To see the other types of publications on this topic, follow the link: Proměna statusu.
Journal articles on the topic 'Proměna statusu'
Create a spot-on reference in APA, MLA, Chicago, Harvard, and other styles
Consult the top 50 journal articles for your research on the topic 'Proměna statusu.'
Next to every source in the list of references, there is an 'Add to bibliography' button. Press on it, and we will generate automatically the bibliographic reference to the chosen work in the citation style you need: APA, MLA, Harvard, Chicago, Vancouver, etc.
You can also download the full text of the academic publication as pdf and read online its abstract whenever available in the metadata.
Browse journal articles on a wide variety of disciplines and organise your bibliography correctly.
1
Žebeljan, Darko. "BUDUĆI PRAVCI RAZVOJA E-TRGOVINE I ULOGA POŠTANSKIH ORGANIZACIJA." Zbornik radova Fakulteta tehničkih nauka u Novom Sadu 36, no. 06 (June 7, 2021): 1106–9. http://dx.doi.org/10.24867/13ds04zebeljan.
Full textAbstract:
Uloga informacionih tehnologija, promenila je tradicionalni način poslovanja licem u lice i stvorila je pogodne uslove za dalji razvoj e-trgovine (engl. e-commerce), kao jednog od oblika e-poslovanja (engl. e-business). Rad je kao svoj osnovni predmet izdvojio sagledavanje veza između e-trgovine i poštanskih usluga, jer je osnovna delatnost poštanske mreže obavljanje tradicionalnih poštanskih usluga. Međutim, pojava globalnih promena na tržištu, izvršila je promene i u poštanskom sistemu, zahtevajući od njega, modernizaciju i proširivanje. U cilju razvoja poštanskog sistema, rad je pružio osvrtna digitalnu platformu .POST, navedeni su primeri svetskih poštanskih operatora, koji su unapredili svoje poslovanje, sagledan je status poštanskog sektora u Srbiji i sagledane su mogućnosti za budući razvoj poštanskog sistema.
2
Radovanović, Dragana. "Employment status in case of status change, i.e. change of employer." Pravo i privreda 58, no. 2 (2020): 297–310. http://dx.doi.org/10.5937/pip2002297r.
Full text
3
Mršević, Zorica. "Poces promena pravnog statusa lezbejki i homoseksualaca u Evropi i kod nas." Glasnik Advokatske komore Vojvodine 72, no. 8-9 (2000): 96–107. http://dx.doi.org/10.5937/gakv0003096m.
Full textAbstract:
Proces širenja nekog domena ljudskih prava neophodno je praćeno određenim političkim zbivanjima. Proglašenje Manifesta ILGA-e od 7. decembra 1998. kao dela proslave Manifesta ILGA-e kojim se proglašavaju lezbejska i prava homoseksualaca kao ljudska prava, deo je takvih zbivanja. 1. "Čovečanstvo je bilo u stanju da ukine ropstvo i da uspostavi određena osnovna prava za žene u gotovo svim delovima sveta. Mi smo razvili demokratske institucije i građanska društva. Poštovanje ljudskih prava i osnovnih sloboda je preduslov uspešnog društvenog i političkog života u svim društvima. Mi moramo priznati jednakost i poštovati raznolikost. Na 50-tu godišnjicu Univerzalne deklaracije ljudskih prava mi pozivamo na jačanje pažnje ljudi i društava na ljudska prava. Osnovni deo ljudskih prava u naše vreme je priznavanje prava pojedinca na razvitak svojih ličnih identiteta i seksualnosti, slobodne od prinude i diskriminacije. Mi osuđujemo diskriminaciju i opresiju koja nastavlja da postoji u mnogim državama, uključujući krivične sankcije i čak i smrtnu kaznu. Zahtevamo punu jednakost pred zakonom za homoseksualne muškarce, lezbejke i biseksuace kao i transseksualne osobe. Zahtevamo kraj ma kakvom krivičnom kažnjavanju. Zahtevamo zakone koji zabranjuju diskriminaciju. Zahtevamo puno i potpuno jednako priznanje odnosa. Nadamo se uključivanju jednakih prava bez obzira na seksualnu orijentaciju u nacionalno zakonodavstvo i međunarodne instrumente ljudskih prava. Mi zajedno možemo da učinimo život boljim jedne za druge i za buduće generacije. Prava lezbejki i homoseksualaca su ljudska prava."
4
Hrustić, Hasiba. "Tax treatment of status change of commercial companies." Glasnik Advokatske komore Vojvodine 90, no. 5-8 (2018): 274–90. http://dx.doi.org/10.5937/gakv1808274h.
Full text
5
Milenković, Dejan. "Adaptive changes of motoric status of preschool children under influence of physical education." Sinteze, no. 13 (2018): 41–49. http://dx.doi.org/10.5937/sinteze7-16929.
Full text
6
Glođović, Danica. "IZMEŠTANJE STATUSA GRAFIČKOG OBLIKOVANJA U DVADESETOM I DVADESET PRVOM VEKU." Srpska politička misao, Specijal 2019 (December 2, 2019): 209–22. http://dx.doi.org/10.22182/spm.specijal2019.13.
Full textAbstract:
U ovom radu, autorka istražuje promenu paradigme grafičkog oblikovanja, od početka dvadesetog veka do danas, kao i transformaciju oblikovnih praksi od tradicionalnih, ka digitalnim medijima. S obzirom na to da je tema ovog rada izmeštanje statusa grafičkog oblikovanja u istraživačkom postupku prilikom obrade predložene teme primeniće se set teorijsko epistemoloških tehnika što podrazumeva raznovrsne interdisciplinarne i transdisciplinarne metode. Radi isticanja i problematizovanja ključnih oblasti teorije grafičkog oblikovanja, preglednom tehnikom će se mapirati i predstaviti opus oblasti i aktuelnih teorija vizuelne kutlure koje pokriva predložena tema, prema zadatim kriterijumima koje ovaj rad predlaže. Kriterijumi za mapiranje i kritičku analizu oslanjaće se na savremene teorije grafičkog oblikovanja, a i društva kao krajnjeg korisnika dizajniranih proizvoda i usluga. Grafičko oblikovanje ne može se tumačiti kao samo jedan od estetskih pojmova, radnje ili proizvoda, već je reč o svojevrsnom političkom fenomenu. Dizajn je, kako estetsko oblikovanje, tako i autentično političko oblikovanje znakovnog i materijalnog sveta. Oblikuje svakodnevni život ljudi, i široko je ugrađen u mehanizam vladanja društvenim prostorom, kao i životom uopšte, u urbanom prostoru i nad telima koja taj prostor zauzimaju. U modernim vremenima, kroz podršku tehnologije, kao i transdisciplinarnom pristupu kroz konvergenciju različitih poslovnih i društvenih praksi, grafičkom oblikovanju pružaju se neke nove mogućnosti, što svakako izmešta ciljeve i dostignuća ove oblasti. Može se reći da je uloga grafičkog oblikovanja da na mikro nivou proizvodi subjektivnost, a na makro nivou uspostavlja fizičke i virtuelne ograde upravljanja, kontrole i kretanja. Stvarni svet više nije uočljiv, jer ono što subjekt vidi i zna o stvarnom svetu, jeste ono što mu govore proizvedeni znakovi. U ovom radu autorka obuhvata i problematiku autorstva u odnosu na dizajnerske prakse. Ovim istraživanjem autorka želi da mapira kluljčne tačke u promeni paradigme grafičkog oblikovanja, kao i da čitaocima predstavi put razvoja dizajnerskih praksi kakve danas poznajemo.
7
Raičević, Nebojša. "Exceptions to the cessation of refugee status due to a change of circumstances in the country of origin." Zbornik radova Pravnog fakulteta Nis 57, no. 80 (2018): 85–100. http://dx.doi.org/10.5937/zrpfni1880085r.
Full text
8
Musa, Anamarija. "Politički savjetnici između politike i uprave." Hrvatska i komparativna javna uprava 18, no. 4 (December 31, 2018): 585–615. http://dx.doi.org/10.31297/hkju.18.4.4.
