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English to Croatian: DISCOVERY OF THE E. COLI RESOLVASE General field: Medical Detailed field: Accounting
Source text - English DISCOVERY OF THE E. COLI RESOLVASE
The key observation which led to the identification of the E. coli Holliday junction resolvase was made by Bernadette Connolly while she was a postdoc in this laboratory. Using recombination intermediates made in vitro with RecA protein, Connolly identified a weak activity in fractionated cell extracts that appeared to produce recombinant products (9). The results would not have been convincing but for the fact that the same fractions resolved small synthetic Holliday junctions into nicked duplex products. Cleavage occurred by the introduction of symmetrically related nicks at the site of the junction, as expected of a resolvase on the basis of studies of the bacteriophage T4 gene 49-encoded resolvase (endonuclease VII). Gel filtration studies indicated that the E. coli resolvase was a small protein with a molecular mass of approximately 20 kDa. To identify its gene, extracts were prepared from various recombination-defective mutants, and one extract prepared from a ruvC strain was found to be deficient in cleavage activity (8). These observations, in combination with bacterial genetics,have since led to the discovery of a number of proteins that interact with Holliday junctions. The first ruv mutants were identified by Otsuji et al. (29) in 1974 and exhibited phenotypes characterized by increased sensitivity to UV light and to mitomycin and an associated cell division defect which led to the formation of nonseptate filamentous cells after DNA-damaging treatments. In certain genetic backgrounds, ruv mutants also showed defects in recombination (22, 46). Subsequent studies showed that the ruv operon, which maps at 41 min on the E. coli linkage map, actually encodes three genes and that either ruvA, ruvB, or ruvC mutants display the same phenotypic properties as the original mutant (7, 40, 45). Otsuji’s mutation is now known to map within ruvB.
IDENTIFICATION OF THE RUVA AND RUVB PROTEINS AS CATALYSTS OF BRANCH MIGRATION
The identification of RuvC as the E. coli Holliday junction resolvase led to new efforts being exerted toward a study of the ruvA and ruvB gene products. The RuvA (22-kDa) and RuvB (37-kDa) proteins had been purified by Hideo Shinagawa and colleagues; Iwasaki et al. (17) showed that RuvB was a DNAdependent ATPase and Shiba et al. (44) showed that its activity was stimulated by RuvA. They also found that the combined action of RuvAB resulted in the reabsorption of an extruded cruciform structure from plasmid DNA (44). Since cruciforms are structurally related to Holliday junctions, the possibility that cruciform loss occurred by a process akin to branch migration was discussed. Irina Tsaneva, a visiting scientist in my laboratory, used recombination intermediates to show directly
that RuvA and RuvB were able to promote ATP-dependent branch migration in vitro (51, 52).
Translation - Croatian OTKRIĆE REZOLVAŽE E. COLI
Ključno promatranje koje je dovelo do identifikacije rezolvaže Holliday-evih unakrsih veza Escherichiae coli izvršila je Bernadette Connolly za vrijeme njenoga postdoktorskog istraživanja u ovoj laboratoriji. Koristeći rekombinacijske intermedijere koji su načinjeni in vitro sa RecA proteinom, Condolly je identifikovala slabu aktivnost u dijelovima razdjeljene ćelije koji su naizgled uzrokovali rekombinantne tvorevine (9). Ovi rezultati ne bi bili značajni da nije bilo činjenice da ti isti djelići nisu razlagali malene sintetičke Holliday-eve veze na urezane duplekse. Rascjep se dogodio uvođenjem simetrički povezanih ureza na mjestu vezivanja, što je i bilo očekivano kada je u pitanju rezolvaža na osnovu istraživanja bakteriofaga T4 gena 49 - kodirana rezolvaža (endonukleaza VII). U istraživanjima uz pomoć gel-filtracije se navodi da je rezolvaža E. coli bila maleni protein sa molekularnom masom koja otprilike iznosi 20kDa. Da bi se njezin gen identificirao, pripremljeni su uzorci različitih mutanta nastalih uslijed neuspješne rekombinacije, te je otkriveno da jedan ekstrakt pripremljen iz iz linije ruvC-a manjka u činjenju razdvajanja (8). Ova opažanja, zajedno sa bakterijskom genetikom su potom dovela do otkrića znatnog broja proteina koji uzajamno dejstvuju sa Holliday-evim vezama. Prve ruv mutante je 1974. godine otkrio Otsuji i njegov tim (29), kada su se ispoljili fenotipi čije je obilježje bila povišena osjeljivost na UV svjetlost, osjetljivost na mitomicin, te oštećenje pri podjeli ćelije, što je dovelo do formiranja vlaknastih nepregrađenih ćelijskih jedinica nakon postupka oštećenja DNK stanica. U određenim slučajevima kada je u pitanju genetika, ruv mutanti također pokazuju oštećenja prilikom rekombinacije (22,46). Naredne studije su pokazale da operona ruv-a, koja se mapira u 41. minuti na genetičkoj mapi Escherichiae coli, zapravo kodira tri gena, te da bilo koji od tri, ruvA, ruvB ili ruvC manifestuju iste fenotipske osobine kao izvorni mutant (7, 40, 45). Za Otsujievu mutaciju se zna da se mapira u okviru ruvB-a.
