MEDICINSKI PODMLADAK
naučni časopis za studente i mlade istraživače

MEDICAL YOUTH
scientific journal for students and young researchers

Glavni i odgovorni urednik Editor-in-Chief Lektor za srpski jezik Reader

Brankica Dimitrijević Nataša Ognjanović

Zamenik glavnog i odgovornog urednika Deputy Editor Lektor za engleski jezik Reader

Smiljana Mihailović Ivana Katić

Redakcija Editorial Board Kontakt Contact

Zlatko Pravdić Medicinski podmladak
Marko Svetel Medicinski fakultet Univerziteta u Beogradu
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Saradnici redakcije Editorial Assistants ISSN-YU 0369-1527
UDK 61 (05)
Stefan Bošković
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Ana Marija Milanović
Ivana Jovanović Medicinski fakultet Univerziteta u Beogradu
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Izdavački savet Advisory Board
Štampa Press
Nebojša Lalić, MD, PhD
Tatjana Simić, MD, PhD Sprint, Beograd
Petar Bulat, MD, PhD
Tiraž Circulation
Vesna Bjegović Mikanović, MD, PhD
Slobodan Savić, MD, PhD 300 primeraka
Aleksandra Isaković, MD, PhD
Marina Svetel, MD, PhD Naslovna strana Cover
Vera Pravica, MD, PhD
Marija Plješa Ercegovac, MD, PhD Aleksandar Bubalo
Nađa Marić Bojović, MD, PhD Stevan Stojanović
Maja Ćupić, MD, PhD
Silvio de Luka, MD, PhD Posebna zahvalnost Acknowledgement
Darija Kisić Tepavčević, MD, PhD
Nela Puškaš, MD, PhD Izdavačkom savetu, za vreme, strpljenje i savete
prof. dr Draganu Simiću i dr Nebojši Markoviću,
Brankica Dimitrijević, glavni i odgovorni urednik za svesrdnu pomoć sa rekvizitima za naslovnicu i materijali-
Smiljana Mihailović, zamenik urednika ma za izradu postera
Ratko Radeta, predsednik CSNIRS-a prof. dr Ivani Novaković, za pomoć u organizaciji
Nebojša Prijović, član CSNIRS-a naslovne stranice broja

ispitivanja signalnih puteva koji modifikuju nakupljanje alfa-sinukleina do kliničkog pristu- pa i ishoda embrioredukcija kod multifetalnih trudnoća. ono što već decenijama čini srž našeg časopisa. Nadamo se da ćemo vas kroz ova tri pregledna rada zainteresovati da saznajete još više o kardiovaskularnom sistemu i izazovima sa kojima se sreću lekari opšte prakse. Aktuelna statistika kaže da svakih 90 sekundi jedan čovek na planeti umre usled bolesti srca. lekari specijalisti i naučnici. jesu originalni radovi studenata osnovnih studija. časopisa za stu- dente i mlade istraživače. martovski broj. genetskih polimorfizama koji se dovode u vezu sa aniridijom. kardiovaskularne bolesti su jedan od vodećih uzroka smrtnosti. potrudili smo se da i u ovom broju izdvojimo najzanimljivije zagonetke koje su rešavali studenti doktorskih studija pripremajući svoje disertacije. lečenju jedne od najučestalijih bolesti srca – okluzije koronarnih ar- terija. Ponosni smo na studente i njihove prve naučne korake. glukokortikoidnoj terapiji autoimunskih bolesti CNS-a i ulozi neurosteroida u patogenezi hepatičke encefalopatije. Srdačno.Uvodna reč foreword Dragi čitaoci. U savremenom svetu. te i u ovom broju možete pročitati najinovativnije naučnoistraživačke radove koje su realizovali studenti biomedicinskih fakulteta uz iskusno vođstvo svojih mentora. ovo je samo prvi od četiri broja koje za vas spremamo ove godine. Neinvazivnu dijagnostiku i minimalno invazivne terapijske pristupe u lečenju ishemijske bolesti srca slikovito smo prikazali na posteru koji smo zajedno sa našim nastavnicima pripremili za vas i prilažemo ga uz broj. Teme njihovih istraživanja bile su najrazličitije od: ispitivanja antimikotičnog dejstva različitih esencijalnih ulja. Kao što smo i najavili. Takođe. koračajte i dalje kroz svet nauke zajedno sa nama. Upravo iz tog razloga. Či- taćete o aktuelnom pristupu antitrombotičnoj terapiji ishemijskog moždanog udara kod pacijenata sa atrijalnom fibrilaci- jom. Pred vama se nalazi prvi broj 67. Do sledećeg broja. Brankica Dimitrijević glavni i odgovorni urednik . izdanja „Medicinskog podmlatka“. preko nakupljanja gvožđa u organima pacova sa starenjem. Sigurni smo da vam ništa manje intrigantan neće biti ni mini pregledni članak o „NANO“ pristupu u osteologiji i o tome šta određuje lomljivost kostiju. zahvalni smo svim mentorima. Redakcija je odlučila da ovaj. recenzentima i profesorima koji svakodnevno doprinose unapređivanju našeg časopisa. posveti autonom- noj kontroli rada srca. krvi i gvožđu kao nezaobilaznom delu priče o kardiovaskularnom sistemu i na kraju. Nakon što smo u prethodnom broju započeli sa objavljivanjem mini preglednih članaka.

.29 Višnja Pađen.......................................Sadržaj contents PREGLEDNI RADOVI Review articles Savremene indikacije za perkutanu koronarnu revaskularizaciju (PCI) kod bolesnika sa hroničnom totalnom okluzijom (CTO) koronarnih arterija...... Marija Mostarica Stojković Antithrombotic therapy in acute ischemic stroke patients with atrial fibrillation....35 Dušan Mladenović. Branko Beleslin Metabolizam gvožđa i laboratorijska dijagnostika pokazatelja metabolizma gvožđa.14 Branislava Medić MINI PREGLEDNI RADOVI Mini review articles “GOING NANO” in the field of osteology: Is bone fragility already determined at the level of bone mineralized matrix?..................................................6 Vesna Ćorić The role of autonomic control in cardiovascular system: summary of basic principles.25 Željka Stanojević.................. Siniša Stojković..........................................................................................20 Petar Milovanović...1 Stefan Juričić.................................................... Olivera Stanojlović.................... Ljiljana Beslać Bumbaširević The role of neurosteroids in the pathogenesis of hepatic encephalopathy........................... Marija Đurić Srejić Different response to glucocorticoid therapy in autoimmune diseases of CNS........... Tatjana Radosavljević .........

.........................41 Dimitrije Cvetković................................58 Roberto Grujičić.............................................. Vera Jovanović................ Stefan Zaharijev.................................. Ratko Radeta............................. Silvio De Luka The influence of multifetal pregnancy embrioreduction in high-risk pregnancies – clinical management and outcome. Jelena Ćuk...........................Sadržaj contents ORIGINALNI RADOVI Original articles PAX6 gene analysis in Serbian patients with aniridia.............. Maja Jovanović....................................................... Željko Miković ................. Ivana Novaković Antimikotična aktivnost esencijalnih ulja matičnjaka..............................44 Đorđe Jovanović....... manuke i vratića na uzročnike otomikoza................... Marija Dulović Promena sadržaja gvožđa u organima starih pacova...................... Bogdan Bjelica................... Sanja Mitrović Uloga protein kinaze aktivirane adenozin monofosfatom (APMK) u dejstvu modifikovanih oblika rekombinantnog vanćelijskog α-sinukleina in vitro......51 Marija Jeremić.....................................63 Petar Zlatanović........

Workshops. Academic Lectures. Courses and a rich Social Programme will adorn our conference. | Broj 1 | Izdanje 67 . the third Global Students’ Conference of Biomedical Sciences is going to take place in Belgrade. Deadline for Abstract Submission: 10th of July 2016 Place: Deanery of Faculty of Medicine University of Belgrade VI  Mart 2016. Medicinski podmladak Partneri From 20th to 23rd October 2016. Our goals: • introducing students to research fields • presenting results • acquiring and sharing knowledge and skills • creating friendships and ideas in future life and work • implementation of contemporary knowledge of the biomedical sciences The aims of our conference are to bring undergraduate and postgraduate students of biomedical sciences together and offer them an opportunity to present the results of their investigations and to discuss them with colleagues and leading experts. the capital of Serbia. We also welcome and encourage students of PhD programs to present their research findings at our congress. Medical education in scientific research and as well as the acquisition of scientific working skills are of great importance to academic institutions of biomedical sciences on the globe.

 1 . ili postojanje rasponom od 0 do 3 koji se odnosi na nivo protoka krvi odgovorne arterije koja nije arterija koja se procenjuje u kroz koronarnu arteriju. b) verovatan (klinički potvrđen): akutni infarkt rija (TIMI 0 koronarni protok u okludiranom segmentu) miokarda u zoni distribucije okludirane arterije.2. Symptomatic patients have significant CTO imaju značajno pobošljanje kvaliteta života. bez kontrastnog bojenja) dokaz ili velika verovatnoća trajanja okluzije ≥ 3meseca sa stabilnim nepromenjenim simptomima angine ili ne- (1). coronary artery bypass grafting.2 1 Klinika za kardiologiju.com Sažetak Abstract Revaskularizacija hroničnih totalnih okluzija (CTO) Percutaneous coronary intervention (PCI) of Chronic perkutanim putem je sve veći izazov u interventnoj kardiolo. (LV) systolic function. pre prethodnog koronarno-arterijski bajpas grafta (engl. thrombolysis in tyocardial infarction. it has been shown to improve left ventricular frakcije. reduces the need for late bypass surgery kao i na smanjenje mortaliteta u dugotrajnom praćenju. TIMI . c) moguć (neodređen): CTO sa TIMI protokom tromboliza u infarktu miokarda) protok kroz okludirani 0 i angiografskom anatomijom koja sugeriše dugotraj- segment jednak 0. koronarni sindrom ili pogoršanje angine bez druge mo- senzusnom dokumentu EuroCTO kluba. Branko Beleslin1. revascularization of CTO. Ukoliko postoje slučajeva. TIMI protok je široko prihvaćen sistem gradiranja. 1 | March 2016. Univerzitet u Beogradu. do 33% bolesnika sa značajnom koronarnom bolešću (2. simptomi (angina pektoris) ili objektivni dokazi ishemije Volume 67 | No. Klinički centar Srbije 2 Medicinski fakultet. ili posle Hronična totalna okluzija (CTO) koronarnih arte. sa mom ishemijom tokom poslednjih meseci. akutnog infarkta miokarda. Uprkos ovome i dalje nije jasno da li i kada treba re- a) siguran (angiografski potvrđen): u malom broju vaskularizovati ove lezije i na koji način. Also. slučaju skorašnjih akutnih ishemijskih epizoda (akutni Procena dužine trajanja okluzije vrlo često nije infarkt miokarda. In this ed- ovom radu dajemo pregled savremene definicije i indikacija za itorial. akutni koja traje duže od 3 meseca. a učestalost varira od 10 koji je uzrokovao okluziju. i kada postoji angiografski ili klinički nu okluziju (razvoj kolaterala. CABG) . lezije se mogu guće odgovorne arterije bar tri meseca pre aktuelnog an- da se klasifikuju kao hronične totalne okluzije kada je giograma TIMI (engl. npr.juricic@gmail. stvo TIMI protoka 0 bar 3 meseca pre planirane procedu- Uvod re. ment sugeriše tri nivoa sigurnosti (1): 3). we review the current definition and indication for the revaskularizaciju CTO.Medical Youth Review articles Savremene indikacije za perkutanu koronarnu revaskularizaciju (PCI) kod bolesnika sa hroničnom totalnom okluzijom (CTO) koronarnih arterija Contemporary indications for percutaneous coronary revascularization (PCI) in patients with Chronic total occlusion (CTO) of coronary arteries Stefan Juričić1. smanjenje potrebe za hirurškom revaskularizacijom. Euro CTO konsenzusni doku. Simptomatski bolesnici nakon uspešne revaskularizacije ventional cardiology. U odsustvu serijskih angiograma vreme trajanja noznih simptoma). Hronično okludirana koronarna arterija je čest na- kih podataka koji se tiču vremena kada se desio događaj laz na koronarnoj angiografiji. Siniša Stojković1. Perkutana improvement in quality of life after successful revascularization revaskularizacija CTO značajno utiče na poboljšanje ejekcione of CTO. Srbija Kontakt: stefan. total occlusion (CTO) is one of the major challenges in inter- giji. nestabilna angina ili pogoršanje angi- moguća. Prema poslednjem kon. kada postoji angiogram koji potvrđuje prisu. okluzije može da se proceni na osnovu dostupnih klinič. rija definiše se kao kompletna okluzija koronarnih arte. U as well as the reduction of the long term mortality.

re. (13) 40 meseci 6 pobošljanja regionalne kontraktilnosti u odnosu na PCI non-CTO CTO. (11) 180 meseci 6 Pobošljanje LVEF kod CTO pacijenata revaskularizovanih sa DES Baks i sar. Poboljšanje kva. dokazanim faktorima koji do- što znači da kolaterale nisu dovoljne (4). kao i redukcijom rizika od maligne aritmije (8). 2) bolesnici kod kojih je PCI bila neuspešna verovat- ralne funkcije adenozin stres testom je. Procena kolate..Stentovi sa osobađanjem leka. ronarnog protoka i miokardne perfuzije odražava se na dija je pokazalo da u prisustvu hronične totalne okluzije poboljšanje ventrikularne funkcije i smanjenje rizika od kolateralna cirkulacija može da uspostavi snabdevanje maligne aritmije miokarda krvlju i tako sačuva vijabilnos. PCI. i studija Suera i sa- ha sa pacijentima kod kojih PCI CTO nije bio uspešan. međutim. postoji vrlo malo podataka o dovoljstvu lečenjem.hronična totalna okluzija. Nekoliko metaanaliza koje su istraživale efekat procedurom primali različitu farmakološku terapiju. objašnjenja (smanjenje mortaliteta): U tabeli 1 su prikazane još neke studije koje poka- zaju benefit nakon uspešne revaskularizacije CTO. navode i da je bolja prognoza nakon uspešne procedure litetu (6. kao i nižu stopu nika pokazala je da prisustvo hronične totalne okluzije preživljavanja bez infarkta miokarda. Opservacione studije koje su pokazale benefit nakon uspešne PCI CTO Broj bolesnika Praćenje Rezultat Chug i sar. CABG i MACE. stabilnosti angine. no imaju težu formu koronarne aterosklerotične bolesti sa zala abnormalnu rezervu protoka u preko 90% slučajeva fibrozom i kalcifikacijama. Slično prethodnim studijama. “Seattle Angina Questionnaire” (SAQ) je u Do danas nije objavljena ni jedna prospektivna savremenoj literaturi apsolutno prihvaćen i validiran za randomizovana studija koja bi poredila PCI CTO sa opti. Do sada je procena kvaliteta života proceni kvaliteta života kod bolesnika sa CTO. U savre. MI. bolesnika sa neuspešnom procedurom. ispitanici u grupama sa uspešno i neuspešno izvedenom litet (5). Slično prethodnim metaanalizama. | Broj 1 | Izdanje 67 . Ovaj upitnik je zasnovan na pet ra- liteta života je stoga jedna od najznačajnijih indikacija za zličitih domena: fizičkom ograničenju. Autori nje mortaliteta i u kratkoročnom i u dugoročnom morta. (12) 27 meseci 5 Pobošljanje volumena leve komore i LVEF procenjeno MRI Uspešna PCI CTO povećava prokrvljenost miokarda i dovodi do ranog Cheng i sar. tre. revaskularizaciju CTO kod Izabranih bolesnika. kod CTO bolesnika pomoću SAQ korišćena u četiri studi- 2  Mart 2016. pokazala je značajno veći mortalitet u grupi bolesni- Hronična totalna okluzija ka sa neuspešnom procedurom (3) . međutim.infarkt miokarda. (10) 75 meseci 6 Pobošljanje LVEF I segmentne kinetike kod pacijenata bez ranijeg MI Nakamura i sar. percepciji bolesti i za- menoj literaturi. Tabela 1. može da bude potencijalni . poka. procenu kvaliteta života bolesnika sa koronarnom arte- malnom medikamentoznom terapijom. na neinfarktoj arteriji kod bolesnika koji se manifestuju Autori objašnjavaju razliku u preživljavanju time što su akutnim infarktom miokarda značajno povećava morta. 1) direktna dobit od ponovnog uspostavljanja ko- ba uzeti u razmatranje rekanalizaciju CTO.perkutana koronarna intervencija Hronična totalna okluzija i Procena kvaliteta života je važna mera uspešnosti revaskularizacije kod bolesnika sa koronarnom arterij- procena kvaliteta života skom bolešću. LVEF. DES. tivne registrе koji su poredili grupe proceduralnog uspe.confounding factor“. Medicinski podmladak Pregledni radovi ili vijabilnosti u zoni distribucije okludirane arterije. prinose proceduralnom neuspehu i većem mortalitetu (3).ejekciona frakcija leve komore. koja PCI CTO na preživljavanje pokazale su značajno smanje. i kardiovaskularni ishod U studiji Hoja (Hoye) i saradnika rezultati pokazu- ju značajno nižu stopu 5-godišnjeg preživljavanja u grupi Metaanaliza O'Konora S.A. Nekoliko stu. učestalosti napada angine pektoris. (O'Conor SA) i sarad. studija Prasada i sar. rijskom bolešću (15). Analizirane studije u ovim metaanalizama u verovatno povezana sa poboljšanjem funkcije leve komo- većini slučajeva predstavljaju restrospektivne ili prospek. radnika pokazala je značajno veće 10-godišnje preživlja- Za ovakav klinički rezultat postoji više ponuđenih vanje u grupi proceduralnog uspeha (9). 7).

koju snabdeva okludirana arterija u slučaju vijabilnog mi- Volume 67 | No. PCI Uspešna vs. TS . PL .percepcija bolesti. AS . U obe grupe je došlo do sličnog po- snika sa dijagnostikovanim CTO (18). bolesniku sa hronično okludiranom koronarnom ar- opremi i tehnikama perkutanih revaskularizacionih pro- terijom (26): bolesnici koji imaju CTO i simptome.Rose Dyspnoea .Score. sa saradnicima je ob- CTO lezija (uspešnost oko 98%)(2. EQ5D DP.zadovljstvo terapijom. CABG. AF . QOL . onda se ide na evaluaciju vijabilnosti u zoni ronarnu bolest (21. Galasi (A Gallasi). Tokom protekle javio algoritam po kojem bi trebalo pristupati svakom decenije došlo je do značajnog napretka u tehnologiji. ne-CTO leziji (19). segment leve komore u zoni okludirane arterije. PCI . naročito bolesnika koji nemaju višesudovnu ko- gde je CTO. RDS . 17). PCI CTO neuspešna PCI CTO CTO samo OMT Značajno poboljšanje Značajno poboljšanje Podjednako Uspešna u svim procenjivanim u svim procenjivanim poboljšanje u svim revaskularizacija parametrima u parametrima u procenjivanim CTO (PCI ili CABG) Rezultati uspešnoj PCI grupi uspešnoj PCI parametrima kod povezana je sa nasuprot neuspešnoj. PL. (16) Borgia i sar. (19) Wijeysundera i sar. Procenjivani parametri AF. 1 | March 2016. godine A. praćenja. neuspešna Uspešna vs.Medical Youth Review articles je. Dve od ovih studija su poredile rezultate SAQ kod bo. skora u SAQ domenima nakon jednogodišnjeg perioda lesnika kod kojih je uspešno rekanalisan hronično oklu. bošljanja u svim domenima SAQ tokom šestomesečnog čena medikamentoznom terapijom nisu uočene promene praćenja. dok je kod bolesnika kod kojih je urađena re- dirani krvni sud sa bolesnicima sa neuspešnim PCI CTO vaskularizacija u CTO teritoriji uočeno pobošljanje u 3 i dokazale značajno pobošljanje SAQ skora kod bolesnika domena SAQ (fizičkom ograničenju.8 Metoda procene SAQ SAQ-UK SAQ SAQ AF. postoje udvojena mišljena o mogućno- ti su za revaskularizaciju. Ukoliko bolesnik ima ili nema sti lečenja ovih bolesnika hirurškom revaskularizacijom simptome ali postoji akinezija ili diskinezija u segmentu (CABG).kvalitet života.hronična totalna okluzija.5 dimenzija Kliničke indikacije za pokušaj Kod bolesnika sa CTO koronarnih arterija odluka revaskularizacije hronične totalne okluzije o daljem lečenju prolazi kroz tri koraka: a) evaluaciju simptoma. PCI CTO vs. b) procenu veličine ishemije i/ili Terapijske nesigurnosti jednim delom vode pore- c) evaluaciju vijabilnosti miokarda miokarda u klo od tehnički kompleksnih procedura revaskularizacije zoni okludirane arterije. TS RDS. PL. Četvrta studija je poredila kva- je poredila revaskularizacione strategije (PCI vs CABG) litet života posle urađene PCI na CTO nasuprot PCI na sa optimalnom medikamentoznom terapijom kod bole. DP . TS. PL. Tabela 2. CTO . AS. 24. PCI non- Poređenje grupe non-CTO. U grupi koja je le. QOL.20). EQ5D Praćenje mesec 1 godine 4 meseci 6 godina 1 PCI CTO.7 78. Treća studija ne i percepciji bolesti). čija je uspešnost do skoro bila 60-70%. (17) Saffley i sar. što je značajno niže u odnosu na revaskularizaciju non- aktuelni Evropskog CTO kluba. učestalosti angi- sa uspešnom rekanalizacijom CTO (16. PL. CTO pomoću PCI. kandida- Drugim delom.učestalost angine. grupi nasuprot uspešno lečenih pobošljanjem kvaliteta neuspešnoj bolesnika u obe života grupe CABG . QOL AF. Studije čiji je cilj istraživanja bila procena kvaliteta života posle revaskularizacije CTO Grentham i sar. 22. sa cedura za lečenje CTO koje uz sve veće iskustvo operate- normalnom funkcijom leve komore ili hipokinetičan ra. (18) Broj bolesnika kojima 125 302 167 46 je rađena PCI CTO uspešnosti % 55 78 84.Seattle Angina Upitnik.stabilnost angine.Evropski kvalitet života . 25).fizičko ograničenje. AF. SAQ .koronarno-arterijski bajpas graft. EQ5D .perkutana koronarna intervencija.  3 . dovode do proceduralne uspešnosti oko 90% (21-23). U avgustu 2015.

vaskularizaciju iz 2014. Literatura okarda indikovana je revaskularizacija. Medicinski podmladak Pregledni radovi Algoritam 1. 2012 udruženja kardiologa prvi put pomenut 2004. PCI je kao pre. Konstantinidis NV. | Broj 1 | Izdanje 67 . ishemije i ukoliko ona zahvata 10 ili više procenata leve ko. Cheema AN. Fefer P. Tabela 3. 2. Christiansen EH. 1. Appropriateness of percutaneous revascularization of coronary chronic total occlusions: an overview. Hildick-Smith D. Tomasello SD. perspectives on coro¬nary chronic total occlusions: the 4  Mart 2016. Wright GA. EuroCTO Club . Yalonetsky S. što može da objasni otpor mnogih inter- teritoriji miokarda i/ili smanjenjem ventnih kardiologa da se oprobaju u ovoj zahtevnoj pro- simptoma angine ceduri (26). radi se evalucija ned J. Po May 15. Papafaklis MI. Weisbrod M.godine lečenje CTO perkutanim Bierstone D. Ukoliko bolesnik nema simptome. Oshe- rov AB. Preporuke za miokardnu revaskularizaciju (ESC/ Iako iskusni operatori imaju približno isti procenat EACTS. European Heart Journal. Recanalisation Razvojem pristupa u lečenju CTO. Karlas A. Galassi et al. Reifart N. more indikovana je revaskularizacija CTO. Di Mario C. skuplja oprema potre- Preporuke Klasa Nivo bna za ove intervencije i veća izloženost zračenju predsta- vljaju dva glavna limitirajuća faktora da ova metoda bude šire prihvaćena. Current pristupom ima IIa klasu. Aug 2015.8(1):139-45. Knudtson ML. Samuel M. Drugi glavni razlog zašto PCI CTO ima Perkutanu rekanalizaciju CTO treba samo IIa klasu preporuke u evropskim i američkim vo- razmotriti kod pacijenata sa očekivanom dičima je nedostatak randomizovanih studija za lečenje redukcijom ishemije u odgovarajućoj IIa B ovakvih lezija. EuroIntervention. Galassi AR. god. Esca- re ili hipokinetičan segment u zoni CTO. Gannot S. Galbraith PD. 2014) (27) uspeha PCI CTO i nonCTO lezija. Sparkes JD. Preuzeto iz: Alfredo R. Gershlick A. poslednjim Evropskim preporukama za miokardnu re. Strauss BH. Sianos G1. of chronic total coronary occlusions: 2012 consensus do- poručeni vid revaskularizacije u vodičima Evropskog cument from the EuroCTO club. Carlino M. Werner GS. nivo dokaza B (27). a ima normalnu pokretljivost leve komo.

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Zbog učešća u redoks-reakcijama.feritin. Oslobađanje iron is regulated by hepcidin that reduces the levels of ferropor- uskladištenog gvožđa iz ćelija reguliše hepcidin. heme and non-heme iron (Fe+2). Ključne reči: gvožđe. and ferritin within the cells. razloga se u organizmu vezuje za proteine. metabolizam. Njegova toksičnost se ispoljava kroz redoks-reakcije gvožđa se nalazi u okviru mišićnih vlakana (mioglobin). tako da apsorpcija gvožđa Zbog toga se svega 0. FPN1 . ROS). gvožđe predstavlja veoma toksičan me.transferin. gvožđa prisutnog u ljudskom organizmu koristi se u eri- U isto vreme. ferroportin. skrecije gvožđa iz organizma. iron possesses toxic properties. može naći kao fero. mora da bude izbalansirana da bi odgovarala njegovim laciji (2).(Fe+2) ili ferijon (Fe+3). Gvožđe se u organizmu proteina (4) (slika 1).com Sažetak Abstract Kod zdravih osoba se dnevno apsorbuje i izluči oko 1-2 Under normal circumstances. Due to its ability to participate gvožđe predstavlja veoma toksičan oligoelement. pa se u cirkulaciji Therefore. laboratorijski pokazatelji laboratory testing Spisak najvažnijih skraćenica: DMT1 . TF . TFR1 . iron is usually found coupled with proteins: transportuje u sastavu transferina. ciklusa. znojenjem i menstrualnim krvarenjem. u kojima učestvuje. troblastima za sintezu hemoglobina. Oko 80% zmu kao kofaktor mnogih proteina u čiji sastav ulazi. dnevnim gubicima. (ii) proteine koji organizam.feroportin 1. apsorpcija gvožđa mora da iron absorption matches iron losses through bleeding. S obzirom na to da ne postoji fiziološki Due to the lack of physiological pathway for iron excretion. razgradnjom tin 1. Ono što je krajnje neobično jeste da tokom mili- vanjem za proteine.transporter dvovalen- tnih metala 1. ukupno unetog gvožđa) i isto toliko izgubi deskvamaci- es. Iron is absorbed in the duodenum. a u ćelijama se skladišti u with transferrin in the circulation. 6  Mart 2016. odgovornih za om epitela sluzokože digestivnog trakta i epiderma kože. Fe+3 – ferijon. oksidativno oštećenje važnih bioloških makromolekula Kod žena se gvožđe dodatno gubi tokom menstrualnog (2). tako i u vanćlijskoj kao i u drugim tkivima organizma. kako u ćelijskoj. poput hidroksil-radikala (OH•). put ekskrecije gvožđa iz organizma. sweat odgovara njegovim gubicima deskvamacijom crevnog i kožnog and sloughing of epithelial cells. transferin. sadrže hem i (iii) enzime koji sadrže gvožđe (3). Fe+2 – ferojon.1% gvožđa nalazi slobodno u cirku. koji transportuje gvožđe iz ćelije. Preostalih 15-20% tal. froportin. Key words: iron. Razmenom U ljudskom organizmu se uravnoteženom ishra- elektrona gvožđe može da učestvuje u proizvodnji veoma nom dnevno apsorbuje oko 1-2 mg gvožđa (10-15% od reaktvnih kiseoničnih vrsta (engl. feritin. Ono ima ključnu ulogu u ljudskom organi. an iron transporter from the cell feroportina 1. Gvožđe se apsorbuje u duodenumu. svi ostali proteini koji sadrže gvožđe mogu se podeli- Gvožđe predstavlja važan oligoelement za ljudski ti na: (i) gvožđe-sumpor (Fe-S) proteine. FT . te se gvožđe transportuje kroz ćeliju on godina evolucije priroda nije razvila fiziološki put ek- i kroz cirkulaciju i skladišti u kompleksu sa proteinima. Medicinski fakultet Univerziteta u Beogradu Kontakt: drcoricvesna@gmail. u sastavu pomenutih sredini bogatoj kiseonikom (1). Medicinski podmladak Pregledni radovi Metabolizam gvožđa i laboratorijska dijagnostika pokazatelja metabolizma gvožđa Iron metabolism and laboratory testing for iron status Vesna Ćorić Institut za medicinsku i kliničku biohemiju. Extracellular export of stored epitela. as ili kao nehem gvožđe (Fe+2). vezano za hem leaves the body each day. 1–2 mg of iron enters and mg gvožđa. reactive oxygen speci. Iz tog in redox-reactions. metabolism. Ova toksičnost gvožđa se prevazilazi njegovim vezi.transferinski receptor 1 Uvod Pored proteina koji učestvuju u metabolizmu gvo- žđa. transferrin. ferritin. kompleksu sa feritinom. | Broj 1 | Izdanje 67 .

Distribucija gvožđa u ljudskom organizmu Slika 2. Apsorpcija gvožđa: DMT1 . Fe+2 – ferojon. HCP1 – engl.duodenalna citohromna b-reduktaza.transporter dvovalentnih metala 1. Fe+3 – ferijon. Volume 67 | No. PCFT – engl. DCBR . Heme-carrier protein 1. Proton coupled folate transporter.  7 .Medical Youth Review articles Slika 1. 1 | March 2016.