Full textAbstract:
Politički savjetnici obilježje su suvremene državne uprave. Predstavljaju osobe koje temeljem imenovanja prema nedefiniranim kriterijima pružaju savjete ministru ili premijeru u pogledu javnih politika, političkih aspekata i koordinacije ili odnosa s javnošću. Dolaze i odlaze s ministrom, a njihov je status u pravilu nejednako reguliran u pojedinim državama, ako je uopće i reguliran. Javljaju se zbog promjena u upravljanju, potrebe za jačanjem koordinacije, vodstva i strategije kao i odgovora na intenziviranje komunikacija. Dovode do promjena u odnosu između politike i uprave u državnoj upravi, a osnovne zamjerke i nepovjerenje javnosti proizlaze iz netransparentnosti, izostanka regulacije i nedostatka odgovornosti. U radu se razmatraju razlozi zbog kojih politički savjetnici postaju neizostavan dio ministarskih kabineta, analiziraju se pojam, vrste i funkcije političkih savjetnika te opisuju posljedice i ključni elementi koncepta. Naglašava se važnost uređenja statusa političkih savjetnika i transparentnosti radi osiguravanja ostvarivanja javnog interesa i povjerenja javnosti u institucije.
9
Pauwels, Patrick, Torben Steiniche, Muriel Gazin, Jan Van de Velde, Ellen Bellon, Evelien Rondelez, Liesbeth Vandenbroeck, et al. "The Idylla MSI Test (CE-IVD) multi-center concordance study: Microsatellite instability detection in colorectal cancer samples." Journal of Clinical Oncology 37, no. 15_suppl (May 20, 2019): e15112-e15112. http://dx.doi.org/10.1200/jco.2019.37.15_suppl.e15112.
Full textAbstract:
e15112 Background: To validate the Idylla MSI Test to detect microsatellite instability (MSI) in colorectal cancer (CRC) samples in comparison with Promega MSI Analysis System v1.2 (Promega MSI) and in conditions similar to normal use. Methods: The study was performed on residual formalin-fixed and paraffin-embedded (FFPE) samples obtained from routine diagnostics by two centers, University Hospital Aarhus and University Hospital Antwerp. Samples originated from CRC patients (all stages). Both centers performed the Idylla MSI Test (with 7 novel homopolymer deletion markers) at their premises on 150 and 180 samples, respectively. The comparator method, Promega MSI, was performed for all 330 samples by University Hospital Antwerp. Results: Seven (n=7) of 330 samples were excluded from the concordance analysis due to invalid or error (failed) results for Idylla MSI Test and/or Promega MSI. For the 323 valid results, overall, positive and negative percentage agreement were 99.7% (98.3%-100%;), 98.7% (92.9%-99.8%) and 100% (98.5%-100%), respectively. A higher number of invalids was observed for Promega MSI (2.1%) compared to the Idylla MSI Test (0.6%) taking into account per protocol retesting (18 retests for Promega MSI (6%) and 3 for Idylla MSI Test (1%)). Conclusions: This study validated the Idylla MSI Test with high performance and low invalid rate to discriminate MSI-H from MSS status on a clinical routine set of CRC samples.
10
Petranović, Anamari. "Ugovorne obveze u Lastovskom statutu." Zbornik Pravnog fakulteta Sveučilišta u Rijeci 39, no. 2 (2018): 753–71. http://dx.doi.org/10.30925/zpfsr.39.2.2.
Full textAbstract:
Analizom odredbi Lastovskog statuta (1310. godina) u radu se podastiru pojedine značajke ugovornih obveza, posebice s obzirom na statuirane detalje (prepoznate/potvrđene recentnijim uredbama) koji postaju pokazateljima konkretnih onovremenih naglasaka gospodarskih i životnih prioriteta Lastova. Elementi raščlambe odnose se na restrikcije pravnog prometa (status ugovaratelja, ograničenja i zabrane trgovanja sa strancima, osobitosti trgovanja određenom robom, maksimiranje cijena…), kao i na pojedina pravna rješenja odstupanja od načela konsenzualnosti – javno oglašavanje prodaje, status notarske isprave, upis u komunalnu kancelarijsku knjigu, odnosno sklapanje ugovora „per aptago“. Rad notira statutarnu stilizaciju definiranja pretpostavki valjanosti ugovora koja uključuje i kaznenopravne elemente, budući da posljedicu ništetnosti može pratiti i sankcioniranje nepoštovanja statuiranog načina sklapanja određenih (obvezno)pravnih poslova, upućuje na pojedine odrednice u problematici zaštite prava ovlaštenika na otkup (odnosno prvokup) putem u interesu otočana osmišljenih rokova javne objave, dotiče pitanje javnog nadmetanja kao i režim nevaljanosti potajne i/ili prikrivene prodaje kao dodatnog ograničenja raspolaganja/otuđivanja nekretnina.
11
Stojanović, Tijana, Dragan Perić, Darko Stojanović, and Toplica Stojanović. "EFEKTI PROGRAMA „ŠKOLA U POKRETU“ NA POSTURALNI STATUS UČENIKA MLAĐEG ŠKOLSKOG UZRASTA." Sportlogia 16, no. 1 (December 17, 2020): 82–91. http://dx.doi.org/10.5550/sgia.201601.se.sdss.
Full textAbstract:
Sa ciljem da se utvrde efekti programa „Škola u pokretu“ na posturalni status učenika, sprovedeno je istraživanje na uzorku od 22 učenika mlađeg školskog uzrasta oba pola u trajanju jednog školskog polugođa. Učenicima su bila održana predavanja o ergonomskom riziku i načinima za smanjenje ergonomskih faktora rizika i distribuiran je određeni broj postera i flajera sa predstavljenim ispravnim tehnikama podizanja i nošenja školske torbe/ranca i ispravnog načina sedenja. Ergonomski program „Škola u pokretu“ podrazumevao je ohrabrivanje učenika da se slobodno pokreću na stolici ili da ustanu i istegnu se tokom časova kada osete nelagodnost/bol i uz nekoliko kratkih vežbi istezanja, za vreme sedenja na stolici. Posturalni status kičmenog stuba je procenjen u sagitalnoj i frontalnoj ravni (torakalna i lumbalna skolioza, kifoza i lordoza) instrumentom „Spinal mouse“ (Quantum Health and Wellness Ltd, Wallasay, England). Za statističku obradu podataka primenjena je analiza varijanse za ponovljena merenja (Repeated measures ANOVA). Rezultati pokazuju da je u periodu od 16 nedelja kod učenika došlo do značajnog poboljšanja vrednosti torakalne skolioze (p=0.003) i kifoze (p=0.006), dok kod lumbalne skolioze i lordoze nije bilo značajnih promena. Na osnovu ovih rezultata se može zaključiti da ovakav program može imati značajan uticaj na poboljšanje posture kičmenog stuba, te se može primeniti u praksi školske nastave i kao preventiva narušavanju posturalnog statusa kičmenog stuba učenika mlađeg školskog uzrasta.
12
Kužel, Petr. "Společensko-Ekonomické Proměny Sportovních Spolků a Vznik Fotbalových Klubů v Pražských Městech a Předměstích Před Rokem 1914." Acta Musei Nationalis Pragae – Historia 71, no. 1-2 (2017): 12–20. http://dx.doi.org/10.2478/amnh-2017-0003.
Full textAbstract:
Football, the most popular game all over the world, reached the territory of todayʼs Czech Republic in the last decades of the 19th century. In Prague districts and suburbs especially, many Czech and German sport associations started to engage in this sport activity originally born in Britain. The sudden and long-lasting interruption of a positive development due to the mobilization in summer 1914 along with significant political and social changes following the end of First World War, isolated pre-war events and made of them the unique relict environment which forms the main topic of this paper. Leaving sports results aside, the study describes the period after 1900 in which football clubs were established, the enthusiastic amateur transformed into a professional player, loyalty to different teams stemmed on the basis of nationality and social status and football moved from the suburbs’ playgrounds to newly-built, larger and better-quality arenas.
13
Marpaung, Noveri Lysbetti, Rahyul Amri, Edy Ervianto, and Nurhalim Dani Ali. "Analysis of Controlling Wireless Temperature Sensor for Monitoring Peat-Land Fire." International Journal of Electrical, Energy and Power System Engineering 1, no. 2 (November 5, 2018): 14–19. http://dx.doi.org/10.31258/ijeepse.1.2.14-19.
