natgillin/translations-raw huggingface dataset:
After the blood has been collected, the plasma is quickly isolated by centrifugation for 3 min at 15000 g (or alternatively for 10 min. at 15000 g at 4 °C).
Nakon uzimanja krvi plazma se brzo izolira centrifugom pri 15000 g u trajanju od tri minute (ili centrifugom u trajanju od deset minuta pri 15000 g na 4 °C).
If desired, percent hematocrit can be determined following centrifugation.
Postotak hematokrita može se, po želji, utvrditi nakon centrifugiranja.
The plasma portion is then removed from the microhematocrit tube and stored in a centrifuge tube with 0,13 units of aprotinin (a protease inhibitor) at – 80 °C until determination of vitellogenin can be made.
Plazma se potom uklanja iz mikrohematokritske epruvete i pohranjuje u epruvetu za centrifugu uz 0,13 jedinica aprotinina (inhibitora proteaze) na – 80 °C do mogućnosti utvrđivanja vitelogenina.
Depending on the size of the fathead minnow (which is sex-dependent), collectable plasma volumes generally range from 5 to 60 microlitres per fish (Jensen et al. 2001).
Ovisno o veličini debeloglave gavčice (koja ovisi o spolu), prikupljeni volumen plazme obično se kreće od pet do 60 mikrolitara po ribi (Jensen et al., 2001.).
Alternatively, blood may also be collected by cardiac puncture using a heparinized syringe (1000 units of heparin per ml).
Druga je mogućnost uzimanje krvi punkcijom srca s pomoću heparinizirane šprice (1000 jedinica heparina po ml).
The blood is transferred into Eppendorf tubes (held on ice) and then centrifuged (5 min, 7000 g, room temperature).
Krv se stavlja u Eppendorfove epruvete (koje se čuvaju na ledu) i potom centrifugira (pet minuta; 7000 g, na sobnoj temperaturi).
The plasma should be transferred into clean Eppendorf tubes (in aliquots if the volume of plasma makes this feasible) and promptly frozen at – 80 °C, until analyzed (Panter et al., 1998).
Plazmu treba premjestiti u čiste Eppendorfove epruvete (u alikvotima ako to dopušta volumen plazme) i odmah smrznuti na – 80 °C do analize (Panter et al., 1998.).
Procedure 2A:
Postupak 2.A:
Japanese Medaka, Excision of the Liver in Medaka
japanska medaka, vađenje jetre u medake
Removal of the test fish from the test chamber
Uklanjanje pokusne ribe iz ispitne posude
Test fish should be removed from the test chamber using the small spoon-net.
Pokusnu ribu potrebno je izvaditi iz ispitne posude malom mrežom u obliku žlice.
Be careful not to drop the test fish into other test chambers.
Pazite da ne ispustite pokusnu ribu u ostale ispitne posude.
In principle, the test fish should be removed in the following order:
Pokusne ribe u pravilu je potrebno vaditi sljedećim redoslijedom:
control, solvent control (where appropriate), lowest concentration, middle concentration, highest concentration and positive control.
kontrolna skupina, kontrolna skupina za otapalo (ako postoji), najniža koncentracija, srednja koncentracija, najviša koncentracija i pozitivna kontrola.
In addition, all males should be removed from one test chamber before the remaining females are removed.
Osim toga, iz ispitne posude treba izvaditi sve mužjake prije vađenja preostalih ženki.
The sex of each test fish is identified on the basis of external secondary sex characteristics (e.g. the shape of the anal fin).
Spol pokusnih riba utvrđuje se na temelju vanjskih sekundarnih spolnih obilježja (primjerice oblika podrepne peraje).
Place the test fish in a container for transport and carry it to the workstation for excision of the liver.
Stavite ribu u spremnik za transport i odnesite je do radne stanice radi vađenja jetre.
Check the labels of the test chamber and the transport container for accuracy and to confirm that the number of fish that have been removed from the test chamber and that the number of fish remaining in the test chamber are consistent with expectation.
Provjerite točnost oznaka na ispitnoj komori i spremniku za transport i provjerite je li broj riba koje su izvađene iz ispitne komore i broj riba preostalih u ispitnoj komori u skladu s očekivanjima.
If the sex cannot be identified by the fish's external appearance, remove all fish from the test chamber.