IDENTIFIKACIJA RuvA i RuvB PROTEINA KAO KATALIZATORA PRILIKOM MIGRACIJE GRANE DNK
Identifikacija RuvC kao rezolvaže Hollidayeve veze E. Coli je dovela do novih pokušaja za provedbu istraživanja genetskih produkta ruvA i RuvB. Hideo Shingawa i njegov tim su pročišćavali RuvA (22-kDa) i RuvB (37-kDa) proteine; Istraživanje koje je proveo Iwasaki i njegovi saradnici (17) je pokazalo da je RuvB ATPaza koja ovisi o DNK, te su Shiba i njegov saradnički tim (44) dokazali da njezinu aktivnost simulira RuvA. Oni su također otkrili i to da kombinovano djelovanje RuvAB rezultira ponovom apsorpcijom ispupčene strukture u obliku križa iz plazmida DNK (44). Obzirom da su strukture u obliku križa po strukturi u svezi sa Hollidyevim vezama, razmatrana je mogućnost da dođe do gubitka struktura u obliku križa pri procesu koji je srodan migraciji grane. Irina Tsaneva, gostujući znanstvenik u mom laboratoriju je koristila rekombinaciju intermedijera da pokaže da RuvA i RuvB mogu promicati ATP-ovisnu migraciju grane u in vitro procesu. (51,52)
Serbian to English: Odloženi angioedem tokom primene inhibitora angiotenzin-konvertujućeg enzima General field: Medical
Source text - Serbian Apstrakt
Uvod. Primena inhibitora angiotenzin-konvertujućeg enzima predstavlja vodeći uzrok angioedema izazvanog lekovima, sa incidencijom od 0,1 do 0,2%. Angioedem nije imunske prirode, već nastaje kod predisponiranih osoba kao posledica nagomilavanja vazoaktivnih medijatora čija razgradnja je blokirana.
Prikaz bolesnika. U ovom radu prikazan je 63-godišnji bolesnik sa hipertenzijom i hroničnom opstruktivnom bolešću pluća koji je imao dve epizode angioedema tokom petogodišnje primene enalaprila. Prva epizoda javila se posle tri godine, a druga posle pet godina od početka primene enalaprila. Oba puta bolesnik je bio hospitalizovan, pri čemu je u poslednjem trenutku izbegnuta urgentna traheotomija kao krajnja terapijska mera. Zaključak. Tokom terapije inhibitorima angiotenzin-konvertujućeg enzima može nastati odloženi angioedem, posebno kod osoba sa inflamacijom respiratornih puteva usled infekcije ili hronične iritacije.
Ključne reči: angiotenzin-konvertujući enzim, inhibitori; angioneurotski edem; lekovi, neželjeno dejstvo, sistemi za izveštavanje.