Heme-carrier protein dva atoma Fe+3 i jedan karbonatni anjon (17). nehem gvožđe se redukuje u Fe+2 na dva koja moraju da poseduju specifičan receptor na površini načina: (i) neenzimski. poput oksidacionog stanja gvožđa. kao i transmembranskog lija ciljnih tkiva i obrnuto. Proton coupled folate transporter) (5. sta- Adaptor-related 2) proteinskog kompleksa (7). Oslobađanje gvožđa Novija istraživanja su pokazala da pored TF posto- iz feritina u enterocitima i njegov transport u cirkulaciju je i drugi transporteri gvožđa. U uslovima normalnih u citoplazmu preko DMT1 transportera (11). niska vednost pH prisutna gvožđe od mesta apsorpcije do cijnih ćelija na kojima u želucu. nozin-trifosfatne (ATP) zavisne protonske pumpe. čime je TFR1 odgovoran za proteinskog kompleksa STEAP2 (engl. Divalent metal U novonastalom endozomu. dolazi do pada pH uslaskom H+ preko ade- za apsorpciju drugih dvovalentnih metala poput kobalta. kalcijumovih kanala na kardiomio- čega transportovano gvožđe oksiduje u Fe+3 dejstvom dve citima (20) i izoferitina (21). (ii) monoferitransferin. Mehanizmi ćelijskog preuzimanja slo- duodenuma i proksimalnog jejunuma. bodnog gvožđa i dalje nisu do kraja razjašnjeni. Hephaestin) (12). Ćelije su u mogućnosti da ferooksidaze: (i) ceruloplazmina (Cp). homeostazu gvožđa na nivou ćelije (18). u enterocitima iz apsorbovanog hema dejstvom hemoksi. nakon koje se oslobođeno (slika 1). 14). se nalazi transferinski receptor (slika 1). što cinka i kadmijuma. cije transferina i pH sredine (3). se prevodi iz Fe+3 u Fe+2 oblik aktivnošću metaloredukta- Sudbina Fe+2 nakon apsorpcije je dvojaka. usled prisustva transportera označenog kao Gvožđe vezano za hem nalazi se u mesu kao sastav. the prostate 3).6). dejstvom duodenalne cito. čiji inače niski nivoi mogu da porastu usled intravasku- Pored oksidacije. | Broj 1 | Izdanje 67 . DMT1) koji je. „Zrt-Irt-like protein 14“ (16). Gvožđe se bilnost gvožđa vezanog za TF zavisi od nekoliko faktora. 8  Mart 2016. pored gvožđa. gvožđe je prisutno u dve forme: i koštane srži (22). DMT1 omogućava posledično otpuštnje gvožđa (2. filtracijom na ovaj način oslobođenog hemoglobina kroz glomerulsku membranu bubrega. samo prema ćelijskoj membrani. proteinske konforma- genaze (3) oslobađa kao Fe+2. Veziva. pod dejstvom želudačne kiseline i svojih ćelija. Oslobođeno gvožđe apsorpciju gvožđa po principu simporta sa H+ (11). i (ii) hephestina (engl. gvožđa. Koristeći gradijent protona. Postojanje 1) i PCFT (engl. hephestin doprinosi i samom trasportu larne hemolize. 3). bez vezanog se apsorbuje kroz luminalnu membranu enterocita en. TFR1 dovodi do stvaranja TF-TFR1 kompleksa koji će- ni Fe+2 je bolje rastvorljiv i pogodan za apsorpciju preko lija preuzima procesom endocitoze zavisne od klatrina. đe vezuje bakar. Six-transmembrane epithelial antigen of njem gvožđa za apoferitin u enterocitima nastaje feritin. gde se izdvaja iz kompleksa i mali deo apsorbovanog gvožđa će se transportovati kroz oslobađa nazad u cirkulaciju (3). zajedno sa proteolitičkim enzimima varenja. Pored toga.10). Preostali apotransferin ostaje u kompleksu sa TFR1 i usmerava se potreba za gvožđem i adekvatnog unosa hranom. Gvožđe uneto hra. odgovoran i kompleks. u ovim istim ćelijama se (i) slobodno gvožđe i (ii) gvožđe u sastavu transferina odvija i razgradnja eritrocita. U toku Transferin (TF) je β1-globulin koji transportuje želudačne faze varenja hrane. Six-transmembra. Redukova. proteina koji tako- preuzmu i gvožđe prisutno u cirkulišućem hemoglobinu. Naime. bazolateralnu membranu enterocita. poput H-proteina i njego- omogućen je zahvaljujući feroportinu 1 (FPN1). koji po. najverovatnije uz Fe+3 i (iii) diferitransferin ili holotransferin. S druge strane. koji vezuje pomoć dva proteina: HCP1 (engl. prisutne u endozomu (10) i transportuje oblik u kome se gvožđe skladišti. ze STAEP3 (engl. gvožđa u slučaju povećane apsorpcije. Hem može naći u tri forme. transferinski receptor 1 (TFR1). kao: (i) apotransferin. poput askorbin. protein haptoglobin Transport gvožđa vezuje cirkulišući hemoglobin i transportuje ga do mono- cita i makrofaga retikuloendotelnog sistema slezine. Ne-hem gvožđe je prisutno u hrani kao Fe+3. jetre U cirkulaciji. niske koncentracije serumskog gvožđa povećava se nivo ske kiseline (8) ili (ii) enzimski. ekspresije transferinskog receptora na membranama će- hrom b-reduktaze (DCBR) (9). gvožđe koristi za ponovnu sintezu hemoglobina. ni deo hemoproteina: hemoglobina i mioglobina. koji u sebi sadrži TF-TFR1 transporter 1. hepatociti. kako bi se izbegao gubitak gvožđa gvožđa na bazolateralnoj membrani (13. iako se nom može se apsorbovati na dva glavna načina: (i) kao smatra da najveći afinitet prema ovoj formi gvožđa imaju gvožđe vezano za hem i (ii) kao nehem gvožđe (slika 2). transportera dvovalentnih metala 1 (engl. U uslovima redukujućih supstanci prisutnih u hrani. nakon vog receptora (19). U cirkulaciji se odgovorna je za izdvajanje hema iz hemoproteina. za posledicu ima konformacionu promenu TF i TFR1 i stoji na nivou membrane enterocita. koji vezuje jedan atom docitozom posredovanom receptorom. ove tri forme transferina sprečava moguće toksične efekte Feritin prisutan u hrani apsorbuje se preko AP2 (engl. Vezivanje TF za ne epithelial antigen of the prostate 2) (9. Da bi Gvožđe se u sastavu TF prenosi do ciljnih tkiva se apsorbovalo. Medicinski podmladak Pregledni radovi Apsorpcija gvožđa Slobodno gvožđe se u cirkulaciji najčešće nalazi kao gvožđe-citrat ili gvožđe-acetat u vrlo niskim kon- Apsorpcija gvožđa se odvija prevashodno na nivou centracijama (15).

zivanjem gvožđa za protein apoferitin. Ovaj mehanizam je pre svega zastupljen male količine gvožđa predaju skoro potpuno zasićenom u ćelijama tokom eritropoeze i zasniva se na direktnom transferinu. Nastaje ve- ra metala na svojoj membrani (32). kako na potencijalom (1) i (iii) mehanizmom pod nazivom „kiss. predmet istraživanja velikog broja najnovijih studija o kva služi kao dobar pokazatelj stanja depoa gvožđa u or.Medical Youth Review articles Skladištenje i korišćenje gvožđa u ćelijama žđe se u mitohondrijama koristi za već pomenutu sintezu hema i Fe-S proteina. (19). metabolizmu gvožđa i smatra se vrlo važnom u njegovoj ganizmu. nekih drugih ćelija (33). S druge strane. lizozomi su takođe bogati ovim oligoelementom tina 1. nivou enterocita. oslobođeno iz endozoma. stvom ćelijskog šaperona za gvožđe. znojenje i menstrualno krvarenje. Oslobađanje gvožđa iz FT je strogo kontrolisan poput makrofaga i monocita. sadrže relativno visoke nivoe bude transportovano iz ćelije preko transportera feropor- gvožđa. Nakon toga tinom 1. Apsorpcija gvožđa samim tim mora Mitohondrije su odgovorne za homeostazu gvožđa na da bude tako regulisana da odgovara dnevnim gubici- nivou ćelije. predaju se apoferitinu posredstvom heterogene nu. Va- agregata i. Kao Oko 80% gvožđa prisutnog u ljudskom organizmu što je već pomenuto. Regulacija nivoa feroportina 1 FT može da se nađe i u nukleusu i u mitohondrijama. gvožđe se transportuje u gvožđa predaje se apotransferinu u cirkulaciji. Poly rC binding protein 1-4) (24). S druge strane. Upravo u ovim organelama vrši se sinteza ma gvožđa kroz deskvamaciju crevnog i kožnog epitela. regulaciji. Svi ovi me- branom mitohondrija. enterocita gubi se njihovom deskvamacijom. kako na sistemskom. plazmatskoj membrani makrofaga i skladištenje gvožđa u jezgru feritina (25). žan deo tih mehanizama obuhvata kontrola pre svega Gvožđe se sporo oslobađa iz hemosiderina (26). Ljudski organizam poseduje nekoliko mehaniza- Hemosiderin je delimično deproteinisan feritin u obliku ma odgovornih za regulaciju metabolizma gvožđa. hema. kroz relativno ne. pre svega u tkivima i organima koji vrše štenju gvožđa zbog ekspresije membranskih transporte- sintezu hemoglobina i drugih hemoproteina. tako i u drugim ćeijama. gvožđa iz organizma. Hemosiderin je drugi Regulacija metabolizma gvožđa oblik uskladištenog gvožđa. gvožđa u organizmu gotovo sva količina apsorbovanog Nakon izlaska iz endozoma. ne postoji fiziološki put ekskrecije koristi se u eritroblastima za sintezu hemoglobina (4). Skorašnja istraživanja ipak sugerišu da postoji mo- Feritin (FT) je glavni protein odgovoran za skla- gućnost da i sami lizozomi mogu da učestvuju u skladi- dištenje gvožđa. U slučaju nedostatka su elektrona na respiratornom lancu mitohondrija (27). nerastvorljiv je u vodi. predstavljaju glavne ćelije proces. dok se veoma and-run“ (28). dolazi do oksidacije gvožđa u Fe+3 pod dejstvom teških Feroportin 1 se nalazi na bazolateralnoj membrani duo- polipeptidnih lanaca feritina. transporterom specifičnim za gvožđe (slika 1). nom proteolizom posredstvom receptora NCOA4 (engl. dok laki lanci omogućavaju denalnih enterocita. a uskladiš- mitohondrije na nekoliko načina: (i) indirektno. S obzirom na (ii) može da bude iskorišćeno u metaboličkim putevima to da autofagocitovani makromolekuli i organele. hanizmi su veoma strogo regulisani. Gvo- Volume 67 | No. kao i većine Fe-S proteina koji učestvuju u preno. koji su u mogućnosti da vežu do 4500 atoma gvožđa u samom Transport gvožđa iz ćelije jezgru proteina (23).  9 . transportuje preko proteina mitoferina 1 i 2 (29). Atomi Fe+2. od strane malog peptidnog hormona hepcidina bila je Izvesna količina FT može se naći i u cirkulaciji i kao ta. Pored citosola. Preostalo gvožđe u mitohondrija- ma skladišti se u formi mitohondrijalnog feritina (30). nog gvožđa se nalazi vezano sa FT. poput (poput sinteze hema u mitohondrijama) ili (iii) može da feritina i mitohondrija. za razliku od feritina. Pod normalnim okolnostima. dok se preostalih 5% nalazi u obliku hemosiderina (2). Apoferitin se sa- stoji od 12 teških i 12 lakih polipeptidnih lanaca. Hepatociti i ćelije retikuloendotelnog sistema. a zatim i skladištenja i reciklaže gvožđa. u ćeliji ima trojaku sudbinu: Nuclear receptor coactivator 4) (19) i proteazomalnom (i) može da bude uskladišteno u sastavu proteina feritina. proteina PCBP1 (25) u uslovima viška gvožđa dolazi do intenzivnijeg skladi- ili direktno. propusnu unutašnju membranu mitohondrija gvožđe se tako i na ćelijskom nivou. apsorpcije. Transport gvožđa u vanćelijski prostor je veoma klearne familije ribonukleoproteina označenih kao PCBP značajan za homeostazu gvožđa i posredovan je feropor- 1-4 (engl. Višak gvožđa koje se nalazi u sastavu feritina kontaktu membrane endozoma sa spoljašnjom mem. koji se pre svega odvija ciljanom autolizozomal- za skladištenje gvožđa u ljudskom organizmu. 95% uskladište. (ii) vođeno mitohondrijalnim membranskim štenja gvožđa usled povećane sinteze feritina. posred. koji nastaje kada se preva- ziđe kapacitet feritina za vezivanje i skladištenje gvožđa. Gvožđe. teno gvožđe se oslobađa iz svojih depoa. razgradnjom pod posebnim uslovima (31). oslobođeni iz endozo- ma. 1 | March 2016.

kao što su koncentracija gvožđa u serumu. kod transkripcije i (ii) posttranslacionih modifikacija (polia- dece. S druge strane. koncetracija transferina. HAMP. Nekoliko studija je pokazalo da povećan sadržaj gvo. iron response protein/iron response ele- ment) odgovoran za stepen degradacije iRNK neophodne za sintezu proteina koji učestvuju u ćelijskom preuzima. SL- karakteriše kongenitalnom i hipohromnom anemijom. gvožđa prevashodno u mozgu. kao i porasta žđa u organizmu. infekcija i zapaljenjski proces poveća. Poremećaji metabolizma gvožđa grubo mogu da se TIBC). Anemija u hro- rajuće mikro-RNK. kao i kod denilacija. Smatra se. 38). pokazano je za gene štenog gvožđa (37). njegovom skladištenju (FT) i gvožđa transportu iz ćelije (FPN1) (42).HFE pored nakupljanja gvožđa i niskog serumskog nivoa tran- protein. Laboratorijski pokazatelji metabolizma nju gvožđa (DMT1 i TFR1). 48). hipoksija. udruženo sa retinalnom fikovano je nekoliko proteina na površini hepatocita koji degeneracijom. SL- enterocitima ili uskladištenog u drugim ćelijama (slika C40A1) (43). unsaturated iron binding capacity. Najveću pažnju istraživača privukao je sferina. ceptora.sideropenijska anemija. Ovaj tip poremećaja metabolizma gvožđa može da kiselina koji se uglavnom sintetiše u jetri ekspresijom se klasifikuje na genetske (hemohromatoza. smatraju „senzorima za gvožđe“ (he. Medicinski podmladak Pregledni radovi Sistemska regulacija metabolizma gvožđa Poremećaji metabolizma gvožđa usled povećanog sadržaja gvožđa u organizmu Jedan od glavnih regulatora metabolizma gvožđa je hepcidin. Aceruloplazminemija vaju ekspresiju HAMP gena za sintezu hepcidina (37. od izuzetne važnosti za ćeliju je IRP/ vanjem gvožđa nakon parenteralne upotrebe (46. IRIDA) (46). total iron binding capacity. što dovodi do povećane sinteze hepcidina i posledično većeg broja mesta za inicijaciju transkripcije. | Broj 1 | Izdanje 67 . atransferinemija i druge) i stečene (primanje FPN1. injekcije gvožđa. Osim toga. IRE sistem (engl. nastalih transkripcijom ključ- procesa. nastaje usled mutacije na CP genu odgovornom za sin- dok nedostatak gvožđa. C40A1 (za ekspresiju FPN1 transportera gvožđa iz ćelije) niskim procentom saturacije transferina. udruženo do smanjene apsorpcije i smanjenog otpuštanja uskladi- sa postojanjem različitih promotora. U cilju laboratorijske dijagnostike poremećaja u metabolizmu gvožđa mogu se koristiti različiti pokazate- Poremećaji metabolizma gvožđa lji. Sideropenijska Pomenuti mehanizmi obuhvataju regulaciju (i) inicijacije anemija se najčešće javlja kod žena u premenopauzi. acerulopla- HAMP gena (34). ukupni ka- pacitet vezivanja gvožđa (engl. vezivanja gvožđa (engl. alternativno isecanje IRP/IRE sistem. TFR2. identi. nivoi transferinskog re- statka gvožđa. hemojuvelin. iron-refrac- nih za regulaciju gena uključenih u metabolizam gvožđa. peptidni hormon sastavljen od 25 amino. da hepcidin uzrokuje degradaciju FPN1 tako što hemohromatoza je najčešći tip naslednog poremećaja indukuje njegovu brzu ubikvitinaciju (36). Poremećaji metabolizma gvožđa usled nedostatka gvožđa Ćelijska regulacija metabolizma gvožđa Ovaj tip poremećaja metabolizma gvožđa takođe se može klasifikovati na stečene (anemija usled niskog ni- Ćelijska regulacija metabolizma gvožđa podrazu- voa serumskog gvožđa . anemija i eritropoeza tezu ceruloplazmina. Prisustvo 6). IRIDA je genetski poremećaj koji se poput DMT1 (za ekspresiju DMT1 transportera). 40). engl. UIBC). zumacija suplemenata koji sadrže gvožđe). žđa u okviru parenhima vitalnih organa. Usled mu- se. human hemochromatosis protein . slabim odgovo- i TFR2 (za ekspresiju proteinskog „senzora za gvožđe“) rom na oralnu upotrebu gvožđa i abnormalnim iskorišća- (3). u kome dolazi do formiranja depozita gvo- 1). Hereditarna naime. ja gvožđa u organizmu i (ii) na poremećaje usled nedo. što za posle. usled čega dolazi do nakupljanja smanjuju njegovu ekspresiju (39. tory iron-deficiency anemia. saturacije serumskog transferina. koji se smatra i glavnim regulatorom ek- spresije HAMP gena (41). zaduženog za izlazak gvožđa iz enterocita. ograničena proteoliza) koje pre svega ničnim bolestima nastaje kod dugotrajnih zapaljenjskih utiču na stabilnost iRNK. 10  Mart 2016. procenat saturacije transferina. makro. metabolizma gvožđa usled mutacije gena za proteine koji dicu ima smanjeno preuzimanje gvožđa apsorbovanog u učestvuju u homeostazi gvožđa (HFE. tacije na genu za transferin nastaje atransferinemija koju. kod stanja sa povećanim gubicima krvi. feritin i drugi (tabela 1). nekodi- nedovoljnog unosa gvožđa ishranom (47). karakteriše i visok nivo serumskog feritina (45). ataksijom i demencijom (44). kada se proizvode visoki nivoi interleukina 6 (IL- nih gena uključenih u metabolizam gvožđa. anemija meva nekoliko složenih mehanizama specifično zaduže- u hroničnim bolestima) i na genetske (engl. TFR2). mojuvelin. brojnih transfuzija krvi. pored TFR1. povećana kon- faga i drugih ćelija u vanćelijski prostor (35). slobodni kapacitet podele na (i) poremećaje nastale usled povećanog sadrža. Hepcidin reguliše nivoe transportera zminemija.

metabolizma gvožđa analiza stolice. Procenat saturacije transferina (%) se izračunava po sledećoj formuli: (100 x sadržaj gvožđa u uzorku) / TIBC. elektroforeza hemoglobina. unsaturated iron binding capacity. trudnoće.engl. za procenu metabolizma gvožđa mogu da utiču mnoga fi. UIBC . Tabela 2. hepatitisa.engl.Medical Youth Review articles Laboratorijski parametri kao pokazatelji meta. broj retikulocita prisustvo hemosiderinurije.engl. aspiracija kostne srži TIBC . merenje sadržaja retikulocitnog hemoglobina. akutnih i hroničnih inflamatornih stanja i dr. UIBC . 1 | March 2016. U cilju postavljanja dijagnoze specifičnih prisustvo hemoglobinurije. Na nivoe navedenih laboratorijskih parametara injekcija gvožđa. nekih oralnih kontraceptiva.   11 . oralnih suplementata koji sadrže gvožđe. unsaturated iron binding capacity Volume 67 | No. poput: menstrualnog ci- bolizma gvožđa u različitim oboljenjima prikazani su u klusa. Laboratorijski pokazatelji u stanjima poremećaja metabolizma gvožđa Koncentracija % saturacije Oboljenje/Pokazatelj serumskog TIBC UIBC Feritin transferina gvožđa Hemohromatoza Visoka Nizak Nizak Visoka Visoka Trovanje gvožđem Visoka Normalan Nizak Visoka Normalan Anemija usled niskog nivoa Niska Visok Visok Niska Nisko serumskog gvožđa Anemije u hroničnim Nizak/ Normalan/ Niska Nizak Niska bolestima Normalan Visok Normalan/ Nizak/ Hemolitička anemija Visoka Visoka Visok Nizak Normalan Normalna/ Normalan/ Nizak/ Sideroblastna anemija Visoka Visok Visoka Nizak Normalan TIBC . Total iron binding capacity. tabeli 2. Laboratorijski pokazatelji metabolizma gvožđa Laboratorijski pokazatelj metabolizma gvožđa Značaj testa za procenu metabolizma gvožđa Kompletna krvna slika Koncentracija hemoglobina - Veoma slab pokazatelj sadržaja gvožđa u Koncentracija serumskog gvožđa organizmu TIBC Mera kapaciteta vezivanja gvožđa za transferin Procenat saturacije transferina Mera ukupnog transferina koji nije zasićen UIBC gvožđem Koncentracija transferina Diferencijalna dijagnoza anemija Pokazatelj potrebe za gvožđem (npr. povišen Transferinski receptor kod sideropenijske anemije) Mera depoa unutarćelijskog gvožđa i najbolji Feritin pojedinačni pokazatelj sadržaja gvožđa Ostalo Periferni razmaz krvi. Procenat saturacije transferina (%) se izračunava po sledećoj formuli: (100 x sadržaj . ziološka i patofiziološka stanja koja su udružena sa pore- Tabela 1. mećajem metabolizma gvožđa.gvožđa u uzorku) / TIBC.engl. merenje nivoa α2 oboljenja udruženih sa poremećajem i fetalnog hemoglobina. Total iron binding capacity.

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lung. ganglionic neuron. which activates adrenergic re- the ANS is divided into two separate divisions . exercise is the stimulus to increase vagal tone (3). columbar region of the spinal cord specifically at T2 to tions.. thermal stress) (2). The nicotinic receptors are located at synapses vessels. ceptors on the peripheral target tissues. In the sympathetic nervous system. It has been shown that only regular dynamic specific cardiac receptors and vascular receptors (1). ).heart. The heart is innervated by vagal and sympathetic mitter from the parasympathetic nervous system. it is exocrine glands. kliničku farmakologiju i toksikologiju. pathetic innervations regarding its role in cardiovascular linergic. in both fibers. on the other ing outflow to the heart and blood vessels. The activation pathetic and sympathetic. of target tissue receptors causes the effects associated The parasympathetic system in human body ori. The autonomic nervous system (ANS) is predomi. the long postganglionic tral receptors (e.and post-ganglionic neurons of the sympa- efferent (exception are some afferent fibres that inner. excit- sinus and aortic arch and have a role in control of HR.g. etc. Its effects include control of heart rate (HR) and force of Responses from these receptors are excitatory and rela- heart contraction. The actions of autonomic nerves are It should be pointed out that is vagal tone declines mediated by the release of neurotransmitters that bind to with aging. the preganglionic and postganglionic neurons. atory responses. urinary bladder. system. thetic and parasympathetic pathways. Nicotinic receptors vate the baroreceptors and chemoreceptors in the carotid in contrast to muscarinic receptors produce rapid.parasym.. nerves and at the ends of cholinergic sympathetic fibers. blood pressure etc. Acetylcholine is the predominant neurotrans. liver. important to repeat the role of parasympathetic and sym- The parasympathetic postganglionic fibers are cho.g. stress (e. 14  Mart 2016. From there. Muscarinic receptors are located in the membranes of ef- nantly an efferent system which transmits impulses from fector cells at the end of postganglionic parasympathetic the central nervous system (CNS) to peripheral organs. When we know the basics of anatomy of ANS. The short postganglionic neurons are lo- cated near effector organs (e. constriction and dilatation of blood tively slow. Nervus vagus (or 10th cranial nerve) represents the cranial preganglionic out- flow of the parasympathetic nervous system (PNS) and Parasympathetic and sympathetic receptors the spinal cord parts of S2-S4 sacral spinal nerves serve in cardiovascular system as sacral part.g.). Most of the fibers in autonomic system are between pre. exercise. The medulla is a major site in the brain for regulat. The nucleus tractus solitarius (NTS) of the medulla L1. intestines. | Broj 1 | Izdanje 67 . with the sympathetic system (5). trachea. preganglionic neurons originate from the thora- bodies for both sympathetic and parasympathetic sec. leased norepinephrine. Medicinski podmladak Pregledni radovi The role of autonomic control in cardiovascular system: summary of basic principles Uloga autonomne kontrole kardiovaskularnog sistema: osnovni principi Branislava Medić Institut za farmakologiju. gins from craniosacral division. baroreceptors and chemoreceptors). The hypothalamus and higher centers modify the activity While the preganglionic neurons of both the sym- of the medullary centers and are particularly important pathetic and parasympathetic system secret acetylcholine in stimulating cardiovascular responses to emotion and (ACh) which is why they are referred to as cholinergic. and travel to a ganglion where synapse with a post- receives sensory input from different systemic and cen.com Introduction Acetylcholine can bind to two types of choliner- gic receptors called nicotinic and muscarinic receptors. Medicinski fakultet Univerziteta u Beogradu Kontakt: brankicamedic@gmail. the majority of the postganglionic sympathetic fibers re- Regarding anatomical and functional differences. It contains cell hand. neurons extend across most of the body (4). etc. kidney.

α1 receptor activation. There- velocity through the atrioventricular node. and on atrial and ventricu- lar coronary circulation). including the tion. norepinephrine) are all involved is dilatation of the vessels. The predominate effect of M3 receptor activation pamine (and ultimately. node. This is achieved by slowing the rate of However. fore. α1 adrenergic receptors are expressed in vascular pathetic efferent nerves are present throughout the atria smooth muscle proximal and stimulate vasoconstric- (especially in the SA node) and ventricles. α1. tion) and positive bathmotropic effect (increase in excit- ANS and cardiovascular system (adapted from reference 2) ability of heart). 6). notropic effect). which in- dial contractility (negative inotropic effect). epinephrine and do- lium. the activation of M2 receptors reduce the overall tivates only β1-adrenergic (mainly in the heart) and cardiac output and lead to decrease in force of myocar.Medical Youth Review articles Currently. acti- receptors are expressed in the heart. nephrine. for the purpose of blood pressure regulation (3. especially in nodal vation of sympathetic nerves causes vasoconstriction and and atrial tissue. Volume 67 | No. Stimulation by the sympa- thetic nervous system results in the following effects on the heart: positive chronotropic effect (increase in heart rate). positive dromotropic effect (enhancement of conduc- Graph 1. 1 | March 2016. The binding of acetylcholine to M2 re. AV node. and α2 adrenergic receptors. positive inotropic effect (increase of contractility). M3 receptors vasopressin. Sympathetic nerve fibers tonically release norepi- conduction system of the heart (2). to inhibition of AV node conduction velocity (negative α2 adrenergic receptors are expressed in vascular dromotropic effect) and decrease in excitability of heart smooth muscle distal from sympathetic nerve terminals. as well as by reducing the conduction finity for epinephrine than for norepinephrine. There are also some low expressions of β2 and α1 receptors in cardiomyocytes. while production by vascular endothelial cells. In the cardio- vascular system there are β1. Released from the adrenal gland. crease cardiac output and cause vasodilation. by stimulating nitric oxide in the initiation of the “fight-or-flight” response. In contrast to the heart. β-adrenergic receptors show greater af- depolarization. Sym. The actions of catechol- amines are mediated by α and β receptors. renin. and atrial impact afterload and vascular resistance which can again natriuretic peptide are all involved in water reabsorption alter cardiac output and blood pressure. which activates α1-adrenergic and β2-adren- In relation to the cardiovascular system the para. The M2 β2-adrenergic receptor distribution in the arteries. however. at least in the atrium (1. β2-adrenergic (mainly in vessels) receptors. but the is also influenced by numerous endocrine hormones. Beside that. M3 receptors are mainly expressed in vascular endothe. while the left vagus innervates the AV node. ergic receptors on blood vessels thereby providing basal sympathetic nervous system has two different kinds of vascular tone. angiotensin. Schematic representation of connection between CNS. emotional stress. increases the systemic vascular resistance primarily via ceptors leads to decreasing in heart rate (negative chro. Since there is greater α1-adrenergic than muscarinic receptors: the M2 and M3 receptors. β2. it is a matter of debate whether para- sympathetic stimulation can exhibit negative inotropic effects. β1 adrenergic receptors are expressed in the heart (in the SA node.   15 . (negative bathmotropic effect). while cap- illaries receive no innervation. as the vagus nerve does not directly innervate car- diomyocytes other than that of the sinoatrial and atrio- ventricular nodes. recent in vivo studies may suggest otherwise. cardiovascular function body do not have parasympathetic innervation. It is important to notice that most blood vessels in In addition to the ANS. but their role is The right vagus nerve primarily innervates the SA not yet fully clarified (7). Addition. 8). circulating epinephrine at low concentrations ac- ally. aldosterone. Their activation also elicits vasoconstriction. Enhanced sympathetic activity is particularly important during exercise. and during hemorrhagic shock. respective- stimulation of the parasympathetic system contrivutes ly. most vessels (arteries and veins) only receive sympathetic innervation.

demonstrated that endothelial dys- In addition.- Increased force of contraction: ß1 (and ß2) Inotropy (contractility) Decreased force of contraction - +++ Dromotropy (conduction Increased conduction velocity ++ Decreased conduction velocity . dilatation (ß2) +++ 0 Venous constriction Constriction (α1). The perfusion deficit is sufficient to cause alters endothelial function in animal studies (13). A similar finding was reported by Lemit. reduced nerve blood flow occurs very early after diabetic Conversely. system. modulates neuronal activity are unclear. the precise mechanism has not creased the release of norepinephrine from sympathetic been revealed. Hijmering tion. The inhibitory for an effect of the ANS on vascular function. NO is one important agent that appears to mod- duced sympathetic stimulation on NO metabolites in rat ulate sympathetic nervous system activity during blood heart tissue (10). periments showed that removal of the endothelium in- ated NO release. other aspects of endothelial function (15.- velocity) Bathmotropy (excitability) Increased excitability ++ Decreased excitability . 12). flow-me. reports et al. function may predispose patients with diabetes to impair- aggerated sympathetic nervous activity modifies ments in peripheral neural conduction (21). biochemical changes in experimental diabetic neuropa- al dysfunction in patients with hypertension (14). Although the mechanisms by which NO In contrast. These thy (20). particularly potent nonbaroreflex sympathoexcitatory stimulus where. pressure control. su.- Blood Vessels Arterial constriction Constriction (α1).. a acids (18). however. | Broj 1 | Izdanje 67 . 16). it has been demonstrated that ANS denervation induction (19). suggesting that endoneurial hypox- In humans. Medicinski podmladak Pregledni radovi Table 1. ia may produce many of the observed morphological and tor agonists has also been shown to improve endotheli. research suggests diated dilation in the femoral artery was not modified by that NO alters neuronal responses to excitatory amino sympathetic activation induced by the cold pressor test. Plater et al.. demonstrating a vascular contribution to regulation of fycantly impairs the flow-mediated dilation response the ANS are relatively limited compared with the evidence via an alpha-adrenergic mechanism (9). Another report showed that NO acts as a sym- findings suggest a contribution of sympathetic nervous patho-inhibitory substance within the central nervous tone on the baseline condition of vascular function. Experiments in animal models. Although the ANS may impact on vascular func- inating from the ANS on vascular function. However. experimental data suggest that ex. endoneurial hypoxia. the streptozotocin-induced diabetic rat. nerve terminals in rabbit carotid artery (17). the reverse may also be true. dilatation (ß2) +++ 0 The Effects of ANS on Vascular Function Effect of Vascular Function on the ANS Several reports have demonstrated an effect orig. The effects of sympathetic and parasympathetic system on heart and blood vessels (adapted from reference 2) Sympathetic system Parasympathetic system Heart Chronotropy (rate) Increased heart rate (HR): ß1 (and ß2) +++ Decreased heart rate (HR) . demonstrated that sympathetic stimulation signi. who demonstrated an inhibitory effect of exercise-in. have shown that as modification did occur in older healthy subjects (11. ANS modulation by α2-adrenorecep. 16  Mart 2016. in young healthy volunteers. Animal ex- effect of sympathetic activation is limited to shear medi.

et al. ical and clinical sense. 66: 1088-94. Hijmering ML. derstanding of these principles represents the basis for sequently developed deficits in lower limb nerve conduc. The cardiovascular system responds to not only Am J Cardiovasc Drugs 2006. Effects of the cold pressor test on muscle sympathetic nerve activity in Conclusion humans. reuptake. Vaseghi M. Interaction between en- imal experiments. et al. Other studies generally failed to show a beneficial effect 2006. Myers J. brachial flow-mediated dilation among patients with 9. or receptor sensitivity requires further investigation (22). Hypertension 1987. Thijssen DH. The role of autonomic nervous system on cardio- lowed for 3 years. whereas losartan improved VF. The role of the autonomic ner- Interventions to Modify ANS Pathways: vous system in sudden cardiac death. 10. 291: H3122-9. J Immunol 1991. J Am Coll Cardiol 2002. Cikim AS. flow-me- Thus. this paper is an attempt to gather in nervous system contributes to the age-related impairment of flow-mediated dilation of the superficial femoral artery. 8. Local mechanisms of blood flow control by It should be pointed out that cardiovascular system perivascular nerves and endothelium. et al. In an- 4. 11. 6. 279: R951-9. Hadley DM. Bonow RO. 10th vascular function (27). Freeman JV. acute but also chronic changes in blood pressure and ho. de Groot P. in subjects with Res 2007. Prog Cardiovasc Dis. 6: 343-8. In addition. Coutinho GC. ed a decline in circulating endothelin. 54: 690-7. one place just basic principles regarding primarily con- Am J Physiol Heart Circ Physiol 2006. Cardiovascular Physiology Concepts 2nd The most popular pharmacological agents used to ed. but is also heavily determined by 14. type 2 diabetes and hypertension. Assessment of sympathetic cardiovascular drive of beta-adrenergic receptor blockade (25. Miyauchi T. vation markedly reduces endothelium-dependent. Matsuda et al. Zipes DP. Victor RG. 3. further learning and research. Cikim K. ed. Yang Z. men treated with atenolol demonstrat. Strosberg AD. 7. Catecholamines stimulate the IFN-gamma-induced class II MHC expression on bovine brain capillary endothelial cells. The effect of moxoni- the autonomic nervous system as well as the endocrine dine on endothelial dysfunction in metabolic syndrome. cal and pathophysiological. In contrast. Iemitsu M. the effect of these agents crine control of cardiovascular function. Autonomic and endo- tor blockade agents. 15. References: Pharmacological or Therapeutic 1. Intense exercise causes decrease in expression of both endothelial NO syn- vestigation. Olijhoek J. Pernow J. 2014. et al. Cardiovasc administration in humans. For instance. Autonomic nervous system interaction with the car- endothelial function and decreased oxidative stress (24). In contrast. Sympathetic acti- ischemic heart disease (28). function remains controversial and requires further in. meostasis. Braunwald’s der-specific beneficial effect of beta-blocking agents on Heart Disease: Textbook of Cardiovascular Medicine. Beta-blocking agents 50(6): 404-19. 8: S95-106. 1 | March 2016. modify the activity of the ANS are beta-adrenergic recep. 7(4): 204-14. Gwathmey JK. nection between ANS and cardiovascular function. Dewey FE. Maeda S. Grassi G. the direct effect of the endotheli. Mann DL. Shivkumar K. Durieu-Trautmann O. 147: 2525-9. 39: 683-8. Böhm F. diovascular system during exercise. 2012.Health Sciences ed that treatment with carvedilol for 4 months improved Division. 2. Stroes ES. Circ Res 1990. tion. Prog Cardiovasc Dis. J Hypertens Suppl is controlled and influenced by not only a unique intrin. The importance of endothelin-1 for there are contradictory reports for beta-blocking agent vascular dysfunction in cardiovascular disease. However. Tschudi M. also demonstrat. Topal E. However. Hypertension 2009. Burnstock G. sic conduction system. Klabunde RE. et al. Leimbach WN.Medical Youth Review articles In recently diagnosed patients with diabetes fol. not only in the physiologi- tion velocity. 12. Lüscher TF. beta-adrenergic receptor blockade dothelin-1 and endothelium-derived relaxing factor in hu- may protect endothelial cells from the negative effect of man arteries and veins. Seals DR. et al. higher von Willebrand factor levels vascular function is significant and undeniable and un- predicted a subgroup of patients with diabetes who sub. Libby P. Volume 67 | No. Philadelphia.   17 . Gordan R. the effect of beta-blocking agents on vascular diated vasodilation. Am J Physiol Regul effect of α-adrenergic blocking agents on vascular func. World J Cardiol. atenolol did not im. Integr Comp Physiol 2000. 5. system. Philadelphia: Lippincott Williams&Wilkinns. in human hypertension: achievements and perspectives. Xie LH. little has been reported about the thase and tissue NOx level in hearts. on vascular function has not been fully clarifyed. but also in the pharmacolog- um on alterations in neurotransmitter release. 1990. Froelicher prove endothelial function. 48(5): 342-62. heightened sympathetic nerve activity (23). 26). 9: 429-36. suggesting a gen. Kooijman M. 13. 76: 8-18. Pennsylvania: Elsevier . Sympathetic In conclusion.

Deng LY. 35: 284-91. 25: 699-703. Mifflin SW. on endothelial function and oxi- 18. Matsuda Y. Interactions between autonomic Willebrand factor in hypertensive middle-aged men. 136: 17. Vasc Res 1992. Wiesli P. 39: 336-43. Effects of dox- nerve function. Pettersson K. Role of nitric oxide in central 26. Kaplan J. Patel KP. 226: sin I-converting enzyme inhibition and beta-blockade for 814-24. Born GV. 140: 753-9. Skantze HB. Diabetologia 1996. Hermann F. Atherosclerosis 1998. Schmelzer JD. beta-blockers and diuretic agents on reactive hyper- perimental diabetic neuropathy. Colombari E. Weisbrod RM. onists. azosin and atenolol on circulating endothelin-1 and von 22. compared with atenolol. Hypertension 1995. modulators on vascular adrenergic neurotransmission. 23: 569-72 beta1-adrenoceptor activation. Carvedilol im- Med 2004. 23. Schiffrin EL. Nakagawa K. J Am Coll Cardiol 2000. Dias AC. Psychosom 28. Cohen RA. Brain 1984. 18  Mart 2016. factor antigen predicts deterioration in diabetic peripheral 27. Exp Biol Med (Maywood) 2001. Tuck RR. et al. 66: 153-64. Higashi Y. 284: H234-40. Recent evidence for the involvement of catechol. et al. Low PA. Ford I. Arnesen H. calcium antag- and oxygen tension in the sciatic nerves of rats with ex. Hirooka Y. Effect of losartan. Am Heart J 2000. Sasaki S. stress causes endothelial injury in cynomolgus monkeys via Ann Med 1991. Flammer AJ. J 24. | Broj 1 | Izdanje 67 . Plater ME. NTS. A comparison of 19. 20. Elevated von Willebrand tients. for the development of cardiovascular disease. 107 (Pt 3): emia in patients with essential hypertension: a multicenter 935-50. et al. 34: 584-8. study. et al. Akita H. Harris KF. Effect of nitric oxide dative stress in patients with type 2 diabetes and hyperten- on excitatory amino acid-evoked discharge of neurons in sion. Tesfamariam B. Andersen P. Seljeflot I. et al. proves endothelium-dependent dilatation in patients with coronary artery disease. Psychosocial amines and of macrophages in atherosclerotic processes. J nervous system activity and endothelial function: a model Cardiovasc Pharmacol 1999. et al. J Hypertens 2007. Comparison of effects of angioten- sympathetic outflow. Dent MT. Matthews KA. Li YF. Cyclic GMP 153-61. Medicinski podmladak Pregledni radovi 16. 2 years on function of small arteries from hypertensive pa- 21. 25: 785-91. Terashima M. Am J Physiol Heart Circ Physiol 2003. 25. 29: 396-404. Endoneurial blood flow angiotensin-converting enzyme inhibitors.