Ako se spol ne može utvrditi iz vanjskog izgleda riba, izvadite sve ribe iz ispitne komore.
In this case, the sex should be identified by observing the gonad or secondary sex characteristics under a stereoscopic microscope.
Spol je u tom slučaju potrebno utvrditi promatranjem gonade ili sekundarnih spolnih obilježja pod stereoskopskim mikroskopom.
Excision of the liver
Vađenje jetre
Transfer the test fish from the container for transport to the anaesthetic solution using the small spoon-net.
Premjestite pokusnu ribu iz spremnika za transport u otopinu anestetika malom mrežom u obliku žlice.
After the test fish is anaesthetised, transfer the test fish on the filter paper (or a paper towel) using tweezers (commodity type).
Nakon anestezije pincetom (običnom) premjestite pokusnu ribu na filtarski papir (ili papirnati ručnik).
When grasping the test fish, apply the tweezers to the sides of the head to prevent breaking the tail.
Pri hvatanju pokusne ribe pincetom obuhvatite glavu kako ne bi došlo do kidanja repa.
Wipe the water on the surface of the test fish on the filter paper (or the paper towel).
Obrišite vodu s površine pokusne ribe filtarskim papirom (ili papirnatim ručnikom).
Then make a small transverse incision partway between the ventral neck region and the mid-abdominal region using dissection scissors.
Potom škaricama napravite mali poprečni rez od leđnog dijela vrata do područja srednjeg trbuha.
Insert the dissection scissors into the small incision, and incise the abdomen from a point caudal to the branchial mantle to the cranial side of the anus along the midline of the abdomen.
Umetnite škarice u mali rez i zarežite trbuh od točke sa stražnje strane škržnog plašta do prednje strane anusa duž središnje linije abdomena.
Be careful not to insert the dissection scissors too deeply so as to avoid damaging the liver and gonad.
Pazite da škarice ne umetnete preduboko kako ne bi došlo do oštećenja jetre i gonade.
Conduct the following operations under the stereoscopic microscope.
Obavite sljedeće postupke pod stereoskopskim mikroskopom.
Place the test fish abdomen side up on the paper towel (glass Petri dish or slide glass are also available).
Položite pokusnu ribu trbuhom prema gore na papirnati ručnik (pri ruci moraju biti i staklena Petrijeva zdjelica ili stakalce).
Extend the walls of the abdominal cavity with precision tweezers and exteriorise the internal organs.
Proširite stijenke trbušne šupljine preciznom pincetom i izvadite unutarnje organe.
It is also acceptable to exteriorise the internal organs by removing one side of the wall of the abdominal cavity if necessary.
Prihvatljivo je, po potrebi, i vađenje unutarnjih organa uklanjanjem jedne strane stijenke trbušne šupljine.
Expose the connected portion of the liver and gallbladder using another pair of precision tweezers.
Otkrijte spojeni dio jetre i žučnog mjehura drugom preciznom pincetom.
Then grasp the bile duct and cut off the gallbladder.
Potom uhvatite žučni kanal i odrežite žučni mjehur.
Be careful not to break the gallbladder.
Pazite da ne dođe do trganja žučnog mjehura.
Grasp the oesophagus and excise the gastrointestinal tract from the liver in the same way.
Uhvatite jednjak i na isti način razdvojite probavni trakt od jetre.
Be careful not to leak the contents of the gastrointestinal tract.
Pazite da ne dođe do izlijevanja sadržaja probavnog trakta.
Excise the caudal gastrointestinal tract from the anus and remove the tract from the abdominal cavity.
Odvojite stražnji dio probavnog trakta od anusa i izvadite ga iz trbušne šupljine.
Trim the mass of fat and other tissues from the periphery of the liver.
Uklonite mast i ostala tkiva s vanjskog dijela jetre.
Be careful not to scratch the liver.
Pazite da ne ogrebete jetru.
Grasp the hepatic portal area using the precision tweezers and remove the liver from the abdominal cavity.
Preciznom pincetom uhvatite jetru i uklonite je iz trbušne šupljine.
Place the liver on the slide glass.
Položite jetru na stakalce.
Using the precision tweezers, remove any additional fat and extraneous tissue (e.g., abdominal lining), if needed, from the surface of the liver.
Po potrebi, s površine jetre preciznom pincetom uklonite višak masti i okolnog tkiva (primjerice trbušnu ovojnicu).