Full textAbstract:
The problem of forest and peatland fires in Riau has become main topic to prioritize its revamping. Land and forest fires in Riau until reached 3,700 hectares area, happened in some last years,.Forest and peatland fires occured in the majority of districts in Riau Province, from low category of 4 hectares until 2,800 hectares. This researchdiscusses aboutanalysis of controlling wirelessTemperature Sensor for monitoring Peat-Land fire by using LM35 Temperature Sensor, Transmitter-Module (HC-12), Receiver-Module (HC-12), Arduino-ProMini to produce outputs on LED, LCD, Buzzer. Detector of Peat-Land fire works when it is burning, so heat will spread through aluminium stalk and be read by Temperature-Sensor1 upto Temperature-Sensor4. Output of each sensor is sent to each control block of Arduino-ProMini in Transmitter-Module(FU1–FU4). From Arduino-ProMini, it is sent toReceiver-Module(FU5). Receiver-Module only receives one data from Transmitter-Module in one time. Every data is received by Receiver-Module goes to Arduino-ProMini, processed to produce outputs on LCD that shows ID of FU1–FU4, Peat-Land Status, temperature of Peat Land. If LED is Green, Peat-Land not burned, LED shows SAFE Condition, Buzzer Off. If LED is Yellow, Peat-Land burned underground, LED shows BE-CAREFUL Condition, Buzzer On. If LED is Red, Peat-Land burned on the ground, LED shows DANGER Condition, Buzzer On. Temperature 0°C–21.9°Cis SAFE Category because Peat-Land not burned. Temperature 22°C–28.4°Cis BE-CAREFUL Category because Peat-Land burned underground. Temperature 28.4°C–50°Cis DANGER Category because Peat-Land burned on the ground. This equipment works as its frame work.
14
Mandić, Marinela, Josipa Krković, Dario Lasić, Maja Budeč, Danijel Brkić, Jasna Bošnir, Jozica Raljević, Anna Pierobon, and Paula Sliva. "Praćenje promjena kemijskih parametara meda ovisno o načinu njegova skladištenja." Journal of applied health sciences 6, no. 2 (October 7, 2020): 239–47. http://dx.doi.org/10.24141/1/6/2/6.
Full textAbstract:
Med je po definiciji namirnica kojoj se ništa ne smije dodavati niti oduzimati kako bi zadržala svoja izvorna karakteristična svojstva. Podaci Food Fraud networka govore kako je riječ o hrani koja podliježe sve češćim prevarama potrošača, odnosno krivotvorenju. Med je specifičnog kemijskog i nutritivnog sastava koji mu daju status visokovrijedne hrane. Taj sastav, osim krivotvorenjem sastojaka, može se ugroziti i njegovim neadekvatnim skladištenjem ili pogrešnom manipulacijom tijekom prerade i prodaje. Cilj je ovoga projekta pratiti promjene osnovnih parametara kvalitete meda kao što su kiselost, elektrovodljivost, aktivnost dijastaze i količina hidroksimetilfurfurala (HMF), koji se mijenjaju s duljinom skladištenja, temperaturom i izloženošću meda Sunčevoj svjetlosti. Istraživanjem je utvrđeno da različiti uvjeti skladištenja meda povezani s promjenom temperaturnog režima imaju utjecaj na promjenu kvalitete meda. Ukupno je analizirano osam uzoraka meda (pet uzoraka meda bagrema i tri uzorka cvjetnog meda), a dobiveni rezultati prikazuju prosječne vrijednosti svih analiziranih uzoraka. Najveće promjene utvrđene su kod dijastaze i HMF-a. Med je skladišten na temperaturi od –20 °C, 0 °C, 20 °C te na temperaturi od 40 °C i 60 °C. Rezultati ukazuju da izlaganje meda višim temperaturama utječe na smanjenje aktivnosti dijastaze ispod 8DN. Jednako tako, rezultati ukazuju da vrijednosti HMF-a pri visokim temperaturama rastu i ako je izloženost meda visokim temperaturama dulja, vrijednosti HMF-a prelaze maksimalne vrijednosti (40mg/kg). Stoga je od velike važnosti da se pri distribuciji i skladištenju meda vodi računa o izloženosti meda utjecaju direktne svjetlosti i temperature okoliša u kojem se med skladišti kako bi se sačuvale prirodne vrijednosti meda. Potrebno je radi zaštite potrošača med skladištiti izvan dohvata Sunčeve svjetlosti i visokih temperatura, a sve u cilju zaštite zdravlja ljudi.
15
Bando, Hideaki, Wataru Okamoto, Takafumi Fukui, Takeharu Yamanaka, Kiwamu Akagi, and Takayuki Yoshino. "Clinical utility of quasimonomorphic variation range (QMVR) on the determination of microsatellite instability (MSI) status in Japanese patients (pts) with colorectal cancer (CRC): GI-SCREEN-CRC-MSI sub-study 01." Journal of Clinical Oncology 35, no. 4_suppl (February 1, 2017): TPS808. http://dx.doi.org/10.1200/jco.2017.35.4_suppl.tps808.
Full textAbstract:
TPS808 Background: The current analysis for the determination of Microsatellite Instability (MSI) status in tumor requires matched normal DNA as a reference. The Promega MSI panel included five monomorphic markers (NR-21, BAT-26, BAT-25, NR-24, and MONO-27) which were known to have few variant alleles in Caucasian as well as Japanese. Therefore, these markers might be able to determine the MSI status without matched normal DNA, although it is conceivable that Quasi-Monomorphic Variation Range (QMVR) in each ethnic group could be different. In our pilot study, the QMVR in 149 Japanese healthy individuals were 98.4-104.4 base pair (bp) in NR-21, 111.4-117.4 bp in BAT-26, 121.0-127.0 bp in BAT-25, 129.5-135.5 bp in NR-24, and 149.9-155.9 bp in MONO-27, respectively. Methods: This clinical evaluation study is to establish the QMVR in Japanese colorectal cancer (CRC) patients (pts) and to evaluate the clinical utility of the QMVR on the determination of MSI status without matched normal DNA. The primary endpoint is the concordance of MSI status between the standard method using DNA from tumor plus matched normal samples and testing method using DNA from only tumor samples. The new MSI kits including the Promega MSI panel were manufactured under the Quality Management System (QMS) for in vitro diagnostics (IVDs). As the decision algorithm, tumors exhibiting 2 or more markers outside the QMVR were classified as MSI-High, cases with 1 marker outside the QMVR were classified as MSI-Low, and those without any marker outside the QMVRs were classified as microsatellite stable (MSS). Planned enrollment will be 400 pts. Clinical trial information: UMIN000024144.
16
Goljenkova, Zinaida. "Basic tendencies in the change of social structure of the Russian society in the conditions of globalization and transformation." Zbornik Matice srpske za drustvene nauke, no. 118-119 (2005): 7–15. http://dx.doi.org/10.2298/zmsdn0519007g.
Full textAbstract:
U radu se, u najkracim crtama, izlazu osnovne tendencije promena drustvene strukture u Rusiji, koje se odvijaju u slozenim uslovima globalizacijske transformacije tog drustva. Na pocetku rada se konstatuje da nastanak nove socijalne strukture u Rusiji prati i promena naucne paradigme, koja se sastoji u napustanju marksistickog teorijsko-metodoloskog pristupa i usvajanju teorija socijalne stratifikacije. Autorica se oslanja na teorijsko-metodolosku zamisao P. Sorokina, koja se sastoji u prihvatanju sledecih kriterijuma raslojavanja u nekom drustvu: ekonomski, profesionalni i politicki. Na osnovu tih kriterijuma pristupa se socijalnoj stratifikaciji kao nejednakosti drustvenih grupa koja pociva na materijalnom bogatstvu nejednakom statusu (onih koji upravljaju i onih kojima se upravlja) i na drustvekom prestizu (koji nastaje kao posledica razlike u nivou obrazovanja i profesionalne pripreme), ali i na stepenu prilagodjavanja transformacionim procesima. Transformacioni procesi, od pocetka 90-ih godina 20. veka uslovili su prepoznatljivu osobenost ruskog drustva koja se ocituje u njegovoj polarizaciji na siromasne i bogate. Ta polarizacija je zahvatila sve sfere drustvenog zivota Rusije, ukljucujuci i sferu kulture (dostupnost kulturnih dostignuca, mogucnost ostvarivanja kulturnih potreba, stil zivota nacin koriscenja slobodnog vremena i sl). Sve to govori da je socijalna baza konfliktnosti ruskog drustva veoma siroka, sto se odrazava na spori razvoj gradjanskog drustva i adaptaciju gradjana na nove drustvene uslove. Socijalna struktura ruskog drustva ima sve bitne osobenosti klasnoslojne diferencijacije. Osnova te diferencijacije se nalazi u materijalnom polozaju ljudi. Na osnovu tog kriterijuma moguce je u savremenom ruskom drustvu primetiti sve ostriju granicu izmedju sledecih grupa nejednakosti ("resursnih grupa"): grupa siromasnih (koja obuhvata oko 65%), grupa malo imucnih (14%) grupa srednje imucnih (21%) i grupa visokoimucnih/bogatih (ostalo). Takva struktura ruskog drustva u fazi transformacije javlja se u okviru protivrecnih procesa globalizacije, sto jos vise utice na narastanje broja marginalnih delova stanovnistva i utice da to drustvo poprima bitne osobine "brazilizacije" ("brazilifikacija").