  19 . 1 | March 2016.Medical Youth Partners Volume 67 | No.

tion of bone and try to understand what it really is. jasno je da glavni uzrok poveća.do structural and compositional characteristics of the aged bone nano-nivoa oslikava svu složenost koštane hijerarhijske orga. recent studies se usredsredile na ispitivanje kosti na nanometarskom nivou i focusing on bone at nano-scale revealed distinctive features of otkrile konkretne parametre koštanog matriksa čije poznavanje the bone matrix that could provide additional explanation for doprinosi boljem razumevanju povećane koštane fragilnosti the increased bone fragility in advanced age. In particular. Majority of those rare hours. with predi. Budući da se kosti mlade osobe u after low-energy trauma. fracture. nezavisno od starosnih promena na drugim age-related effects at other levels of bone hierarchical structure. How- ever. Marija Đurić Srejić Institut za anatomiju. from macro. this actually means that the appear at the femoral neck (hip fractures) (2). it is clear ne koštane lomljivosti potiče od strukturnih i fizičko-hemijskih that the main cause of easy bone fracturing originates from the karakteristika same stare kosti. mineral crystals lection for female sex (3) and occur due to a low-energy Why should we investigate bones at all? trauma (usually a fall from the standing height) (4). mineralni kristali Key words: Bone. Despite a few hours on bone microscopic slides viduals. ganized structure (5). aging. Ispitivanje kostiju od makro. are spent in Anat. nano. if we really want to omy where only the macroscopic appearance of bones is understand why bones do break. micro-level. one in youth and the other scales: at the macro-level. especially in elderly indi- presented. itself. during which medical stu.com Sažetak Abstract Prelomi kostiju su česti kod starijih osoba čak i nakon Bone fractures frequently occur in elderly persons even dejstva sile niskog intenziteta. Posebno su značajne skorašnje studije koje su ous determinants of bone strength. not too different from a piece Short overview of bone structure (bone of stone or a wooden stick. just an anchor- age point for the muscles. Given that the young individuals takvim uslovima ne bi slomile. It becomes more interest. nano. independent from kod starijih osoba. would not sustain a fracture under such conditions. we have to reject common macroscopic percep- in Histology and Pathology. considering that Studies of Medicine do not put much focus on bones. Observing bone. young bone would not break in such conditions. nivoima koštane organizacije.to nano-level. a fall is not sufficient to break the hip. Medicinski fakultet Univerziteta u Beogradu Kontakt: drpmilovanovic@gmail. starenje. | Broj 1 | Izdanje 67 . the characteristics of the bone itself. which means that it has differ- ing when one checks statistics of bone fractures: there are ent organization and appearance at different length actually two incidence peaks. er bones as something simple and dead. Hence. fraktura. Ključne reči: Kost. most of the students consid. In simple terms. identifying and treating a bone fracture is probably one of the first and key Bone is a very complex and hierarchically or- associations to bones in general. Medicinski podmladak Mini pregledni radovi “GOING NANO” in the field of osteology: Is bone fragility already determined at the level of bone mineralized matrix? “NANO” u osteologiji: Da li je lomljivost kosti već određena na nivou mineralizovanog koštanog matriksa? Petar Milovanović. The fact that bones can break hierarchical organization) does not change such an impression. So. Still. Fractures in aged individuals commonly (figure 1). bone looks quite different depending on the scale of ob- 20  Mart 2016. shows us the nizacije i ukazuje na različite faktore od kojih zavisi otpornost complexity of bone hierarchical organization and reveals vari- kosti na prelome. the main cause of easy bone fracturing must originate from dents acquaint themselves with bones. and nano-level in senescence (1).

(B) Section of the femoral neck showing cortical (outer) and trabecular (inner) bone compartments. 1 | March 2016. strength. Main properties of the bone mineral component Mineral characteristic Parameter Method of detection Quantitative backscatter Degree/distribution of mineralization Calcium weight-percentage electron imaging Chemical Degree of carbonate substitution Carbonate-to-phosphate ratio Raman. Therefore. and nano-level (E-F). (A) A view on the proximal half of a human femur (white line shows the location of the section in B). (D) Microscopic view (backscatter SEM) of cortical bone showing numerous osteons (white circle) with Haversian canals (white asterisk) in the middle. and the most numerous osteocytes) (7). AFM – Atomic force microscopy. XRD – X-ray diffraction. (F) Transversal section of collagen fibrils (schematic). (E) Longitudinal view (schematic) on collagen fibrils covered by mineral crystals. Non-collagenous proteins are not shown here for simplicity.Medical Youth Mini review articles Figure 1. FTIR characteristics Type of hydroxyapatite Calcium-to-phosphorus ratio EDS Shape and size of mineral crystals Crystal length SEM. Note that mineral crystals lie both between and inside the collagen fibrils (interfibrillar and intrafibrillar mineral). Most of the bone volume is occupied by (cancellous or spongy) compartment consisting of a net. AFM. Volume 67 | No. XRD Crystal perfection Crystallinity XRD. (C) Microscopic view (SEM) of trabecular bone showing multiple interconnected trabeculae. microscopic (C-D). servation. to profoundly understand what work of interconnected bony plates or rods (trabeculae) bone really is and to comprehend what determines bone that fills the bone interior (5. one has to consider bone features at various hi. Going down to the microscopic level. it exposes erarchical levels.   21 . EDS – Energy dispersive X-ray spectroscopy. extensively interconnected and intensive- of cortical and trabecular compartments (figure 1B-D). bone as a living tissue composed of cells with specific First. FTIR Morphological characteristics Roughness Surface roughness AFM-PSD Structural complexity Fractal dimension AFM-PSD Abbreviations: Raman – Raman spectroscopy. extracellular matrix (bone material or bone matrix). teoclasts. 7) (figure 1B-C). AFM-PSD – AFM-based power spectral density. Bone structural organization: Macroscopic (A-B). It is frequently visible with a na. Table 1. Raman. SEM – Scanning electron microscopy. in contrast to the porous trabecular organism (8-10). macroscopic observation shows the shape functions (bone-forming osteoblasts. bone-resorbing os- and the size of the bone. These ked eye after making a cross-section that bones consist cells are active. FTIR – Fourier transform infrared spectroscopy. ly communicating to maintain or adapt bone structure to Cortical (or compact) bone is the outer bony layer with a the local mechanical and global metabolic needs of the very low porosity (6).

Moreover. 11. 20). ing of bone at nano-level could not be possible without terial or matrix level) reveals that bone matrix is a na. light or electron beams and system of lenses to obtain the ever. 11). Clearly. cently applied to bones (11. A number of chemical and mainly bare collagen fibrils. advanced the knowledge on bone nanostructure. For ment to fibrils.D). | Broj 1 | Izdanje 67 . Atomic force microscopy (AFM) is another powerful tool eral crystals. Bone min. understand. while fracture surface of bo- morphological properties can be analyzed to describe vine trabecula exposed mineralized collagen fibrils (23). and contribute to bone instance. Collagen different material properties and allow mechanical char- type I – the main constituent of theorganic phase (90%) acterization of materials in addition to imaging (AFM (5-6) – is organized in fibrils that are reinforced by min. In contrast to light or electron microscopy that use Bone mineral is carbonated hydroxyapatite. especially hardness of the outer surface of human trabecular bone showed and strength of bone (15-16). uniform shape and size in the same bone (22). 14). 21). applying AFM on bovine vertebral trabecu- toughness (5. probe to physically “touch” the specimen and provide a eral is organized in particles of various sizes (commonly 3D image of the specimen’s surface topography (figure termed “mineral crystals” irrespective of their true degree 2). Analysis mines bone mechanical properties. and for characterization of nanomaterials that has been re- water (5. bone mineral component (table 1). organic phase (mostly collagen fibrils). crystals and cells.B) and with bone fracture (C. image of the specimen. Scale bar = 200 nm. The remaining 10% of organic First AFM studies were mainly qualitative and phase are non-collagenous proteins that provide attach.C) and corresponding Phase images (B. mostly of plate-like shape (13).D): AFM Topography (A. AFM allows great spatial resolution without the need of excessive sample prepara- Nanostructure of bone tion. Medicinski podmladak Mini pregledni radovi Going further down to the nano-level (bone ma. 22  Mart 2016. eral crystals (figure 1E-F). AFM can distinguish between the areas of of crystallinity). nanoindentation) (11. its exact physicochemical characteristics are com. how. lae showed that interfibrillar mineral crystals are not of It is believed that the mineral part mainly deter. the use of advanced technology (table 1) (9. no-composite material (figure 1E-F) composed of min. Cortical bone nanostructure at the femoral neck of an elderly woman without bone fracture (A. 17-19). the AFM uses a sharp mechanical plex and still attract research attention (12-13). The mineralized fibrils detected on fracture surfaces led to the assumption that the mineral-to-mineral interface Figure 2.

Draughn RA. Liu B. hip fractures (21). Jobke B. Dennison EM. et al. Milovanovic P. 6. et al. Van Staa TP. McCreadie BR. Fratzl P. Melton LJ. that 2008. Bone fects at other levels of bone hierarchical structure. compared to young women (30). bone fracture. Sacrificial bonds and hidden length material differs across age (21). 16. Bone 2014. J. same rate in all individuals.47(1):41-54. Nat Mater 2005. femoral neck revealed an increased size of mineral crys. Weaver JC. aged bone matrix dissipate energy as mineralized fibrils separate during showed less elastic behavior in the elderly. Dennison E. provided direct evidence that the quality of bone H. (27) (Figure 2). Currey J. Cooper C. provide new insights into the problem of bone fragility. sensitivity. Osteocytes subjected to fluid ological approach with AFM – proved useful for explain.Medical Youth Mini review articles is the weakest link in bone and that fractures are mostly References initiated there (23).1(6):624-49. 18. hence. AFM studies in animal bone elegant- ly showed collagen fibrils after removal of the mineral 1. editor. Morris MD. Osteoporosis epidemiolo- of age-related effects on the bone mineral (21.3-22. editors.7(9):7542-51. Wiley Interdiscip Rev Nanomed Nanobiotechnol 2009. Hip Fracture Incidence in Relation to Age. Cooper C. Namely. Birkedal women. larger particles have a decreased material strength (31).42(3):456-66. N. The many adaptations of bone. Apart from increased crystal size. 2.: Princeton University Press. Spectral Density analysis (PSD) – another novel method. These findings sug- Bone mechanics handbook. 2000. Boca Raton: CRC Press. Meno- Recent AFM studies made a step forward by in. et al. 3. p5/1-5/33. Bonucci E. Thurner PJ. PLoS Med 2009. Conversely. and proved AFM as a powerful tool for the assessment 4. ACS Nano 2013. Amling M. decreased fractal di.1-68.59:37-43. Semeins CM. Bone treatment with collagenase allowed better visualization 2001. femoral neck cortex in the fracture group. Busse B. Hassenkam T. Roddam brils (15. Djonic D.29(6):517-22. particles by means of EDTA or NaF (24-26). Balkwill A. trabeculae in the elderly denoted their reduced structural 12. but not at the book. Bone mechanics hand- Crystal size increases generally with aging. Fantner G.4:612. structures composed of and the bone–implant interface. Beral V. Namely. Hahn M. these nano-structural differences (30) contribute 7. Boca Raton: CRC Press. Hip fractures in of bone minerals revealing that 70% of mineral is placed the elderly: A world-wide projection. Bone min- es of the matrix aging. complexity and surface roughness.6(11):e1000181. HRTEM Study of the min- fragility in aged persons (29). eral phases in human cortical bone. eralization density distribution in health and disease. nano-structural evaluation of the 2003.2(6):285-9. Currey JD. J Biomech Taken together. Curr Rheum Rep 2008. Sudhaker Rao D. Cooper C.   23 . independent from age-related ef. Paschalis EP. Xin R. Cole Z. 15. Jee WSS. Integrated bone tissue physiology: Anatomy and to increased fragility of the femoral neck in aged females. The external cortical surface 8. Osteoporos Int around collagen fibrils (interfibrillar or extrafibrillar min- 1992. de Vries TJ. In: Cowin SC. editor. Djuric M. the 9. Princeton. In: tals in aged. In: Cowin SC. Everts V. Bone mineralization.10(2):92-6. ing. 1 | March 2016. Milovanovic P. another factor leading to works: Morphological Implications for Altered Mechano- increased brittleness and impaired resistance to fracture. the nanometer scale (29). which in turn leads to increased brittleness and bone 13. Osteocytic Canalicular Net- degree of mineralization (27). nate-osteoclasts: Changes in osteoclast morphology and tained a hip fracture displayed larger mineral crystals function induced by antiresorptive nitrogen-containing than in age-matched women without skeletal diseases bisphosphonate treatment in osteoporosis patients. our AFM study of the trabecular bone at the 5. Finally. Pechenik L. ing differential trabecular bone fragility across the age at Bone 2007. 26). Mechanical testing of bone of thumb in materials science.12(9):B552–B7. p. Widjaja E. 17. Roschger P. Campion G. Bisphospho- of the femoral neck in postmenopausal women who sus. Namely. A. Leufkens HGM. Reeves GK. showed higher Püschel K. Boskey AL. Boca Raton: CRC Press. Tan SD. Chen T-c. et al. Epidemiology of fractures in England and Wales. Atomic force microscopy and indenta- mension of the interfibrillar mineral of the femoral neck tion force measurement of bone. Klaushofer K. and Age at Menopause: Prospective Analy- troducing quantitative analysis of bone nanostructure sis. Klein-Nulend J.36(10):1487-95. As a rule An YH. Finney WF. Zimmermann EA. eral) (26). bone matrix revealed particular mechanical consequenc. J Mater Sci perimental insights into the determinants of age-related 2012. gest a decreased ability to dissipate energy during load. while the rest is located inside the collagen fi. Basic composition and structure of bone. Physiology. Currey JD. in young and elderly 14. recent nano-scale mechanical assessment 2010. Bone tissue compositional Volume 67 | No. Adv Eng Mater. pausal Status. 2001. flow inhibit osteoclast formation and bone resorption. Bones: structure and mechanics. Kindt JH. p. 2001. adding new ex. Wang N. 2002. Banks E. The structure and mechanics of bone. of the femoral neck trabeculae. Application of Fourier-transform based Power 10. 27-30). gy update. Leng Y. 11. Kuijpers-Jagtman AM.41(5):745-51.

Imaging of bone ultrastructure using atomic force microscopy. Kreutz C. et al. Jungmann R. Riedel C. Bone 2006. Thurner PJ. Thurner P. tors. et al. J Mater Sci Mater Med 2002. Neskovic O. Milovanovic P. et al. J Anat 2012. Potocnik J. Microscopy and Microanal- 22. Taniguchi M. High-resolution AFM imaging of intact superolateral femoral neck in elderly hip fracture patients and fractured trabecular bone. 21.66(9):1202-8. Exp Gerontol 2014. py. Biomaterials 27. aged.18(13):135102. Oroudjev E. Hansma PK. duced hydroxyapatite–collagen detachment on bone frac.7(9):3446-51. Schitter G. Compos Sci Technol S. Milovanovic P. vs. Djuric M. Rabinovych O. Djuric M. et al. Finch MM. Modern research and educational topics in microsco- ture. Nikolic sist fracture on several length scales. Neskovic O. Zivkovic V. 30. Kindt ter 2011. mineralized matrix in AFM topography images: Potential 23. Hassenkam T.45(0):46-55. compositional and 20. et al. Díaz J. Rakocevic Z. Hansma PK. Nanotechnology chanical behavior of CaP and CaP/polymer composites for 2007. Potocnik J. Djonic D. bovine femoral cortical bone at the collagen fibril-mineral cyte network reveal trends in quality of young. Milovanovic P. bone apposition during aging. Sasaki N. Schitter G.7(1):16-30.37-48. Bone 2004. ysis 2013. Milovanovic P. cations for bone fragility in elderly women. et al. Goto T. Scheidt Av. Fantner GE. Lauer ME. edi- 24  Mart 2016. Kindt correlates of bone tissue age and bone fragility in female JH. tion of trabecular bone in the lateral femoral neck: Impli- 24. teoporotic and antiresorptive-treated bone.55:19-28. Atomic force microscopy characteriza- bone mechanical properties at material level: Nanoinden. Badajoz: Formatex. level. 31. Acta Biomater Fantner GE. Hahn 2015. Hierarchical interconnections in the femoral neck trabeculae. A new technique for imaging mineralized fibrils on bovine 29. Weaver JC. A review of the me- ture surfaces by atomic force microscopy. Milovanovic P.35(1):4-10. p. Nakata M. Cutroni JA. 2011. Zivkovic 28. Nikolic S. | Broj 1 | Izdanje 67 . 26. Schitter G. 19. derly women: potential nanostructural traces of periosteal Exp Gerontol 2012. Acta Bioma- 25. Nano-structural. Thurner PJ. Milovanovic P. Medicinski podmladak Mini pregledni radovi differences in women with and without osteoporotic frac. Thurner PJ. Wagoner Johnson AJ.13(3):333-7. Multi-level characterization chi K. healthy aged controls. applications in bone replacement and repair. Age-dependence trabecular bone fracture surfaces by atomic force micros. Nikolic S. Mater Res Soc Symp Proc 2005. et al. tion of the external cortical bone surface in young and el- tation study of the femoral neck in women by using AFM. Tagami A. et al. Morse micro-architectural signs of cortical bone fragility at the DE. Djonic D. nano-composite material bone: Fibrillar cross-links re. os. Potocnik V. Krause M. of power spectral density and fractal dimension of bone copy. Djonic D. Bosma BL. Kindt JH.874:59-65. In situ observation of fluoride-ion-in. Age-related deterioration in trabecular J. Herschler BA. Djuric M. M. JH. Hiki- Herzog L. Fantner GE. In: Méndez-Vilas A.19(5):1341-9.221(5):427-33. 2007. Stoiljkovic M. Fantner GE. Nanostructure and mineral composi- 2006. Kindt JH. Zimmermann EA. Schitter G. Rakocevic Z. Atomic force microscopic studies on the structure of of human femoral cortices and their underlying osteo.39(6):1190-5.47(2):154-9. Djonic D. Nikolic S.

stanojevic@med. metilprednizolon. Pod istim uslovima je inhibitory effect of the MP on production and expression of značajno više izraženo inhibitorno delovanje MP na proizvodnju the genes for IFN-γ. IL-17 in mitogen-stimulated lymph node cells (LNC) as well sintetski glukokortikoid. zani sa najnovijim saznanjima o efektima GK na Th17 ćelije a synthetic glucocorticoid. i Th17 limfocita koji su glavni ćelijski izvori IFN-γ. citokina koji nificantly more pronounced. IL-17. methylprednisolone.ac. susceptible laboratory animals. the same phenomenon proizvodnju IL-17 i IFN-γ zapažen je i na ćelijama izolovanim was observed: the proportion of IFN-γ producing. 1 | March 2016. lekova koji se primenjuju kao terapija inflamatornih i effect of glucocorticoids (GC) on expression and production autoimunskih oboljenja. Ključne reči: autoimunost. Nije precizno definisano ni mesto action of GC is not precisely defined. under the same conditions i antigenom na dozno zavisan način. Različita osetljivost Th1 achieves its effects not only in the peripheral lymphoid tissues. Razumevanje molekulskih effect of the GC has been observed in other animal models and mehanizama koji leže u osnovi relativne otpornosti Th17 in human disease. još of IL-17 has not been thoroughly tested yet.Medical Youth Mini review articles Different response to glucocorticoid therapy in autoimmune diseases of CNS Različit odgovor na glukokortikoidnu terapiju primenjenu u autoimunskim oboljenjima centralnog nervnog sistema Željka Stanojević. Different sensitivities of Th1 and Th17 IL-17 na delovanje GK zapažena je na drugim animalnim cells that are major cellular sources of IFN-γ or IL-17 in the modelima i u bolestima kod ljudi. Th17 cells and interleukin (IL-17). eksperimentalni autoimunski Key words: autoimmunity. This paper presents the dejstva GK.rs Sažetak Abstract Th17 ćelije i njihov glavni citokin interleukin 17 (IL-17). Understanding the molecular mechanisms ćelija na delovanje GK od ključnog je značaja za razvoj novih underlying the relative resistance of Th17 cells on the operation strategija u lečenju onih oblika autoimunskih i hroničnih of GC is very important for the development of new strategies bolesti koje su rezistentne na delovanje glukokortikoida. koji su pove. odnosno but also in target tissue. Medicinski fakultet Univerziteta u Beogradu Kontakt: zeljka.bg. their signature cy- imaju ulogu u patogenezi autoimunskih oboljenja centralnog tokine. have the main role in the pathogenesis of autoimmune nervnog sistema kao što su multipla skleroza i eksperimentalni diseases of the central nervous system such as multiple sclerosis autoimunski encefalomijelitis (EAE). in the treatment of those forms of autoimmune and chron- ic diseases that are resistant to the effect of glucocorticoids. experimental autoimmune enceph- encefalomijelitis. čime je takođe of IL-17 cells were reduced in cells isolated from MP treated pokazano da MP ostvaruje svoje efekte ne samo u perifernom rats in comparison to control rats which indicates that MP limfnom tkivu. The (GK). Isti obrazac delovanja MP na ekspresiju i effects of MP applied in vivo in EAE. Interestingly. na ekspresiju i proizvodnju IL-17. induced in IL-17 na modelu EAE indukovanog kod pacova. inhibira in vitro proizvodnju IL-17 as in myelin basic protein (MBP)-stimulated draining LNC in od strane ćelija limfnog čvora pacova stimulisanih mitogenom dose. Marija Mostarica Stojković Institut za medicinsku i kliničku biohemiju. However. but not all iz kičmene moždine pacova obolelih od EAE. Th17 Volume 67 | No.   25 . IL-17. Also. Th17 alomyelitis. a cytokine that TH1 cells generate.dependent manner. Efekat glukokortikoida and experimental autoimmune encephalomyelitis (EAE). when we analyzed stvaraju Th1 ćelije. is sig- i ekspresiju gena za interferon gama (IFN-γ). inhibit in vitro production of kod ljudi i na animalnim modelima. main results of the Doctoral thesis devoted to studies of GC ske disertacije posvećene ispitivanju dejstva GK na proizvodnju on the production of IL-17 in the model of EAE. Metilprednizolon (MP). the site of uvek nije detaljno ispitan. već i u ciljanom tkivu. U ovom radu su prikazani glavni rezultati doktor. Methylprednisolone (MP).

The lack of deepen- these effects with corresponding effects on IFN-γ. IL-6. our results showed that MP inhibited IL-6 production in LNC population devoid of Glucocorticoids and central nervous system T lymphocytes. a synthetic glucocorticoid fore. although it was acute relapses in MS (10). IL-21 and IL-22 (8). convincingly demonstrate that MP inhibits IFN-γ pro- entially regulate the production of Th1 and Th2 cyto. Indeed. decreased IL-17 and IFN-γ expression and production in GF-β). IL-18 (12). | Broj 1 | Izdanje 67 . It seems that the direct effect of MP and they have been shown effective in the treatment of on T cells is crucial for IFN-γ inhibition.T. MP expression of anti-inflammatory cytokines (e. the same phenomenon was not observed Glucocorticoids (GC) are used to treat a wide with IFN-γ. (Con A) and determined by measuring the expression of toimmune pathogenesis (1). Experimental autoimmune IL-17 and IFN-γ mRNAs in cultivated cells by PCR and encephalomyelitis (EAE). as IL-1. tion of IL-12. ing IL-17 inhibition by the addition of exogenous IFN-γ ined results partially contributed better understanding of is unexpected but it might be explained by saturating ef- glucocorticoids on of Th-17 effects in health and disease. an organ-specific autoimmune concentration of the cytokines in culture supernatants by inflammatory disease. conducted through inhibition of to be mediated by myelin-specific CD4+ T lymphocytes. inflammato. The observed inhibition as its experimental model (2). allergic and autoimmune diseases T cells as on LNC.Results Since ample evidence indicates that GCs differ. require IL. they still drug on IL-17 expression and production. the aim of my thesis (13) was to analyze effects antibody eliminated such an inhibitory effect (14). cytokines. induced in susceptible laboratory ELISA. Namely. fect of IFN-γ that remained upon MP treatment. but interestingly.g. shows significant similarities to MS in both clin. demyelinating disease of the CNS with a putative au. remained after MP action is still adequate to inhibit IL-17 ic inflammatory diseases frequently treated with GCs. T cells is supported by recent findings that GCs directly mune responses. these production of IL-17. as well as of EAE (11). production since the addition of anti-IFN-γ-neutralizing Therefore. We can 26  Mart 2016. the effect of MP was studied in an in vitro system consisting of rat lymph node cells (LNC) stim- Multiple sclerosis (MS) is a chronic. at least in mice. It suggests that cells. potency of the direct effect of MP on IFN-γ production in since GC down-regulate both innate and adaptive im. it seems that although MP inhibits IFN-γ. respectively (3. the influence of more specifically by Th1 and Th17 cells and their signa. Multiple convincingly demonstrated that MP inhibit the produc- mechanisms are proposed to explain glucocorticoid ther. production (14). as well as in cells isolated from the tion of regulatory cells (12). while enhancing the on lymphocytes from immunized rats was analyzed. at least partially. both direct influence on T cells and indirect influence 23 for survival and expansion (7) and secrete a profile of on other LNC populations contributing to T cells IL-17 inflammatory cytokines including IL-17 and IL-17F. MP is less effective if the drug is applied to purified T ture cytokines. TNF-α. a shift in the cells from lymph nodes draining the site of immunization population of Th cells from Th1 to Th2 and the propor. irrespec- kines (12) it is important to know how GCs affect the tively of experimental setting used. IFN-γ and IL-17. production of various proinflammatory cytokines and Similar results were obtained when the MP effect mediators (e.g. However. MP decreased expression and production of both animals. The apeutic efficacy in autoimmune damage to the CNS (12). Obta. ulated with polyclonal T cell mitogen. and compare cooperate to limit IL-17 generation. TNF-α). RorγT transcription factor. IL-6. T-cell apoptosis and redistribution. TNF-α. Concanavalin A ry. such main feature of their patophysiology is neuroinflamma. the essential transcription factor of lymphocyte proliferation and lymphocyte expression and IFN-γ-producing cells (16). IL-6.4). necessary for Th1 differentiation (15). However.6). tory reaction (9). In line with these data. Additionally. IL-1β. Both diseases are assumed was. its effect was less pronounced ical and pathological aspects and therefore is widely used on IL-17 than on IFN-γ (14). develop from naive CD4+ T cells action of MP on IL-17 generation in rat LNC includes under the influence of TGF-β and IL-6 (5. Th17 cells than to mixed population of LNC. GC have been shown to inhibit interfere with T-bet. a cytokine accused to be critically results present evidence that small production of IFN-γ involved in the pathogenesis of autoimmune and chron. with encephalitogen. It is thus expected that MP affects IL-17 inflammation production in LNC more potently than in purified T cells (14). as MP had almost equal inhibitory effect on range of inflammatory. IL-10. including dendritic cells and tory functions of these cytokines suggest their important macrophages which produce numerous cytokines that role in pathogenesis of both MS and EAE because the are important for the stimulation of IL-17 in T cells. spinal cords of rats which had developed EAE (14). There- of methylprednisolone (MP). Medicinski podmladak Mini pregledni radovi Introduction First. The proinflamma. MP was shown to affect activ- GM-CSF. duction more potently than IL-17 production. ity of accessory LNC cells.

Gao W. Moreover. Th17 cells and how these mechanisms might be used for rier (BBB). the selection of optimal therapy of autoimmune and oth- Fewer infiltrating cells within the CNS of MP treated rats er chronic inflammatory diseases. hydrocortisone inhibition (23) and severe forms of al- ferent response to glucocorticoid therapy. Korn T. Stromnes IM. In our experiments. The most recent data demonstrate were inhibited (19). IL-23 drives a pathogenic T Our results demonstrating low sensitivity of Th cell population that induces autoimmune inflammation.   27 . the drug might affect Th17 cells and their cytokines Nature 2006. Sedgwick JD et al.205:1535- 41. Helms.14:337-42. Goverman When we analyzed effects of MP applied in vivo JM. that although Th17 cells are refractory to glucocorticoid hibition of encephalitogenic cell infiltration into the CNS suppression at a genome-wide level. but not of IL-17 producing 4. explaining the clinical efficacy of the filtration might be explained by increased apoptosis and latter in the treatment of diseases with steroid resistant weaker activation of encephalitogenic cells in lymphoid form (26). However larger studies will be required to un- organs. ings that T lymphocytes derived from MS patients with duction. among pop. as previously suggested (20). Linington C. Elson CO et al. while ation of pathogenic effector Th17 and regulatory T cells.and IL-23-modulated T cells induce distinct types of comparison to control rats (19). O'Quinn DB. which have 6. Brain 2006. Sospedra M. Transforming growth fac- tor-β induces development of the TH17 lineage. Oukka M. through its effect on CNS resident cells. the number of may underlie steroid hyporesponsiveness (25). Harrington LE. Bettelli E.201:233-40. WS. 2. Exp Med 2005. The proportion of cells EAE based on histology. Martin R. plentiful data convinc.441:235-8.129:1953-71. Carrier Y. Liggitt D.441:231-4. by decrease in expression of adhesion molecules derstand the mechanisms underlying steroid resistance of on leukocytes and endothelial cells of the blood brain bar. J Exp Med 2008. Recent evidence the inhibitory actions of steroids but also that these cells from EAE induced in mice suggests that the major targets are actively and specifically selected for within mixed T of GC action are peripheral rather than CNS-residing T cell cultures upon exposure to steroids (25). both cytokines. lergic asthma associated with Th17 response are poorly ulations of autoreactive T cells. Nat Med 2008. This could be a consequence of the in. portion of IFN-γ producing. It is also cells isolated from the spinal cord of MP-treated animals interesting to speculate. both in vivo and in vitro. Conclusion 7. in accordance with recently described 5. CNS chemokine profile. Andjelkovic AV. IL- cells was reduced in cells isolated from MP treated rats in 12.23:683-747. there is a reciprocal that has been previously reported following treatment of sensitivity of this subset of cells to calcineurin inhibition EAE with glucocorticoids (20). we could not detect increased cell death rate under the influence of MP in EAE rats (19). Mattson J. Strom TB. immunity by Th1 and Th17 cells.14. Differential regulation of central nervous system auto- in EAE. Langrish CL. Mangan PR. Reciprocal developmental pathways for the gener- that the influence of MP on Th1 might be direct. extend current notions of steroid resistance in Th17 ingly demonstrate that GC directly influences cells within cells by identifying a distinctive subset of Th17 cells that the target tissue (18). which is in ac. These data lymphocytes (17). On the contrary. high levels of BCL-2 (21). with cyclosporin A . previously been shown to modulate IL-17 production. and re- isolated from the spinal cord of diseased rats expressed sponse to cytokine inhibition. Harris RA. as well as by increased integrity of BBB (2). Segal BM. purposely the site of action. Blumenschein WM. 1. the same phenomenon was observed: the pro. Understanding patho- (17). Immunology of multiple sclerosis. Volume 67 | No. Gold R. Most recent data demonstrate that both murine and genesis and therapy of multiple sclerosis via animal mod- human Th17 cells are resistant to glucocorticoid-induced els: 70 years of merits and culprits in experimental autoim- apoptosis and the undelying mechanism is ascribed to mune encephalomyelitis research. (12) there is a diverse controlled by corticosteroids (24). Carlson TJ. The limitation of the in. J 17 cells. Chen Y. It is tempting to speculate et al. 3. it has been response to glucocorticoid therapy depending on the en. an enhanced Th17-like phenotype are less sensitive to Besides various mechanisms that underlie the dif.19) have been later confirmed by find- less sensitive to the influence of MP than the IFN-γ pro. to the inhibitory ac. 1 | March 2016. Bullard DC.. recently shown that not only Th17 cells are refractory to vironment. might also be a consequence of increased rate of apopto- sis of these cells in the CNS. cordance with the recent investigation performed in mice Annu Rev Immunol 2005. that steroid treatment of auto- immunized with encephalitogen was significantly lower immune patients may directly enrich for precisely the than the number of cells isolated from control diseased proinflammatory T cell subsets linked with inflammation rats and their IFN-γ and IL-17 expression and production and drug resistance. Kroenke MA. Lassmann H. Basham B. Cerretti LM. References Surprisingly. findings in DA rat EAE (22). Nature 2006.Medical Youth Mini review articles conclude that the expression and production of IL-17 is tion of MP (13.