Measure the liver weight with 1.5 ml microtube as a tare using an electronic analytical balance.
Izvažite jetru s pomoću elektroničke analitičke vage i mikroepruvete od 1,5 ml kao tare.
Record the value on the worksheet (read: 0,1 mg).
Zabilježite vrijednost u obrazac (očitanje: 0,1 mg).
Confirm the identification information on the microtube label.
Potvrdite podatke o identifikaciji na naljepnici mikroepruvete.
Close the cap of the microtube containing the liver.
Zatvorite poklopac mikroepruvete u kojoj se nalazi jetra.
Store it in a cooling rack (or ice rack).
Pohranite je u stalak za hlađenje (ili stalak s ledom).
Following the excision of one liver, clean the dissection instruments or replace them with clean ones.
Nakon vađenja jedne jetre instrumente očistite ili ih zamijenite čistima.
Remove livers from all of the fish in the transport container as described above.
Izvadite jetre svim ribama u spremniku za transport kako je prethodno opisano.
After the livers have been excised from all of the fish in the transport container (i.e., all males or females in a test chamber), place all liver specimens in a tube rack with a label for identification and store it in a freezer.
Nakon vađenja jetara svim ribama u spremniku za transport (odnosno svim mužjacima ili ženkama u ispitnoj komori) stavite sve uzorke jetara u stalak za epruvete s identifikacijskom oznakom i pohranite ih u zamrzivač.
When the livers are donated for pre-treatment shortly after the excision, the specimens are carried to the next workstation in a cooling rack (or ice rack).
U slučaju uzimanja jetara za preliminarno tretiranje ubrzo nakon vađenja primjerci se do sljedeće radne stanice nose u stalku za hlađenje (ili stalku s ledom).
Following liver excision, the fish carcass is available for measurement of secondary sex characteristics.
Nakon vađenja jetre truplo ribe podvrgava se mjerenju sekundarnih spolnih obilježja.
Specimen
Uzorak
Store the liver specimens taken from the test fish at ≤ – 70 °C if they are not used for the pre-treatment shortly after the excision.
Pohranite uzorke jetara pokusnih riba na ≤ – 70 °C ako se one ne podvrgavaju preliminarnom tretiranju ubrzo nakon vađenja.
A cut is made just anterior to pectoral fins with scissors.
Škarama se načini rez neposredno ispred prsnih peraja.
The midline of abdomen is incised with scissors to a point approximately 2 mm cranial to the anus.
Škarama se razreže središnja linija abdomena do točke približno 2 mm od kranijalne strane anusa.
The abdominal walls are spread with forceps for exposure of the liver and other internal organs.
Pincetom se trbušne stijenke šire radi vađenja jetre i ostalih unutarnjih organa.
(Alternatively, the abdominal walls may be pinned laterally).
(Druga je mogućnost bočno pričvrstiti trbušne stijenke).
The liver is bluntly dissected and excised using forceps.
Jetra se izrezuje i vadi s pomoću pincete.
The intestines are gently retracted using forceps.
Crijeva se nježno povlače pincetom.
Testis 6
Slika 6.
Both ends of the intestines and any mesenteric attachments are severed using scissors.
Škarama se režu oba kraja crijeva i sva mezenterička vezivna tkiva.
Testis 7 (female)
Slika 7.
The procedure is identical for the female.
(ženka) Postupak je jednak za ženke.
Testis 8
Slika 8.
The completed procedure.
Kraj postupka.
Procedure 2 B:
Postupak 2.B:
Japanese Medaka (Oryzias latipes), Liver Pre-treatment for Vitellogenin Analysis
japanska medaka (Oryzias latipes), preliminarno tretiranje jetre za analizu vitelogenina
Take the bottle of homogenate buffer from the ELISA kit and cool it with crushed ice (temperature of the solution: ≤ 4 °C).
Uzmite bocu homogenizacijskog pufera iz kompleta za analizu ELISA i ohladite je smrvljenim ledom (temperatura otopine: ≤ 4 °C).
If homogenate buffer from EnBio ELISA system is used, thaw the solution at room temperature, and then cool the bottle with crushed ice.
U slučaju upotrebe homogenizacijskog pufera iz sustava EnBio ELISA otopite otopinu na sobnu temperaturu, a potom bocu ohladite smrvljenim ledom.