17
Fatić, Aleksandar. "The question of 'corrective change' and the criminal justice system." Glasnik Advokatske komore Vojvodine 68, no. 9 (1996): 386–400. http://dx.doi.org/10.5937/gakv9610386f.
Full textAbstract:
In recent criminological discourse, considerable emphasis has been placed on initiatives for deinstitutionalization, deformalization and liberalization of the regime of criminal justice institutions and policies. This initiative, which is largely contemporary, follows an earlier trend, which insisted on segregation of "deviants", on the centralization of the criminal justice apparatus, and on a certain "cold realism" regarding the real possibilities for inducing positive behavioural changes in the populations of "deviants" by restrictive and particularly penal measures. This older view thus implied that, although punishment and other restrictive practices did not in fact "reform" offenders, or encourage them in any way to become more legitimate members of society, it was the best and safest way for handling criminal deviance society could possibly come up with. These two paradigms, or views, are sometimes labelled "the first" and "the second correctional change". The first correctional change characterized the nineteen and the beginning of the twentieth century. This is the older view mentioned above. The second correctional change, insisting on liberalization and deformalization, is a product of this day. Apart from presenting the two theoretical paradigms and some elements of the background of labelling them "correctional changes", this paper attempts to show that the real constructive potential of the "second correctional change" depends on traditional values and virtues nurtured in the informal community, and that it does not appear to stand in any positive correlation with contemporary values and organization of the system. The reason for this is that, even if institutions could be significantly reduced, which is far from being uncontroversially acceptable, the real impact of informal procedures in place of formal ones will depend on the status of inter-relationships of trust in the informal community, namely the trust between the two sides of penal policy, including the consensual majority of the democratic public on the one, and the dissensual minority of "deviants", on the other hand. The paper argues that the de facto level of this trust is unsatisfactory, given that in liberal social arrangements interest is what governs social policy, and the interests of the majority of the consensual public stand largely opposed to the interests of the dissensual minority. This means that there is likely to be a great deal of reluctance on the two sides of the "control divide", and particularly on the two sides of the penal divide (the convicted and penalized offenders and the administrators of criminal justice) to invest trust in the belief that this structural opposition which leads to confrontation and "the control effort" can be eliminated and informal mechanisms of overriding cooperation and optimization put in its place. In any particular case, with any particular society, the paper argues, the success of informal strategics which many abolitionists are proposing these days, within the paradigm of "second correctional change", always depends on how vital the older, informal virtues and values are within the institutional texture of the system, and how easily and quickly they can be brought back into operation. This question, the paper concludes, is to a considerable extent independent of the question of the virtues and vices of the system itself.
18
Heubner, Martin Leonhard, Rainer Kimmig, Winfried Siffert, and Frey Ulrich. "The haplotype of three polymorphisms in the SATB1 promoter-region and impact on survival in breast cancer patients." Journal of Clinical Oncology 31, no. 15_suppl (May 20, 2013): 584. http://dx.doi.org/10.1200/jco.2013.31.15_suppl.584.
Full textAbstract:
584 Background: Special AT-rich sequence binding protein 1 (SATB1) has regulatory effects on gene expression and appears to play an important role in tumor progression. We screened the promoter region of the SATB1 gene for polymorphisms, evaluated the corresponding haplotypes regarding alterations in promoter activity in vitro and analyzed the impact of these haplotypes on the clinical course of breast cancer patients. Methods: 241 caucasian breast cancer patients who had been treated were enrolled in this retrospective analysis. The median follow up time was 93 months (4-155 months). PCR products from DNA of 10 healthy unrelated volunteers were analyzed to identify new polymorphisms within the promoter region. Genotyping was conducted using restriction length polymorphism and pyrosequencing. PCR constructs with the respective alleles from the four most frequent haplotypes were cloned into the vector pGEM-Teasy (Promega Corporation, Madison, WI, USA) and then transferred into the luc2-containing reporter vector pGl 4.10 Vector (Promega) for transfection of HEK293 cells. The pGl 4.73 Vector (Promega), containing hRluc, was used for norming the transfection rates. Results: Sequencing the region -3807bp to -2828 upstream from ATG of ten healthy blood donors, we found three single nucleotide polymorphisms SNPs consisting of base exchanges, -3600T>C, -3363A>G and -2984C>T.The SATB1 -3600T/-3363A/-2984C haplotype had a lower promoter activity than all other constructs in vitro and showed a significant association with the nodal status (p=0.049). Kaplan-Meier survival analysis revealed a significantly better survival for homozygous SATB1 -3600T/-3363A/-2984C haplotype carriers compared with heterozygous or the other haplotypes (p=0.033). Conclusions: The SATB1 -3600T/-3363A/-2984C haplotype is associated with lower promoter activity and appears to impact upon survival in breast cancer patients.
19
Pejin, Jovan. "KRUG OKO NAJNOVIJEG ZAHTEVA DZVH ZA MESTA U VLADI I SKUPŠTINI SRBIJE." Politička revija 66, no. 4/2020 (January 5, 2021): 419–25. http://dx.doi.org/10.22182/pr.6642020.20.
Full textAbstract:
Politički Beograd i Srbija izloženi su višedecenijskim pritiscima, tačnije rečeno, spolja velikih sila i unutrašnjim raznih stranaka, stranaka nacionalnih manjina i nevladinih organizacija. Uoči puta u Vašington vodećih političara zemlje radi razgovora sa najvišim i najznačajnijim ljudima jedne od vodećih sila u svetu o pitanju uključenja NATO nezavisnog protektorata Kosovo uspostavljenog na okupiranom delu Srbije u ekonomski razvoj i saniranja štete izazvane terorističkom pobunom Arbanasa u Pokrajini Kosovo i Metohija, desilo se preuzimanje vlasti u Bujanovcu i Preševu, nakon izbora 2020, od predstavnika samo arbanaške nacionalne manjine. Zatim, Srbija se suočila sa zahtevom Hrvatske nacionalne zajednice i Demokratske zajednice Hrvata u Vojvodini da promene svoj status u pokrajini i Republici ulaskom njihovih članova u vladu i skupštinu.
20
Correia, Arícia Fernandes. "REGULARIZAÇÃO FUNDIÁRIA URBANA: PRETENSOS CONTRIBUTOS PARA REGULAMENTAÇÃO DA LEI FEDERAL N. 13.465/2017 E PARA A CONSTRUÇÃO DO PLANO DIRETOR DA CIDADE DO RIO DE JANEIRO 2021." Geo UERJ, no. 36 (February 14, 2020): e47270. http://dx.doi.org/10.12957/geouerj.2020.47270.
Full textAbstract:
A Lei Federal n. 13.465/2017 expressamente dispensa a intervenção do legislador local para a aplicabilidade imediata de seus preceitos, tornando despicienda em tese a atuação do legislador local. Ocorre que a lei federal traça normas gerais (i) cujas especificidades, em matéria de regularização fundiária para a população de baixa renda, já estão disciplinadas pelo parlamentar municipal carioca e (ii) cujas potencialidades, em matéria de regularização fundiária para a população de média e alta renda, podem ser exploradas diante das especificidades locais e dos princípios constitucionais de direito urbanístico que o informam, de modo que torna sua regulamentação local, por lei formal, um valioso instrumento de Política Urbana. A regulamentação municipal da nova lei federal sobre regularização fundiária urbana passa a ser um desafio também para que (i) o planejamento urbano ex ante não seja convertido em convalidação da ilegalidade urbana ex post e (ii) se promova uma regularização fundiária urbana sustentável capaz de gerar uma cidade supostamente menos iníqua e desigual, já que sua aplicação acrítica tende a provocar concentração fundiária, o que concorreria justamente para o quadro fático oposto ao que dela se espera, e (iii) se promova a mudança do status quo ante e não sua mera legitimação estatal. Por outro lado, nada obsta que desde logo se verifique como o anteprojeto de novo Plano Diretor da Cidade do Rio de Janeiro deve cuidar do tema da regularização fundiária urbana, diante das diretrizes gerais ditadas pelo marco regulatório nacional.
21
Diković, Marina, and Marlena Plavšić. "Vrijednosti obrazovanja i nastavnički identitet iz učeničke, roditeljske i nastavničke perspektive." Metodički ogledi 26, no. 1 (2019): 7–31. http://dx.doi.org/10.21464/mo.26.1.4.