McCombe PA.BCL-2 protects human and mouse 13. Neurosci kin-17 and interferon-gamma expression by both naive Lett. Shin S.12840 experimental autoimmune encephalomyelitis. Brain Behav Immun Dhanda AD et al. Dong mune System. Wüst S. peutic target for steroid-insensitive asthma. Proc Natl Acad Sci U S A. 17. flammation. IL-17 expression and production by cells infiltrating cen- 9. Ramesh R. tal autoimmune encephalomyelitis treated with dexameth- Mechanisms of glucocorticoids in the control of neuroin. An inflammatory review of 26. Low sensitivity to glucocorticoid HM. Peripheral T cells are the ther-apeu. TJ. et al. Ohtsuka S. Mangan PR. Miljkovic Z. Momcilović M.180:8434-43. 20. Alvarenga R et al. Schleimer RP. McPherson KG. and primed T cells. Holsboer F et al. Kuchroo VK. alomyelitis.. 2014. Quintero MA. Tucker. van den Brandt J. Kasahara TM. Adv Exp Med Biol. Ramić Z et al. Toscano M. Miljković Z.21:259-72. Momcilovic M. 2012. School of 22.148:209-18. 2008.42:71-104. 21.1111/all. 2012. Stanojević Ž.et al. Goodin DS. Refojo D. Matsuyama M.Molecular mechanisms of glucocorticoids in the inhibition of in vitro Th17-related cytokine production in control of inflammation and lymphocyte apoptosis. to glucocorticoids. 2013. sis of T lymphocytes and macrophages in the central and Handb Clin Neurol. FASEB J. University of Belgrade. Banuelos J. Schewitz-Bowers LP. 25:821-52. Li S.112:4080-5. Wu W. Carlson mann JP. L.2013:609395.Glucocorticoid treatment of multiple sclerosis. J Neuroinflammation. Popadić D. Reichardt ros PO. Pender MP. Kleiman A. Hatton RD. Methylprednisolone inhibits interleu. Budinger GRS. DOI: 10. Mostarica-Sto- family cytokines and the expanding diversity of effector T jkovic M. 2008. Methylprednisolone inhibits IFN-gamma and cell lineages. 24.. Schweingruber N. Sav. Xin J.214:1177-88. asone. Using EAE to better under. popolysaccharide levels.9:47.Clin Immunol. Rangachari M.. tral nervous system in experimental autoimmune enceph- stand principles of immune function and autoimmune pa. Th17 cells from glucocorticoid-induced apoptosis. Lu NZ. Bar- 15. McKevitt K. C. Reichardt HM.211:89–104.6:37. expression and production in active experimental auto- ić E.BMC Immunol. Tischner D. Bittencourt VC. Sorrells SF.447:148-52. Kleiman A. 28  Mart 2016. Djuretic IM. Th17-associated cytokines as a thera- hibits the transcription factor T-bet by direct protein-pro. Kinetics of IFN-gamma and IL-17 14. 2014. J Neuroendocrinol. The activated glucocorticoid receptor in. Popadić D. Copland DA. Nguyen KB. Crit multiple sclerosis patients is related to elevated plasma li- Rev Clin Lab Sci 2005. Annu Rev Immunol 2007. Reichardt SD. Lait PJ. Kawaguchi M et al. 25. glucocorticoid actions in the CNS.872:217-33. Rein T. Glucocorticoid-resistant Th17 cells are 2007. immune encephalomyelitis in Dark Agouti rats. Morales-Nebreda 12. Cato AC. munol. IL-17 19. Zhou L. thology. 2013. J Exp Med. 16. Druker J. 2015. Miljković Z. Morishima Y. starica-Stojković M. Ano S. 23.J Immunol 2008.122:455-64.56:58-69. Effects of Glucocorticoids in the Im. Gold R. peripheral nervous systems of Lewis rats with experimen- 11. Sapolsky RM.24:174-82. Miljkovic D.45:31-9. Effects of Methylprednisolone on expres. 2009. Liberman AC. | Broj 1 | Izdanje 67 . Lingen MW. Bochner BS. Marković M.Clin Dev Im- tein interaction. Pro-inflammatory human Th17 tic targets of glucocorticoids in experimental autoimmune cells selectively express P-glycoprotein and are refractory encephalomyelitis. 2015. Chen P. Ishii Y. 18. selectively attenuated by cyclosporine A. Ferreira TB. Harrington LE. 2007. Allergy sion and production of interferon - and interleukin 17 in 2016. J Autoimmun. Lühder F. Oppong E. Miljković D. Weaver CT. Kozhaya L. Cao Y. 2013. Medicinski podmladak Mini pregledni radovi 8. Tuckermann JP. Teixeira B. Mo- Medicine.Increased apopto- 10. J Neuropathol Exp Neurol 1997. Momcilović M.

although bleeding risk is primenjuje kod bolesnika koji su prethodno bili na terapiji VKAs slightly raised. agulacije mogu da pomognu u cilju identifikacije onih bolesnika The use of OAC is recommended for stroke prevention. For secondary stroke prevention all AF patients should sa AF indikovano je uvođenje OAC. Stroke prevention is central to the management of AF. The aim of this article was to provide an overview on current elni pristup u prevenciji i lečenju AIMU udruženih sa AF. be performed in order to influence modifiable risk factors for Procenu rizika od nastanka krvarenja treba vršiti u cilju uticaja bleeding. va za stratifikaciju faktora rizika. Data regarding safety of IVT used in patients on ukoliko im je vrednost međunarodnog normalizovanog odnosa NOAC are very scarce. Ona podrazumeva ili stroke prevention in AF patients. prevention. iako je could help to identify those patients who might be eligible for tada rizik od pojave krvarenja malo viši. Njena upotreba se može razmatrati u određenim slučaje- vima kada je antikoagulisanost nedovoljna. if patients are fully anticoagulated. This includes either vitamin antagoniste vitamina K (VKAs) ili nove oralne antikoagulanse K antagonists (VKAs) or novel oral anticoagulants (NOACs). Ljiljana Beslać Bumbaširević2 1 Klinika za neurologiju. Klinički Centar Srbije 2 Klinika za neurologiju. tor lošeg ishoda AIMU. ishod.7. Podaci o primeni IVT thrombolysis. then IVT is contraindicated. tion. INR) manja od 1. stroke. International normalized ratio. Posledično. all others are indicated for antitrombotična terapija. kod bolesnika sa NOAC su oskudni. Medicinski fakultet Univerziteta u Beogradu Kontakt: visnja.padjen@hotmail. some coagulation parameters (engl. it may be značaja i za njeno sprovođenje se savetuje upotreba OAC.   29 . it is also a significant predictor of stroke’s poor outcome. ali određeni parametri ko. kontraindikovana je primena IVT. approach in AF associated stroke prevention and treatment. by any Prevencija AIMU kod bolesnika sa AF je od ključnog medication.7. ministrated safely in patients given VKAs if the internation- ska trombolitička terapija (IVT) verovatno može da se bezbedno al normalised ratio is less than 1.com Sažetak Abstract Atrijalna fibrilacija (AF) predstavlja nezavisni faktor Atrial fibrillation (AF) is an independent risk factor that koji pet puta povećava rizik od nastanka akutnog ishemijskog increases the risk of acute ischemic stroke by five-fold. udar. prevencija. tion is subtherapeutic. outcomes. Intravenous thrombolysis (IVT) can probably be ad- na one faktore rizika koji mogu da budu modifikovani. U primarnoj prevenciji AIMU. tromboliza thrombolysis Volume 67 | No. akutni ishemijski moždani Key words: atrial fibrillation. Klinički Centar Srbije. Intraven. U considered in some circumstances if the level of anticoagula- akutnoj fazi AIMU. however. 1 | March 2016. In the koji bi bili pogodni za primenu trombolitičke terapije. In primary prevention. korišćenjem skoro. The assessment of bleeding risk should also ji AIMU indikovano je uvođenje OAC za sve bolesnike sa AF. U prevenciji AIMU kod bolesnika sa AF preporučuje se Oral anticoagulant therapy (OAC) is recommended for oralna antikoagulantna terapija (OAC). ukoliko su bolesnici adekvatno antikoagu. potrebno je identifikovati AF clinicians should identify low-risk AF patients who do not bolesnike sa „niskim rizikom“ kojima nije potrebna nikakva require antithrombotic therapy. Ona predstavlja i značajan predik. In addi- moždanog udara (AIMU). lisani primenom bilo kog OAC. However. (NOAC). Ključne reči: atrijalna fibrilacija. svim ostalim bolesnicima OAC. U sekundarnoj prevenci. be offered OAC.Medical Youth Mini review articles Antithrombotic therapy in acute ischemic stroke patients with atrial fibrillation Antitrombotična terapija kod pacijenata sa akutnim ishemijskim moždanim udarom i atrijalnom fibrilacijom Višnja Pađen1. Cilj ovog članka je da se analizira aktu. by using risk stratification schemes. acute stroke phase.

Medicinski podmladak Mini pregledni radovi Introduction Table 1. (22) Various scores have been proposed. while CHA2DS2-VASc score ≥ 2 is a high risk. (8) Moreover. CHA₂DS₂-VASc Score it increases the stroke risk by 5-fold. is the fact that it classifies a stroke. CHA2DS2-VASc score = 1 represents moderate not need antithrombotic therapy. | Broj 1 | Izdanje 67 . they are more likely to lead to disability (11). CHA2DS2-VASc score has been shown to have score value of 0 represents low risk for thromboembol. Vascular disease 1 tors are: heart failure. (1) Around 2% Congestive heart failure 1 of the world‘s population has AF and it is predicted that its prevalence will rise for over 2. approximately 0. 30  Mart 2016. large portion of patients into the intermediate risk cat- while in the case when the CHADS2 score ≥ 2 it is a high egory (C statistic. es. recent guidelines have moved toward initial identification Current recommendations state that all patients with of low-risk patients first (because these patients do not CHADS2 and CHA2DS2-VASc score ≥ 2 are indicated need any antithrombotic therapy).15) More. hypertension. particular value in identifying low-risk patients who do ic event. Prevention stratification schemes for thromboembolic risk Atrial fibrillation (AF) is the most prevalent sus- tained heart rhythm disorder. to increase costs (12) and extended hospital care. (6) Subsequent studies have confirmed AF to be an inde- pendent risk factor for stroke. CHADS₂ Score sequences that include heart failure. (13) Diabetes mellitus 1 Stroke risk factors and Risk stratification schemes Stroke/TIA 2 Most common reported AF related stroke risk fac. (14. BLED score has been validated in multiple independent cohorts (table 2). (10) Fur. it is import- 17). Age 65-74 years 1 over. diabetes. reduced qual- ity of life. while in cases of identifying high-risk patients. Hypertension 1 thermore. This study has shown that individuals with AF are Diabetes mellitus 1 five times more likely to have a stroke. (7) AF is estimated to be responsible for approximately 15-20% of all strokes. (16) Most commonly used are CHADS2 and CHA2DS2-VASc score (table 1). various stroke risk factors in AF have been used to formulate several stroke risk prediction tools to help risk Sex (female gender) 1 stratify patients with AF. 21) Hence. 19) Never- risk for throboembolic events. AF is the most important single cause of Congestive heart failure 1 ischemic stroke in the elderly. (5) Age >75 years 1 About 20 years ago. Stroke/TIA 2 junction with hypertension or congestive heart failure. in comparison with those without AF. by estimating that. cases in which thromboprophylaxis is being considered. associated with severe con. compared Age >75 years 2 to non-AF strokes. especially on clinical risk factors. overall. (9) AF-associated strokes are more severe with a higher mortality rate.5 times by 2050. (20. (16. (17) CHA2DS2-VASc theless. that are based ant to evaluate bleeding risk in AF patients. CHADS2 score value of 0 represents low risk of pecially for CHADS2 score. these stratification schemes. age and pri- or stroke/transient ischemic attack (TIA). but only HAS- (18) It is argued that the most important limitation. currently averaging between 75 and 85 years.6). poor mental health and death. (2-4) Hypertension 1 The average age of patients with this condition is steadily rising. This incidence is even higher in con. more risk. In addition to stroke risk assessment. est predictive value for identifying high risk patients. aspirin or none (5. (16) moderate risk therapy can be: OAC. CHADS2 score = 1 represents moderate risk. have been showing weak or mod. the Framingham study has point- ed out AF as an independent risk factor for the ischemic stroke. stroke. (17). rather than focus on for oral anticoagulant therapy (OAC). However.

blood of nearly half time. (31) Current estimations are that only 1/5 of all AF patients actually receive adequate AF diagnosing and effective anticoagulation at any point of time. In contrast recognized as a priority for stroke prevention. (16) With VKAs. (23) as well as in use of novel to treat. op.. although some patients’ characteristics.Medical Youth Mini review articles Table 2. since there is still limited experience with these agents. the strict ad- Studies have shown that the risk of stroke in AF herence to approved indications is recommended. the use of these medications Stroke prevention does not require INR monitoring. which block the formation of multiple active estimated that about one half of AF patients remain un.g. rivaroxaban. compliance. However. dabig- recommend that. portunistic screening for AF by pulse palpation. current recommendation is that the use of Abnormal renal/liver function 1 or 2 antiplatelet therapy (as aspirin–clopidogrel combination therapy or—less effectively—aspirin monotherapy for Stroke 1 those who cannot tolerate aspirin–clopidogrel combina- tion therapy) for stroke prevention in AF should be lim- Bleeding 1 ited only in cases of patients who cannot or refuse to take any form of OAC. since it reduces the risk of stroke Hypertension 1 almost two times more compared to aspirin.   31 . There patients is reduced by the use of antithrombotic therapy. is scape in AF-stroke prevention in recent years. Compliance and than warfarin. (5) VKAs. to withhold OAC. detected. in the primary stroke focus on correcting the potentially reversible factors that prevention. so patients would be left without Volume 67 | No.7% per year (number needed to treat. (25. 1 | March 2016. (28) Moreover. is insufficient evidence to recommend one NOAC over (27) All major guidelines emphasize the role of OAC for another. aban have twice daily dose regimens). Drugs or alcohol intake 1 or 2 Vitamin K antagonists are associated with an ab- solute RR of 2.g. 37) HAS-BLED score has been shown to be predic. should be in clinical trials have shown inferiority compared with considered for the early detection of AF. consistently limiting the num- ber of ICH. for embolism in patients with no history of prior stroke tive of intracranial haemorrhage risk. tolerability and cost may be important con- Contrary to popular belief that aspirin is safer siderations in the choice of the agent.0). tend to have inefficient or unsafe warfarin levels in the excessive alcohol intake. with a target INR of 2.g. (29) Con- sequently. particularly in the gastro. a high HAS-BLED score is not a reason in therapeutic range [TTR]. in patients aged 65 years or over. 12) for patients with a history of prior stroke (sec- oral anticoagulants (NOACs) (16). studies have shown that aspirin actually adherence to treatment is crucial (dabigatran and apix- increases the risk of bleeding. uncontrolled hypertension. followed apixaban and edoxaban). is about 54-61 %. atran) and oral factor Xa inhibitors (e. mainly because symptoms are considered to be IX and X). These drugs have a intestinal tract. to predict bleeding (primary prevention) and 8. VII. (5) Moreover. vitamin K-dependent coagulation factors (factors II. None of NOACs so far tested by recording of an ECG to verify diagnosis. these drugs block the activity of one single step unimportant. but it can be used to highlight patients’ However. e. by reporting that warfarin is significantly superior. Prevention stratification schemes for bleeding risk compared aspirin with warfarin in AF-associated stroke prevention have shown that the efficacy of stroke preven- HAS-BLED Score tion with aspirin is weak. (16) Labile INR 1 Current recommendations are that thrombopro- phylaxis in AF patients can be obtained with vitamin K Elderly (>65 years) 1 antagonists (VKA.4% per year (number needed during bridging therapy. (30) and that these patients contribute to bleeding (e. studies have shown that the proportion of pa- potential risk of bleeding and to point out to the need to tients with AF treated with warfarin. the quality of anti- values ≥ 3 indicate that there is a high risk of bleeding coagulation control is essential (as reflected by the time (17). studies that have directly relatively short half-life. warfarin) or a non-VKA oral an- ticoagulant (NOAC).26) Therefore.0-3. (5) NOACs consist of two groups of med- of underlying AF. However. current guidelines ications: the oral direct thrombin inhibitors (e.g. a stroke is the first sign in coagulation. HAS-BLED score ondary prevention). (24) It is to VKAs. labile INRs). drug prevention of AF-associated strokes. For many patients. with better safety. (31) The introduction of NOACs has changed the land- Diagnosing AF before first complications occur.

reported a benefit J Med 2002.7. Am al studies have. 114:119-25. et al. and implications on the projections for es. 34-39) Data about efficacy of IVT in the be eligible for thrombolysis. as well as identification of low risk patients (who Intravenous thrombolysis in AF-associated do not need any antithrombotic therapy). many AF patients cannot be treated of renal function (by CrCl) is mandatory. (32. silent missed. Furthermore. an activated partial history and a laboratory test reliably suggest the absence thromboplastin time (aPTT) can be used although the of an anticoagulant effect. for patients with acute ischemic stroke deficit after 3 months. rhythm management and stroke prevention: the AnTico- tically significant difference in the occurrence of sICH. incidence of atrial fibrillation in Olmsted County Minne- derly patients. Gersh BJ. or last well effect of these agents reliably and rapidly and to estimate known time. of IVT administration in AF-associated stroke patients. (42) Observation. Circulation 2006. of AF in asymptomatic patients(especially older than 65 years). as well as those with previous stroke/TIA should be offered Intravenous thrombolysis (IVT) is the only ap. Currently. Hart CL. et al. Hole DJ. especially dabigatran. et al. since only few randomised controlled trials have analyzed this References specific subpopulation. within the time frame of 4. Moreover. treatment of AF-associated strokes is still scarce. Thus. all of these drugs have a degree of infarcts or microbleeds. apy with a baseline INR > 1. in these cas- to check the presence of an anticoagulation effect. the prevalence of subclinical brain chang. | Broj 1 | Izdanje 67 . (32. the symptoms onset and it significantly increases the The challenge for clinicians evaluating and considering proportion of patients who are left without neurological treatment options. some coagulation pa- CT showing no haemorrhage or well-established acute rameters could help to identify those patients who might infarct. 113:359-64.46) Furthermore. es. Barnes ME. future prevalence. (46) Another issue is the renal excretion. (iii) non-contrast IVT use in these patients. Philips KA. in practice. reported opposite results that the risk of sICH in these 4. with OAC (either VKAs or NOAC). (ii) an acute neurologic deficit expected to re.33) Indications of IVT are: (i) who are taking NOACs. clearly defined 4. JAMA 2001. assessment fact that. have elapsed since the most recent dose of the NOAC (in trations.54) Moreover. et al. and have reported conflicting re- sults: with a non-significant trend in favour of placebo 1. (55) (PT) and this might be used as a rough estimate of an anticoagulation effect. in majority of cases. 82:2N-9N symptomatic haemorrhagic transformation (sICH) in 3. data regarding the occurrence of population-based estimates. such as white matter changes. Stewart S. Subsequently. almost no data are available for the safety of sult in significant long-term disability. is high. Hylek EM. (5) None of the novel OACs has a specific In recent years. strokes. be given. particularly at higher concen. Prevalence. as AF occurs in el. may pre.5 hours from is less than 1. Benjamin EJ. ing the NOACs may also present with an acute ischemic There are non-specific coagulation tests that can be used stroke. Prevalence of diag- thrombolyzed patients with AF-associated stroke are also nosed atrial fibrillation in adults: national implications for conflicting: some observational studies found no statis.7. in- cidence and predisposing conditions for atrial fibrillation: (43 . 1980 to 2000. Secular trends in patients is higher.5 hours before IVT will the potential increased risk of symptomatic hemorrhage. strokes patients with at least 1 additional stroke risk factor. Stud. IVT can probably proved therapy for the treatment of patients with acute be safely administered in patients on VKAs if the INR ischemic stroke. Current recommendations state that. 285(18):2370-5. A population-based in ECASS III (40). but the management of bleeding is considered been registered in the field of prevention of AF-associated to be largely supportive. given that these drugs have a rel. (5) For the Factor Xa inhibitors a Conclusion better anticoagulant estimate can be provided by an an- ti-Xa assay. sota. Go AS. Am J Cardiol 1998. tion: 20-year-follow-up of the Renfrew/Paisley study. a substantial improvement has antidote. in favour of IVT in IST-3 (41) and study of the long-term risks associated with atrial fibrilla- no effect of the treatment in NINDS. Wolf PA. Current guidelines recommend early detection atively short half-life (5 to 17 hours). or until at least two half-lives correlation is not linear. Miyasaka Y. agulation and Risk Factors in Atrial Fibrillation (ATRIA) (47-50) while other studies (including 2 meta-analyzes) Study. (5) Rivaroxaban prolongs the prothrombin time patients with normal renal function). is to determine the anticoagulant acute ischemic stroke symptoms with onset. however. although bleeding risk is slightly raised. thrombolysis should only be initiated if the clinical ies have shown that for dabigatran. Kannel WB. patients tak- ic coagulation test (in contrast to the INR for VKAs). associated with cognitive impairment or not. The NOACs with IVT because of on-going oral anticoagulation ther- do not require dose adjustment on the basis of a specif. Medicinski podmladak Mini pregledni radovi any anticoagulation protection if more than one dose is dispose to bleeding. (51. 2. 32  Mart 2016.

2013. 7. Lamassa M. Document Reviewers. Omran H. 2012 and thromboembolism in atrial fibrillation. 160:41-6. a consensus conference organized by the German Atrial 14. Die- 9. et al. Camm AJ. Jovanovic DR. Lip GY. Wolf PA. European Heart management of atrial fibrillation--developed with the spe. Fang J. Lip GY. ters for trials in atrial fibrillation: recommendations from Stroke 1996. Inde. Marin F. Benjamin EJ. Thromb Stroke 1991. 1049. role of baseline CHADS2. Gladstone DJ. Giralt-Steinhauer E. aspirin in nonvalvular atrial fibrillation: an lation: when. et al. Lip GY. Andreotti F. Neurology. systematic review. Thrombolysis with a CHADS2 score 0-1: a nationwide cohort study. Comprehensive risk reduction in Comparison between CHADS2 and CHA2DS2-VASc patients with atrial fibrillation: Emerging diagnostic and score in a stroke cohort with atrial fibrillation. fibrillation: an update of the 2010 ESC Guidelines for the 22. Brandes A. 23. Arch Inter Med 1987. endorsed by the European Society of independent risk factor for stroke: the Framingham Study. Hacke W. Ghali WA.Meta-Analysis of CHADS2 33. ment with intravenous alteplase and outcome in stroke: an Volume 67 | No. Leys D et al. Heart Rhythm Association. from the 3rd AFNET/EHRA consensus conference. Stroke prevention in atrial fibrillation: a for the management of atrial fibrillation.2013. ogy/American Heart Association Task Force on Practice lation in Europe: data from a multicentre multinational Guidelines and the European Society of Cardiology Com- hospital-based registry (the European Community Stroke mittee for Practice Guidelines (Writing Committee to Re- Project). 30. 2012. boembolism in atrial fibrillation patients independent of date of the ESC Guidelines for the management of atrial anticoagulation. von Kummer R. et al. Abbott RD. Cannon DS. Europace 2007. Ezekowitz M. Cardiology [ESC]Working Group on Thrombosis. 98(10): 946-652. Rübenacker S. 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Stroke 2006. 79:31-38. Pancioli A. Weimar C. 2014. Hacke W. 40. De Keyser J. | Broj 1 | Izdanje 67 . Jovanovic D. 48. et al. Zhang JB. plinary Working Groups: the American Academy of Neu. Padjen V. 34  Mart 2016. Rivolta J. Jung YH. Bogousslavsky J. warfarin therapy entails an increased bleeding risk after can Stroke Association. Padjen V. Sacco RL. Kim YD. Thrombolytic and harms of intravenous thrombolysis with recombinant therapy for ischaemic stroke in patients usingwarfarin: a tissue plasminogen activator within 6h of acute ischaemic systematic review and meta-analysis.0-4. Fulton R. Council on Cardiovascular Radiology and Intervention: 46. 2014. Sandercock P. Subgroup acute stroke in patients taking novel oral anticoagulants. 298:101–105. Stroke rt-PA Stroke Study Group: Tissue plasminogen Subtherapeutic warfarin is not associated with increased activator for acute ischemic stroke. Wardlaw JM. 52. Ding ZY. 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established. hepatic encephalopathy. oksidativni stres. deficit and alterations in sleep-wake cycle. glycinergic. Cilj sion. manganese and proinflammatory cytokines. mitohondrijska disfunkcija i energetski deficit. This review aimed to summarize current knowledge of ovog preglednog rada je bio da prikaže dosadašnja saznanja o the role of neurosteroids in the pathogenesis of HE. Univerzitet u Beogradu 2 Institut za medicinsku fiziologiju . Hepatic Encephalopathy (HE) represents a neuropsy- jatrijski sindrom uzrokovan akutnom ili hroničnom insufici.3). induces cytotoxic brain edema due to the A HE has a rapid course with deterioration of mental swelling of astrocytes. holinergičkih. Hy- jencijom jetre. hepatična encefalopatija. This may potentially lead to the fatal outcome due to the brainstem herniation in the foramen magnum. neuroinflama.Rihard Burijan‘‘. Ammonia. Mehanizmi kojima amonijak izaziva razvoj HE su through modulation of neurotransmission. komponentom multi. of serotoninergic. Neuroste. to the devel- chronic liver failure (1). convulsions and the loss of conciousness. Additional potential nim poremećajima u HE. 1 | March 2016. After release from rona.Medical Youth Mini review articles The role of neurosteroids in the pathogenesis of hepatic encephalopathy Uloga neurosteroida u patogenezi heapatične encefalopatije Dušan Mladenović1. type B The pathogenesis of HE is complex and not fully (caused by porto-systemic shunt without liver disease). stimulate neuro- tornim citokinima stimuliše sintezu neurosteroida ushodnom steroid synthesis by up-regulation of translocator protein. Olivera Stanojlović2. Cholesterol serves as mitohondrije astrocita. signs and symptoms of chronic HE (usu- ally type C) may vary from subtle cognitive dysfunction Hepatic Encephalopathy (HE) represents a com.ac. Ključne reči: neurosteroidi. allopregnanolone and tetrahydro-deoxycorticoste- drodeoksikortikosteron pojačavaju GABA-ergičku transmisiju rone potentiate GABAergic transmission by positive allosteric pozitivnom alosternom modulacijom GABAA receptora. Key words: neurosteroids. amonijak. glutamatergičkih opioid receptor activities. Neurosteroids contribute significantly to increased roidi značajno doprinose povećanoj GABA-ergičkoj aktivnosti GABAergic tone in HE. kao i modulaciju genske ekspresije. Tatjana Radosavljević1 1 Institut za patološku fiziologiju. poremećaji neurotransmisije. lone and tetrahydro-deoxycorticosterone. On pressure (4). Glavnu ulogu u patogenezi HE ima hiperamo. Holesterol služi kao supstrat za sintezu a substrate for the synthesis of neurosteroids allopregnano- neurosteroida alopregnanolona i tetrahidrodeoksikortikoste. astrocytes. and attention deficits.   35 . Type other toxins. jor forms: type A (caused by acute liver failure). Univerzitet u Beogradu Kontakt: dmladen@med.rs Sažetak Abstract Hepatična encefalopatija (HE) predstavlja neuropsihi. Nakon oslobađanja iz astrocita alopregnanolon i tetrahi. as well as modulation of gene expres- i opioidnih receptora. Medicinski fakultet. chiatric syndrome caused by acute or chronic liver failure. ammonia. along with and type C HE (associated with liver cirrhosis) (2). opment of hepatic coma in the stage 4 of HE (2. and toninergičkih. neurotransmisija neurotransmission Introduction the other hand. in combination with u HE.. glicinergičkih. a regulacijom translokatorskog proteina. mechanisms of neurosteroid action in HE include modulation stva neurosteroida u HE uključuju modulaciju aktivnosti sero. component of multiprotein complex that stimulate cholesterol proteinskog kompleksa koja stimuliše transport holesterola u transport into astrocytic mitochondria. oxidative stress. gy deficit. It may be presented in three ma. ulozi neurosteroida u patogenezi HE. which cannot be detected by the plex neuropsychiatric syndrome caused by acute or standard neurological exam (minimal HE). glutamatergic. mitochondrial dysfunction. In type A HE hyperammonemia. Volume 67 | No. Amonijak u kombinaciji sa manganom i proinflama. and ener- cija. thus contributing to cognitive doprinose poremećajima ciklusa budnost-spavanje i kognitiv. čime modulation of GABAA receptor. with subsequent rise in intracranial functions. Dodatni potencijalni mehanizmi dej.bg. cholinergic. perammonemia plays a pivotal role in the development of HE nijemija. neuroinflammation.

)(24). androstane neurosteroids (androstanediol. | Broj 1 | Izdanje 67 . Deoxycorticosterone. Cholesterol is then converted to neurosteroids through a series of enzymatic reactions (TSPO. 5α-DHPROG. Voltage-dependent anion channel. Allopreg- Synthesis of neurosteroids nanolone (ALLO) and Tetrahydro-deoxy-corticosterone (THDOC) in series of enzymatic reactions. which occur Neurosteroids have been initially defined as steroid in the smooth endoplasmic reticulum and mitochondria compounds synthesized within brain. serotoninergic complex on outer mitochondrial membrane. This re. ticosterone).23). type B and type C HE ed in the central nervous system after ablation of periph- are not characterized by prominent brain edema. DOC.25) (figure 1).16). Medicinski podmladak Mini pregledni radovi Opposite to the acute form.9). but eral sources (21). 36  Mart 2016. Neurosteroids contribute significantly intermembrane space (24). Alterations in neurotrans. Peripheral-type benzodiazepine receptor. Translocator pro- tein. (22. etiocholanolone) and sulphated neurosteroids (preg- ic mechanisms involved in the development of chronic nenolone sulphate. TSPO associ- mission have an important role in the pathogenesis of ates with Voltage-dependent anion channel (VDAC) and types B and C HE and include changes in glutamatergic Adenine nucleotide carrier (ANC) to form a heteromeric (14). into three major groups: pregnane astrocytosis (large pale nuclei with prominent nucleoli neurosteroids (allopregnanolone. Pregnen- olone is further converted to progesterone. cholinergic (15. of neurosteroids in the pathogenesis of HE. that remain elevat. ANC. cytoplasmic vacuolization). They may be classified. (24. which al- (17). 5α-dihydro-progesterone) (modified according to Ahboucha et al. by the energy deficit (10). After transfer cytochrome to changes in neurotransmission found in HE. a rate-limiting step in neurosteroid synthesis. Neurosteroidogenesis starts with uptake of cholesterol into astrocytic mitochondria mediated by TSPO.6). Adenine nucleotide carrier. according to the the dominant pathological change is Alzheimer type II structural features. tetrahydro-deoxy-cor- frequently occuring in pairs. dopaminergic (18). neuroinflammation (11) and mito. Figure 1. Tetrahydro-deoxycortico-sterone. P450 cholesterol side chain cleavage enzyme (P450scc) view aimed to summarize current knowledge of the role catalyzes the conversion of cholesterol to pregnenolone. GABAergic (14). the transport of cholesterol into the mitochondria. action of Translocator protein (TSPO) formerly known as chondrial dysfunction (12.13). oxidative stress (8. Synthesis of neurosteroids in astrocytes. found in both grey and white matter (5. dehydroepiandrosterone sulphate) forms of HE are closely interrelated and include: alter. VDAC. The initial step in neurosteroid synthesis includes ations in neurotransmission (7). Pathogenet. THDOC. adrenergic (19) and purinergic lows the transfer of cholesterol through the hydrophilic transmission (20). 5α-dihy- drodeoxycorticosterone. 5α-DHDOC.

neurosteroids may modulate brain oxida- es are followed by both an increase in mean voltage of tive stress and neuroinflammation in HE (16. lowest increase in the striatum (38. which motor activity. from cirrhotic patients who died Volume 67 | No. and prove the course of TAA-induced HE and to reduce mor.24). the ma- in pregnenolone and ALLO was also found in autopsied jority of data suggest that the up-regulation of TSPO by brain tissue from cirrhotic patients who died in hepatic hyperammonemia and other toxins plays a pivotal role in coma (32). with of GABAergic transmission was the fact that “endoge- the highest increase in the cerebellum and pons. may have beneficial effects in HE (48). as well as tality in rats (33). astrocytes with pathophysiologically relevant concentra- son’s disease.Medical Youth Mini review articles Neurosteroids were found to exhibit some funda. els of HE (46). TSPO expression was found to be increased in a ing the role of benzodiazepine-like ligands in alterations region-selective manner in the PCA model of HE. synaptic organization binding sites in all experimental models (35. creased blood-brain transfer of lipophylic neuroactive ste- genesis of HE was first suggested by Zaman (30). EEG bands and a decrease in delta band proportion in total power density. increased neurosteroid synthesis in HE may be also mediated by in- The involvement of neurosteroids in the patho. Region-selective brain and cerebrospinal fluid of HE patients (49). How- have found increased binding site densities of the iso. but with highest increase tors of changes in GABAergic transmission in HE. Radiometric assays in manganese in chronic forms of HE (24). prevents the development of hepatic coma. Increased TSPO expression in the brain rosteroid site on GABAA receptor and potentiate GAB- is mediated by hyperammonemia in combination with Aergic transmission in HE. that administration of flumazenil. epilepsy and stroke (23). These up-regulation of TSPO was also found in humans with findings shed light on neurosteroids as potential media- different etiologies of cirrhosis. Ahboucha et al. Neurosteroids and HE Apart from the up-regulation of TSPO. Alzheimer’s disease. late GABAergic transmission in HE (47).50). these findings point out the sig- nificant role of neurosteroids in the pathogenesis of HE. Finasteride pretreatment ameliorates the modulation of intracellular receptor activities. preserves vital reflexes and completely can regulate gene expression (genomic effects) (14. In product octadecaneuropeptide.34). (35). the role of „endogenous benzodiazepines“ has been quinoline ligand [3H]PK11195 and the peripheral benzo. have also been postulated accordance with the supposed role of neurosteroids in to stimulate neurosteroid synthesis (44). partly caused by the up-reg. Further oppos- mice. who antagonist. Furthermore. Additionally. ALLO and THDOC in experimental HE patients (45) and in brain extracts from animals mod- models of HE induced by thioacetamide. such as diazepam binding inhibitor and its processing azepines„ mediate increased GABAergic tone in HE. as an roids (43). by its ‘‘endogenous ligands’’ alternative to the hypothesis that “endogenous benzodi.42). pallidum and known that ALLO and THDOC bind to a specific neu- putamen (40). Peripheral sources of neurosteroids may gether with liver ischemia caused by portacaval anasto. Similar changes occur after treatment of cultured (29) and may also exert neuroprotective effects in Parkin. since neuroprotective effect of diazepine ligand [3H]Ro5-4864 in brains of TAA-treated flumazenil in HE was not consistent (24). as well (26-28). N-methyl-D-aspartate (NMDA). An increase these additional mechanisms cannot be excluded.39). This is evident autopsied brain tissue. brain levels of neurosteroids correlate with the severity of Mechanisms of neurosteroid action in HE include HE. thus suggesting ulation of TSPO on astrocytic mitochondrial membrane. ever. Together. 1 | March 2016.All findings correspond to mild stages Neurosteroids and GABAergic transmission of HE (33. as positive allosteric modula- proinflammatory cytokines (IL-1β. serotoninergic receptors (nongenomic effects). body temperature. Although mosis (PCA) and hepatic artery ligation (24). mission have been most extensively studied. whenever hyperammonemia de- sexual behavior.33). benzodiazepine site This was. cluding GABAA. tions of manganese (41) and proinflammatory cytokines (24. was found to im.35-37) have shown that neuro. The effects of neurosteroids on GABAergic trans- Various studies (24. Motor chang. (TAA) (31) to.   37 . Activation of TSPO. Earlier stud- steroid levels in HE are elevated due to increased neuros. with the nous benzodiazepines„ were not elevated in the plasma. also contribute to the pathogenesis of HE (44). the blockage of neurosteroid synthesis modulation of various membrane receptor activities. blood pressure velops. by increased densities of [3H]PK11195 or [3H]Ro5-4846 mental effects on the myelination. anxiety. 5α-reductase inhibitor. in- by finasteride.38). els were found to be elevated in the cerebrospinal fluid of el of pregnenolone. later studies have confirmed the increased brain lev. TNF-α) in acute.24. Both peptide lev- HE. as in human HE (40). (24) have demonstrated that the increased synthesis of neurosteroids in HE. firstly demonstrated by Itzhak et al. seriously questioned. and tors of GABAA receptors (39. They participate in the regulation of food intake. It is in right dorsolateral prefrontal cortex. ies postulated that “endogenous benzodiazepines“ modu- teroid synthesis consistently.

negative al. on the course of the role of neurosteroids in regulating the expression of HE. Further. Progesterone. manganese and not been fully established. but the contribution of these tylcholinesterase activity was found to rise in the thala. it increases oxidative lipid damage in the thala- mus in TAA-induced liver failure. to motor and cognitive changes in HE should be further investigated. astrocytic genes (39. The significance of these effects in the pathogenesis of HE should be further investigated. Genomic effects of neurosteroids have been postulated to hydroepiandrosterone sulphate (DHEAS). Recent studies revealed a reduced synthesis of de. evident as a dose-dependent AL. DHEAS. Hyperammonemia. in HE (53). since ALLO and proges- summary. aquaporin IV. including Glial sites. glycine (GLYT-1) and gluta- and benzodiazepine binding site has been further con. It is known that neurosteroids apart from GAB- AA affect the function of various receptors in the brain. the vestigated. they just modulate both sites on GABAA receptor fibrillary acidic protein (GFAP). Chole- increased serotoninergic transmission. tors and change the expression of GFAP and aquaporin IV. neurosteroids modu- lation of serotoninergic transmission in HE. based on an im. Conclusion including serotonin 5-HT3 receptors. Neurosteroids also act in synergy with ammonia at neurosteroids bind to progesterone or pregnane X recep- the benzodiazepine site. | Broj 1 | Izdanje 67 . it is clear that neurosteroids rosteroids and attenuates the effects of ALLO on GA. It is suggested that HE is accompanied with increased cholesterol uptake into mitochondria. Among 5-HT3 receptor antagonist ondansetron (17). Although benzodiazepine partial inverse agonist. glycine receptors and opioid receptors (54).33) effect that can be impeded by finasteride. this losteric modulator of GABAA receptor. However. worsens HE and increases serotonin concen. tration in synaptic clefts in PCA rats (56). the most important seems to be the modu- more.24.60). contribute to astrocyte swelling in HE (59). In hypothesis remains controversial. in a region-selective manner. administration of venlafaxine. glutamate. mate (EAAT-2).54. Ace. potentiate GABAergic transmission and contribute in oxidative brain injury in HE (16). increased synthesis of ALLO and THDOC.51). Interactions between neuros. its metabolites and possibly other (52). Inhibitors of neurosteroid synthesis (16.58). in HE (14. as positive allosteric modulators of GABAA receptors. neurosteroid or neurosteroid site antagonists at GABAA receptor (44) synthesis inhibitor (16). This increase potentiate GABAregic transmission and contribute to is associated with increased level of ALLO (57). as well as enzymes Monoaminooxidase firmed by consistently beneficial effects of Ro15-4513. thus enabling studied. serotonin reuptake lation of GABAergic transmission. such as traumatic brain injury (24. Prooxidative effect of Other potentially relevant effects of neurosteroids neurosteroids in the cortex could be mediated by reduced in HE catalase activity (16). effects to the pathogenesis of HE should be further in- mus and caudate nucleus in TAA-induced HE in rats. these effects. pretreatment reduces lipid peroxidation in the cerebral wake cycle in HE (14).44). GLUT-1 and MAO-A gene expression may the presence of ammonia (39). cognitive dysfunction and alterations in sleep-wake cycle gesting that neurosteroids may be involved in the modu. thus sug. transport- complex (39. The interaction between neurosteroids ers for glucose (GLUT1). inhibitor. Neurosteroids may have a dual region-specific role tor. negative allosteric modulator of GABAA recep. the Neurosteroids play an important role in the patho- relevance of these effects in the pathogenesis of HE has genesis of HE. Additionally. could change the expression of several genes (genomic BA-induced chloride currents in hippocampal neurons effects). While finasteride to the cognitive dysfunction and alterations in sleep.39. which are then released from astrocytes and exert serotonin antagonist methysergide (55) and selective nongenomic and genomic effects in neurons. proinflammatory cytokines up-regulate TSPO in outer teroids and serotoninergic transmission have been most mitochondrial membrane of astrocytes. these proteins is still blurred. ALLO and THDOC. A (MAO-A) and nitric oxide synthase (39. terone were found to reduce brain edema in other patho- positive allosteric modulators and reduced synthesis of logical states. Medicinski podmladak Mini pregledni radovi in hepatic coma. as well as the expression of key neuronal and transmission in HE. Ro15-4513 reduces the modulatory activity of neu.58). setrol serves as a substrate for the synthesis of neuroste- provement of HE after administration of nonselective roids. late the activity of serotonin. glycine. LO-induced increase in the binding of (3H)flumazenil in Additionally. NMDA receptors. 38  Mart 2016. be altered by activation of glucocorticoid receptors (24). The contribution of this effect could have beneficial effects on the course of HE. have shown that neurosteroids do not HE is associated with altered expression of sever- change neither GABA nor benzodiazepine recognition al key astrocytic and neuronal proteins. opioid Neurosteroids may also modulate cholinergic receptors. cortex. However.