Calculate the volume of homogenate buffer for the liver on the basis of its weight (add 50 μl of homogenate buffer per mg liver weight for homogenate).
Izračunajte volumen homogenizacijskog pufera za jetru na temelju njezine mase (dodajte 50 μl homogenizacijskog pufera po mg mase jetre za homogenat).
For example, if the weight of the liver is 4,5 mg, the volume of homogenate buffer for the liver is 225 μl.
Primjerice, ako masa jetre iznosi 4,5 μl, volumen homogenizacijskog pufera za jetru iznosi 225 μl.
Prepare a list of the volume of homogenate buffer for all livers.
Sastavite popis volumena homogenizacijskog pufera za sve jetre.
Preparation of the liver for pre-treatment
Priprema jetre za preliminarno tretiranje
Take the 1,5 ml microtube containing the liver from the freezer just before the pre-treatment.
Neposredno prije tretiranja iz zamrzivača izvadite mikroepruvetu od 1,5 ml u kojoj se nalazi jetra.
Pre-treatment of the liver from males should be performed before females to prevent vitellogenin contamination.
Preliminarno tretiranje jetre mužjaka potrebno je obaviti prije ženki kako bi se izbjegla kontaminacija vitelogenina.
In addition, the pre-treatment for test groups should be conducted in the following order:
Osim toga, preliminarno tretiranje ispitnih skupina potrebno je obaviti sljedećim redoslijedom:
The number of 1,5 ml microtubes containing liver samples taken from the freezer at a given time should not exceed the number that can be centrifuged at that time.
Broj mikroepruveta od 1,5 ml s primjercima jetara izvađenih iz zamrzivača u određenom trenutku ne smije premašiti broj mikroepruveta koje je moguće istovremeno centrifugirati.
Arrange the 1,5 ml microtubes containing liver samples in the order of specimen number on the ice rack (no need to thaw the liver).
Poredajte mikroepruvete od 1,5 ml s primjercima jetara prema redoslijedu broja primjerka na stalku s ledom (jetre ne treba otapati).
Operation of the pre-treatment
Postupak preliminarnog tretiranja
Addition of the homogenization buffer
Dodavanje homogenizacijskog pufera
Check the list for the volume of the homogenate buffer to be used for a particular sample of liver and adjust the micropipette (volume range: 100-1000 μl) to the appropriate volume.
Provjerite popis kako biste utvrdili koji je volumen homogenizacijskog pufera potrebno upotrijebiti za određeni uzorak jetre i podesite mikropipetu (raspon volumena: 100 – 1000 μl) na odgovarajući volumen.
Attach a clean tip to the micropipette.
Na mikropipetu stavite čisti vrh.
Take the homogenate buffer from the reagent bottle and add the buffer to the 1,5 ml microtube containing the liver.
Uzmite homogenizacijski pufer iz boce s reagensom i dodajte pufer u mikroepruvetu od 1,5 ml u kojoj se nalazi jetra.
Add the homogenate buffer to all of 1,5 ml microtubes containing the liver according to the procedure described above.
Dodajte homogenizacijski pufer u sve mikroepruvete od 1,5 ml u kojima se nalazi jetra prema prethodno opisanom postupku.
There is no need to change the micropipette tip to a new one.
Vrh mikropipete nije potrebno zamijeniti novim.
However, if the tip is contaminated or suspected to be contaminated, the tip should be changed.
Međutim, vrh je potrebno zamijeniti ako je kontaminiran ili se sumnja na kontaminaciju.
Homogenisation of the liver
Homogenizacija jetre
Attach a new pestle for homogenisation to the microtube homogeniser.
Na homogenizator mikroepruveta stavite novi tučak za homogenizaciju.
Insert the pestle into the 1,5 ml microtube.
Stavite tučak u mikroepruvetu od 1,5 ml.
Hold the microtube homogeniser to press the liver between the surface of the pestle and the inner wall of the 1,5 ml microtube.
Držite homogenizator mikroepruveta tako da jetru pritišćete između površine tučka i unutarnje stijenke mikroepruvete od 1,5 ml.
Operate the microtube homogeniser for 10 to 20 seconds.
Uključite homogenizator mikroepruveta na 10 do 20 sekundi.
Cool the 1,5 ml microtube with crushed ice during the operation.