Full textAbstract:
U Hrvatskoj i svijetu manji je dio nastavničkoga kadra koji smatra da je nastavnička profesija cijenjena. Većina se autora i autorica slaže da i jačanje nastavničkoga identiteta moglo pomoći podizanju vrijednosti obrazovanja. Ciljevi istraživanja bili su: ispitati na koje načine obrazovanje doprinosi kvaliteti života i koje su prednosti nastavničke profesije, ispitati doživljaj statusa nastavničke profesije i promjena u obrazovnom sustavu te utvrditi motivaciju za nastavničku profesiju. Uzorak je obuhvaćao 74 učenika i učenica osnovnih i srednjih škola, 73 roditelja, 38 studenata i studentica nastavničkih studija te 35 osnovnoškolskih i srednjoškolskih nastavnika i nastavnica. Rezultati se interpretiraju u kontekstu Korthagenovog modela te pokazuju da nastavnici i nastavnice više spominju obilježja nastavničkoga identiteta koja se dotiču temeljnih vrijednosti, dok roditelji i učenici više spominju obilježja ponašanja i kompetencija. Zadaća je inicijalnoga obrazovanja nastavnika i nastavnica poticati mlade na promišljanje o vrijednostima nastavničke profesije jer će time cijeniti i poticati razvijanje vrijednosti nastavničkoga identiteta.
22
Višnjić-Jevtić, Adrijana, and Ivana Visković. "Roditeljstvo u vrijeme pandemije COVID-19." Metodički ogledi 28, no. 1 (July 16, 2021): 11–38. http://dx.doi.org/10.21464/mo.28.1.4.
Full textAbstract:
Uslijed pretpostavljenih promjena načina života zbog pandemije Covid-19, tijekom lockdowna u prvom valu epidemije istražena je samoprocjena roditeljstva kod roditelja djece rane i predškolske dobi. Cilj je bio istražiti kvalitetu roditeljstva kroz procjenu učestalosti zajedničkih aktivnosti s djecom te načina ponašanja prema djeci. Relativna većina sudionika istraživanja procjenjuju da nisu značajno promijenili pojedine aktivnosti s djecom (razgovor, pripovijedanje, fizički iskazi ljubavi), dok četvrtina roditelja procjenjuje da s djecom priča, uči i iskazuje ljubav češće nego prije. Gotovo polovica roditelja procjenjuje da se više igraju s djecom nego prije lockdowna. Obiteljski odnosi izdvojeni su kao najznačajnija dimenzija roditeljstva. Životna dob, razina formalnog obrazovanja i bračni status sudionika istraživanja, kao prediktorski sklop objašnjava tek 2,3 % varijabiliteta samoprocjene roditelja pa je opravdano pretpostaviti postojanje drugih, značajnijih prediktora kvalitete roditeljstva. Sakupljeni podaci ukazuju na prednosti (velik uzorak i brza provedba) te nedostatke (specifičnost uzorka) online istraživanja. Veličina uzorka ukazuje na angažiranost roditelja na društvenim mrežama.
23
Smyth, Elizabeth Catherine, Sanna Hulkki Wilson, Matthew Guy Nankivell, David Gonzalez de Castro, Andrew Wotherspoon, Alicia Frances Clare Okines, Ruth E. Langley, Sally Patricia Stenning, and David Cunningham. "Prognostic and predictive effect of microsatellite instability (MSI) in MAGIC." Journal of Clinical Oncology 33, no. 3_suppl (January 20, 2015): 62. http://dx.doi.org/10.1200/jco.2015.33.3_suppl.62.
Full textAbstract:
62 Background: MSI is prognostic for survival in diverse cancers and predicts resistance to fluoropyrimidine chemotherapy in colon cancer. We examined the interaction between MSI and patient (pt) characteristics and survival for pts randomised to surgery alone or perioperative ECF chemotherapy in the MRC MAGIC trial. Methods: Tumor and normal control DNA was extracted from resection FFPE. MSI status was determined using the Promega MSI Analysis System (NR-21, BAT-26, BAT-25, NR-24, MONO-27). Tumors were classified as MSS when all markers were stable, MSI-L when only one marker was unstable and MSI-H with minimum of instability in two markers. MSI status was correlated with pt characteristics and survival. Results: MSI results were available for 303 pts (66% resected pts). Twenty (6.6%) and 2 (0.6%) pts were MSI-H and MSI-L. All pts with MSI-H had stomach tumors (vs.GEJ/esophagus). KRAS mutation was more common in MSI-H tumours (30% vs. 4%, p-value <0.001); rates of BRAF, PIK3CA and TP53 mutations were comparable. Median overall survival (OS) from surgery was greater for pts with MSI-H tumours treated with surgery alone (1.69y for MSS vs. not reached for MSI-H) (Table 1), whereas patients with MSI-H tumors treated with chemotherapy plus surgery had inferior outcomes compared to MSS stable pts (median OS 0.8y vs. 1.89y respectively). An interaction test for MSI-H vs. MSS status and treatment arm was positive (p=0.007). Conclusions: In MAGIC, MSI-H status is associated with a positive prognostic effect in pts treated with surgery alone, and a negative predictive effect in pts treated with chemotherapy. As pt selection biomarkers for perioperative chemotherapy are lacking, validation of this finding in an independent dataset is warranted. [Table: see text]
24
Ossege, A. L., and Marie Togashi. "Mapeamento genético e dignidade humana: um panorama da legislação brasileira atual frente à prospecção axiológica." Revista Brasileira de Bioética 14, edsup (April 12, 2019): 157. http://dx.doi.org/10.26512/rbb.v14iedsup.26677.
Full textAbstract:
O objetivo deste trabalho é verificar o status da legislação sobre dados genéticos vigente no Brasil, em comparação com as legislações internacionais de vanguarda que contemplam os dados genéticos humanos: a americana, a alemã e a francesa, tendo como foco a área laboral para dimensionar a realidade normativa do país ante as tendências sociais de reconhecimento das diferenças e a abertura jurídica prospectiva. Método: Trata-se de uma revisão da literatura e pesquisa documental, cujo escopo é o diálogo da Bioética na interface com a Medicina do Trabalho e a Genética, em vista do referente comum: a dignidade humana. Resultados: Há tendência a um consenso legalístico mundial quanto à admissão do mapeamento genético de trabalhadores em áreas como pesquisa e tratamento médico. No Brasil, apenas o Código Civil brasileiro a contempla dentro da perspectiva culturalista que o envolve. Conclusão: Faz-se necessária uma estrutura legal consistente e segura que atenda a demanda moderna e que acompanhe o desenvolvimento biotecnológico e, que sobretudo, promova o acatamento à dignidade humana. As legislações internacionais podem ser um excelente campo de pesquisa e servir de referência para construção das leis brasileiras.
25
Kawazoe, Akihito, Kohei Shitara, Masaaki Noguchi, Shota Fukuoka, Yasutoshi Kuboki, Hideaki Bando, Wataru Okamoto, et al. "Clinicopathological features of microsatellite instability-high (MSI-H) metastatic colorectal cancer (mCRC) in Japanese patients." Journal of Clinical Oncology 34, no. 4_suppl (February 1, 2016): 629. http://dx.doi.org/10.1200/jco.2016.34.4_suppl.629.
Full textAbstract:
629 Background: Recently, anti-programmed death 1 (PD-1) antibody has shown promising efficacy in a phase II trial for patients with microsatellite instability-high (MSI-H) tumors, especially in those with metastatic colorectal cancer (mCRC). Currently, several clinical trials of anti-PD-1 antibody for patients with MSI-H mCRC are ongoing. However, little is known about the frequencies and clinicopathological features of MSI-H mCRC in Japanese patients. Methods: Patients with histologically confirmed adenocarcinoma of mCRC were eligible for this observational study. MSI status was analyzed in tumors using the MSI Analysis System (Promega) composed of 5 mononucleotide markers. KRAS, NRAS, BRAF and PIK3CA mutations were also evaluated using the multiplex kit. Results: A total of 232 patients were enrolled until August 31, 2015, of which MSI-H was detected in 5 patients (2.1%). Among five patients with MSI-H mCRC, median age was 45 years (28-75), four had tumors on right-sided colon, five had moderately differentiated adenocarcinoma, one had poorly differentiated adenocarcinoma, and three had stage IV disease at presentation. Overall survival for patients with MSI-H mCRC from the start of first-line systemic chemotherapy ranged from 8.1 to 62.0 month. All five patients with MSI-H mCRC had RAS wild-type tumors and two had BRAF V600E mutation. One patient with MSI-H mCRC was enrolled in a phase II trial of anti-PD-1 antibody. Conclusions: MSI-H mCRC is rare in Japanese patients. A large scale nationwide cancer genome screening project together with MSI status is required to evaluate more detailed clinicopathological features and facilitate the enrollment of patients with MSI-H mCRC for clinical trials with anti-PD-1 antibody.