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Medical Youth
Original articles

PAX6 gene analysis in Serbian patients with aniridia

Ispitivanje sekvence PAX6 gena kod pacijenata sa aniridijom

Dimitrije Cvetković1, Jelena Ćuk1, Ivana Novaković1,2
1
Medicinski fakultet Univerziteta u Beogradu
2
Mentor: Institut za humanu genetiku, Univerzitet u Beogradu

Kontakt: cvetkovic.d@outlook.com

Sažetak Abstract

Uvod: Aniridija je retka kongenitalna anomalija oka, Introduction: Aniridia is a rare congenital eye anomaly
praćena različitim oftalmološkim tegobama. Uzroci ovog obo- associated with various ophthalmological problems. Some of
ljenja su mutacije u PAX6 genu (engl. paired box 6, gen sa blo- the causes of this disease are mutations in the PAX6 gene or
kom koji kodira parni domen) ili delecija lokusa na kratkom deletion of loci on the short arm of chromosome eleven, which
kraku jedanaestog hromozoma koja može da bude parcijalna ili can be complete or partial. In this study, PAX6 gene sequence
kompletna. U ovoj studiji je ispitivana sekvenca PAX6 gena kod was analysed in four unrelated patients.
četiri nesrodna bolesnika sa aniridijom. Aim: Examination of the entire coding sequence of the
Cilj: Ispitavanje celokupne kodirajuće sekvence PAX6 PAX6 gene in our patients in order to detect mutations in this
gena kod naših bolesnika u cilju otkrivanja mutacija u ovom gene.
genu. Material and Methods: Patients' DNA was isolated
Materijal i metode: Iz uzoraka periferne krvi izolovana from samples of peripheral blood. All exons and flanking in-
je DNK bolesnika. Nakon toga je metodom polimerazne lanča- tronic sequences of the PAX6 gene were amplified using PCR
ne reakcije (PCR metodom) izvršena amplifikacija kodirajućih and screened for mutation with direct DNA sequencing by
PAX6 egzona i graničnih regiona i dati uzorci su direktno se- Sanger method.
kvencirani metodom po Sangeru. Results: In one patient, the insertion type of mutation
Rezultat: Kod jednog bolesnika je nađena mutacija (c.342-2insC) was found in the second intron of the PAX6 gene.
po tipu insercije (c.342-2insC) u drugom intronu PAX6 gena. Other three patients had normal PAX6 sequence although they
Kod preostala tri bolesnika nije nađeno odstupanje u sekvenci had phonotypical expression of aniridia.
PAX6 gena. Conclusion: These are first results of PAX6 gene anal-
Zaključak: Ovo su prvi rezultati analize PAX6 gena ysis in Serbia. Apart from most common mutations, deletions
u Srbiji. Osim najčešćih mutacija, delecija različitog obima u of various extent in coding regions of PAX6 gene, other types
kodirajućem regionu ovog gena, mogući uzroci aniridije su i of mutations are also possible causes of aniridia. The insertion
druge vrste mutacija. Insercija pronađena u ovoj studiji nije found in this study has not been reported before. It is also pos-
prijavljena ranije. Moguće je i da su mutacije u drugim genima sible that mutations in other genes are associated with iris mal-
povezane sa malformacijama dužice, što može da objasni nalaz formation, which can explain the finding of normal PAX6 gene
normalne sekvence kod tri bolesnika sa aniridijom. sequence in three patients with aniridia.

Ključne reči: aniridija, PAX6 gen, kongenitalne anomalije oka Key words: aniridia, PAX6 gene, congenital eye anomaly, Serbia

Introduction In the human genome, PAX6 gene is located on
the short arm of chromosome 11 at position 11p13 and
PAX6 gene belongs to the family of Pax genes and consists of 14 exons (2). During embryonic development
it plays an important role in the formation of tissues and PAX6 proteins activate genes involved in the formation
organs, in the course of embryonic development. Mem- of the eye, olfactory regions of the brain and spinal cord.
bers of this gene family are also involved in maintaining Mutations in the gene PAX6 are primarily associated
the function of cells after birth. Pax genes maintain its with malformations of the eye and from these disorders,
role through various proteins (transcription factors) that the most notable is aniridia (1).
bind to target places of the regulatory DNA, thus control Aniridia is a structural eye disorder, which is char-
the expression of many other genes (1). acterized by complete or partial absence of the iris of the

Volume 67 | No. 1 | March 2016.   41

Medicinski podmladak
Originalni radovi

eye. Glaucoma, cataract, strabismus and foveolar hypo- PCR conditions followed touch down protocol:
plasia are often associated with this disorder (3). Also, 1. initial denaturation 950 °C 10 min.; 10 cycles
the deletion of a large scale in 11p13 region can lead to (steps 2-4):
WAGR syndrome, which is characterized by Wilms' tu- 2. denaturation 950°C 1 minute
mor, aniridia, genitourinary malformations and mental 3. primers’ annealing 650°C 1 minute
retardation (4). 4. elongation 720°C 1 minute; 20 cycles (steps 5-7)
The aim of this study was to examine whether there 5. denaturation 950°C 1 minute
is a change in the sequence of PAX6 gene in four patients 6. primers’ annealing 650°C to 550°C (decreasing
with aniridia. temperature 0,50C per cycle) 1 minutes
7. elongation 720°C 1 minute; 10 cycles (steps 8-10)
Material and methods 8. denaturation 95°C C 1 minutes
9. primers’ annealing 550°C 1 minute
The study included four unrelated patients diag- 10. elongation 720°C 1 minutes
nosed with aniridia. Molecular genetic analysis was con- 11. final elongation 720°C 8 minutes
ducted in genetic laboratories of the Department of Neu- After purification of PCR products by ExoSap meth-
rology, Clinical Center of Serbia, Belgrade. od, we performed direct sequencing of PAX6 fragments by
Entire coding region of the PAX6 gene (exons Sanger method, using BigDye Terminator chemistry. Cap-
3-14), including exon - intron boundaries, has been an- illary electrophoresis for DNA sequencing was done on
alyzed. After the DNA extraction, we performed Poly- ABI 3500 Genetic Analyzer (Life Science, USA), together
merase chain reaction (PCR), in order to amplify target with data analyzed by Sequencher software.
PAX6 regions. Depending on used primers, products of
PCR amplification showed different size. Results
PCR reaction mix was consisted of: In one patient, the insertion of one base pair was
Dream Taq 10x PCR buffer: 2,5 µl, found in the second intron (c.342-2insC) of PAX6 gene.
10mM dNTPs: 0,5 µl, This change in sequence has led to different processing of
primer fw and rv (10 pmol): 0,5 µl each, primary mRNA transcript. By Sequencher software pre-
Dream Taq polymerase (5U/ µl): 0,2 µl, diction, it is, most probably, responsible for the phenotyp-
DNA (10ng/ µl): 2 µl, ical change.
H2O to 25 µl. Other patiens had no mutation in the PAX6 gene,
although they had phenotypic expression of aniridia.

Table 1. Primers’ sequences used for PAX6 gene analysis
Primer Sequence

Primer 3f CCA-TGG-ACG-TAT-GCT-GTT-GA

Primer 4f ACC-TCG-GTT-GGG-AGT-TCA-G

Primer 5f CTC-TTC-TTC-CTC-TTC-ACT-CTG-C

Primer 6 + 7f CAG-TAA-GTT-CTC-ATA-CCA-TTG-AAG-G

Primer 8f TGG-GTG-ACT-GTG-TCT-TCA-GG

Primer 9f AGA-CTA-CAC-CAG-GCC-CCT-TT

Primer 10f AGG-TGG-GAA-CCA-GTT-TGA-TG

Primer 11 + 12f GGG-CTC-GAC-GTA-GAC-ACA-G

Primer 13f GTG-GCT-GTG-TGA-TGT-GTTCC

Primer 14f TTC-CAT-GTC-TGT-TTC-TCA-AAG

42  Mart 2016. | Broj 1 | Izdanje 67

In our study. MolPathol. Pediatrics PAX6 gene sequences. Zumkeller W.56:180–183. Gramer E. M. According to PAX6 mutation da. Molecular In the Polish study. Wang Y. Petersen. patients with aniridia. Sikora A. Krawczynski MR. other and Total Aniridia. Reiter C. ing regions of the PAX6 gene as the main cause of aniridia. the normal sequence of the analysis of the PAX6 gene for congenital aniridia in the PAX6 gene was found in three patients. in order to be able to uncover the origin of this rare disease. a clinical study. 2012. Acta Ophthalmol. studies performed in China (8) of aniridia in Sweden and Norway. Three novel PAX6 mutations formed in Europe. as well as in three Korean population: identification of four novel mutations. Kim MS. This has been in northern China. 10. b) insertion of 1BP within the consensus sequence AG at the turn of introns and exons Discussion References: PAX6 gene sequences analysis has not been done 1.342-2in. Crolla JA. 7. the prevalence of this 3. but the evidence for this is scarce. van Heyningen V. congenital aniridia and glaucoma (10). Edward DP. but at different locations. Kim Y. Mol Vis.77: 409–420. Zang X.   43 . this change has not been described previously. This is just one piece of the puzzle and it is clear that further research is necessary. bia. insertion type of mutation (c. (7). Chae H. while in Northern European countries 1 : 72000 (6). the frequency of aniridia in Serbia. (2005). Kokotas H. Chen P. Park SH. Glaucoma and frequency disorder in the general population is 1 : 50000-100000 of ocular and general diseases in 30 patients with aniridia: (5). Gramer G. Edén U. Abu-Amero KK. 2.116(4):984–988. supported by results of the study which showed that mu.110.1169–77. Clinical and molecular aspects of in our country and these studies are generally rarely per. Epidemiology On the other hand. Am J Hum Genet. Gal A. 2003. 2015. 8.54:34551. Ophthalmic Genet.71:1138–49. mations in these patients may be due to mutation in an.86:727–9. Apart from the most common mutations. Luis CA. Kuszel L. Riise R. Electropherogram of the border region of intron 2 and exon 3 in PAX6 gene: a) a reference sequence. Wawrocka A. Sun D. Wang Y. and Poland (9). Lewis J. Orth U. 2012.22(1): 104 . 2013.18:488–94. Although there are no recent data on in patients with aniridia. tabase. aniridia. et al. Sika M. other gene that has a role in the formation of the eye. aberrations revealed by molecular cytogenetic studies in served. highlight deletions of different scope in cod- 2008. sC ) is determined in non-coding ( intron ) part of the WAGR syndrome: a clinical review of 54 cases. Kim M. 5. 2002. Fischbach BV. 4. Trout KL. Volume 67 | No. Al-Shahwan S. three intronic mutations were ob. Eur J Ophthalmol.36:1 89-91. deletions A Novel CYP1B1 Mutation with Congenital Glaucoma of various extent in coding regions of PAX6 gene. and Mutation analysis of paired box 6 gene in inherited aniridia thus aniridia is phenotypically present (9). Clinical Genetics 2010. Iggman D. J Appl Genet. 6. types of mutations are possible causes of aniridia. of our patients(9). 2013. 11p13 deletions can be more frequent than the PAX6 gene point tations in the CYP1B1 gene can lead to the appearance of mutations in Polish patients with aniridia.Medical Youth Original articles a) b) Figure 1. The conclusion is that the iris malfor. 9. These are first results of PAX6 gene analysis in Ser. Frequent chromosome In the Korean study. 1 | March 2016. Tornqvist K. Alzuhairy S. Zhao X.

isolated from swabs taken from the sa znacima i simptomima otitis externe. i Aspergil. antimikotična aktivnost parium. One of the therapeutic ap- staje aktuelan u lečenju otomikoza je primena esencijalnih ulja proaches that became topical in treatment of otomycosis is the za koja je pokazano da imaju različita biološka svojstva.2 1 Medicinski fakultet Univerziteta u Beogradu 2 Mentor: Institut za mikrobiologiju i imunologiju. i vratića od 1. Antigljivična aktivnost external ear canal of patients with signs and symptoms of otitis esencijalnih ulja matičnjaka. izuzev u slučaju except in the case of a strain of Aspergillus niger. Key words: essential oil. including the antimicrobial effect. Materijal i metode: Ispitivana je osetljivost 20 sojeva Materials and Methods: In this study. Univerzitet u Beogradu Kontakt: doriangrey92@gmail. manuke i vratića na sojeve Candida spp.39- rasponu od 0. Cilj: Cilj istraživanja je ispitivanje antimikotičnog dej. and Aspergillus spp. Tanacetum vulgare. manuke i vratića ispitivana je mi.39 do 50 μL/mL 50 μl/ml. Vera Jovanović1. Melissa officinalis. we investigat- Candida spp. Antifungal activity of essential oils of lemon balm. manuka and tansy to the causative agents of otomycosis Đorđe Jovanović1. Ključne reči: esencijalno ulje. Rezultati: Vrednosti MIC matičnjaka kretale su se u Results: MIC values of lemon balm were in the range of 0. ma- krodilucionom metodom. Tanacetum vulgare. Sanja Mitrović1. tolerant of the lemon balm essential oil. oil activities of lemon balm. Retko dovode do komplikacija. Melissa officinalis. Otomikoze Otomikoze su površne gljivične infekcije kože predstavljaju globalni problem. određivanjem vrednosti minimalnih nuka and tansy was tested by using the microdilution method inhibitornih (MIC) i minimalnih fungicidnih koncentracija by determining the value of the Minimum inhibitory concen- (MFC). među application of essential oils. Conclusion: The investigation has shown that essential Zaključak: Ispitivanjem aktivnosti esencijalnih ulja ma. no ulje matičnjaka. Karakteristični simptomi su nelagodnost cija korelira sa klimatskim uslovima i kreće se u opsegu u uvu (koja može da varira od pruritusa do ozbiljnih bolova 5-25% ukupnog broja bolesnika sa znacima i simptomima koji se pogoršavaju pri žvakanju) i otoreja (2). izolovanih iz briseva spoljašnjeg ušnog kanala bolesnika spp. Aim: The objective of the research is to examine antimy- stva esencijalnih ulja matičnjaka (Melissa officinalis). Najčešći izazivači otomikoza su gljive 44  Mart 2016. Leptospermum sco- mum scoparium. tracije su bile 2-4 puta veće od vrednosti MIC. što dovodi do parcijalnog gubitka sluha i tinitusa. tration (MIC) and Minimum fungicidal concentration (MFC). manuke cotic effects of essential oils of lemon balm (Melissa officinalis). manuka and tansy to the strains tičnjaka. gare) on the causative agents of otomycosis. of Candida spp. Leptosper. manuka of 0.56-50 μL/mL. which is shown to have different kojima je i antimikrobni efekat. The MFC values were 2-4 times higher than the MIC values. Minimalne fungicidne koncen. externa. i Aspergillus spp. mikrodilucionom metodom pokazano je da navedena method. and Aspergillus spp. esencijalna ulja imaju antimikotični efekat. (Leptospermum scoparium) i vratića (Tanacetum vulgare) na manuka (Leptospermum scoparium) and tansy (Tanacetum vul- uzročnike otomikoza. Zapaljenje otitis externe (4). biological properties. | Broj 1 | Izdanje 67 .39-50 μL/mL and tansy of 1.39 do 50 μL/mL. have antifungal effect. izabranih metodom slučajnog ed the sensitivity of 20 randomly selected strains of Candida izbora. Prevalen- srednjeg uva (1). Jedan od terapijskih pristupa koji po.. using the microdilution lus spp. Medicinski podmladak Originalni radovi Antimikotična aktivnost esencijalnih ulja matičnjaka.com Sažetak Abstrast Uvod: Otomikoze su površne gljivične infekcije kože Introduction: Otomycosis is а superficial fungal in- spoljašnjeg ušnog kanala.56 do 50 μL/mL. fection of the external ear canal. antifungal activity Uvod može da uzrokuje oticanje spoljašnjeg ušnog kanala. manuke od 0. prašnjavo okruženje doprinosi da otomikoze preovla- kada dolazi do perforacije bubne opne sa zahvatanjem davaju u tropskim i suptropskim predelima (3). which was jednog soja Aspergillus niger koji je bio tolerantan na esencijal. s tim što vlažno. toplo i spoljašnjeg ušnog kanala. manuke i vratića na uzročnike otomikoza Antifungal activity of essential oils of lemon balm.

8 cineola. raceae. antivirusnu. definisana kao najniža koncentracija ispitivanog esencijalnog β-tujona. sponu od 50 do 0. reklom sa Novog Zelanda i jugoistoka Australije. uzročnike otomikoza. kao najmanja koncentracija esencijalnog ulja koja je spre- čavala porast nakon presejavanja. Najzastupljeniji sastojci esencijalnih ulja su je za mikologiju i parazitologiju Medicinskog fakulteta terpeni. Identifikacija do nivoa stepen antimikrobne aktivnosti. antiholineste. Dosadašnja is. linalol i selinen. je određivana minimalna inhibitorna koncentracija (MIC). geraniol.5 McFarland i podešavan na finalnu koncen- Glavni sastojci esencijalnog ulja matičnjaka su citrali (ge. manuke (Leptospermum scoparium) jeva. 1. Mucor dejstva esencijalnih ulja matičnjaka. osveženi presejavanjem na SDA. seskviterpeni. komplek- sna organska jedinjenja koja karakteriše snažan miris i Materijal i metode predstavljaju sekundarne metabolite aromatičnog bilja.. dr Ane vidualnim varijacijama koncentracija i proporcija ovih Džamić sa Biološkog fakulteta Univerziteta u Beogradu. antihelmintsku. 1 | March 2016. osim nekih mediteranskih ostr. pri čemu njihova koncentra. Dosadašnja Minimalna fungicidna koncentracija (MFC) odre- istraživanja esencijalnog ulja vratića pokazuju njegovu đivana je presejavanjem po 10 µL sadržaja iz bunarčića značajnu antiinflamatornu. antiinflamatornu. prirodna. (15. Ove veoma složene smeše sadrže 20-60 komponenti u Istraživanje je sprovedeno na kliničkim izolatima različitim koncentracijama koje određuju njihova biolo. limonen. raznu. Citotoksični efekat esencijalnih ulja zasnovan je na Kulture gljivica su čuvane na Saburovom dekstroznom prooksidativnoj aktivnosti njihovih komponenti i čini ih agaru (Sabouraud dextrose agar. po. sa preovladavanjem određenih kompo. esencijalnih ulja i gljiva). tj. bez porasta na SDA i 24-časovnom inkubacijom na 37°C. i Aspergillus spp. (5). U ispitivanje svakog soja značajnu antimikrobnu. gljivica. tometrijskom metodom (Dinko Instrument Colorimetar. kontrola rasta) i negativna kontrola (bez dodatih Vratić je višegodišnja zeljasta biljka iz familije Aste. U tu svrhu su napravljene serije dvostrukih raz- krobnu. ulja. terpenoidi. ploče za mikrotitraciju i Miler-Hinton (Müller-Hinton) noterpene. leptospermon. traciju inokuluma od 103 do 104 CFU/mL ranial + neral). i korišćenjem diferencijalne hromogene Za ispitivanje antimikotičnog dejstva na uzročni. manuke i vratića.09 µl/mL. i Rhizopus spp. va. nenti u zavisnosti od hemotipa biljke (18). komponenti (8). tzv. kamfora. manuke i vratića na spp. Matičnjak pripada familiji Lamiaceae i široko je Barcelona). sa sezonskim i indi. nakon čega evropskog kontinenta. i kultivisanjem na 37°C. Dosadašnja istraživanja esencijalnog ispitivana je modifikovanom mikrodilucionim metodom ulja matičnjaka pokazuju da ono ima značajnu antimi. ke otomikoza odabrana su esencijalna ulja matičnjaka U daljem radu su korišćene 24-časovne kulture so- (Melissa officinalis). ka. metilcinamat. dizovan na 0. Ispitivana je osetljivost 20 so- na jedinjenja. Esencijalno ulje vratića predstavlja smešu α-tujona. izabranih metodom (6). koje su se kretale u ra- Manuka je cvetna biljka iz familije Myrtaceae. vrste u okviru rodova Candida i Aspergillus vršena je na stavu esencijalnih ulja biljaka sa različitih lokacija mogu osnovu mikroskopskih i kulturalnih karakteristika. Indi. Svaki bunarčić ploče u koji je do- kariofilen. seskviterpene.   45 . Antifungalna aktivnost esencijalnih ulja matičnja- β-kariofilen-oksid i germakren D. hranljive podloge CHROMagar (HiMedia. proapoptoznu i antitumorsku aktivnost (8-13). Hemijski sastav esencijalnih ulja može da varira u slučajnog izbora. Aktivne Za izvođenje mikrodilucione metode korišćene su komponente esencijalnog ulja manuke obuhvataju mo. Rasprostranjena je širom Zasejane ploče su inkubirane 48 h na 37°C. a inokulum je pripreman u destilovanoj vodi denzi- i vratića (Tanacetum vulgare). aromatična i alifatič. uzročnika otomikoza. poreklom iz Evroazije. 23). Esencijalna ulja su isparljiva. antiinflamatornu. blaženja za svako od esencijalnih ulja u rastvaraču. India). pri čemu je broj mikroorgani-zama standar- rasprostranjen u Centralnoj i Južnoj Evropi i Maloj Aziji. davano po 90 μL podloge i 10 μL odgovarajućeg razblaže- cija u ulju zavisi od hemotipa biljke (14). 24 h pre pravljenja ino- vidualne komponente esencijalnih ulja pokazuju različit kuluma. ka. Sojevi su odličnim antiseptičkim i antimikrobnim agensima. dobijenih ljubaznošću doc. Candida spp. kanala bolesnika sa znacima i simptomima otitis externe. antimikrobnu i antiulceroznu aktivnost (7. bujon sa 2% glukozom. 19-22). nja esencijalnog ulja zasejavan je sa 10 μL podešenog ino- traživanja esencijalnog ulja manuke pokazuju da ono ima kuluma ispitivanog soja gljiva. a za da budu uzrok promenljivog antimikrobnog dejstva (7). davanona i ulja koja sprečava vidljiv rast mikroorganizma (odsustvo zr- hrizantenona. U retkim slučajevima je Cilj našeg istraživanja je ispitivanje antimikotičnog utvrđeno da su izazivači bolesti Penicillium spp. izolovanih iz briseva spoljašnjeg ušnog zavisnosti od geografskog porekla i perioda berbe bilja. iz kolekcije Laboratori- ška svojstva. antioksidativnu. Volume 67 | No.Medical Youth Original articles iz roda Aspergillus i Candida su. smeši etanol-voda (70% etanol). β-kariofilen. mirtenola. SDA) na 4°C. nastog ili homogenog taloga na dnu bunarčića). koje karakteriše niska molekularna masa jeva Candida spp. antivirusnu i bile su uključene pozitivna (bez dodatog esencijalnog miorelaksantnu aktivnost (15-17). citronelal. Univerziteta u Beogradu. zbog čega varijacije u sa.

56 3.39 C.5 50 50 >50 * 3.5 >50 * 1. krusei 12.25 6.56 25 25 C.39 0.56 0.78 1.56 25 A.78 C.25 50 50 C.56 C.56 1.5 3. 20%) i Aspergillus ispitivanih sojeva u tabeli 1.25 12.niger 25 25 25 25 3. dok je 7 (35%) esencijalnog ulja manuke bila je veća od 50 μL/mL kod identifikovano kao Aspergillus spp.13 C. veća od 50 μL/mL u slučaju jednog izolata.56 1.56 6.fumigatus 25 50 25 >50 * 3.tropicalis 6. dok je MFC ulja matičnjaka bila okviru roda Candida su Candida krusei (5.25 6.78 3. krusei 3.5 3.13 >50* A.25 25 6.13 50 C.tropicalis 6. Opseg variranja minimalnih inhibitornih (a) i minimalnih fungicidnih (b) koncentracija esencijalnog ulja matičnjaka na uzročnike otomikoza 46  Mart 2016. krusei 6.13 * > od najvece testirane koncentracije a) b) Grafikon 1. 13 0.56 C.niger 25 50 12.tropicalis 6.25 25 25 A.13 3.25 25 6.25 6.56 1. 15%). krusei 6.13 25 A. Od 20 ispitivanih uzoraka. Identifikovane vrste u četiri ispitivana izolata. Aspergillus spp. krusei 6.25 25 1.39 1.13 25 A. Tabela 1.13 C.78 25 1.5 12. krusei 6. 40%).78 0.13 12.5 0.fumigatus 3. Rezultati anti- dida tropicalis (8.13 3.13 0.25 25 0.tropicalis 50 50 25 25 3.09 do 50 μL/mL.13 25 C.tropicalis 1.25 1.niger 12.56 1.25 50 0.56 25 A..56 1. krusei 50 50 0. krusei 6. u okviru ispitivanih koncentracija od skopskih karakteristika.78 1. Medicinski podmladak Originalni radovi Rezultati Ulja biljnih vrsta Melissa officinalis. nu aktivnost prema testiranim sojevima Candida spp.56 50 1.niger 12.25 25 6.56 C.56 1.5 1.56 C.5 50 25 50 1.56 C. Leptospermum scoparium i Tanacetum vulgare su pokazala antigljivič- Ispitivani izolati uzročnika otomikoza identifiko. Minimalna fungicidna koncentracija (65%) je identifikovano kao Candida spp. 25%) i Can. dok su u okviru roda Aspergilus gljivične aktivnosti su prikazani pojedinačno za svaki od pripadale vrstama Aspergillus niger (4.39 0.5 50 50 >50 * 3.fumigatus 25 25 12.39 0.56 1. Rezultati antifungalne aktivnosti ispitivanih esencijalnih ulja na uzročnike otomikoza Ispitivani sojevi Tanacetum vulgare Leptospermum scoparium Melissa officinalis gljiva MIC MFC MIC MFC MIC MFC A. fumigatus (3.13 50 1.25 12.25 1. i vani su do nivoa vrste na osnovu kulturalnih i mikro.25 50 6.56 0. | Broj 1 | Izdanje 67 .

MFC50 i kao i varijacije MFC u istom opsegu. kao i vrednosti MFC. 25 µL/mL 25 µL/mL 50 µL/mL Candida spp. 50 µL/mL 12. a) b) Grafikon 3. 25 µL/mL 50 µL/mL 50 µL/mL Candida spp. Najveću aktivnost ovo u odnosu na izolate C. MIC90. tropicalis. 50 µL/mL 1. 50 µL/mL 25 µL/mL 50 µL/mL Tanacetum vulgare Aspergillus spp. Vrednosti MIC50. do 50 μL/mL. dok se minimalne fungicidne koncentracije Minimalne inhibitorne koncentracije vrednosti nalaze u opsegu 0. 1 | March 2016. U odnosu na vrstu gljivica. Vrednosti MIC matičnjaka za ispitivane sojeve nalaze se u opsegu od 0. što je prikazano u tabeli 2.78-50 μL/mL. Esencijalno ulje biljke Tanacetum vulgare je pokaza. Vrednosti MIC MIC i MFC ispitivanih sojeva prikazan je na grafikonu 1. MIC90. esencijalno ulje matičnjaka je najveću aktivnost koza.39 esencijalno ulje matičnjaka je najveću aktivnost pokazalo do 50 μL/mL. U odnosu na vrstu MFC90 ispitivanih esencijalnih ulja na uzročnike otomi- gljivica.Medical Youth Original articles a) b) Grafikon 2.5 µL/mL 50 µL/mL Leptospermum scoparium Aspergillus spp.5 µL/mL 50 µL/mL Volume 67 | No. tljivosti određivane su vrednosti MIC50.   47 .56 µL/mL 50 µL/mL Melissa officinalis Aspergillus spp. MFC50 i MFC90 ispitivanih esencijalnih ulja na uzročnike otomikoza Esencijalna ulja Izolati gljivica MIC90 MFC50 MFC90 Candida spp. krusei.56-50 μL/mL. Opseg variranja vrednosti ulje je pokazalo prema izolatima C. esencijalnog ulja manuke kretale su se u rasponu od 0. krusei. 50 µL/mL 12. i MFC ispitivanih sojeva prikazane su na grafikonu 3. Na osnovu konstruisanih kriva kumulativne ose- lo varijaciju MIC vrednosti u intervalima 1. Opseg variranja minimalnih inhibitornih (a) i minimalnih fungicidnih (b) koncentracija esencijalnog ulja vratića na uzročnike otomikoza Minimalne inhibitorne koncentracije esencijalnog ulja pokazalo u odnosu na izolate C. Opseg variranja minimalnih inhibitornih (a) i minimalnih fungicidnih (b) koncentracija esencijalnog ulja manuke na uzročnike otomikoza Tabela 2.39 i MFC za ispitivane sojeve prikazane su na grafikonu 2.