Tijekom postupka hladite mikroepruvetu od 1,5 ml smrvljenim ledom.
Lift up the pestle from the 1,5 ml microtube and leave it at rest for about 10 seconds.
Izvadite tučak iz mikroepruvete i odložite je na približno deset sekundi.
Then conduct a visual check of the state of the suspension.
Potom vizualnim pregledom utvrdite stanje suspenzije.
If pieces of liver are observed in the suspension, repeat the operations (3) and (4) to prepare satisfactory liver homogenate.
Ako su u suspenziji vidljivi djelići jetre, ponovite postupke (3) i (4) kako biste pripremili zadovoljavajući homogenat jetre.
Cool the suspended liver homogenate on the ice rack until centrifugation.
Ohladite suspendirani homogenat jetre na stalku s ledom do centrifugiranja.
Change the pestle to the new one for each homogenate.
Mijenjajte tučak za svaki homogenat.
Homogenise all livers with homogenate buffer according to the procedure described above.
Homogenizirajte sve jetre homogenizacijskim puferom prema prethodno opisanom postupku.
Centrifugation of the suspended liver homogenate
Centrifugiranje suspendiranog homogenata jetre
Confirm the temperature of the refrigerated centrifuge chamber at ≤ 5 °C.
Provjerite je li temperatura rashlađene komore za centrifugiranje ≤ 5 °C.
Insert the 1,5 ml microtubes containing the suspended liver homogenate in refrigerated centrifuge (adjust the balance if necessary).
Stavite mikroepruvete od 1,5 ml u kojima se nalazi suspendirani homogenat jetre u rashlađenu centrifugu (po potrebi prilagodite ravnotežu).
Centrifuge the suspended liver homogenate at 13000 g for 10 min at ≤ 5 °C.
Centrifugirajte suspendirani homogenat jetre deset minuta pri 13000 g na ≤ 5 °C.
However, if the supernatants are adequately separated, centrifugal force and time may be adjusted as needed.
Međutim, u slučaju pravilne separacije supernatanta centrifugalnu silu i vrijeme moguće je prilagoditi po potrebi.
Following centrifugation, check that the supernatants are adequately separated (surface: lipid, intermediate: supernatant, bottom layer: liver tissue).
Nakon centrifugiranja provjerite jesu li supernatanti pravilno separirani (površina: lipidi, sredina: supernatant, donji sloj: tkivo jetre).
If the separation is not adequate, centrifuge the suspension again under the same conditions.
U slučaju nepravilne separacije suspenziju ponovno centrifugirajte u istim uvjetima.
Remove all specimens from the refrigerated centrifuge and arrange them in the order of specimen number on the ice rack.
Iz rashlađene centrifuge izvadite sve uzorke i poredajte ih prema broju uzorka na stalku s ledom.
Be careful not to resuspend each separated layer after the centrifugation.
Pazite da nakon centrifugiranja ne dođe do ponovne suspenzije separiranih slojeva.
Collection of the supernatant
Vađenje supernatanta
Place four 0,5 ml microtubes for storage of the supernatant into the tube rack.
U stalak za epruvete stavite četiri mikroepruvete od 0,5 ml za pohranu supernatanta.
Collect 30 μl of each supernatant (separated as the intermediate layer) with the micropipette and dispense it to one 0,5 ml microtube.
Mikropipetom izvadite 30 μl svakog supernatanta (separiranog kao srednji sloj) i stavite u jednu mikroepruvetu od 0,5 ml.
Be careful not to collect the lipid on the surface or the liver tissue in the bottom layer.
Pazite da ne zahvatite lipid na površini ili tkivo jetre iz donjeg sloja.
Collect the supernatant and dispense it to other two 0,5 ml microtubes in the same manner as described above.
Izvadite supernatant i stavite ga u druge dvije mikroepruvete od 0,5 ml na prethodno opisani način.
Collect the rest of the supernatant with the micropipette (if feasible: ≥ 100 μl).
Mikropipetom izvadite ostatak supernatanta (po mogućnosti: ≥ 100 μl).
Then dispense the supernatant to the remaining 0,5 ml microtube.
Potom ga stavite u preostalu mikroepruvetu od 0,5 ml.
Close the cap of the 0,5 ml microtube and write the volume of the supernatant on the label.
Zatvorite poklopac mikroepruvete od 0,5 ml i na naljepnicu zapišite volumen supernatanta.