26
Franke, Aaron J., Maira Gaffar, Michael Feely, Jason Scott Starr, Hiral D. Parekh, Sanda Tan, Atif Iqbal, Carmen Joseph Allegra, and Thomas J. George. "Results from an institutional universal reflex testing program for MSI/MMR in colorectal cancer." Journal of Clinical Oncology 36, no. 4_suppl (February 1, 2018): 796. http://dx.doi.org/10.1200/jco.2018.36.4_suppl.796.
Full textAbstract:
796 Background: In colorectal cancer (CRC), clinical guidelines and immunotherapy treatment selection requires knowledge of tumor DNA mismatch repair gene deficiencies (dMMR) or accumulation of microsatellite repeats through genomic errors (MSI-H). Tumors harboring dMMR/MSI-H are found in 15- 20% of early stage CRC while the prevalence is ~5% in metastatic disease. Reflex testing of CRC to identify these important subsets has been proposed as a system-solution to improve identification. We present a large, single-institutional database of CRC reflexively profiled for MSI/MMR status at the University of Florida (UF). Methods: Beginning in 2009, stage IV CRC underwent reflex testing for MSI/MMR status. Earlier stage CRC testing began in subsequent years. For all new CRC diagnosed at UF, concurrent testing for dMMR by IHC (MLH1, PMS2, MSH2, MSH6), MSI by PCR (Promega MSI kit) and NGS is performed with appropriate positive and negative controls. IHC protein loss is confirmed by second GI pathologist. MSI is not performed if inadequate adjacent normal tissue. We conducted a retrospective analysis of all CRC samples analyzed between 2009 and 2017. Clinical data was collected from the EMR. This study was approved by the UF IRB. Results: A total of 388 new CRC cases were reflex tested (16% Stage I, 23% Stage II, 35% Stage III, and 25% Stage IV). Median age at diagnosis was 63 yrs (range: 17-98 yrs), 51% male and 79% white. Both MMR and MSI were performed in 244 (63%) tumors with 100% concordance when concurrently tested. dMMR/MSI-H incidence (20% in overall population) decreased with stage: I/II (31%), III (18%) and IV (~9%) and was associated with BRAF V600E mut in 36/76 cases (47%). Importantly, in pts < 50 yrs, 13% of stage IV patients were dMMR/MSI-H. Conclusions: Reflex testing of MMR/MSI status in CRC is feasible with concordant results. Routine testing results in identification of dMMR/MSI-H at or higher than published rates. Notable findings include the high prevalence in those with sporadic CRC and/or younger than 50 yrs. Continued impact analysis of this approach is warranted to maximize IO therapy offering.
27
Dentz, René Armand. "Por uma teopoética do perdão a partir de Paul Ricoeur." TEOLITERARIA - Revista de Literaturas e Teologias 9, no. 18 (September 3, 2019): 355–86. http://dx.doi.org/10.23925/2236-9937.2019v9n18p355-386.
Full textAbstract:
O nome de Deus adquire significado por meio de relações diferenciais nas quais sempre aparecem Deus/humano, Deus/mundo, Deus/deuses, infinito/finito, eterno/temporal, etc. Seja qual for o poder nominativo desse nome, e o status real de seu referente - o nome “infinito” é finito, o conceito “inconcebível” é concebível, e o nome “inefável” é bastante exprimível em palavras. Assim, é permitida a abertura à dimensão da promessa. A promessa, ela sim, inscreve-se na linguagem, à qual estamos sempre respondendo. Não se trata de um ato particular de fala, mas a promessa da própria linguagem. Não existe aqui salvação que salve ou prometa salvação. Mas o fato de que não exista necessariamente determinado conteúdo na promessa feita ao outro, e na linguagem do outro não torna menos indisputável a abertura da fala a algo que se pareça com messianismo, soteriologia ou escatologia. Trata-se de abertura estrutural, de messianismo, sem o qual o messianismo em si não seria possível. A linguagem dada a nós agita-se com diversos dons, e se inquieta com palavras de força donativa, como “dom”. A indeterminação é justamente o que permite a promessa, que, por sua vez, possibilita o perdão. A inquietação descreve os gemidos da história e da linguagem para produzir o evento da vinda do totalmente outro, do futuro imprevisível. A história e a linguagem movimentam-se no ambiente da promessa, do espaço aberto entre o passado e o futuro.
28
Campagna, Desirée, and Daniela Angelina Jelinčić. "Indikatori interkulturizma u lokalnim kulturnim politikama." Hrvatska i komparativna javna uprava 18, no. 1 (March 28, 2018): 47–71. http://dx.doi.org/10.31297/hkju.18.1.4.
Full textAbstract:
Iako se o utjecaju multikulturalnosti i interkulturizmu sve više raspravlja, razlike između tih dvaju pojmova te njihov utjecaj na gradske kulturne politike ne istražuju se dovoljno. Vijeće Europe nastoji promicati Intercultural Cities Index (Interkulturni indeks gradova), no taj instrument sagledava samo neke aspekte kulturnih politika iz interkulturne perspektive. S obzirom na to, u radu se predlažu sveobuhvatniji indikatori koji bi mogli procijeniti razinu interkulturizma u lokalnim kulturnim politikama. Indikatori slijede analitičku shemu kojom se nastoji utvrditi kako su dva moguća pristupa kulturnoj raznolikosti (multikulturalnost i interkulturizam) ugrađena u tri dimenzije lokalnih kulturnih politika (dimenzije diskursa, upravljanja i kulturnih sadržaja). Indikatori se odnose na eksplicitniju dimenziju kulturnih politika koja se bavi kulturnim nasljeđem i oblicima umjetničkog izražaja, no mogu se također primijeniti na implicitnu definiciju kulture kao načina života. Predlaže se dvanaest indikatora pomoću kojih su autori usporedili razine interkulturizma u tri hrvatska grada (Rijeci, Osijeku i Puli) tijekom razdoblja njihova statusa grada kandidata za Europsku prijestolnicu kulture. Ovaj europski program nudi uvjete za usporedbu promjena gradskih kulturnih politika pod istim uvjetima i pritiscima. Kao prvo, pregled tih triju gradova naglašava snažnu međusobnu povezanost različitih dimenzija lokalnih kulturnih politika. Čak i kada je nedvojbeno prisutan u dimenziji diskursa, interkulturizam se ne može u potpunosti postići bez upravljačke dimenzije koja potiče stvaranje zajedničkih kulturnih sadržaja. Kao drugo, podaci govore o povratku gastronomije kao sektora koji eksperimentira s ultikulturnim i interkulturnim pristupima. Ključna uloga hrane u promicanju kulturnih susreta ističe potrebu za razmatranjem širega spektra načina života, tradicija i običaja prilikom proučavanja multikulturalnosti i interkulturizma u lokalnim kulturnim politikama.
29
Mandic, Marko. "UTICAJ POLITIKE USLOVLJAVANJA EVROPSKE UNIJE NA SPOLJNU POLITIKU REPUBLIKE SRBIJE." Politička revija 66, no. 4/2020 (January 5, 2021): 363–82. http://dx.doi.org/10.22182/pr.6642020.16.
Full textAbstract:
Politika uslovljavanja Evropske unije predstavlja mehanizam koji EU praktikuje kao deo sopstvene politike proširenja kako bi uticala na unutardržavne političke i ekonomske reforme zemlje kandidata za članstvo. Pomenuta politika doživela je veliki uspeh u procesu proširenja EU na zemlje Centralne i Istočne Evrope. Međutim, ovakav uspeh nije ponovljen u procesu pristupanja Srbije. Srbija odbija da harmonizuje svoju spoljnu politiku sa Zajedničkom spoljnom i bezbednosnom politikom EU (ZSBP EU) u okviru pregovaračkog poglavlja 31, najviše zbog nespremnosti uvođenja sankcija Ruskoj Federaciji. Podsticaji koje članstvo u EU u vidu demokratizacije, vladavine prava, jedinstvenog tržišta nudi državi kandidatu nisu garant usaglašavanja ako su na pregovaračkom stolu teme vezane za nacionalne interese države koji nisu u skladu sa spoljnopolitičkim stavovima EU. Centralno pitanje ove istraživačke analize je zašto se u slučaju Republike Srbije politika uslovljavanja ne pokazuje kao uspešan mehanizam usaglašavanja sa ZSBP EU. Prvi deo posvećen je konceptu politike uslovljavanja EU i kriterijuma koji su preduslov njene uspešnosti. Drugi deo rada posvećen je analizi usklađenosti spoljne politike Republike Srbije sa ZSBP EU. Treći deo rada bavi se ograničenjem politike uslovljavanja EU i njene moći da utiče na preoblikovanje spoljne politike Srbije. Bez obzira na podsticaje koje EU nudi kroz svoju politiku uslovljavanja, troškovi promene spoljne politike Srbije bili bi veliki za obe strane, kako za vladajuće strukture u Srbiji koji temelje spoljnopolitički kurs na evrointegracijama i bliskim odnosima sa Rusijom, tako i za EU koja usled zamora proširenja veći akcenat stavlja na fleksibilnije uslovljavanje u cilju očuvanja bezbednosti i status quo na Balkanu.