Candida MIC ovog izolata bila veća od 16. Danas postoji opravdano povećano interesovanje bolesnika sa sistemskim mikozama. rezi- mogu da dovedu do komplikacija. kao i značajnim miorelak- santnim efektom (17). Malassezia furfur. niger je pokazao toleranciju na antivirusni efekti na širok spektar patogena: Staphylococ- esencijalno ulje matičnjaka. Escherichia coli. Candida albicans. pri čemu je vrednost odnosa MFC/MIC rok spektar dejstva. Pokaza- umerene klime. Pored toga. Matičnjak čajeva. Staphylococcus koje su u svojoj studiji pokazali Mimica-Dukić i saradnici aureus. 30). ulja vratića ukazuju na njegov širok spektar dejstva. Trichophyton spp. Potencijalni neže- esencijalnih ulja i njihove veće primene u lečenju različi- ljeni efekti ove grupe lekova i povećanje broja rezistentnih tih infekcija.. ali se gaji ekstrakata i esencijalnih ulja. | Broj 1 | Izdanje 67 . preovladava u to- na je i antitumorska aktivnost na linije humanih malignih plim klimatskim uslovima (28). Vrednosti MIC ovog ulja za gljive roda Aspergillus su hyton mentagrophytes (7. radu su pokazali da je Candida spp. poseduje izraziti antioksidativni efekat koji se pripisuje fe- Ovakav rezultat je donekle očekivan s obzirom na to da nolnim komponentama (10). Značajan problem za ispitivanje aktivnosti mnogih biljnih ekstrakata i esen- predstavljaju i površne mikoze. Proteus mirabilis. 16). što može da koristi u mikoza. pri čemu je vrednost MFC/ cus aureus. utvrđena su i antivirusna bile nešto više i za najveći broj izolata iznose 3. u obliku komercijalnih Manex preparata (26). Rezultati su. Esencijalno ulje matič- nih reakcija (27). ćelija. Medicinski podmladak Originalni radovi Određivanjem odnosa MFC/MIC ispitivanih ćelije ovo ulje pokazalo izraženo citotoksično dejstvo u esencijalnih ulja utvrđeno je da među ispitivanim izola.. Vratić je rasprostranjen širom evropskog konti- pokazali in vitro osetljivost ispitivanih sojeva na esenci- nenta. pri čemu izostaje citotoksični efekat na kontrolnu U ovoj studiji je ispitivan relativno mali broj sojeva grupu normalnih ćelijskih kultura (13). izolovan u 35 % slučajeva. Zelanda i jugoistoka Australije. Esencijalno ulje manuke ima širo- Diskusija ku primenu na Novom Zelandu. izolovana u 65% slu- Epidermophyton spp... Fusarium spp. Candida parapsilosis i Trichop- (8). Trichophyton spp. Dosadašnja farmakolo- Tolerantni sojevi se karakterišu smanjenom osetljivošću ška ispitivanja esencijalnog ulja manuke ukazuju na ši- na antimikotike. Samo jedan soj A. posebno pojedinih vrsta.56 μL/mL. HSV-1 i zali toleranciju na ovo ulje jer je MFC/MIC iznosila 2-4. Candida albicans. Ovo dejstvo je dokazano i ispitivanjem in vitro pitivana ulja kao efikasnije na izolate roda Candida. Ostali izolati nisu poka. U Srbiji raste sporadično. svojstva ovog ulja prema tipu 1 Herpes simplex virusa dok vrednosti MFC iznose 25 μL/mL. Ove vrednosti su znatno niže od vrednosti na vrste Escherichia coli. Dosadašnja istraživanja esencijalnog jalna ulja ispitivanih biljaka. Brojna ulja i antimikrobnih lekova imaju svoje prednosti: siner- farmakološka ispitivanja esencijalnog ulja matičnjaka gistički ili aditivni efekat i smanjenje toksičnosti i neželje- ukazuju na širok spektar dejstva. Derivati azola predstavljaju značajnu grupu anti- trebalo bi razmotriti mogućnost intenzivne proizvodnje gljivičnih lekova u lečenju otomikoza. dok Aspergillus spp. kontrolnoj grupi normalnih ćelija (25). da se Candida spp. najčešći izazivači oto- holinesterazna aktivnost ovog ulja. pri citotoksičnog efekta esencijalnog ulja na vrstu Schistoso- čemu je MIC najvećeg broja izolata 0. (8. U Esencijalno ulje biljke matičnjaka pokazalo je narodnoj srpskoj medicini vratić se koristi kao antihel- značajniju antigljivičnu aktivnost u odnosu na ostala is- mintik. kao i mogućnost terapijske primene. Kombinacije esencijalnih kao lekovita sirovina za farmaceutsku industriju. Upoređivanje dobijenih rezultata sa rezultatima že. s obzirom na to iz literature je otežano s obzirom na to da je u večini dru- da je prilikom ispitivanja citotoksičnog efekta na maligne 48  Mart 2016. tima uzročnika otomikoza nisu nađeni tolerantni sojevi Manuka je endemska biljka. ali i ekonomskih razloga (6). najčešće izoluje u okviru potencijalnom lečenju Alchajmerove bolesti (11). dok je Aspergillus spp. Ovi rezultati su u (HSV-1) (21). i Candida spp. Neophodna su dodatna ispitivanja stepena korelaciji sa rezultatima pokazanim u sličnim radovima resorpcije esencijalnog ulja vratića preko kože i sluzoko- (29. Pored širokog antimikrobnog dejstva 1.13 μL/mL. Microsporum spp. njaka ispoljava antifungalnu aktivnost u odnosu na brojne Rezultati identifikacije uzročnika otomikoza u patogene: Candida spp. 23). tropicalis. od kojih je najčešća stencije mikroorganizama.78 μL/mL i MFC ma mansoni (20). Mnoge lekovite biljke se već vekovima koriste u Gljivična oboljenja predstavljaju veliki problem tradicionalnoj medicini u lečenju i prevenciji mnogih današnjice zbog sve većeg broja imunokompromitovanih bolesti. poreklom sa Novog na esencijalna ulja vratića i manuke (MFC/MIC = 2-4). poput otomikoza koje cijalnih ulja zbog toksičnih efekata mnogih lekova.. Dokazani su njegovi antimikrobni i > 16. gljiva. 24). Budući da naši prostori poseduju izuzetno bogatu floru. Dokazana je izražena anti- su Aspergillus spp. HSV-2 (15. međutim. tj. sojeva uzročnika otomikoza opravdano su povećali inter- Matičnjak je biljka široko rasprostranjena u central- es za ispitivanje antimikotične aktivnosti mnogih biljnih noj i južnoj Evropi. pa i u Srbiji. otitis media. odlikuje se izuzetnim antiin- flamatornim svojstvima (15).

growing in Slovakia. Mikulašo. Bent J. Iako se u statističkim razmatranjima određivanje lata iznose 25 μL/mL.25 μL/mL i MFC 25 μL/mL. Idaomar M.Medical Youth Original articles gih radova korišćen disk-difuzioni metod koji je manje Značaj rezultata ovog rada je u karakteristikama precizan i ne daje vrednosti MIC i MFC. kao i o relativ- roda Candida (7). Vrabec JT. International Journal of Pediatric Otor- za Candida spp. Za razliku od ostalih izolata kod 1. stentniji u odnosu na sojeve izolovane iz spoljašnje sredi- ćeg broja izolata 6. izazivača otomikoza. a Literatura MFC je bila veća od maksimalne ispitivane koncentracije ovog ulja (> 50μL/mL). gde je obuhvaćen relativno mali broj izolata (N = 20). Topical antibiotic in- razlikuju (tabela 2). Jackman A. prikaz rezultata i u ovakvim istraživanji- Esencijalno ulje manuke se. Ho T. 2004. Alnamer R. U slučaju jednog izolata A. 2006. kojih odnos MFC/MIC nije bio veći od 2-4. s obzirom na to da se radi o hu- Esencijalno ulje vratića se pokazalo efikasnijim manim izolatima. April M. manuke i vratića na sojeve Candida spp. Ovakav effects of essential oils – A review.69:857-60. nažalost. Mycoses. Journal of agricultural and food 50%. Averbeck S. kod pome- 2.46:446-75. kao i prethodna ulja. korišćenih sojeva gljiva. što bi trebalo da. 2008. Aspergillus spp. sojeva gljiva. Coker NJ. Vrednosti MIC ovog ulja za gljive iz no malom broju ispitanih sojeva. Razlike u vrednostima MIC50 i MIC90 duced otomycosis. Otomycosis: prevalence. Vaverková Š. sential oils from Tanacetum vulgare L. Prabhu prema esencijalnom ulju matičnjaka. Kotigadde S. Mimica-Dukic N.135:787-91. Richardson MD. Yoo D. razlikuju se najviše za dvostruko Vivo Potential Anti-Inflammatory Activity of Melissa offi- razblaženje esencijalnih ulja. Iste vrednosti za 5. Hajjaj G. Ovakav in vitro rezultat ukazuje na po- kao i Liz-Balkin (Lis-Balchin) i saradnici (15. variraju za dvostruku ili četvorostruku 6. Cherrah Y. ma.. Kurnatowski P. tencijalnu primenu esencijalnih ulja navedenih biljaka u Značajni su i podaci koji se odnose na tolerantne lečenju otomikoza. Mikulášová M. pokazano je da navedena esencijalna ulja imaju an- mL). dok vrednosti MFC iznose 50 μL/ kumulativnih vrednosti MIC50 i MIC90. tj.. što je krite. 32). Otomycosis: clinical fe- nutog izolata je odnos MFC/MIC veći od 16. u zavisnosti od esencijalnog ulja. Averbeck D. 2014. koji su rezi- prema gljivama iz roda Candida. sojeva (MIC50 i MIC90) za ova dva roda gljiva značajno se 4. 9. s obzirom na to da tolerancija mikroorgani. Naši rezultati su u suprotnosti sa rezultatima studije MFC90 uglavnom smatra reprezentativnim samo ukoliko Džonsa (Jones) i saradnika. Ovakav nalaz ukazuje na postojanje tolerancije ovog soja 3. In ne razlikuju značajno. Bounihi A. ukoliko se dokaže odsustvo 7. tivanih esencijalnih ulja koja su delovala fungicidno na (Lamiaceae) essential oil. therapeutic procedure. Food and Chemical To- nalaz ukazuje na veliku razliku u osetljivosti pojedinih xicology. In- su delovala inhibitorno na 50%. Vred- nosti MFC ovog esencijalnog ulja su izrazito varijabilne Zaključak i kreću se u opsegu od 0. odnosno 90% ispitivanih ternational Journal of Microbiology. crobial and antioxidant activities of Melissa officinalis L. Nova Biotechnol. and Salvia officina- utiče na korišćenje visokih koncentracija ulja u eventual. Filipiak A. Sokovic M. nim preparatima za lečenje otomikoza. ne. hteva dodatna ispitivanja. 2001. Otolaryngology--Head rijum za procenu tolerancije na antimikrobno sredstvo. P et al. Slične rezultate su pokazali Čen (Chen) i saradnici. Simin N. izazivačima infekcije uha. 8. Thada ND. i Aspergillus Aspergillus u većini slučajeva nije bilo moguće odrediti jer spp. nisu potpuno uporedljivi va (Mikulášová) i saradnici su takođe pokazali umerenu sa rezultatima iz literature. Primary Otomycosis in the Indian Subcontinent: Vrednosti MIC ispitivanih esencijalnih ulja koja Predisposing Factors. Antimicrobial effects of es- citotoksičnog efekta esencijalnog ulja na ćelije domaćina. Zellou A.   49 .. koji su negirali antifungalnu je ispitivanjem obuhvaćen veliki broj sojeva mikroorgani- aktivnost ovog ulja (31). odnosno 90% ispitivanih sojeva (MFC50 i MFC90) chemistry.13 μL/mL.9:161-5. zama na antimikrobna sredstva predstavlja početni korak ka rezistenciji. Ward R. 2008. tigljivični efekat. zama (N > 100). niger pokazano je da je MIC esencijalnog ulja matičnjaka 3. Dobijeni rezultati. Vrednosti MFC za ka. Shekhar M. kao i MFC50 i mL. 1 | March 2016. s obzirom na to da se radi o antifungalnu aktivnost ovog esencijalnog ulja na gljive iz različitim vrstama u okviru roda Candida. Bozin B. pri čemu je MIC najve. čemu je MIC najvećeg broja izolata 6. Warnock DW. 2005. 2009. Fungal Infection: Diagno- sis and Management: Wiley. Bakkali F. clinical symptoms. pokazalo efikasnijim prema gljivama iz roda Candida.52:2485-9. Antimi- Zapaža se da se vrednosti MFC koncentracija ispi. and Neck Surgery. and Classification. Sistemska primena ovih supstanci za- sojeve gljiva. atures and treatment implications. Biological koncentraciju u zavisnosti od esencijalnog ulja. hinolaryngology. roda Aspergillus su bile nešto više i za najveći broj izo. Volume 67 | No. lis L. razli. pri može da bude koristan i ilustrativan. kuju se za 8-32 puta veću koncentraciju. Vrednosti MIC ovog ulja za gljive iz roda Aspergillus bile su nešto više i Ispitivanjem aktivnosti esencijalnih ulja matičnja- za najveći broj izolata iznose 25μL/mL.39 do 50 μL/mL.25 μL/mL. Prasad SC. Microbiology. bujon mikrodilucionom meto- su prevazilazile najveću ispitivanu koncentraciju (>50 μL/ dom. pre svega citotoksičnog efekta.2014:1-9.44:472-9.

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nitrated and dopamine-modified ASYN) statistički značajnog pada vijabiliteta SH-SY5Y ćelija huma.com Sažetak Abstract Uvod: Glavni patohistološki supstrat Parkinsonove bo. Ratko Radeta1. violet assay. Za ispitivanje ćelijskog vijabiliteta korišćen je dopamine-modified ASYN). led to substantial decrease in number of viable differentiated nog neuroblastoma diferenciranih u neuronski fenotip. diferenciranoj u Materials and methods: All experiments were con- neuronski fenotip all-trans retinoičnom kiselinom. povećanog preživljavanja ćelija. nitrated and njenim oblikom. modified recombinant wt ASYN (oligomeric. AMPK. Maja Jovanović1. Pharmacological activator of AMPK farmakološkog aktivatora AMPK dovela je do značajnog pora. što je SH-SY5Y cells. AMPK. together with an AMPK i nishodne kinaze Raptor. nase (AMPK) signaling have been shown in several neurode- Cilj: Cilj ovog rada je bio da se ispita uloga AMPK u generation models. Cell viability was assesed using crystal na imunoblot tehnikom. Posttranslational modifications. Marija Dulović1. accompanied by inactivation of AMPK/Raptor praćeno smanjenjem aktivnosti protein kinaze aktivirane adeno. nitrozilacija) (PD) pathogenesis. Farmakološki aktivator AMPK (AICAR) dovodi do tective role of AMPK in neurotoxicity of extracellular ASYN. autofagija Volume 67 | No. can increase aggregation of ASYN. including PD. kao i povećanim prelaskom increase in the conversion of LC3-I to LC3-II. Rezultati: Analiza ćelijskog vijabiliteta je ukazala da Results: Treatment with modified recombinant wt sva tri oblika rekombinantnog vanćelijskog ASYN dovode do ASYN (oligomeric. što sve zajedno upućuje na neu- roprotektivno dejstvo AMPK u neurotoksičnosti izazvanoj delo- vanjem modifikovanih oblika vanćelijskog wt ASYN. Univerzitet u Beogradu Kontakt: marijaj26@yahoo. Ključne reči: modifikovani oblici rekombinantnog vanćelijsk. Activation of AMPK and Raptor. difikovanih oblika rekombinantnog ASYN. Primena with recombinant ASYN. dejstvu modifikovanih oblika rekombinantnog vanćelijskog wt Aim: The aim of this study was to investigate the role of (engl. nitrated and dopamine-modified rekombinantnog vanćelijskog α-sinukleina (ASYN) dovode do ASYN) induced decrease in number of viable differentiated smanjenog preživljavanja ćelija humanog neuroblastoma. kristal violet test. Ove ćelije ducted in all-trans retinoic acid-differentiated human neu- su tretirane modifikovanim oblicima rekombinantnog vanće. uključujući i PB. indicating a pro- autofagije. including ASYN pospešuju njegovu agregaciju u neuronima.Medical Youth Original articles Uloga protein kinaze aktivirane adenozin monofosfatom (APMK) u dejstvu modifikovanih oblika rekombinantnog vanćelijskog α-sinukleina in vitro The role of adenosine monophosphate-activated protein kinase (AMPK) in the action of modified recombinant extracellular wt (wild-type) ASYN in vitro Marija Jeremić1. autophagy og wt α-sinukleina. ASYN on differentiated SH-SY5Y cells. Alterations of adenosine monophosphate-activated protein ki- delima neurodegenerativnih bolesti. using immunoblotting. the presence of all 3 types of modified recombinant wt ASYN. što je SH-SY5Y cells. signalling pathway and autophagy induction. Conclusion: Modified forms of recombinant wt (wild- Zaključak: Oligomerni. kao i konverzija proteina LC3-I u LC3-II formu praće. (AICAR) significantly increased the number of viable cell in sta vijabiliteta ćelija koje su gajene u prisustvu pomenutih mo.   51 . were monitored Raptor. Introduction: Alpha-synuclein (ASYN) accumulation lesti (PB) predstavlja nagomilavanje proteina ASYN. upon treatment proteina LC3-I u LC3-II oblik vezan za autofagozome. Western blot analysis showed decreased levels bilo praćeno smanjenjem nivoa aktivne (fosforilisane) forme of AMPK and its downstream kinase Raptor. wild-type) ASYN na diferencirane SH-SY5Y ćelije. nitrovanim i dopaminski izme. Pharmacological zin monofosfatom (AMPK)/Raptor signalnog puta i indukcijom activation of AMPK improved cell survival. nitrovani i dopaminski oblik type) ASYN (oligomeric. roblastoma SH-SY5Y cells. dok je promena u aktivnosti enzima AMPK i together with conversion of LC3-I to LC3-II. Pokazano is considered to be one of the key factors in Parkinson’s disease je da posttranslacione modifikacije (oksidacija. Key words: modified recombinant wt ASYN. SY5Y ćelijskoj liniji humanog neuroblastoma. treated with 3 different types of lijskog wt ASYN: oligomernim. Promene u oxidation and nitration. 1 | March 2016.2 1 Medicinski fakultet Univerziteta u Beogradu 2 Mentor: Institut za medicinsku i kliničku biohemiju. AMPK in the action of modified recombinant wt (wild-type) Materijal i metode: Svi eksperimenti su rađeni na SH. aktivnosti signalnog puta AMPK pokazane su u različitim mo.

Medicinski podmladak Originalni radovi Uvod spešuje agregacija (14.6-tetrahidropiridin) (17). Rigiditet. Pokazano je da aktivacija AMPK u in vitro mo- teolitička sistema (6 . silent information regulator in oslobađa posredstvom egzozoma (12). cilj ovog rada je bio pokazala da je oligomerni oblik toksičan za neurone (13).3. Do povećanog nakupljanja ASYN u nizama u ćeliji. nitrovani i oblici izmenjeni dopaminom) (5). 4). Studije pokazuju da za patološke doga. a posledica su odumiranja dopaminergičkih U patogenezi PB značajnu ulogu ima oštećenje neurona u pars compacta substantiae nigrae mezencefa.8). rađeni na SH-SY5Y ćelijskoj liniji humanog neurobla- gičke neurone posebno osetljivim na toksično delovanje stoma. specifičnog biomarkera u dijagnozi i praćenju PB (12). Na- kupljanje različitih oblika ASYN. ASYN podleže različitim posttranslacio. European collection of animal cell ASYN. nabavljenoj od Evropske kolekcije animalnih će- ASYN (14). koje prekomerno proizvode ASYN. toksičnom modelu PB indukovanom rotenonom. a kao posledica metabolizma dopamina može da cultures. 6). čime se po. 9). Takođe je pokazano da 1. i da na taj na. što otvara mogućnost korišćenja ASYN kao karakteristični simptomi neophodni za dijagnozu obole.2. sa posledičnim padom unutarćelijske koncentracije Patološki supstrat Parkinsonove bolesti predsta. iz Velike Britanije). ili zbog smanjenog kapaciteta za razgradnju ovog prote. AMPK (6). ali se pretpostavlja da u nastanku njenja kapaciteta za sintezu adenozin trifosfata (ATP). u uzorcima tkiva mozga. oligomerni. ovaj prote. lona (2). rekombinantnog vanćelijskog wt ASYN u kulturi dife- nim modifikacijama. već da se promene u koncentraciji ovog neurodegenerativno oboljenje ljudi. AMPK. gde je uron (11). te neurološke bolesti praćene promenama u aktivnosti đaje u PB može da bude odgovorna disfunkcija oba pro. uključujući i razgradnju proteina putem ćelijama može doći usled njegove prekomerne sinteze i/ autofagije (7). Kultura ćelija i reagensi dirani. Kao rezultat degeneracije mitohondrija. kao i na drugom neuro- fibrilarni ASYN može da se prenese sa neurona na ne. Kao potvrda toga. Iako je fibrilarni Imajući u vidu oskudne podatke o ulozi AMPK u ASYN glavna komponenta Levijevih tela. kao što je spo- do oštećenja sistema za razgradnju proteina. 2). odmah posle Alc. modelu Hantingtonove bolesti kod miša. proteina mogu otkriti i u cerebrospinalnoj tečnosti obo- hajmerove bolesti (1). 2) preko aktivacije autofagije (8). kao i da poboljšava preživljavanje ćelija u peronima i makroautofagija) (6. Neuroprotektivna konformaciju usled čega lako dolazi do stvaranja razli. što može da uzrokuje aktivaciju (ASYN) unutar struktura nazvanih Levijeva tela (1. nom delovanjem neurotoksina MPP+ (1-metil-4-fenil- protofibrilarni i filbrilarni (10). kao i da se iz ćelija humanog neuroblastoma protektivna uloga rezveratrola posredovana signalnim SH-SY5Y. čini dopaminer. dolazi do sma- potpunosti razjašnjeni. | Broj 1 | Izdanje 67 . mitohondrija. istraživanja su dejstvu modifikovanih oblika ASYN. ECACC. Dva glavna proteolitička sistema u neuronima koja nju energetskog statusa ćelija i njihovog preživljavanja u učestvuju u fiziološkoj razgradnji i uklanjanju proteina uslovima energetskog stresa (8). putem AMPK-SIRT1 (engl. oksi. koji se ak- dopaminergičkim neuronima u fiziološkim uslovima. iako noviji podaci daju prednost delu Alchajmerove bolesti inhibira nastajanje amiloidnih lizozomalnim mehanizmima (autofagija posredovana ša. najnovija istraživanja pokazuju da ASYN nije isključivo unutar- Parkinsonova bolest (PB) je drugo najučestalije ćelijski protein. O ulozi AMPK Pokazano je da ASYN ima nestabilnu osnovnu u Parkinsonovoj bolesti se malo zna. zahvaljujući 52  Mart 2016. AMPK i u neuronima ima značajnu funkciju u održava- ina. koje mogu da budu uzrok njegove renciranih ćelija humanog neuroblastoma SH-SY5Y. Etiologija i patogeneza bolesti još uvek nisu u glavnog energetskog generatora u ćeliji. lih od PB. kao i interakcija ovog Za potrebe ovog istraživanja svi eksperimenti su proteina sa metabolizmom dopamina. ATP-a i porasta unutarćelijske koncentracije adenozin vlja citoplazmatsko nakupljanje proteina α-sinukleina monofosfata (AMP). Protein kinaza aktivirana adenozin monofos- Alfa sinuklein je protein koji ostvaruje svoju funkciju u fatom je glavni metabolički energetski senzor. koji dovodi karakteristike dopaminergičkih neurona. su ubikvitin -proteazni sistem i lizozomalna razgradnja Dosadašnja istraživanja ukazuju da su različi- – autofagija (5. plakova (16). sobnost sinteze dopamina i noradrenalina. dobijenim nakon smrti obolelih od PB. ali tivira u uslovima povećanja AMP/ATP odnosa i reguliše njegovo nagomilavanje dovodi do degeneracije neurona metaboličku homeostazu kontrolom većeg broja meha- i pojave PB (1. da se ispita uloga AMPK u dejstvu modifikovanih oblika Pored ovoga. povećane agregacije. 15). tremor i bradikinezija su lelih od PB. Ove ćelije poseduju nastane dopaminski izmenjen oblik ASYN. uloga AMPK je pokazana na in vitro modelu PB izazva- čitih oblika ASYN kao što su: monomerni. Kao i u ostalim ćelijama našeg organizma. Uz sve navedeno. bolesti učestvuju genetski činioci i faktori sredine (3). čin ASYN može da pređe u zdrave ćelije. uočeni Materijal i metode su proteinski agregati u kojima se nalaze modifikovani oblici ASYN nastali posttranslaciono (fosforilisani. Dopamin stabilizuje protofibrilarnu formu lijskih kultura (engl.

15 i 20 mg/ml. koja se dobija pomoću goveđeg se- korišćen farmakološki aktivator AMPK: 5-aminoimida. Kako bi Modifikovani oblici rekombinantnih wt ASYN su se sprečilo nespecifično vezivanje antitela za membranu. 22). Jeddah. Test vijabiliteta Imunoblot je korišćen za vizuelizovanje promene u aktivnosti AMPK i Raptor-a u SH-SY5Y ćelijama di. SAD) u potvrđeno je pomoću oligomernog ASYN ELISA testa i mleku (5% u rastvoru TBS-T).   53 . Za ovu analizu. ženi su kao procenat živih ćelija u tretmanu u odnosu na koji se specifično inkorporira u membranu autofagozo. u uzorku upotrebom odgovarajućih denzitometrijskih tehnika. Velika Britanija) na 570 nm u prvih Modifikovani oblici rekombinantnog alfa-sinukleina sat vremena. 21. ja proteina u svakom uzorku metodom po Bradfordu. LC3 (1:900) inskih protofibrila. rumskog albumina (engl. Modifikovani oblici rekombinan-tnog wt dodato je sekundarno antitelo. koja menja intenzitet u prisustvu proteina (od Vijabilitet ćelija je određivan kristal violetnom me. 15 minuta na 4ºC. tehnikom imunoblota je ćelijama. Raptor (1:1000). 40. dobijeni su kvalitativni je Ag). nitrovani (n-ASYN) i ASYN izmenjen gelu. (1:1000). Jednake količine proteina iz svakog uzorka razdvo- fikovane forme rekombinantnog wt ASYN: oligomerni jene su pomoću SDS-PAGE na 10% poliakrilamidnom oblik (o-ASYN). Marnes-la-Coquette. 2 x plave boje proporcionalan je koncentraciji proteina u uzor- 104 ćelija u svakom bunaru. Univerziteta King Abdel Aziz. a potom je izvršen transfer na nitrocelulozne mem- dopaminom (DA-ASYN) (5). Jackson IP nakon čega je rađen test vijabiliteta.Medical Youth Original articles ekspresiji tirozin hidroksilaze i dopamin β-hidroksilaze zirane na ledu 30 minuta puferom za liziranje (engl. Ova metoda se koristi i za kvantifikaciju proteina podaci o prisustvu određenog proteina. brane (Bio-Rad. tech. u toku vremenskog intervala od 6 dana centrifugirane su na 14000 g. sa Me. Nakon njegovog ispiranja imunoblotom. 1 mM fenilmetilsulfonilfluorid i inhibitori proteaza). Diferencijacija ćelija u neuronski fenotip vršena je buffer) (30 mM Tris-HCl pH 8. gde se koriste At specifična za uzorak koji signala na rendgenskom filmu. nitrovanog i oligomernog ASYN. 150 mM NaCl. ku. U ovom radu je na standardnu krivu. p-Raptor (1:1000). Coomesie Brilli- finalnoj koncentraciji 0. Ova boja se vezuje za proteine u živim indukcija procesa autofagije. Ćelije su gajene na 37ºC u vlažnoj atmosferi koja čega je pokupljen supernatant. cije 20 mM (17). 1:5000. braon do različitih nijansi plave boje). anti-rabbit IgG-HRP At. Ovom prilikom je science. SAD) na nitrocelulozne membrane i vizuelizacijom titelo (Ag-At).all-trans“ retinoične kiseline (RA). Membrane su stajale tokom noći potopljene u (20. 1% NP- pomoću .A. Kao kontrolu dobijenih imunobloto- va koristili smo određivanje nivoa proteina aktina. 10. sadrži 5% ugljen-dioksida (CO2) u medijumu RPMI Nakon liziranja ćelija određivana je koncentraci- (Roswell Park Memorial Institute) 1640. bovine serum albumin. Intenzitet dobijene todom nakon zasejavanja ćelija u ploče sa 96 bunara. Velika Britanija). lysis (18). netretirane (kontrolne) diferencirane ćelije (Ø) za koje je ma i autofagolizozoma (23). vijabilitet arbitrarno postavljen na vrednost od 100%.05% Tween 20 (TBS-T) (1 sat na sobnoj tem- difikovanih oblika ASYN opisan je u ranijim studijama peraturi). obogaćenom fe. a zasniva se na reakciji antigen-an. Kristal violet metoda je korišćena za ispitivanje će- ferenciranim u neuronski fenotip. 1 | March 2016. ćelije su li. datkom 0. rastvoren u medijumu u korišćena boja kumasi sjajnoplava (engl. Nalivanjem hemiluminescent- Imunoblot tehnika je metoda koja služi za otkriva. Volume 67 | No. talnim goveđim serumom (10%). rastvorenom u Tris-puferovanom rastvoru (TBS) sa do- Saudijska Arabija. Kako bi se potvrdila lijskog vijabiliteta. Dobijeni rezultati predstavljaju srednju le-associated protein 1A/1B-light chain 3) u LC3-II oblik vrednost triplikata iz tri nezavisna eksperimenta i izra- proteina koji je konjugovan sa fosfatidil-etanolaminom. ant Blue). membrana je inkubirana u 5% nemasnom mleku u prahu dicinskog fakulteta. koncentraciji od 5. kako Tehnika imunoblota (Western blot) bismo utvrdili da je na gelove nanošena jednaka količina proteina za svaki uzorak. SAD). AMPK Formiranje dopaminski izmenjenih alfa-sinukle. BSA) u zol-4-karboksiamid ribonukleotid (AICAR) (Tocris Bio. Apsorbanca je očitana na čitaču za mikrotitarske plo- če (TECAN Sunrise. rastvor primarnog antitela za: p-AMPK (1:1000). Za potrebe ovog istraživanja korišćene su modi. Laboratories. nog supstrata za peroksidazu (Amersham Pharmacia Bio- nje proteina u uzorku.0. koje se vezuje za perok- ASYN su korišćeni u koncentraciji od 5 µM u toku 48h. Francuska).. microtubu. broju živih ćelija. sidazu (engl. a nakon (19).125 mM. tako da je intenzitet plave boje proporcionalan ispitivana konverzija solubilnog LC3-I (engl. El-Agnafa. koncentra. i β-actin (1:5000) (Cell Signaling Technology. L-glutaminom (2 mM) Koncentracije proteina u uzorcima se računaju u odnosu i smešom antibiotik/antimikotik (1%). Nakon očitavanja apsorbanci napravljena je standardna kriva sa koje su očitane koncentracije proteina. Proces dobijanja i karakterizacije mo. dobijeni ljubaznošću dr Omara M. Bristol.

ali je poboljšao (p < 0. nosu na kontrolu. nakon čega je kristal violet testom određen vijabilitet ćelija. Sva tri modifikovana oblika rekombinantnog van- Statistička analiza ćelijskog wt ASYN dovela su do smanjene zastupljenosti aktivnog (fosforilisanog) oblika AMPK (pAMPK). Odliven je metanol i dodat je ras. tj. U svaki bunar je do- dato po 50 μL 33% rastvora sirćetne kiseline. Rezultati Modifikovani oblici rekombinantnog vanćelijskog wt ASYN uzrokuju pad vijabiliteta ćelija humanog neuroblastoma SH-SY5Y Sva tri modifikovana oblika rekombinantnog van- ćelijskog wt ASYN dovela su do statistički značajnog pada vijabiliteta ćelija humanog neuroblastoma diferenciranih Slika 1. u od- tvor boje kristal violet (1:10 u fosfatnom puferu (PBS-u)). kod su oprane su puferovanim fiziološkim rastvorom (PBS) i koga je vijabilitet ćelija pao na 58. netretirane diferencirane ćelije hu- po 50 μL u svaki bunar. Farmakološka aktivacija AMPK poboljšava preži- ćelijskog wt ASYN na vijabilitet SH-SY5Y ćelija. ćelije su isprane u posudi sa vodom. Apsorbanca je očitana na čitaču za mikrotitar. AICAR. kao i Za poređenje rezultata korišćen je Studentov t-test aktivnog (fosforilisanog) oblika nishodne kinaze Raptor za dva mala nezavisna uzorka. Raptor signalnog puta ske ploče (TECAN Sunrise. (AMPK.73% dok je najmanje toksičan bio ASYN koji je doveo izvršena fiksacija ćelija. Farmakološki aktivator AMPK. Rezultati su izraženi kao srednja vrednost triplikata iz 3 repre- zentativna uzorka ± S. Zastupljenost ukupnih formi proteina (statistički veoma značajno). i na ovaj način je 1. Velika Britanija) na 570 nm. modifikovanih oblika vanćelijskog ASYN njenog).5 ± 10 minuta stajale na sobnoj temperaturi. Diferencirane ćelije su gajene 48 sati u prisustvu modifikovanih oblika vanćelijskog ASYN (oligomernog. Kristal violet Modifikovani oblici rekombinantnog vanćelijskog se pod dejstvom sirćetne kiseline rastvara i rastvor postaje wt ASYN dovode do smanjene aktivacije AMPK/ plave boje. nog neuroblastoma SH-SY5Y. Medicinski podmladak Originalni radovi Nakon isteka inkubacije.59% (Grafikon 1). Uticaj modifikovanih oblika rekombinantnog vanće- lijskog wt ASYN na aktivnost AMPK/Raptor signalnog puta. Posle 10 minuta odlivena je boja i manog neuroblastoma SH-SY5Y. one u neuronski fenotip. ćelijama je odliven medijum. Raptor) u svim analiziranim uzorcima ostala je nepromenjena (slika 1). nakon čega je imunoblot analizom otkri- vena aktivnost AMPK/Raptor signalnog puta. Ploče su potom ASYN koji je uzrokovao smanjenje vijabiliteta na 65.05 u poređenju sa netretiranim veo do statistički značajnog smanjenja vijabiliteta di- diferenciranim ćelijama (Ø)) ferenciranih SH-SY5Y ćelija.05 (statistički značajno) i p < 0. Kao kriterijum značajnosti (pRaptor) kod diferenciranih SH-SY5Y ćelija humanog razlike uzeto je p < 0.01 neuroblastoma. Diferencirane vljavanje diferentovanih diferenciranih ćelija huma- ćelije su gajene 48 sati u prisustvu modifikovanih oblika van. | Broj 1 | Izdanje 67 .D. Uticaj modifikovanih oblika rekombinantnog van.05 u poređenju sa netretiranim diferenciranim ćelijama (Ø)) Grafikon 1.05) preživljavanje diferenciranih SH-SY5Y ćelija u dvostru- 54  Mart 2016. zatim o- dodato je po 50 μL metanola u svaki bunar.55 ± 1. nitrovanog i dopa- minski izmenjenog). Najtoksičniji je bio DA-ASYN.69 ± 1. (*p < 0. do pada vijabiliteta na 69.12%. nije do- Rezultati su izraženi kao srednja vrednost triplikata iz 3 repre- zentativna uzorka ± SD (*p < 0. gajenih u prisustvu ćelijskog ASYN (oligomernog. nitrovanog i dopaminski izme.

Uticaj modifikovanih oblika rekombinantnog vanćelij- skog wt ASYN na prelazak LC3-I u LC3-II oblik proteina. otporni nju dvostruko ASYN+AICAR tretiranih diferenciranih ćelija sa na hidrolizu proteinazom K. Rezultati su izraženi kao srednja vrednost triplikata iz 3 repre- ke.05. Prekomerno na- Grafikon 2. primenjeni samostalno (u odsustvu AICAR). mar- kera autofagije. SY5Y gajenih u prisustvu a) oligo-mernog b) nitrovanog i c) Karakteriše ga nestabilna osnovna konformacija. Ovaj rezultat ukazuje na potencijalni značaj autofagije u dej- stvu modifikovanih oblika ASYN (slika 2). Uticaj farmakološkog aktivatora AMPK (AICAR) kupljanje ovog proteina u neuronima i njihovom okruže- na vijabilitet diferenciranih ćelija humanog neuroblastoma SH. doveli do statistički značajnog (p < 0.Medical Youth Original articles kom tretmanu sa svim ispitivanim oblicima modifikova- nog vanćelijskog ASYN koji su. Diskusija c) Alfa sinuklein je protein koji igra centralnu ulogu u funkciji dopaminergičkih neurona (3).D.05. što je verovatno ključno za ćelijama tretiranim samo modifikovanim oblicima ASYN) formiranje Levijevih tela i neurodegeneraciju (24). nitrovanog i dopaminski izmenjenog). nastalih solubilnih nastaju stabilni oligomeri. gde su određene vrste oligomera nosioci toksičnosti zentativna uzorka ± S. Prethodna istraživanja su pokazala da od početno netretiranim diferenciranim ćelijama (Ø) **p < 0. (*p < 0.   55 . Pored toga što se za oligomerni oblik ASYN smatra da je nosilac Volume 67 | No. b) Slika 2.05) smanjenja broja živih diferenciranih SH-SY5Y će- lija. svojstvenog autofagiji (27). u poređenju sa kontrolnim (10). 1 | March 2016. tako da dopaminski modifikovanog oblika rekombinantnog vanćelij- lako prelazi iz monomernog u različite oligomerne obli- skog wt ASYN. u poređe. što je posebno izraženo u tretmanu dopaminskim oblikom ASYN. Modifikovani oblici rekombinantnog vanćelijskog ASYN dovode do povećane zastupljenosti marke- ra autofagije u diferenciranim SH-SY5Y ćelijama humanog neuroblastoma Modifikovani oblici rekombinantnog vanćelijskog a) ASYN u diferenciranim SH-SY5Y ćelijama humanog ne- uroblastoma dovode do porasta u prevođenju LC3-I u LC3-II oblik. nju je važan etiopatogenetski faktor u nastanku PB (1). nakon čega je imunoblot analizom detektovana konverzija LC3-I u LC3-II. Ćelije su gajene 48 sati u prisustvu modifikovanih oblika vanćelijskog ASYN (oligomernog. u kulturi ćelija humanog neuroblastoma SH- SY5Y.