Then immediately cool the microtubes on the ice rack.
Mikroepruvete potom odmah ohladite na stalku s ledom.
Change the tip of the micropipette to the new one for each supernatant.
Mijenjajte vrh mikropipete za svaki supernatant.
If a large amount of lipid becomes attached to the tip, change it to the new one immediately to avoid contamination of the liver extract with fat.
Ako se na vrhu nakupi velika količina lipida, odmah ga promijenite kako ne bi došlo do kontaminacije ekstrakta jetre mašću.
Dispense all of the centrifuged supernatant to four 0,5 ml microtubes according to the procedure described above.
Stavite sav centrifugirani supernatant u četiri mikroepruvete od 0,5 ml prema prethodno opisanom postupku.
After dispensing the supernatant to the 0,5 ml microtubes, place all of them in the tube rack with the identification label, and then freeze them in the freezer immediately.
Sve mikroepruvete od 0,5 ml u kojima se nalazi supernatant stavite u stalak za epruvete s identifikacijskim oznakama i potom ih odmah zamrznite u zamrzivaču.
If the VTG concentrations are measured immediately after the pre-treatment, keep one 0,5 ml microtube (containing 30 μl of supernatant) cool in the tube rack and transfer it to the workstation where the ELISA assay is conducted.
U slučaju mjerenja koncentracija VTG-a neposredno nakon preliminarnog tretiranja ostavite jednu mikroepruvetu od 0,5 ml (u kojoj se nalazi 30 μl supernatanta) da se hladi u stalku za epruvete i odnesite je do radne stanice gdje se provodi test ELISA.
In such case, place the remaining microtubes in the tube racks and freeze them in the freezer.
U tom slučaju stavite preostale mikroepruvete u stalke za epruvete i zamrznite ih u zamrzivaču.
After the collection of the supernatant, discard the residue adequately.
Nakon vađenja supernatanta ostatke zbrinite na odgovarajući način.
Storage of the specimen
Čuvanje uzorka
Store the 0,5 ml microtubes containing the supernatant of the liver homogenate at ≤ – 70 °C until they are used for the ELISA.
Mikroepruvete od 0,5 ml u kojima se nalazi supernatant homogenata jetre čuvajte na ≤ – 70 °C do upotrebe za test ELISA.
Procedure 3A:
Postupak 3.A:
Zebrafish, Blood Collection from the Caudal Vein / Artery
zebrica, uzimanje krvi iz repne vene/arterije
Immediately following anaesthesia, the caudal peduncle is severed transversely, and the blood is removed from the caudal artery/vein with a heparinised microhematocrit capillary tube.
Neposredno nakon anestezije korijen repa poprečno se zarezuje i uzima se krv iz repne arterije/vene s pomoću heparinizirane mikrohematokritske kapilarne epruvete.
Blood volumes range from 5 to 15 μl depending on fish size.
Volumen krvi kreće se od pet do 15 μl ovisno o veličini ribe.
An equal volume of aprotinin buffer (6 μgml in PBS) is added to the microcapillary tube, and plasma is separated from the blood via centrifugation (5 minutes at 600 g).
Jednaki volumen pufera aprotinina (6 μgml u fiziološkoj otopini s fosfatnim puferom) dodaje se u mikrokapilarnu epruvetu, dok se plazma centrifugiranjem odvaja od krvi (pet minuta pri 600 g).
Plasma is collected in the test tubes and stored at – 20 °C until analyzed for vitellogenin or other proteins of interest.
Plazma se stavlja u ispitne epruvete i čuva na – 20 °C do analize vitelogenina ili ostalih proteina koji mogu biti od interesa.
Procedure 3B:
Postupak 3.B:
Zebrafish, Blood Collection by Cardiac Puncture
zebrica, uzimanje krvi punkcijom srca
To avoid coagulation of blood and degradation of protein the samples are collected within Phosphate-buffered saline (PBS) buffer containing heparin (1000 units/ml) and the protease inhibitor aprotinin (2 TIU/ml).
Kako bi se izbjeglo zgrušavanje krvi i propadanje proteina, uzorci se uzimaju u fiziološkoj otopini s fosfatnim puferom koja sadržava heparin (1000 jedinica/ml) i aprotinin (inhibitor proteaze) (2 TIU/ml).