30
Carvalho, J. O., V. A. Michalczechen-Lacerda, F. C. Rodrigues, R. Sartori, M. M. Franco, and M. A. N. Dode. "278 METHYLATION STATUS IN THE INTRAGENIC DIFFERENTIALLY METHYLATED REGION OF THE IGF2 LOCUS IN UNSORTED AND SEX-SORTED SPERM." Reproduction, Fertility and Development 23, no. 1 (2011): 237. http://dx.doi.org/10.1071/rdv23n1ab278.
Full textAbstract:
Methylation is the main coordinator of epigenetic inheritance between generations, influencing the regulation of gene expression. Therefore, changes of its pattern in any cell can cause important alterations affecting its function. The methylation pattern of imprinted genes has been reported to be altered by numerous environmental factors such as nutrition, diseases, and drugs. Sexing by flow cytometry exposes the sperm cells to various procedures that can affect sperm quality and could induce methylation changes, resulting in lower fertilization when compared with conventional sperm. Although many studies have related changes in methylation pattern to in vitro culture of oocytes and embryos, no reports have evaluated these changes in sperm caused by the sexing process. The IGF2 imprinted gene, which is predominantly expressed by the paternal allele with the maternal one silenced, is expected to have its intragenic differentially methylated region (DMR) highly methylated in the sperm. Therefore, it became an excellent candidate region to be used for evaluating changes in methylation status of the sperm. The objective of this study was to evaluate the influence of sexing by flow cytometry on the methylation pattern of the DMR located in exon 10 of the IGF2 gene. Frozen–thawed unsorted and sex-sorted sperm samples from 4 Nellore bulls were used (5–10 years old). Each ejaculate was separated into 3 fractions: nonsexed (NS), sexed for X-sperm (SX), and sexed for Y-sperm (SY). Then, 1 straw of each treatment/bull was thawed, placed in 40:70 Percoll gradient (GE Bioscience®, Uppsala, Sweden) and centrifuged at 700 × g for 45 min to separate the somatic cells from the sperm. Sperm pellets were then used for DNA extraction. After that, DNA was treated with sodium bisulfite using the EZ DNA methylation kit (Zymo Research®, Orange, CA, USA). Bisulfite-treated DNA was amplified in 2-round PCR strategy (nested-PCR) and the amplicons were purified using GenClean III kit (MP Biomedical®, Solon, OH, USA). The purified amplicons were cloned into the pGEM-T easy vector system (Promega®, Madison, WI, USA) and transformed into Escherichia coli cells (XL-1 Blue). The resulting individual clones were sequenced, using a dideoxy fluorescence terminator system (ABI 3130xl, Applied Biosystems, Foster City, CA, USA). The experiment was performed in 3 replicates per bull. Sequences were analysed using the BiQ Analyzer software with a GenBank sequence (X53553) as a reference. At least 60 clones were sequenced per group. The methylation status of the 28 CpG sites was compared among the groups using analysis of variance and Tukey test in the Prophet software. No differences in DNA methylation were found between NS (97.5 ± 1.7%), SX (95.8 ± 1.8%), and SY (97.3 ± 0.2%) (P = 0.36). Moreover, 100% of analysed sequences in all groups were hypermethylated. Therefore, we can conclude that the process of sexing by flow cytometry does not change the methylation pattern in the intragenic DMR located in exon 10 of the IGF2 gene. Financial support: FAPESP and Embrapa.
31
Parma, Matteo, Elisa Doni, Federica Colnaghi, Arianna Colombo, Elena Elli, Pietro Pioltelli, Elisabetta Terruzzi, and Enrico Maria Pogliani. "Chimerism Status Analysis after Hematopoietic Stem CellTransplantation on Granulocytes and Mononucleated Cells of Peripheral Blood." Blood 112, no. 11 (November 16, 2008): 4604. http://dx.doi.org/10.1182/blood.v112.11.4604.4604.
Full textAbstract:
Abstract Background Chimerism Status (CS) analysis is largely useful for evaluation of donor (D) cells engraftment after allogeneic haematopoietic stem cell transplantation (HSCT). The largest performed method is based on the evaluation of the DNA sequences “Short Tandem Repeats” (STR) and Variable Number Tandem Repeats” (VNTR). Their high polymorphism permits to differentiate donor (D) or recipient (R) origin of the analysed cells. PCR amplification and DNA sequencing permit to quantify D and R amount. Patients and methods CS analysis after HSCT can be conducted on different cell populations in marrow or peripheral blood: whole blood cells, mononucleated cells, or single separated populations. A complete CS (CCS) usually correlates with a favourable outcome of HSCT, conversely a mixed CS may predicts a relapse of disease or a rejection of HSCT, particularly in the condition defined as increasing-mixed CS (IMCS), in which the amount of R origin cells increases along the time. In our study we employed a simple and easily reproducible method to analyse CS on peripheral blood cells at +30, +60, +90 and +120 days after HSCT. CS analysis was conducted separately on granulocytes and mononucleated cells, obtained after double gradient centrifugation. A semi-quantitative method, based on multiplex PCR amplification of 16 STR markers using a commercial kit (PowerPlex® 16 System–Promega), was applied to define the difference between D and R. We assumed the CS in granulocyte population as expression of myeloid engraftment. We also correlated the CS in mononucleated population, with the different leukocyte subpopulations (lymphocytes, monocytes) in peripheral blood count and with the reconstitution of lymphocyte subpopulations (B, T cells and NK cells) at the same time point of analysis, in order to compare the reciprocal course. We evaluated 13 HSCT on 12 patients (pts) (1 pts was submitted to double HSCT for a rejection occurred after the first HSCT), 7 from sibling related donor (SRD) and 6 from matched unrelated donor (MUD). All the pts were affected by neoplastic disease in first (9) or next (3) remission: acute lymphoblastic (5) or myeloblastic leukaemia (4), mielodisplasia (1), chronic myeloid leukaemia (1), multiple myeloma (1). All the conditioning regimens were mieloablative, all the MUD-HSCT and one of the SRD-HSCT received Anti-Thymocyte-Globulin (ATG), 7/8 SRD-HSCT did not received ATG nor other T-depletion procedures. Results: Evaluation on time + 30, +60, +90 and +120 days after HSCT was performed on 13 HSCT, except in two pts who stopped before (+30 and +60) due to their death for relapse or toxicity. A stable CCS in granulocytes was observed in 10/13 HSCT since the first control. 3/13 HSCT presented IMCS followed by relapse (2 pts) or rejection. In mononucleated cells population 10/13 HSCT presented CCS at the first control, 2/10 evolved to CCS in the successive times. The 3 pts who relapsed or rejected presented a IMCS in mononucleated cells too. Considering the different leucocyte populations we observed that the mononucleate cells amount was constituted for only 50% from lymphocytes, the remaining 50% was represented from monocytes. Therefore the analysis of lymphocyte subpopulations showed that T-lymphocytes were virtually absent (either T4 or T8) till to +120 in pts who received ATG, so in these ones CS analysis in mononucleated cells was performed only in monocytes, B lymphocytes and NK cells. Instead pts who did not received ATG presented complete and more rapid reconstitution of all the lymphocyte subpopulations (median at +60 day: T4 200, T8 120, B 60, NK160 × 10^9/l). Conclusions: our method is simple and rapid in analysing CS. Granulocytes CS reflect the engraftment of myeloid lineage. Instead CS in mononucleated cells must be considered in the context of the type of transplantation (MUD or SRD, T-depletion or T repletion) and correlated with the single subpopulations of peripheral blood.
32
Shroff, R. T., M. M. Javle, X. Dong, V. S. Kumar, S. Krishnan, R. A. Wolff, J. L. Abbruzzese, and D. Li. "The prognostic value of polymorphisms in the insulin-like growth factor receptor (IGFR) pathway in patients with locally advanced pancreatic cancer (LAPC)." Journal of Clinical Oncology 27, no. 15_suppl (May 20, 2009): 4500. http://dx.doi.org/10.1200/jco.2009.27.15_suppl.4500.