što je potvrđeno imunohistohemij- se smatralo da je ASYN isključivo unutarćelijski protein i skim analizama neurona (29). a indukuju autofagiju. nitrovanim i autofagozome. modela PB. često korišćenih in vitro drugi neuron. a da oligomerni oblik ASYN je takođe pokazano da se najlakše rezultati ovog istraživanja ukazuju da modifikovani oblici unosi u ćelije ili prenosi iz jedne ćelije u drugu. ličinu aktivnih fosforilisanih formi AMPK i Raptor. dakle. doprinosi neurotoksičnosti modifikovanih oblika van- stanku Levijevih tela. za autofagiju ka- ne SH-SY5Y ćelije tretirane su modifikovanim oblicima rakterističnog. značaj- ubikvitinom. naime. diferencijaciji neurona (28). ulogu aktivacije AMPK. kojima su tokom života transplantirani preživljavanje neurona (30). čime maćina na graft (26). Svi analizirani modifikovani oblici re. Medicinski podmladak Originalni radovi toksičnosti u PB. prelaska LC3-I u LC3-II oblik tipičan za rekombinantnog wt ASYN: oligomernim. nitrovani i dopaminski oblik rekombinantnog Rezultati dobijeni ovim istraživanjem ukazali su vanćelijskog wt ASYN dovode do smanjenog preživlja- na neurotoksični efekat vanćelijskih modifikovanih obli. blastoma SH-SY5Y. Oni ka wt ASYN na diferencirane ćelije humanog neurobla. što govori u prilog indukciji autofagije u dopaminski modifikovanim ASYN u cilju ispitivanja ulo. Rezultati dobijeni ovim is- fetalni mezencefalični dopaminergički neuroni. smo. dalja istraživanja treba da rasvetle vezu između AMPK U ovom istraživanju je pažnja usmerena na ulogu i autofagije u mehanizmu dejstva modifikovanih oblika AMPK i nishodne kinaze Raptor u dejstvu modifikovanih ASYN na diferentovane diferencirane ćelije humanog ne- oblika wt ASYN na diferencirane ćelije humanog neuro. oblika. konstantno je eksprimiran i što ima poseban značaj u širenju bolesti (25). način dovodi do oštećenja i posledične smrti ćelija. Novija istraživanja. ge AMPK u delovanju modifikovanih oblika vanćelijskog Rezultati ovog istraživanja. ukazuju da oli- ASYN. Imajući u vidu da AMPK humanog neuroblastoma SH-SY5Y apoptozom (27). Ove paminski modifikovan ASYN blokira autofagiju i na taj ćelije poseduju karakteristike dopaminergičkih neurona. da svi ispitivani oblici ASYN umanjuju ko- živelom graftu. Zaštitna uloga AMPK je da ne postoje mehanizmi kojima bi prelazio u vanćelijsku pokazana u slučaju neurotoksičnog oštećenja ćelija usled tečnost ili druge neurone. a posebno kiseline (19). što je u skladu sa prethodnim istraživanjima nju neurotoksičnog dejstva modifikovanih oblika ASYN koja su ukazala da nitrovani oblik dovodi do smrti ćelija na diferencirane SH-SY5Y ćelije. tako i primarnih neurona u zahvaljujući ekspresiji tirozin hidroksilaze i dopamin kulturi (31). ćelijskog ASYN. U modelu Hantingtonove bolesti kod miša bijeni na obdukcionim analizama tkiva mozga pacijenata pokazano je da farmakološka aktivacija AMPK pospešuje obolelih od PB. uka- β. Ovim traživanjem odgovaraju podacima iz literature. Osim toga. Pokazali analizama je pokazano da se Levijeva tela nalaze i u pre. je nedvosmisleno ukazano na značaj deaktivacije AMPK lih od PB pokazani su i citosolni agregati različitih oblika u dejstvu modifikovanih oblika wt ASYN. međutim. ovim ćelijama. Rezultati ovog istraživanja. fosforilisanog. što ukazuje na prenošenje bolesti sa do. koja ima brojne ka. Na isečcima moždanog tkiva obole. što dalje sugeriše moguću protektivnu U ovom istraživanju je korišćena ćelijska linija hu. i u skladu je sa dosadašnjim po- manog neuroblastoma (SH-SY5Y). međutim. delovanja 1-metil-4-fenilpiridina (MPP+) (17). što dovodi ASYN smanjuju aktivnost AMPK. diferencira. mehanizma aktivacije ovog signalnog puta u prevazilaže- rane ćelije. dacima iz literature (7). dopaminski mofikovanom. gde su od posebnog značaja rezultati do. neuroblastomskih SH-SY5Y. skorašnja istraživanja su pokazala da se lovima nervnog sistema. otvaraju mogućnost istraživanja detaljnog molekularnog ciranih SH-SY5Y ćelija u odnosu na kontrolne netreti. kao i u govore o mogućem prenosu ASYN iz jednog neurona u slučaju delovanja rotenona (7). Protein kinaza aktivirana adenozin monofosfatom je široko rasprostranjen enzim u svim de- 56  Mart 2016. S druge strane. rakteristike dopaminergičkih neurona i zbog toga se če. dovodi do. Godinama funkcionalno aktivan. i ima značajnu ulogu u razvoju i on najlakše oslobađa iz ćelija i prenosi u susedne ćelije. vanja diferenciranih ćelija humanog neuroblastoma. Ćelijemogu da se diferenciraju i u zuju da tretman diferenciranih SH-SY5Y ćelija svim mo- dopaminergičke neurone pomoću all-trans retinoične difikovanim oblicima vanćelijskog wt ASYN. čime je potvrđeno da smanjena aktivnost AMPK značaj posttraslacionih modifikacija ovog proteina u na. AICAR. i značajnog smanjenja broja živih diferentovanih diferen. delovanja svih ispitivanih modifikovanih oblika ASYN kombinantnog vanćelijskog ASYN doveli su do statistički smanjenje aktivnosti AMPK/Raptor signalnog puta. što ukazuje na ćelija. do oštećenja i smrti susednih zdravih neurona (25). takođe sugerišu da je mogući mehanizam neurotoksičnog stoma SH-SY5Y. kako kao što je sposobnost sinteze dopamina i noradrenalina. pri- ASYN: nitrovanog. | Broj 1 | Izdanje 67 . podaci iz literature ukazuju. Za potrebe ovog istraživanja. uroblastoma. kao i oksidovanog oblika ASYN dobijenog no je poboljšala preživljavanje diferenciranih SH-SY5Y kao posledica metabolizma dopamina (5). Za može da indukovati indukuje proces autofagije (9). da do- sto koristi kao in vitro model Parkinsonove bolesti. obeleženog mena farmakološkog aktivatora AMPK.hidroksilaze (18). gomerni.

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elderly. blood. krv) eksperimentalnih životinja. tissue demonstrated a tendency towards decreasing amounts of Zaključak: U ovom eksperimentu smo dokazali da sta.com Sažetak Abstract Uvod: Gvožđe je esencijalni oligoelement koji je ključan Introduction: Iron is an essential trace element. kao i u krvi (p mice. U tkivu jetre i srca pokazana je tendencija ka opadanju kidney(p < 0. order to determine the iron content in them. hemoglobina i 55 g/mg TM).2 1 Medicinski fakultet Univerziteta u Beogradu 2 Mentor: Institut za patološku fiziologiju . Aging is an žan fiziološki proces koji dovodi do niza štetnih promena. najčešće gvožđa. kidney. blood) in experimental animals. it is shown that bigger fluctuations in its concentration could ličkih smetnji i oštećenja tkiva. Obe grupe su brojale groups: old and young. Nakon dostizanja željene starosti jedinke su After the completion of the experiment. Dvadeset životinja this experiment. as well as imbalances in the amount of trace elements. the distribution of iron in the body. po deset jedinki. so it is necessary to examine the influence of age on gvožđa u organizmu. age causes a highly statistically significant accumulation of iron zuje tendenciju opadanja. jetra. od čega su dve trećine sastavni deo hemo- 58  Mart 2016. testis Uvod mioglobina. Iron imbalance in the elderly is a widespread zbog je neophodno ispitati kako starost utiče na distribuciju problem. jetra. DNK Gvožđe je esencijalni oligoelement koji je bitan sintezu i druge oksidoredukcione procese (1). as da veće promene u njegovoj koncentraciji dovode do metabo. prven. testis.001) porast količine gvožđa u tkivu testisa u grupi starih je. Medicinski podmladak Originalni radovi Promena sadržaja gvožđa u organima starih pacova Determining the level of iron in organs of old rats Roberto Grujičić1. U telu od- za mnogobrojne fiziološke procese u organizmu. bubrega i krvi.Dr Ljubomir Buba Mihajlović“. testis. most often of iron. Koncentracija for numerous physiological processes in the organism. Ključne reči: Gvožđe. how aging affects the distribution of iron in certain tissues bubreg. testis Key words: Iron. we demonstrated that testisa.01). | Broj 1 | Izdanje 67 . a njihova tkiva su tretirana u cilju ficed under general anesthesia and their tissues were treated in određivanja sadržaja gvožđa u njima. Materijal i metode: U eksperimentu su korišćene je. Statistically significant increase was also confirmed in the < 0.001) potvrđen je i u bubregu. dok količina gvožđa u jetri i srcu poka. bubreg.. liver. kidney. Total of twenty animals were divided into two je podeljeno u dve grupe: stare i mlade. Ovakav disbalans gvožđa potencijal.001). kidney and blood. heart. lead to metabolic disturbances and tissue damage. iron. changes. the amount of iron in the testicular tissue in the group of old dinki u odnosu na kontrolnu grupu mladih. lans gvožđa kod starih osoba je široko rasprostranjen problem. a služi i kao kofaktor u mnogim enzimskim reakcijama uključenim u oksidativni metabolizam. starost. rost uzrokuje statistički značajno nagomilavanje gvožđa u tkivu Conclusion: In this experiment. animals (p < 0. Silvio De Luka1. Cilj: Cilj ovog rada je ispitivanje da li i na koji način Aim: The aim of this paper is to examine whether and starenje utiče na distribuciju gvožđa u pojedinim tkivima (srce. Disba. Material and Methods: We used male Wistar rats in dinke muškog pola Vistar (Wistar) pacova. raslog čoveka nalazi se približno 3-5 grama gvožđa (45- stveno za sintezu transportnih proteina. and in blood (p < 0. animals were sacri- žrtvovane u opštoj anesteziji. Rezultati: Ustanovili smo statistički značajan (p < Results: We found a statistically significant increase in 0. Univerzitet u Beogradu Kontakt: robertogrujicic@gmail. srce. Statistički znača. krv. in tissues of the testis.001). Both groups consisted of ten animals.01). kao universal and inevitable process that leads to a series of adverse i disbalansa u količini oligoelemenata. iron in the liver and the heart tends to decrease. Bogdan Bjelica1. compared to the control group of young jan porast (p < 0. (heart. Starenje je univerzalan i neizbe. The liver and heart količine gvožđa. vital za mnogobrojne fiziološke procese u organizmu. while the amount of no može da dovede do oštećenja ovih tkiva. The gvožđa u tkivima mora da bude usko regulisana jer je pokazano concentration of iron in tissues must be closely regulated. liver. This kind of changes can create potentially harmful effects on these tissues.

Dokazano je da nagomilavanje gvožđa do- stvima proizvođača. raznim mehaničkim oštećenjima ili procedurama opisanim u Zdravstvenom vodiču za brigu i oksidativnom stresu (9). Volume 67 | No.   59 . dobijene iz vivarijuma Vojnomedi- Apsorpcija je najvećim delom regulisana hormonima. koje su odgovorne za je Etički komitet za rad sa eksperimentalnim životinja- povećan rizik od morbiditeta i mortaliteta. dok manjak može da dovede do sideropenijske odvojeno od ovog plazmatskog proteina. u opštoj etarskoj anesteziji. Eksperimentalne životinje stemsko gvožđe je regulisano na nivou apsorpcije jer nije regulisana njegova ekskrecija. makrofazi. godine. Poznato je da je nedostatak gvožđa uzrok jedne i mlade.Ca2+. Značajan porast je otkriven u moždanom tkivu Gvožđe je određivano u krvi. Gvožđe vezano za plazmatski Materijal i metode transferin predstavlja 0. pa u potpunosti zavisi od U eksperimentu su korišćene jedinke pacova mu- deskvamacije epitela. F. u sveukupnoj količini vode. a sprovođenje ogleda odobrio akumuliranih u ćelijama i tkivima. Svi protokoli koji se tiču eksperimentalnih životi- goelemenata (Fe. Cilj našeg ogleda je bio da sa. Jedinke su dobijene kada zavisnosti od potreba organizma. pokazale da Određivanje gvožđa u tkivima se nivoi gvožđa u određenim tkivima povećavaju sa sta- renjem. Iako je njegovo žrtvovanja. a javlja se usled čestih hroničnih inflamacija. rosti. Si. slobodno (ad libitum) bile na raspolaganju. U trenutku citi. Oštećenja naj. dolazi i do disbalansa for Care and Use of Laboratory Animals. USA). škog pola Vistar soja. veze koji su sadržali po 4-5 jedinki. K+) i oli. a naročito visoke vrednosti se otkriva- i srcu. Sa starenjem.5).1 % celokupnog gvožđa u orga- nizmu. testisu. duodenumu. ispitamo sadržaj gvožđa u pojedinim organima oglednih koštanoj srži i u manjoj količini u drugim organima (3). se gvožđe u najvećoj meri akumulira prilikom njegove ekscesivne aplikacije (4. Upotrebom gvanidijuma gvožđe je i tkiva. a njihova tkiva su dalje tretira- okultnih gastrointestinalnih krvarenja. jetri zdravih individua. Ljudski or. koncentra. koristeći eksperimentalni model starosti. Jon gvožđa je u svom feriobliku (Fe+3) vodi do povećanog oksidativnog stresa i oštećenja ćelija vezan za transferin. a druga grupa (grupa starih) bila je stara tri cija gvožđa u tkivima mora da bude strogo regulisana (8). decenija dokazano je da se kod starijih osoba poremećaji homeostaze gvožđa javljaju često u poređenju sa ostalim Dizajn eksperimenta oligoelementima. U poslednjih par nja razmotreni su i odobreni od strane Etičkog komiteta. eksperimentalni protokoli su sprovedeni u saglasnosti sa stom (UV) zračenju. sideropenijske Nakon dostizanja željene starosti sve jedinke su žrtvovane anemije. elektrolita (Na+. ma Medicinskog fakulteta Univerziteta u Beogradu. roidnih antiinflamatornih lekova (NSAIL) i peptičkog ulkusa (11). set nedelja. fiziološkog ili patološkog krvarenja. Upravo zbog jene komplekse koji su mereni spektrofotometrijski. I). pored disfunkcije korišćenje eksperimentalnih životinja (engl. koji re. ćelije jetre i placente) (7). Pri eksperimentalnom radu poštovan je Etički Starenje je definisano kao univerzalan i neizbežan kodeks naučnoistraživačkog rada Medicinskog fakulteta fiziološki proces koji je rezultat niza štetnih promena Univerziteta u Beogradu. tivanju ovog oligoelementa. Ni. kao i zbog nekonzistentnosti podataka u literaturi gde ulazi u građu intraćelijskih enzima (2). dok je učestalosti poremećaja homeostaze gvožđa u populaciji ostatak u makrofazima. feroportina. životinja. Mn. prva grupa (grupa mladih) bila je stara de- prisustvo nezamenljivo za ljudski organizam. Podeljene su u dve grupe i smeštane u zasebne ka- gulaciju vrši preko membranskog proteina. hidroksilamina redukovano u ferooblik (Fe+2). bubregu. slezini. ali nije utvrđena uzročno-posledična njem Biosistemskog fotomera tipa BTS-330 prema uput- veza (12-13). Određene studije su. hemosiderina i feritina. Health Guide mnogih organa i organskih sistema. Posle toga Iz svega navedenog vidimo da su promene u kon- je dodat ferozin sa kojim su ferojoni (Fe+2) formirali obo- centraciji gvožđa kod starih uočljive i česte. hepcidin. o ovoj pojavi. Hrana i voda su im eksprimiranog na mnogim ćelijama u organizmu (eritro. u jetri.Medical Youth Original articles globina eritroidnih prekursora i zrelih eritrocita. ju kod degenerativnih oboljenja moždanog tkiva (Alchaj- Ukupno serumsko gvožđe određeno je korišće- merova demencija). od najučestalijih bolesti starije populacije. našu pilot studiju smo zasnovali na ispi- ganizam skladišti gvožđe u obliku proteinskih komplek. Washington DC. u cinske akademije u Beogradu. Co.Poremećaj homeostaze gvožđa se najče- šće manifestuje kao deficit gvožđa u celokupnom organi- Dvadeset životinja je podeljeno u dve grupe: stare zmu (10). hepatocitima i drugim ćelijama. Najbitniji hormon za su imale 8 nedelja i gajene su do dostizanja željene sta- regulaciju apsorpcije je hormon jetre. upotrebe neste- na u cilju određivanja sadržaja gvožđa u njima. Svi češće nastaju kao posledica stalnog izlaganja ultraljubiča. Zn. I prva i druga grupa su sadržale po deset jedinki. 1 | March 2016. a zatim pomoću anemije i drugih štetnih efekata (14). međutim. ali je u kinetičkom smislu najaktivniji pul (6). starih. kao i moguću kauzalnu vezu između ove dve Istraživanja su pokazala da su jetra i slezina mesta gde pojave.

001). p < 0. ** . ali se ovi rezultati nisu pokazali statistički (Institute for Reference Materials and Measurements.19 ± 1. peraturi je stajalo 10 minuta.64 spektrometru „SpectrAA 220“. Vrednosti gvožđa su izražene kao µg Fe po mg osušenog analiziranog tkiva. Belgium). bubrega. Ova činjenica nas je navela da istraživanje pre- usmerimo na druga tkiva u organizmu i ispitamo kakve se promene u količini gvožđa dešavaju u njima prilikom starenja organizma. istraživanja usmerena na urođena Grafikon 1. Nasuprot tome. u našem ogledu je analiziran sadržaj gvožđa u krvi. 34. terijala BCR-186 (Community Bureau of Reference .5 g) tretirano je sa 8 ml azotne kiseline µg Fe/g. verzija 18. Analize su vršene na atomskom apsorpcionom tistički značajnog nakupljanja u populaciji starih (509. i utvrđivanje uzročno-posledične veze između koncen- Dobijeni rezultati merenog gvožđa u ove dve grupe jedin. Diskusija Statistička analiza Koliki je značaj gvožđa govori i podatak o visokoj stopi morbiditeta i mortaliteta koja se javlja kod osoba Statističke analize su korišćenjem softvera SPSS za koje imaju deficit ovog elementa. testisa i jetre su pre. Uticaj disbalansa Rezultati gvožđa na normalno funkcionisanje organa. slezini.. | Broj 1 | Izdanje 67 . Italy) prema uputstvima proizvođača.01).p vrednost < 0. p < 0. pomoću dejonizovane vode prebačeni u normalan sud Merenje u krvi je pokazalo da i ovde dolazi do sta- od 50 ml.r. Starenje je ireverzibilan. Analitički kvalitet kontro. ture o distribuciji gvožđa u pojedinim organima u toku starenja su oskudni i nekonzistentni. Ovaj materijal predstavlja liofilizovani dencija opadanja koncentracije gvožđa u jetri i u srcu kod bubreg svinje korišćen za određivanje elemenata u tragu starih jedinki. 22. značajan porast gvožđa u grupi starih jedinki (106. progresivan fiziološki proces koji.12 ± 27.25 Ciljno tkivo (0.84 ± temperaturi.43 ± 10.97 µg Fe/g) u odnosu na mlade (24. jetri. Merenjem u bubregu pokazan je statistički veoma premljeni mikrotalasnom digestijom (ETHOS TC. (HNO3) i 2 ml hidrogen-peroksida (30% H2O2). što je pokazano visokom prevalenci- jom starijih osoba sa sideropenijskom anemijom.BCR. testisu i srcu na eksperimentalnom modelu sta- renja. preko 15% osoba starijih od 65 godina pati od anemija. kao 0. Podaci iz litera- je Man-Vitni (Mann-Whitney) U test. eksperimentalnih životinja iz grupe starih jedinki postoji U centru istraživanja poslednjih godina bila je po- statistički značajna promena koncentracije gvožđa (p < vezanost između akumulacije gvožđa u tkivu mozga. Tempe. kao i mo- gućnost povezanosti ovog disbalansa sa starenjem pod- U ovom istraživanju smo ustanovili da u tkivima stiče nova istraživanja u ovom pravcu. Eksperimentom smo još pokazali da postoji ten- Brussels. Prema nekim studijama. Geel.001 i p < 0. Medicinski podmladak Originalni radovi Uzorci tkiva iz srca.09 mg Fe/dl) u odnosu na mlade (478. Uzimajući u obzir oskudnost poda- taka o distribuciji gvožđa u pojedinim tkivima.26 µg Fe/g) u odnosu na grupu mladih (77. sa (13-19). Mile.01. a zatim je preneto na 180°C i na ovoj tem.01) u odnosu na kontrolnu grupu mladih.17 ± stone S.p vrednost < 0. značajni. Sorisole. Prikaz količine gvožđa (µg Fe/g tkiva) u različitim (primarna hemohromatoza) i stečena stanja (stečena he- tkivima kod dve grupe eksperimentalnih životinja mohromatoza kod oštećenja koštane srži) koja dovode do ( * . tracije gvožđa i starenja kod neurodegenerativnih proce- ki prikazani su na grafikonu 1.l. nagomilavanja ukupnog gvožđa u organizmu dokazala su da kod jednog dela bolesnika dolazi do povećanog rizi- 60  Mart 2016. bubregu.94 mg le je postignut analiziranjem potvrđenog referentnog ma.001). dovodi do promene u koncentraciji oligoelemenata. Belgium). pored ostalih štetnih promena u organizmu.66 µg Fe/g. Fe/dl. Merenjem u testisu je pokazano statistički veo- raturni režim je sledeći: tkivo je 5 minuta bilo na sobnoj ma značajno nakupljanje gvožđa u grupi starih (75. Za poređenje dve grupe korišćen smatra toksičnim za ćelije i tkiva (8. dok se njegov suficit Windows.001). 12). Uzorci su nakon hlađenja p < 0.0. u pr- vom redu gvožđa. ± 31. koje su u najvišem procentu posledica nedostatka gvožđa (20).

13. World Gastroenterol 2009. Results of long-term iron chelating the- Pretpostavljamo da su uzrok povećanja gvožđa upravo rapy. et al. Ovakve promene posledično dovode do 16. Ovaj podatak nije sues in vivo. koja dovodi do njegovog viška intrace. et al. 1997. Sultzer D. strukturne promene tkiva testisa u starosti (atrofija) i nje. and cellular mechanisms of iron homeostasis and toxicity rumu) (26).15: 4617–4626 su ovakvi rezultati dobijeni zbog čestih komorbiditeta 7. pa čak i do smrti uzrokovane najčešće U ovom eksperimentu smo dokazali da starost srčanom disfunkcijom (12. da ove 4–8. rezultira njegovim prooksidativnim dejstvom sa i srcu pokazuje tendenciju opadanja. dox Signal. Vayenas D. Increased iron and free radical generation in preclinical Alzheimer disea- pokazano da izlaganje eksperimentalnih životinja veli- se and mild cognitive impairment. PapanikolaouG. 1996. takođe. 6. bubrega i krvi. Wilmet S. 2011. ry in healthy aging. Gender and iron genes promena u vidu degeneracije seminifernih tubula. kim količinama gvožđa dovodi do njegovog nakupljanja 2010. Smith MA. dical Sciences. 11. dolazi do značajnog porasta količine gvožđa u tkivu bu- brega. što poten- injected iron 59 and manganese 54 in hypotransferrinemic cijalno može da uzrokuje taloženje gvožđa u ovom tkivu. Osim toga. 21). Moguće je da physiology.196: u skladu sa većim brojem dosadašnjih istraživanja koja 47-57 pokazuju tendenciju ka padu nivoa gvožđa u populaci. larne barijere. Gabutti V. Evaluation of the gastro-intestinal tract in patients with iron. 2002. et al. A.306: 2090-2093 razne kombinovane terapije koji mogu da povise nivoe 8.   61 .36: geneze (27). Ne Eng J U našem ogledu smo uočili značajno povećanje Med. ency and morphological distribution of anemia in elderly ni rast gvožđa u serumu posledica smanjenja cirkulišuće patients. 1375–1384 Volume 67 | No. J InorgBiochem. Ovakav disbalans posledičnim oštećenjem i starenjem ćelije (22). Ovakva kontradiktornost rezultata može da in mammalian cells. Alzheimers Dis.Turkish Journal of Geriatrics. M. 2003. Muñoz M. Hurrell RF. Bartzokis G.5: 557–61 lim studijama nivoi gvožđa u serumu mereni uglavnom 10.51: antioksidativnog kapaciteta. Tabaton M. maligniteti. moguće je da je relativ. Rockey DC. U starenju dolazi i do oštećenja krvno-testiku. Dosadašnja istraživanja su pokazala da nagomila. Iron metabolism and to- toga što u starosti funkcija bubrega znatno slabi. Increased le- značaja.329: 1691-5 koncentracije gvožđa u tkivu testisa kod starih jedinki. Dickinson TK. Concentration of Iron and Distribution of Iron and Rezultati pokazuju značajno povećanje koncentra.Medical Youth Original articles ka od intolerancije na glukozu. its internalization. Çoban. 1 | March 2016. Eur J ClinNutr. Mintz J. što potencijalno može da bude ation of age-related increases in brain iron. Pankhurst Q. 76(3-4): 235-254 tije (23).19: 363–372 u testisu (27). srčane disfunkcije. M. Bartzokis G. dok su u osta. Tuttle MS. 26). Devenyi AG. AJNR Am. In vivo MR evalu- gova disfunkcionalnost. Connor JR. gvožđa potencijalno može da dovede do oštećenja ovih Naši rezultati pokazuju da u grupi starih jedinki tkiva i da uzrokuje poremećaje koji prate starenje. Timuragaoglu. Crichton RR.15: 1129–1138 gvožđa u tkiva koja mu u tom trenutku nisu od vitalnog 14. ošte.202: 199–211 4. Distribution of do smanjenja glomerulske filtracije i sekrecije. ActaHaematol. Powelson J. 5. uzrokuje statistički značajno nagomilavanje gvožđa u gulacija gvožđa.Research and Practice. Zaključak ćenja endokrinih organa. dijabetesa melitusa. Pretpostavljamo da je ovakav nalaz dobijen zbog 3. et al. 2014. 7:131–132 tečnosti. J. atro. E.Antioxid Re- eksperimentu korišćen animalni model. mice.deficiency anemia. The free radical theory of aging. Piga A. Tingus K. Potvrđeno je i da disre. Papanastasiou DA. Nemeth E. Vassilopoulos A. J. J. Literatura vanje gvožđa u bubrezima eksperimentalnih životinja do- vodi do povećanja oksidativnog stresa. tkivu testisa. Akın. Saito H. kao i da 2. Pathology . Hepcidin regulates cellular iron efflux by binding to ferroportin and inducing kod starih ljudi (akutne i hronične infekcije. The frequ- u humanoj populaciji. Neuropsychopharmacology. U prethodnim istraživanjima je 15. dolazi xicity. J Lab Clin Mcd 128: 270-278 U bubregu se sa godinama dešavaju i značajne morfolo. Zhu X. 2004. Pokazano je. An update on iron ji starih ljudi (preko 65 godina) (25. Khan N. Villar I. Lu PH. Aykut. gvožđa u ovom tkivu. kao i do smanjenja 1. vels of magnetic iron compounds in Alzheimer's disease. što bi takođe moglo da objasni taloženje Alzheimers Dis.13: 49–52. Hautot D. may modify associations between brain iron and memo- fije tkiva i poremećaja ili potpunog odsustva spermato. Pantopoulos K. 2000. Cello JP.91: 9–18 bude i posledica različitog dizajna studije jer je u našem 9. tj. Harman D. A. Sciense. 2003. dok količina gvožđa u jetri lularno. Ward RJ. García-Erce JA. Transferrin After Experimental Iron Overload in Rat Tis- cije gvožđa u serumu starih jedinki. Metabolism of iron stores. Nagoya Journal of Me- su potencijalni kofaktor u nastanku dijabetesne nefropa. 12. Repanti ške promene koje dodatno slabe njegovu funkciju (24). promene utiču na lošiju prognozu nekih bolesti. Bioavailability of iron. Dobson J.95: 26–36. 1994. Molecular hepcidina i da utiču na akutno smanjenje gvožđa u se. Toxicol Appl Pharmacol 2005. 2008. Legssyer R. razlog zašto organizam teži da taloži ekscesivne količine Neuroradiol. 1993. fibroznih promena u jetri. što je isto tako poremećaj koji prati starenje.

Girelli D. The effects of intensi- gan. From Benger Laboratories Ltd. Prevalence and causes of 19. Scialo F. prions. Busti F. mer’s. Finch C. systems biology understan. bactericides. era. Alzhei. Renal function in elderly. Biological Trace Element Research. Zhonghong G. Archives of Toxicology.Front Pharmacol. 2010.84: 825-889 25. 2014.. Luran MM. anemia in a geriatric hospitalised population. Golberg. 5:83 mias report of a WHO scientific group. The interplay Manchester between mitochondrial protein and iron homeostasis 62  Mart 2016. 56: 123–134 and Therapeutic Opportunities. Joosten E. 28:111-7 Neurodegeneration. Kell DB. World Health Or.Martin LE.20: 1324-1363 ses Diabetes . Blanc B. 23.160: 368- ding of large-scale cellular death and destruction caused 375 by poorly liganded iron: Parkinson’s. Ayton S. Holmes Chapel. Pelemans W. Iron defi- 2014. Tech. Sanz A. 4051–40 ve and prolonged administration of iron parenterally in 21. Hiele M. revisited in the hepcidin 20. J P Smith. Wanxia G. 2003:40-61 Cheshire. A. Towards a unifying. Ser. Ann Clin Lab Sci. Swati H. in Rats. Singh N. Lei P. Cook JD. chemical toxicology and others 1995. Rep 1998. 27. Campostrini N. Nigral Iron Elevation Is an Invariable Fe. L. | Broj 1 | Izdanje 67 . University of 22. Gerontology ature of Parkinson’s Disease and Is a Sufficient Cause of 1992. Xueli L. Hailing L.. Hallberg L. Martinelli N. Ajai KT. Huntington’s. BioMed Research International. Beutler E. et al. Mallikarjun V. Sriram A. Nutritional anae. Iron Increa- 2014. Antioxid Redox Signal. Brain Iron Homeostasis: and its possible role in ageing.. and the Department of Pathology.Induced Kidney Injury and Oxidative Stress 18. 24. 2014. Overload. Hematology. et al.2014:581 ciency in the elderly population. Hoffbrand AV. Medicinski podmladak Originalni radovi 17. 26. Iron Deficiency and animals.25: 122-133 as examples. Experimental Gerontology From Molecular Mechanisms To Clinical Significance (2014)..

8-37 nedelja).6 nedelja Results: Median gestation age at delivery was 35.6 ( za triplete redukovane uz pomoć MFPR do blizanačkih interquartile range [IQR]. The most frequently pregnancy reduction. cije kod žena sa trihorionskim tripletima.66%).   63 .66%). Rezultati: Srednja gestacijska starost bila je 35. intrakardijalna ili intratoraklna primena kalijum hlorida Aim: The main objective of this study was to evaluate između 11 i 14 nedelje gestacije. blizanačkim trudnoćama. ishod nancy outcome.9%] vs 18 [35. technology (ART) (1). preg. tripleti. Materijal i metode: U ovoj retrospektivnoj studiji all women with trichorionic triplet pregnancies undergoing uključene su sve žene sa trihorionskim tripletima kojima je MFPR to a twin pregnancy. ali i dalje više kod primarnih blizanačkih trudnoća Conclusion: Multifetal pregnancy reduction (MFPR) . manjim rizikom od gubitka trudnoće pre 24 i 32 nedelje gesta- rionic triplet reduction.6%) in the ongoing gubitak trudnoće pre 24 nedelje se desio kod 7 trudnoće nakon triplet group. žene sa tekućim trihorionskim trudnoćama i onic triplet pregnancy and with those with a primary dichori.29%] ). Prevremeni porođaj weeks of gestation occurred less in the reduction group com.7 (11. Preterm delivery before 32 primarnih blizanaćkih trudnoća (6.1-38.66%). 1 | March 2016.7 weeks) for ongoing trip.7 (11. MFPR) se primenjuje kao jedna od applied method is through ultrasound guided transabdominal metoda za smanjenje rizika kod trudnoća sa tri ploda. sa trudnoćama sa tri ploda bez MFPR i blizanačkih trudnoća.3 nedelja) za blizanačke trudnoće (p <0. Multifetal triplets pregnancies MFPR is preformed.1-38.8 to twins.Medical Youth Original articles The influence of multifetal pregnancy embrioreduction in high-risk pregnancies – clinical management and outcome Uticaj multifetalne embrioredukcije kod visokorizičnih trudnoća - klinički pristup i ishod Petar Zlatanović1. statistički značajna razlika između grupa). women with an ongoing trichori. 32. multiple pregnancy. 35. 27. triplets trudnoće. early reduction. rana redukcija Introduction 100000 in 1980 to 153 per 100000 births in the United States.28%) vs 10 tripleta bez redukcije (19. 27. rađen MFPR. Methods of ART that contribute lets and high-order pregnancy has increased from 37 per the most to multifetal pregnancies are in vitro fertilisa- tion (IVF) and controlled ovarian stimulation (COS).2 1 Medicinski fakultet Univerziteta u Beogradu 2 Mentor: Odeljenje za visokorizične trudnoće. perinatal outcomes in triplet pregnancies after transabdominal Cilj: Osnovni cilj ove studije je da proceni uticaj MFPR as compared with ongoing triplet pregnancies and twin transabdominalne MFPR kod trudnoća sa tri ploda u poređenju pregnancies. Ključne reči: MFPR.01 difference between all groups). and 37 weeks ( IQR.com Abstract Sažetak Introduction: To decrease the risks associated with Uvod: Multifetalna embrioredukcija (engl. [35. There were 7 women (p < 0.6-34. Material and methods: In this retrospective study.28%) vs 10 (19.8-37 weeks) for triplets reduced trudnoća (interkvartalni opseg [IQR]. nedelja ( IQR. seems to be associated with a decreased risk of pregnancy loss Zaključak: Multifetalna embrioredukcija je povezana sa both before 24 and 32 weeks of gestation in women with tricho. injection of potassium chloride into the fetal heart or chest cav. pre 32 nedelje gestacije se desio u manjem broju kod MFTR pared with the ongoing triplets (12 [20. 32. 32. Kompletan in the reduction group (12.8 weeks (IQR..3 weeks) for primary twins i 37 nedelja (IQR. Key words: MFPR. Najčešća primenjivana metoda MFTR je ultrazvukom vođena ity at 11-14 weeks of gestation. whi and 4 (6. 32. grupe u poređenju sa grupom bez redukcije (12 [20.9%] vs 18 but still more often than in primary twins .6-34.Narodni front“ Kontakt: petarzlatanović@yahoo.6%) i 4 kod whom not a single fetus survived. Ac- Volume 67 | No. multiple trudnoće. Željko Miković1.66%) in the primary twin group in redukcije (12.01.29%] ).7 nedelja) za tekuće horionske triplete lets. Stefan Zaharijev1. GAK . onic twin pregnancy were included. mainly because of use of assisted reproductive Over the past three decades the incidence of trip. 35.

