the negative control corneas are used to correct the test article and positive control-treated corneal opacity and permeability values in the IVIS calculations.
Poleg tega se vrednosti motnjave in prepustnosti roženic z negativno kontrolno reakcijo uporabljajo za korigiranje vrednosti motnjave in preglednosti predmeta testiranja in pozitivnih kontrolnih vzorcev pri izračunih IVIS.
Preparation of the Eyes
Priprava oči
Corneas free of defects are dissected with a 2 to 3 mm rim of sclera remaining to assist in subsequent handling, with care taken to avoid damage to the corneal epithelium and endothelium.
Nepoškodovane roženice se secirajo tako, da se ohrani 2 do 3 mm širok rob beločnice, kar olajša nadaljnje ravnanje, pri čemer je treba paziti, da se epitelij in endotelij roženice ne poškodujeta.
Isolated corneas are mounted in specially designed corneal holders that consist of anterior and posterior compartments, which interface with the epithelial and endothelial sides of the cornea, respectively.
Izolirane roženice se pritrdijo v posebej za to izdelana držala, sestavljena iz sprednjega in zadnjega dela držala, na katera je roženica pripeta z epitelno oziroma endotelno stranjo.
Both chambers are filled to excess with pre-warmed Eagle's Minimum Essential Medium (EMEM) (posterior chamber first), ensuring that no bubbles are formed.
Oba dela držala se do roba napolnita z ogretim Eaglovim minimalnim osnovnim medijem EMEM (najprej zadnji del držala), pri čemer je treba zagotoviti, da se ne tvorijo mehurčki.
The device is then equilibrated at 32 ± 1 °C for at least one hour to allow the corneas to equilibrate with the medium and to achieve normal metabolic activity, to the extent possible (the approximate temperature of the corneal surface in vivo is 32 °C).
Naprava se nato ekvilibrira pri 32 ± 1 °C najmanj eno uro, da se roženice ekvilibrirajo z medijem in da se doseže čim bolj normalna presnova (približna temperatura na površini roženice znaša in vivo 32 °C).
Following the equilibration period, fresh pre-warmed EMEM is added to both chambers and baseline opacity readings are taken for each cornea.
Po ekvilibriranju se v oba dela držala doda svež, ogret medij EMEM, za vsako roženico pa se zabeležijo izhodiščne vrednosti motnjave.
Any corneas that show macroscopic tissue damage (e.g., scratches, pigmentation, neovascularisation) or an opacity > 7 opacity units are discarded.
Roženice z makroskopskimi poškodbami tkiva (npr. praskami, pigmentacijo, neovaskularizacijo) ali motnjavo, večjo od sedem enot, se zavržejo.
The mean opacity of all equilibrated corneas is calculated.
Nato se izračuna povprečna motnjava vseh ekvilibriranih roženic.
A minimum of three corneas with opacity values close to the median value for all corneas are selected as negative (or solvent) control corneas.
Za negativno kontrolo (ali kontrolo s topilom) se izberejo najmanj tri roženice, katerih motnjava je blizu povprečni motnjavi vseh roženic.
The remaining corneas are then distributed into treatment and positive control groups.
Preostale roženice se nato razdelijo na skupino za nadaljnje testiranje in skupino za pozitivne kontrole.
Because the heat capacity of water is higher than that of air, water provides more stable temperature conditions for incubation.
Ker je toplotna kapaciteta vode večja od zraka, zagotavlja voda stabilnejše temperaturne pogoje za inkubacijo.
Therefore, the use a water bath for maintaining the corneal holder and its contents at 32 ± 1 °C is recommended.
Zato se priporoča, da se držalo za roženico in njegova vsebina hranita v vodni kopeli pri 32 ± 1 °C.
However, air incubators might also be used, assuming precaution to maintain temperature stability (e.g., by pre-warming of holders and media).
Lahko se uporabljajo tudi inkubatorji za vzdrževanje temperature zraka, če zagotavljajo stabilno temperaturo (tj. s predčasnim segrevanjem držal in medijev).
Application of the Test Substance
Uporaba testne snovi
Two different treatment protocols are used, one for liquids and surfactants (solids or liquids), and one for non-surfactant solids.
Vodita se dva različna testna protokola - eden za tekočine in površinsko aktivne snovi (trdne snovi ali tekočine) ter eden za površinsko neaktivne snovi (trdne snovi).
Liquids are tested undiluted, while surfactants are tested at a concentration of 10 % w/v in a 0,9 % sodium chloride solution, distilled water, or other solvent that has been demonstrated to have no adverse effects on the test system.
Tekočine se testirajo nerazredčene, medtem ko se površinsko aktivne snovi testirajo v 10-odstotni koncentraciji w/v in 0,9-odstotni raztopini natrijevega klorida, destilirani vodi ali drugem topilu, za katerega je dokazano, da nima škodljivih učinkov na testni sistem.
Semi-solids, creams, and waxes are typically tested as liquids.
Poltrdne snovi, kreme in voski se navadno testirajo kot tekočine.