Uvod
Primena inhibitora angiotenzin-konvertujućeg enzima (ACE) predstavlja vodeći uzrok angioedema izazvanog lekovima i jedan od najčešćih poznatih uzročnika angioedema uopšte 1–8. Uzevši u obzir procenu da ove lekove koristi više desetina miliona ljudi širom sveta 9, sa stalno prisutnom tendencijom porasta upotrebe koja sledi incidenciju kardiovaskularnih oboljenja, pojava angioedema, kao potencijalno fatalne komplikacije lečenja, iako nije česta, svakako se mora smatrati značajnim zdravstveno-ekonomskim problemom savremenog doba. O značaju pomenutog neželjenog efekta svedoče podaci iz dosadašnjih epidemioloških istraživanja koji, iako nisu konzistentni, ukazuju da je prosečno 1/3 bolesnika lečenih u urgentnim centrima zbog angioedema uzimala ACE inhibitore; čak 20% takvih bolesnika bilo je neposredno životno ugroženo usled opstrukcije disanja i podvrgnuto intubaciji ili hitnoj traheotomiji 1–7, 10–16. U ranijim izveštajima, zabeležena je stopa smrtnosti takvih bolesnika od oko 11% 14, međutim, brža i agresivnija terapija tokom poslednje 1–2 decenije dovela je do znatnog smanjenja broja smrtnih ishoda 1–3, 16.
Angioedem posle primene ACE inhibitora javlja se relativno retko: najveći broj studija procenjuje učestalost kod 0,1–0,2% bolesnika 4, 8, 10, 17, mada stvarna incidencija može biti veća imajući u vidu rastuću upotrebu ove značajne grupe lekova i posebno činjenicu da zbog specifičnosti kliničkog ispoljavanja ovog oblika angioedema, koja se pre svega ogleda u tome da može nastati bilo kada u toku primene leka, nezavisno od doze, znatan broj bolesnika u etiološkom smislu biva u praksi neprepoznat 2, 7, 8, 10, 18–27. U radu je prikazan bolesnik sa odloženom pojavom angioedema posle primene enalaprila.
Translation - English Abstract
Introduction. Angiotensin-converting enzyme inhibitors are leading cause of drug-induced angioedema, with incidence of 0.1 to 0.2%. The angioedema is not of immune nature; in predisposed individuals it is caused by accumulation of vasoactive mediators due to reduced activity of angiotensinconverting enzyme.
Case report. We presented a 63-year old male patient suffering from hypertension and chronic obstructive pulmonary disease, who had developed two episodes of angioedema during a 5-year long therapy with enalapril. The first episode happened after three, and the second after five years of the therapy. On both occasions, the patient was admitted to the hospital and tracheotomy was avoided in the last moment. Conclusion. Long-term therapy with angiotensin-converting enzyme inhibitors could be associated with delayed angioedema, especially in patients with inflammation of airways caused by infection or chronic
irritation.
Introduction
The use of angiotensin-converting enzyme (ACE) inhibitors is a leading cause of drug-induced angioedema and is one of the most common known causes of angioedema in general 1-8. Bearing in mind the assessment that this drug is used by tens of millions of people around the world 9, with the ever-present tendency of the usage increase which follows the incidence of cardiovascular diseases, the appearance of angioedema, as a potentially fatal treatment complication, although being infrequent, is certainly to be considered a significant health and economic problem of the modern ages. The importance of the aforementioned adverse effects is testified by the data collected from the previous epidemiological studies which, although not consistent, indicate that the average 1 / 3 of the patients treated in emergency facilities were taking ACE inhibitors due to angioedema; 20% of these patients were directly at the risk of death due to the breathing obstruction and were subjected to intubation or emergency tracheotomy 1-7, 10-16. In previous reports, the recorded mortality rate of these patients was around 11% 14, however, faster and more aggressive therapy during the last one/two decades has led to a significant reduction in the number of deaths 1-3, 16 .
After the use of ACE inhibitors, the occurence of angioedema appears to be relatively rare: the greatest number of studies estimate their occurence in 0.1 to 0.2% of patients 4, 8, 10, 17 , although the actual incidence may be higher, taking into account the growing use of this important group of drugs and particularily the fact that due to the specific clinical manifestation of this form of angioedema, which is primarily reflected by the fact that it can occur at any time during the drug application process, regardless of dosage, a significant number of patients, in etiological sense,happen to be unrecognized in practice 2, 7, 8, 10, 18-27. This paper presents a patient with a delayed appearance of angioedema after the use of enalapril.
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2020 University of Sarajevo; Faculty of Philosophy in Sarajevo
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2010 – 2012 University of Sarajevo; Faculty of Philosophy in Sarajevo
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