The risk of spontaneous abor. neonatal birth associated with MFPR at 11-14 weeks. including the op. and primary twins) was com- potassium chloride into the fetal heart or chest cavity at pared in terms of gestational age at delivery.7). hypertensive dis.2 ± 0. non-in.05. ate and between three groups 1-way analysis of variance uate the effectiveness of early transabdominal MFPR as (ANOVA) test or Kruskal-Wallis test when the data were compared with ongoing triplet pregnancies and twin not normally distributed. the main objective of this study was to eval. fetal measurements. The course of pregnancy in women with a reduced triplet was compared with the outcome in women with an ongoing triplet pregnancy and with those with a primary dichorionic twin pregnancy. Fifty one women with trichorion- jecting potassium chloride intracardially. with dichorionic twin pregnancy were identified as a con- Women were identified by searching in the ultrasound. there Wilk or Kolmogorov–Smirnov tests. The orders such as preeclampsia and cesarean delivery (5). premature rupture of tion (MFPR) is performed and several approaches have membrane and delivery mode were determined and been described. vasive prenatal testing. delivery less than 32 weeks. restriction. trol group. Baseline characteristics in three groups (triplet pregnan- „Narodni Front“. Triplet pregnancies are at an increased chorionic twin pregnancies were collected using a similar risk of neonatal morbidity and mortality compared with selection procedure as described for the triplet control twin or singleton pregnancy. | Broj 1 | Izdanje 67 . nancy who underwent fetal reduction to a twin pregnan- cluded. pregnancies at the Clinic for Gynecology and Obstetrics table 1.9 weeks. genital anomaly fetus were not included in the study. intrauterine growth high-order pregnancies multifetal pregnancy reduc. Compared maternal and where appropriate neonatal medical re- with women with twin gestation. Results let pregnancies undergoing fetal reduction to a twin pregnancy at the Clinic for Gynecology and Obstetrics Fifty seven women with a trichorionic triplet preg- „Narodni Front“ over the period of 2009-2014 were in. Chicago. All procedures ic triplet pregnancies without reduction and 60 women were performed between 11 and 14 weeks of gestation. The study was limited to trichorionic triplets only. Chi-squared or Fisher’s exact pregnancies in terms of perinatal outcomes. For perinatal mortality women who had all their portunity to carry out chorionic villus sampling . pregnancies without reduction vs dichorionic twin pregnancy). The most frequently applied method is compared. All trichorionic triplet preg- nancies in the period of 2009-2014 with at least 1 ultra- 64  Mart 2016. ongoing triplets. 39% is a consequence form which 3 living fetuses were identified and fetal reduction COS and 18% of them occurs spontaneously (2). and pregnancy outcome were collected. The cases cy who underwent fetal reduction to a twin pregnancy vs triplet in which fetal reduction was preformed because of con. The baseline characteristics of the 3 groups are obstetrics and fetal databases at the unit of high-risk shown in table 1. but was limited to women with 2 living fetuses. including gestational To decrease the risks associated with triplets and diabetes. using Mann–Whitney U test or student t-test as appropri- Thus. In this retrospective study. mainly because of increased group. children or no children surviving were compared. was 12. where MFPR was preformed. All statistical analysis were Materials and methods performed using SPSS 20 (SPSS Inc. those with triplets have cords. chorionicity. including de- a significantly increased risk of maternal morbidity such mographic data ultrasound findings. Data regarding original di- number of fetuses. sound performed between 8 and 15 weeks of gestation in nancies is a result from IVF. placental abruption. The outcome of the 3 groups (triplets reduced through ultrasound guided transabdominal injection of to twins. Medicinski podmladak Originalni radovi cording to the data 43% of triplets and high-order preg. Comparison of con- remains dabate whether multifetal pregnancy reduction tinuous variables between the two groups was conducted improves obstetrics outcome of triplets reduced to twins. The mean gestational age at reduction after local anesthetic by using 20G or 22G needle and in. IL). delivery less 11-14 weeks of gestation (6. as gestational diabetes mellitus (GDM). Maternal and fetal characteristics. The procedure was performed transabdominally cy were identified. However. The was not performed were included in the control group of risk of perinatal mortality and morbidity rises with the ongoing triplet pregnancies. Several advantages are than 24 weeks. weight. risk of preterm birth (3). all women with trip. Sig- nificance was set at P < 0. Normality of the data was tested using Shapiro– lies etc. rates of pregnancy complications. gestational hypertension. test was used for comparison of categorical variables. Data were collected retrospectively by reviewing tion is 15% for triplets and 8% for twins (4). ultrasound screening for anoma. in the event of triplet pregnancy. Chorionic- ity was determined at the first-trimester ultrasound scan.

35.231. duction resulted in more advanced gestation at birth as tween all groups) (table 4).12%) in the ongoing triplet group and 110 of 120 (91.01). table 2. there wasn’t discovered table 4.01. table 5. but this dif- No significant difference was found between three ference wasn’t significant between reduction and twin groups in terms of mode of delivery (table 3).39%) for pregnancies without reduction vs dichorionic twin pregnancy) ongoing triplets and 56 of 60 (93. Pregnancy outcomes by group (triplet pregnancy who any statistically significant difference among three groups underwent fetal reduction to a twin pregnancy vs triplet preg- regarding gestational hypertension. Loss of the complete pregnancy prior to 24 weeks occured in 3 pregnancies after reduction (5.   65 . p < 0.1-38.96%) vs 123 of 153 (77. Perinatal complications among three groups (triplet pregnancy who underwent fetal reduction to a twin pregnan- cy vs triplet pregnancies without reduction vs dichorionic twin pregnancy) The total number of surviving children in the reduced group was 98 of 114 (85. Delivery mode among three groups (triplet pregnancy es surviving that were intended to survive was 50 of 54 who underwent fetal reduction to a twin pregnancy vs triplet (87.7 weeks) for ongoing triplets.33%). 32. difference be.125). Perinatal mortality among three groups (triplet preg- nancy who underwent fetal reduction to a twin pregnancy vs triplet pregnancies without reduction vs dichorionic twin preg- nancy) Median gestation age at delivery was 35.6%) in the ongoing triplet group. 1 | March 2016.3%) vs in 9 ongoing triplets (19.6 ( inter- quartile range [IQR].4 ± 587. 35.8 weeks (IQR. whi and 4 (6.66%).512.6 vs Volume 67 | No.83 ± 533.33%) for primary twins (p < 0. This difference was statisti- cally significant (f = 105. The number of women who had all their fetus- table 3. 32.66%) for primary twins. Preterm delivery before 32 weeks of gestation occurred less common in the reduction group compared with the ongoing triplets ( 12 [20.6%) and 2 primary twins (3.122.3 weeks) for prima- ry twins (p < 0. group (χ2 = 3.94 in the reduc.01 difference between all groups) (Table 2). compared with expectant management in triplets. and 37 weeks ( IQR.022).05 ± 428. 1613.29%] ). The mean neonatal birthweight was 2230. p = 0.893 in the ongoing triplets and tion of 1 fetus in a trichorionic triplet pregnancy.Medical Youth Original articles Regarding complications.9%] vs 18 [35.28%) vs 10 (19. gestational diabetes. There were 7 women in the reduction group (12. 27.98 in primary twins (p < 0.71%) in the reduction group vs 41 of 51 (80. the difference between reduction and triplet group was statistically significant (χ2 = 5.66%) in the primary twin group in whom not a single fetus survived (table 5).8-37 weeks) for triplets reduced to twins. p = 0. Re- 2336. nancies without reduction vs dichorionic twin pregnancy) placental abruption. When compared.04). but still Discussion more often than in primary twins 7 (11. This study showed the effectiveness of the reduc- tion group. intrauterine growth restriction and premature rupture of membrane (table 2).6-34.

D’Ercole C. triplets to twins. Avigidou KA. Katy B. long-term outcome of these children was not studied. Women who had. 66  Mart 2016. this study didn't show neonatal morbidity. Maternal morbidity and infant death specifically mentioned when counseling parents opting in twin vs triplet and quadruplet pregnancies.17: 209-17. However.62:1-72. Serour GI. Curr Opnin weeks of gestation in women with trichorionic triplet re- Obstet Gynecol 2008:386-93. for example. Perinatal and neonatal outcome of triplet ges- In terms of pregnancy complications and delivery tations based on placental chorionicity. Paek BW. Genzel-Boroviczény O. fore should have priority over MFPR. it showed a lower risk of pregnancy loss prior to 24 Mathews.05% which is comparable with Papageorgh- iou et al. Hum Reprod 2006. Amin YM. and this possible risk should be 5. which might restrict the ability to detect differenc.2% in the review of 2011. with triplets and 60 with twins met inclusion criteria for 7. | Broj 1 | Izdanje 67 . Shulman A. Hepp H. sample size was limited. Herson VC. TJ et al. tunately. Unfortunately.) and a higher risk of pregnancy loss 2. 2001. Hughes K. 4. Am J Obstet for MFPR. This parents should be counseled for MFPR. could be that inclusions started to early. 4. Apparently the increased tuses for women with multiple pregnancies. Dodd JM. MFPR seems to be associated with an review. with triplets reduced to twins doing al. Curr Opnin lack of reliable data in our control groups. Borgida AF. versus expectant management: new data and systematic In summary. because it also showed an improvement in the References median gestational age from 33 to 36 weeks (8-10). vs 4. Sebire NJ. tients. Strauss A. Mansour RT. Seidman DS. 11. Papageorghiou AT. Papageorghiou et al. from 15% to 5%. Egan JF. the 10. The data also showed triplets to twins in the first versus early second trimesters a reduction in the delivery rate less than 32 weeks from after detailed fetal screening. Ingardia CJ. In contrast. it is 6. Nico- most as well as nonreduced twins (12).3 vs 8.29% to 21. Fryd- prior to 24 weeks in ongoing triplet pregnancies (19. Management and outcome of triplet pregnancy. Evans MI. Ka- also important to stress that most of the excess loss may mal A. Natl Vital Stat Rep 2013. although primary twins still did duction. Unfor.3%) (11). decreased risk of pregnancy loss both before 24 nad 32 12. or to 24 weeks gestation. How- ever when compared with research from Papageorghiou 1. Pons JC. Britt DW. Crowther CA. Sepulveda W. Evans MI.18:35-8. Sattar MA. Medicinski podmladak Originalni radovi 32. Hamilton BE. a large prospective study is needed to vali. slightly better. Schulze A. when fered for mode of conception and parity.198: 401. only 51 cases Steril 1999. Fetal Diagn Ther the pregnancy loss rate after MFPR in triplet pregnancy 2002. the prevention of multiple gestations there- The baseline characteristics of the 3 groups dif. Sebire NJ.e1-10.8 weeks of gestation. Gestational age at birth is a good 9.26: 587-90. Risk of miscarriage and early preterm birth es. in trichorionic triplet pregnancies with embryo reduction date the results. This in significant reduction in perinatal deaths. Charlemaine C. Ultrasound Obstet Gynecol 35. A possible explanation for this difference Eur J Obstet Gynecol Reprod Biol 1998. Achiron R. and the miscarriage was usually more than 2 weeks. These results 2009. Nold CJ. cation of the technique and analysis of the outcome. Luke B. Lipitz S. Therefore. 3 living fetuses at 12 weeks gestation and a Janssen U. Kozhimannil et al. Martin JA. due to 8. This small num- comparative study of multifetal pregnancy reduction from ber could have influenced the data. Nicolaides KH. Because the interval between the reduction Gynecol 2008. Multifetal pregnancy reduction: modifi- occur several weeks after the procedure. Spencer JV. Nevertheless.21:1912-7. 3. Evans and Britt reported a decrease in perinatal outcome of 112 pregnancies. study is partially in concordance with previous published studies. JK Osterman. Brown MB. Apparently the laides KH. Ventura SJ. Fetal reduction 2008. Multifetal gestation—maternal and miscarriage at 14 weeks gestation. Am J Perinatol mode there wasn't found any difference.8% to 18%.71:380-4. Fetal reduction 2008. Births: final data for weeks from triplets to twins (5. which is a small number. Dubreuil E. gestational age was used.46% man R. Effects of embryo reduction from trichorionic procedure of MFPR itself can cause pregnancy loss pri. A control group . Papiernik E. Aboulghar MA. Reduction of the number of fe- proxy for neonatal morbidity.104:1201-3. could be explained by the relatively small number of pa. Zalel Y. Br J Obstet Gynaecol 1997. Cochrane Da- gestational age among reduced triplets will have reduced tabase Syst Rev 2012. Fertil Moreover. who found a reduction from 28. fact that this was not a randomised controlled trial. Britt DW. Bakoulas V..76:131-9. short-term morbidity and mortality. because of the these pregnancies occur despite adequate precautions. As surrogate the Obstet Gynecol 2008:386-93.10:CD003932.

Medical Youth Partners Volume 67 | No.   67 . 1 | March 2016.

Medicinski podmladak Partneri 68  Mart 2016. | Broj 1 | Izdanje 67 .

1 | March 2016.   69 .Medical Youth Original articles Volume 67 | No.

• Ključne reči / Key words 70  Mart 2016.bg. Naslovnu stranu „Medicnski podmladak“ objavljuje originalne članke 2. mentorom. je rad realizovan.. Naslovna strana radove. bilo kakvih pitanja. studenata doktorskih studija i mladih Roman.5cm (Page Layout- časopis koji objavljuje naučne i stručne radove studenata Margins-Normal). • Zaključak/Conclusion ma želji autora. U tekstu koji sledi. stomatologija. Obim rada ne sme da pređe 3000 reči. Svaki studentski rad mora lo da bude kratak (ne više od 15 reči). (Left. godine. ne uključujući mite svoj tekst kako bi bio uzet u razmatranje za publiko. Sažetak na srpskom jeziku studenata i mladih istraživača na srpskom ili engleskom 3. i 3. Dozvoljeno je da sadrži najviše 300 reči (ne Vrste radova uključujući podnaslove i ključne reči). Nije dozvoljena upotreba podebljanih (Bold) slova u biohemija.ac. autori i mentori potvrđuju originalnost rada i etičnost 2. autori radova mogu biti studenti osnovnih Naslovna strana sadrži samo naslov rada koji bi treba- i poslediplomskih studija. 1. osnovnom tekstu. Prilozi „Medicnski podmladak“ objavljuje studentske naučne 1.). Studentski rad može biti u formi originalnog rada ili u Delovi sažetka/abstract-a su. Dakle. marginama 2. BOLD drugi časopis i koji nisu u procesu recenzije. uredništvu i svojim potpisom (u propratnom pismu). Medicinski podmladak MEDICINSKI PODMLADAK naučni časopis za studente i mlade istraživače MEDICAL YOUTH scientific journal for students and young researchers Dragi autori.. • Rezultati/Results Rad može biti pisan na sprskom ili engleskom jeziku. mora imati: njihovom putu ka uspehu u svetu nauke. | Broj 1 | Izdanje 67 . po navedenom redosledu: formi prikaza slučaja. sažetke na srpskom i engleskom jeziku.rs na papiru formata A4. Tekst rada 5. Sažetak rada/Abstract naučno-istraživačkog procesa. niti bilo „Medicnski podmladak“ uzima u razmatranje i kakve informacije koje bi rad mogle identifikovati sa au- objavljuje samo one radove koji (u delu ili celosti) nisu torima. ni mesto u kojoj se one nalaze. naslovnu stranu. farmacija. njihova zvanja kao ni nazive ustanova u kojoj JCR listi sa impakt faktorom većim od 1. Časopis u kontinuitetu izlazi od 1949. možete nas kontaktirati putem naše Neophodno je da rad bude napisan na sledeći način: mail adrese: medicinskipodmladak@med. po izboru autora. kao ni priloge. sa dvostrukim proredom (Line „Medicnski podmladak“ je međunarodni. Danas težimo da povežemo Struktura rada mlade istraživače širom sveta i budemo prvi stepenik na Rad koji nam šaljete. institucijom ili mestom istraživanja. nalaze se instrukcije kako da pripre. Sažetak na engleskom jeziku jeziku. molekularna biologija. Slanjem rada veličine 12 pt. što nas čini jednim od najstarijih studentskih naučnih časopisa u Evropi.doc ili . Na naslovnoj strani ne sme sadržati imena autora najmanje jedan rad objavljen u časopisu indeksiranom u i mentora. Za font slova odaberite Times New osnovnih. Sažetak rada piše se na srpskom i engleskom jeziku (Abstract). nikako Justify) i svaki novi pasus uvučen 1cm. pre. • Uvod/Introduction • Cilj/Aim Format teksta • Materijal i metode / Material and methods Rad možete predati u . vanje u časopisu Medicinski podmladak. recenzirani Spacing-Double). publikovani ranije niti su poslati na publikovanje u neki Naslov bi trebalo da bude ispisan velikim slovima. jasan i informati- imati mentora. Literatura Uslovi za prijavljivanje 6. Ukoliko imate literaturu.docx formatu. Mentor (kao prvi autor) mora imati van. poravnjanje sa leve strane istraživača u oblasti biomedicinskih nauka (medicina. 4. veličinu slova 12 pt.

no i precizno opisati sprovedeno posmatranje ili ek- Pericardiology: Contemporary Answers to Continuing speriment.. 10) pokazuju... standard- prikazane u tabelama ili ilustracijama. myopathies and congestive heart failure. Obratite pažnju da ukoliko ima više od 6 autora. zašto je preduzeto istraživanje koje se radom predsta. Cardi- no ime leka). New York: McGraw navesti vrstu. Kirshenbaum LA. In: Singal PK. stopni • Uvod sadrži literaturne podatke o problemu koji se Primer: Mnogobrojne studije (5-8) pokazuju. tekstu. Moreno T. Autorovo prezime inicijali. opis. standardni format citiranja je: vlja. starost. Takođe. Funck R. broj (izdanje): stranice. et al. strukturi ili u formi prikaza slučajeva (kada mogu koristite kratku crtu da biste obeležili seriju uzastop- odstupati od navedene forme. sažetak/ moraju biti navedene striktno u skladu sa uputstvima. Navešćemo Vam ovde šta to 4. et al. Navesti broj. Tekst rada podrazumeva. i zaključke koje rezultati ne podržavaju u potpunosti. vratićemo Vam rad kako biste svoju • Prikaz slučaja / Case presentation literaturu prepravili. 1 | March 2016. Susin C. Maisch B. druge karakteristike Hill. a zarez kada brojevi referenci nisu uza- uredništvom). de Castro GD. Redosled reference navesti okruglim zagradama šeg rada jer preko njih i naslova radovi mogu biti pretra. U poslednjem pasusu randomized clinical trial... Harrison’s princi- laciji. Prilozi 5. Naslov: podnaslov. kako bi na osnovu navedenih informacija i Challenges. teristike ispitanika. 2008. u radu obrađuje. nave- • Rezultate bi trebalo predstaviti logičnim redosledom dite prezimena i inicijale prvih 6 autora pa onda napi- u tekstu.. tabelama i ilustracijama. Pregled poznatih činjenica ne sme Mnogobrojne studije (5. Ristic AD. editors.Medical Youth Ukoliko je vaš rad u formi prikaza slučajeva. Prilozi se numerišu onim redom Volume 67 | No. da bude isuviše dugačak. 2000. ples of internal medicins. u koordinaciji sa nih brojeva.. Ukoliko je reč o ekspreimentalnim životinjama. Originalni radovi mogu biti napisani prema takozvanoj IMRaD (Introduction. Jameson JL. editors. „Medicnski podmladak“ Reference su izuzetno važan deo svakog rada. Koristiti merne jedinice SI sistema. p. Svaki od priloga šite et al. Datum publikacije. mora sadržati odgovarajuće tekstualno objašnjenje.   71 . Mesto izdavanja: izdavač. 2008 Aug. abstract se sastoji iz navedenih delova: uniformno i bez odstupanja. Izdanje se osnovne hipoteze i ciljevi rada. Naziv časopisa • Diskusija se sastoji od analize dobijenih rezultata u (skraćen). Dhalla NS. ženi.ovako se navodi literatura (1). broj. Kod navođenja imena lekova. poređenju sa već objavljenim poznatim rezultatima. navode Autorovo prezime inicijali. Hauser SL. Braunwald E. Pored naziva leka navesti i dozu i način ac Remodeling and Failure. kao poslednji pasus u uvodu. starosnu dob. 10) pokazuju. Izbegavajte tvrdnje 35(8):696-704. Results and Discussion) a) Kada se koristi više referenci u tekstu zajedno. Primer: Haas AN. „Medicnski podmladak“ se pridržava vankuverskog sistema navođenja referenci. Oppermann RB. godina. Kasper DL. ali mora jasno da opravda Mnogobrojne studije (5-8. ali poželjno je ne ponoviti u tekstu sve podatke c) Kada citirate članke naučnih časopisa. Reference moraju biti • Ključne reči/Key words numerisane redosledom kojim su prvi put navedene u Za ključne reči pažljivo odaberite 3-5 reči koje se tiču va.35-65. Ukoliko ne budu navedene • Uvod/Introduction prema pravilima. () nakon citata.“ rada i šansu citiranja. b) Kada citirate knjige. Obuhvatiti i ni format je: kratko prikazati samo važne nalaze. 17th ed. Azithromycin as a ad- rezultata. ukoliko se radi o humanoj popu- Longo DL. • Zaključak/Conclusion Maksimalan broj referenci je 50. proizvođač). diskusije iznesite svoj zaključak. pol i druge karak- Primer: Fauci AS.. ispitivanih životinja. Naziv članka. 8. Boston-Dordrecht-London: aplikacije. Navesti imena metoda koje su Ako je u pitanju poglavlje u knjizi: korišćene u toku rada kao i tačne nazive aparature Primer: Maisch B.. drugi istraživači nedvosmisleno mogli ponoviti istra- Ako ima više od 6 autora/urednika: živanje. pol. Pametno izabrane ključne reči povećavaju vidljivost Primer: „. editors. J Clin Preiodontol. Literatura Kada su u pitanju Vaši prilozi. Methods. obrade rezultata. Dilated cardio- (model. koristiti generički naziv (u zagradi navesti registrova- Dixon IMC. komentarišite ih u svetlu rezultata nekih junctive treatment of aggressive periodontitis: 12 months drugih studija i istraživanja. Spodick DH. • Materijal i metode je odeljak u kome bi trebalo detalj- Primer: Seferovic PM. Al- Nemojte u ovom odeljku samo ponoviti opis svojih bandar JM. (ako nije prvo). One dozvoljava do 8 priloga. 2003. 6. Beograd: Nauka. Opisati statističke metode korišćene tokom Kluwer Academic Publishers.

tj. po dogovoru sa urednikom. glavni autor će biti oba- slike može biti 300 dpi. da smo rad primili. sive slike imaju više nijan. ki prilog pošaljite kao poseban fajl u originalnom forma- Širina Vašeg priloga može biti između minimalne širine tu. širina 1. po- priloge u boji. Medicinskog prilog ostane na naznačenom mestu. jednaka širini jedne cele kolone Uz rad. Minimalna rezolucija slike može biti 300 dpi. Tabela 1. uz taj fajl. Slika bi morala da bude „tesno“ vešten putem maila. Slika bi morala da bude „te. jedne i po kolone ili čitave dve kolone. osim što ga pošaljete u samom cije prenose se sa autora na časopis. zelene i plave (red. Redosled kojim su ophodno kako bi u štampanoj verziji časopisa prilog bio navedeni radovi u časopisu. i širine jedne kolone. eventualne sugestije ured- okvirima crne i bele boje (ne i sive). Prilikom prijema • Sive slike u TIFF/JPEG formatu korigovanog materijala. Debljina linije mora da bude u opsegu od uredništvo časopisa angažuje kompetentne stručnjake. Put od predaje rada do publikovanja • EPS format Nakon prijave Vašeg rada. Naslov i opis grafikona i slika trebalo bi da bude ispod grafikona/slike. sva- PowerPoint. rad dobija konačnu odluku o si sive. Vodite računa da na graficima obeležite x i y ose. ili RGB slike sastoje se od crvene. U tekstu naznačite meste Završne napomene gde bi trebalo da stoji prilog (npr. Ovo je ne. dobićete email koji potvrđuje EPS tip slike je baziran na vektorima. Korigovani materijal se potom dostavlja nazad redakciji sno“ isečena (minimalna širina belih margina). blue). Radovi se ne vraćaju. a naslovi tabela iznad tabele. green. zatim skenirano proprat- „Medicnski podmladak“ to podrazumeva: no pismo. Molimo Vas da skenirano propratno pismo ša- minimalna širina priloga = 40 mm ljete kao jedan PDF dokument. ili fajlovi Microsoft Office paketa (Word. Grafik 1. | Broj 1 | Izdanje 67 . graficima. Redakcija Objavljivanje u Medicnskom podmlatku je besplatno. 72  Mart 2016. Bitmap format slike definiše sliku samo u Nakon završenih recenzija. formatu rad predati putem mail-a. Minimalna rezolucija publikovanju. 0. Excel). koje nisu zavisne od piksela i Prispeli rad podleže recenziji. adekvatno prikazan bez gubljenja kvaliteta. ste grafikona.10pt do 1. Za recenziranje radova rezolucije. Za format potom potpisano i pečatirano. tekstu gde bi trebalo da stoji. Medicinski podmladak kojim se pojavljuju u tekstu. • Slike u boji (RGB) u TIFF/JPEG formatu Slike u boji. U procesu recen- • TIFF (bitmap) format ziranja autori neće znati ko su recenzenti njihovog rada. koje omogućava najkomfornije čitanje rada).ac. JPEG. moguće ukoliko format drugačije ne dozvoli. morate nam poslati odvoje. ovih slika može biti 1000 dpi. isečena. na osnovu odluke Izdavačkog Za razliku od bitmap formata. Svi prilozi moraju pravilno obeleženi. napravite odgovarajuću legendu. Minimalna rezolucija nika i recenzenata dostavljaju se autoru radi korekcije. neophodno je poslati i elekstronski popunjeno. teksta. Ovaj tip fajla se takođe može koristiti za različite vr. trebno je da ga sačuvate u Word formatu. saveta časopisa i urednika. će se prilikom pripreme teksta za štampanje potruditi da zahvaljujući podršci našeg osnivača.5 kolone = 130 mm Svoj rad pošaljite na našu mail adresu: širina 2 kolone = 181 mm medicinskipodmladak@med. Ukoliko je rad prihvaćen na publikovanje. „Medicnski podmladak“ zadržava pravo na određivanje no. Koristite Times New Roman slova veličine 11 pt. Svaki rad ima najmanje dva recenzenta. prava distribu- Napomena: Svaki prilog. časopisa. ali manje izmene su fakulteta Univerziteta u Beogradu. ne odražava vrednost rada.5pt. niti recenzenti znaju ko su autori rada kojeg recenziraju. ne samo crnu i belu boju.bg. jer ćete u tom TIFF.) i ubacite priloge na odgovarajućem mestu (na onom mestu „Medicnski podmladak“ posvećen je kvalitetu.rs Postoje određeni uslovi koje biste morali ispuniti za svaki tip priloga koji šaljete. Takođe. kao odvojen fajl u originalnom formatu. PDF. „Medicnski podmladak“ štampa crno-bele priloge i Nakon što ste rad sredili po navedenim uputstvima. hemijskim formulama. Dozvoljeni formati priloga su: EPS. Slika bi morala da bude „tesno“ isečena. O konačnoj odluci. ne svaku stranicu pisma širina 1 kolone = 87 mm odvojeno. redosleda objavljivanja radova.

kada juju tekstove koji bi pomagali studentima u pripremanju je ovaj predmet prvi put i uveden u redovnu nastavu. Nadamo se da ćemo u njemu. još za vreme studija. godine u Medicin­skom Osamdesetih godina obim časopisa uvećan je iz- podmlatku pored rubrike „Originalni radovi“ postoje i davanjem odgovarajućih dodataka – supplementum-a druge rubrike: „Opšti pregled“. i mašina na koje se mladi lekari više nego što je potrebno mladak. bavi donosi beskrajno mnogo uživanja kad je podstican U uvodnoj reči prve sveske Medicinskog podmlat. te je časopis pripadao nje IV interne klinike u Pasterovoj 14. godine piše: „Stručni kongresi su ona služiti i kao podstrek za nova pregnuća. 1 | March 2016. dine. već će skog podmlatka 1969. časopis je tutom Medicinskog fakulteta. a javlja se ideja o integraci- nastao kao izraz volje i težnje studenata medicine da se ji svih medicinskih fakulteta u zemlji. gde po prvi put nastavnici objavl.put ka uspehu - M edicinski podmladak je među­narodni. On treba da bude izraz i simbol stremljenja naših u budućnosti u saradnji i uz razumevanje Škole i njenih studenata. On treba da posluži kao instrument zbliženja institucija uspeti da nađemo rešenje i za ovaj problem. Brojevi su izla. Jedno od poznatijih izdanja bilo je Opšta pa- „Prikazi“. uz nova jnu ulogu jer će iz njihovih redova poteći mnogi naučni i iskustva počeli su i prvi osvrti na probleme. urednik Medicin­ nije samo smotra dosadašnjih rezultata i uspeha. greba. Novog dobrovoljnoj osnovi. pokrenuli naj­bolji studenti Medicinskog fakulteta Uni. kako im omogućiti još bo­lji i masovni- treba da čita ovaj časopis. način. prof. svedoče naše stranice: „Zar nije neophodno da se i na ovaj cenzirani. „Problemi studija i nastave“. Već 1969. nik savremene onkologije u Beogradu. Niša. zdravom ambicijom i lišen koristoljublja. Sve ispita. „Biblio­grafija“ i tologija tumora profesora Vladimira Kanjuha 1980. Kvalitetom se uveliko prevaziš. između ostalog. Ksenofon Ostaje i dalje otvoreno pitanje kako stimulisati studente Šahović. oslanjaju. vizionarski ljude. Prvo sedište redakcije bilo je u prostorijama tadaš­ su širene čitavom Jugoslavijom. re. Otuda pojava Medicinskog podmlatka pratiti kasnije redakcije.“ i saradnje sa ostalim studentima drugih medicinskih. redovno i u kontinu. da se snaga sopstvenog razmišljanja stavlja iznad tehnike verziteta u Beogradu davne 1949.“ Rodonačel. naši najveći ali i jedini izvor originalnih stručnih radova. časopisa. stručni i naučni časopis za stu. Kao što to obično biva. far. znači vaspitavati ka članovi prve redakcije su. student­ski radovi.Medical Youth NAJSTARIJI STUDENTSKI NAUČNI ČASOPIS U JUGOISTOČNOJ EVROPI . Sada. godine.“ Dakle. O entuzijazmu tih generacija studenata medicine intenzivnije su se u saradnju sa Medicinskim podmlat- Volume 67 | No.   73 .“ zapisali: „Naučne grupe u kojima naši drugovi i drug. odigraće veoma znača. Svaki student treba da sarađuje ji pristup klinikama i institutima. Godine su prolazile ne prekidajući kontinuitet rada arice produbljuju stečeno znanje. Ljubljane. stiče ona divna navika da se o dente i mlade istraživače koji ima najstariju svemu razmišlja sopstvenom glavom. ta u Beogradu. Započinje intenzivna saradnja sa Farmakološkim instu- maceutskih i stomatoloških fakulteta. pa već 1954. uključe u naučno­istraživački rad fakulteta. Rijeke i Skoplja. Izdavani su zajednički brojevi sa nizovala redovno publikovanje časopisa. samopregorno i entuzijatski orga. godine i od tada Pod. go- rubrika „Podsetnik“. Pomagati ovakav rad. oplemenjivati ih. Medicinskog podmlatka. koji svakome ko se njime itetu izlazi. Ta redakcija je na studentima medicine Beograda. kako mu skraćeno tepamo. U narednim godinama ideje Pod­mlatka du. Tako je Medicinski podmladak prerastao okvire la prvobitna zamisao da se u njemu štampaju samo Beograda i Beogradskog univerziteta. redakcijama stručnih časopisa medicinara Sarajeva i Za- zili i na svaka dva meseca. da bi ubrzo studentima Medicinskog fakulteta u Beogradu i Novom pospešio i saradnju sa drugim fakultetima u Jugoslaviji. „Preventivna medicina“. do­dao je: „Svaki student Medicinske velike škole na samostalan rad. koji će u radu nastavni kadrovi. Sadu pridružili su se i studenti Stomatološkog fakulte- Osnivač časopisa je Medicinski fakultet u Beogra. da se opšte priznate tradiciju u jugoistočnoj Evropi. Časopis su istine proveravaju i potvrđuju sopstvenim zapažanjima. koje je korišćeno kao udžbenik iz onkologije. Šetalo Jožef.

Tijana Ralić 2009/10 Miloš Maksimović 2011. Gordana Vujanić dakcija. ali 1978. Jovan Kačaki nja. 1981. Ljubiša Rakić zvaničnim udžbenicima. Milica Maksimović 2007/08. 1973. do. Snežana Andrejević 1987. Žarko Martinović Osnovni zadatak koji smo dobili od naših prethod. sa ciljem da podstakne studente medicine na naučnoistraživački rad. Ljubomir Crevar brovoljnim radom. Ljiljana Mirković torstvom svojih učitelja. Zorica Živković 1983/84. Slobodan Imširagić traživačkog i publikacijskog rada studenata. Milorad Radetić za priprema­nje nekih ispitnih pitanja manje obrađenih u 1954. Dragoljub Jovanović pokretanje međunarodnog časopisa studenata medi­cine 1959. Pavle Pavlović traživačkog rada. Brankica Dimitrijević 74  Mart 2016. 1967. 1968. Miroslav Radojković dejalektičara. Miloš Branković 2013. upravo zbog 1960. a pre svega da studente medicine. Ljiljana Planić 1981. izgradnji struč­njaka 1974. Danilo Zdravković u Beogradu i njihovih učitelja. I sve to. Branislav Donfried doprinese vaspitanju mladog lekarskog naraštaja. Nebojša Đurišić 2003/04/05. 1974/75. ali i da 1949. Nebojša Marković 2014/15 Stefan Barišić 2015. 1966. Jožef Šetalo nika trudimo se da ispunjavamo i danas: da Pod­mladak 1969/70/71/72. Slaviša Ćurćić i sa željom da ostavi nove podsticaje generacijama koje 1979/80. Lidija Tasić 1999/2000. stva. Dušan Đurić ih informiše o najnovijim dostignućima iz oblasti bio. tako da je danas Urednici Medicinskog podmlatka: kooperacija na zavidnom nivou. Zoran Vukašinović 1982. Jovan Bijelić Nesrećne okolnosti u našem okruženju sprečile su 1956/57/58. pod men. Aleksandar Anđelković u teškim vremenima socijalnih rastakanja i ratnih zbiva­ 1962. 2016. Ljubomir Erić ugleda našeg Medicinskog podmlatka. To je cilj kome stremi i današnja re. 1950/51. Mirjana Petrović je ovaj časopis i u vremenima sankcija studentima našeg 1963. Danica Sabovljević fakulteta bio svetionik za praćenje savremenih dostignuća 1964/65. Milan Marinković 2012. svesna veličine nasleđa svojih prethodnika. 1976/77. Časopis se održao i 1961. Medicinski podmladak kom uključivali profesori sa svih katedri. | Broj 1 | Izdanje 67 . 1976. Boško Kukolj medicninske nauke i podsticaj za neprekidnost naučnois. Živko Perišić dolaze. Snežana Jovanović 1985/86. uvede u lepote i tajne naučnois. Milorad Grujičić 2005/06. Ljubodrag Mihajlović medicinskih i javnozdravstvenih nauka i pruži literaturu 1952/53. Marija Marinković 1998. Svetlana Anić … 1997. Tako 1963. 1966. 1955. Istraživačka bujica studenata medicine Univerziteta 1962. Ljubomir Stefanović čije sedište je trebalo da bude u Beogradu. progresivno je rasla. Miroslav Vrbanović ranju ličnosti visoke moralne snage. Dejan Nešić 2001/02.