Full textAbstract:
4500 Background: The IGFR pathway is activated in pancreatic cancer and may result in aggressive disease course. The study of single nucleotide polymorphisms (SNPs) involved in this pathway may provide prognostic information and predict response to IGFR directed agents. We investigated IGFR pathway SNPs in patients with LAPC. Methods: We evaluated 39 SNPs from 7 candidate genes in the IGFR pathway (IGF1R, IGF2R, IGF1, IGF2, IRS1, IRS2, IGFBP3) in 105 LAPC patients. DNA extraction from whole blood was performed using the Qiagen Flexigene DNA and Promega Maxwell 16 kits. Genotyping was performed using the Sequenom method. Overall survival was measured from date of diagnosis to date of death or last follow-up. Kaplan-Meier plot, log-rank test, and Cox regression were used to compare survival of patients according to genotype corrected for previously identified prognostic factors, including induction chemotherapy, CA 19–9, albumin, LDH, hemoglobin and Karnofsky performance status (KPS). Results: Median survival time (MST) was 15 months (95% CI 13.3–16.7). Induction chemotherapy, LDH, CA 19–9 level, hemoglobin, and KPS were not significantly associated with survival. Serum albumin and three SNPs of the IGF pathway (IGF1R IVS20–3431A>G, IRS1 G971R, and IGF2 *4352A>G) were significantly associated with prognosis ( Table ). Two of the three genotypes remained as significant predictors for survival in Cox regression analysis when adjusted for clinical factors. A significant combined genotype effect was observed wherein patients with all three deleterious alleles had significantly worse survival than those with only two or one (10 vs. 16.3 vs. 21.3 months, p< 0.0001). Conclusions: These data suggest that SNPs in the IGFR pathway genes may have prognostic value for LAPC patients. This information may identify population subgroups that could benefit from IGFR-targeted agents. [Table: see text] No significant financial relationships to disclose.
33
Kendić, Sulejman, Avdo Šakušić, and Nijaz Skender. "STRES - BOLEST DANAŠNJICE, UTJECAJ NA ZDRAVLJE." Zbornik radova Islamskog pedagoškog fakulteta u Bihaću 1, no. 1 (December 8, 2007): 217–33. http://dx.doi.org/10.52535/27441695.2007.1.217-233.
Full textAbstract:
U psihološkom smislu, «stres je stanje koje nastaje kad su ljudi suočeni s događajima koje ocjenjuju prijetećima za svoju dobrobit ilikoji od njih traže ulaganje s posebnim naporom kako bi udovoljili zahtjevima koji se pred njih postavljaju». Postoji vrlo čvrsta povezanost stresa i bolesti, osobito bolesti srca i krvnih žila. Ukoliko postoji velika količina stresa, a hormoni stresa se ne iskorištavaju u nekoj fizičkoj ili drugoj produktivnoj aktivnosti, dolazi do povećanja rada srca, pojave visoki krvnog pritiska, pada imuniteta, pokretanja brojnih patofizioloških mehanizama nastanka bolesti. Stres dijeluje supresivno na urvnoteženost, adaptaciju organizma, a potpomaže slom i dekompenzaciju fizioloških funkcija. Svi ljudi kvalitativno jednako reagiraju na stres, ali kvantitativno postoje znatne razlike. Isti uzrok stresa (stresor) neće jednako pogoditi svakog pojedinca, jer je pri tome bitna lična procjena i shvatanja ugroženosti. Potvrđena je i znanstveno valorizirana povezanost stresa s imunološkim sistemom i njegovim slabljenjem. Osim fizičkih problema, stres je povezan i s mentalnim problemima, poput poremećaja tjeskobe i depresije. Psihološki stres je poseban odnos između osobe i okoline, koji osoba procjenjuje kao vrlo zahtjevan ili kao odnos koji prelazi njezine mogućnosti i ugrožava njezinu dobrobit. Izvori stresa su najčešći iz društvenih okolnosti i to: brze promjene u našem društvu, promjene političkog sistema, poratna situacija-stanja, socijalne tenzije, promjene u zakonodavstvu vezaneuz radno mjesto, razne reforme, promjene društvenih vrijednosti, slaba plaća, nizak socijalni status što je u neskladu s proklamiranom važnošću djelatnosti, omalovažavajući stav okoline prema poslu koji radite, nasilje. Stres zasigurno dovodi do tjelesnnih i zdravstvenih promjena, a to su: psihosomatske bolesti (čir na želucu, visok tlak, alergije, migrene), probavne smetnje, poremećaji tjelesne težine (debljanje ili mršavljenje), manja otpornost prema bolestima, poteškoće sna i spavanja. Antistresno može se djelovati: usostvaljanjem podnošljivih međuljudskih odnosa, psihoanalizom, određivanjem dostižnih ciljeva, organizacijom poslova i obveza.
34
Lepsius, Oliver. "Protección de los derechos fundamentales en la pandemia del coronavirus." Teoría y Realidad Constitucional, no. 47 (April 29, 2021): 71. http://dx.doi.org/10.5944/trc.47.2021.30707.
Full textAbstract:
¿Ha cambiado la situación de los derechos fundamentales en tiempos de la pandemia provocada por el coronavirus? Lo cierto es que la protección de los derechos fundamentales no es una vía de un solo sentido que comienza con el individuo, sino una promesa de la Ley Fundamental que debe ser implementada por todos los poderes estatales. Todo el ordenamiento jurídico en sí debe ser libre, no solo el estatus legal del individuo. En los últimos meses, incluso la jurisprudencia de los tribunales inferiores y del Tribunal Constitucional Federal ha demostrado ser bastante débil. En opinión del autor, el mérito de haber provocado un cambio fundamental en la cultura de ejecución de toma de decisiones dentro del poder judicial corresponde al Tribunal Supremo Administrativo (en adelante OVG) de Münster, que en su «decisión Gütersloh» (OVG Münster, decisión de 29.06.2020-13 B 940/20.NE-Lockdown Gütersloh) volvió a la aplicación correcta de la prueba de proporcionalidad.Has the situation of fundamental rights changed in times of the corona pandemic? One thing is certain: the protection of basic rights is not a one-way street that begins with the individual, but a promise of the Basic Law that must be implemented by all state powers. The legal system as such, not just the legal status of the individual, should be a free one. In the last few months, even the case law of the lower courts and the Federal Constitutional Court has proven to be rather pale. The merit of having brought about a fundamental change in the executive decision-making culture within the judiciary belongs in the perception of the author to the Supreme Administrative Court (OVG) Münster, which in its «Gütersloh decision» (OVG Münster, decision of 29.06.2020 - 13 B 940 / 20.NE - Lockdown Gütersloh) showed the way back to a normal proportionality test.
35
Antonarakis, Emmanuel S., Jose Maria M. Piulats Rodriguez, Marine Gross-Goupil, Jeffrey C. Goh, Ulka N. Vaishampayan, Ronald De Wit, Tuomo Alanko, et al. "Biomarker analysis from the KEYNOTE-199 trial of pembrolizumab in patients (pts) with docetaxel-refractory metastatic castration-resistant prostate cancer (mCRPC)." Journal of Clinical Oncology 38, no. 15_suppl (May 20, 2020): 5526. http://dx.doi.org/10.1200/jco.2020.38.15_suppl.5526.
Full textAbstract:
5526 Background: In the phase II KEYNOTE-199 study (NCT02787005), pembrolizumab monotherapy demonstrated antitumor activity in pts with docetaxel-refractory mCRPC (n = 258). Here we evaluated the association between prespecified molecular biomarkers and clinical outcomes. Methods: Cohorts 1 (C1) and 2 (C2) enrolled pts with RECIST-measurable PD-L1–positive (combined positive score [CPS] ≥1 using immunohistochemistry) and PD-L1–negative (CPS <1) disease, respectively. C3 enrolled pts with nonmeasurable, bone-predominant disease, irrespective of PD-L1 status. Biomarkers evaluated in this analysis were tumor mutational burden ([TMB; mutations/exome] n = 155), PD-L1 CPS (n = 255), tumor microenvironment–based 18-gene RNA expression profile ([GEP] n = 196), and microsatellite instability ([MSI] as determined by Promega PCR analysis; n = 147). Outcomes evaluated for C1 and C2 (n = 200) were ORR, disease control rate (DCR), and radiographic PFS (rPFS) per blinded, independent central review per PCWG-modified RECIST v1.1. Outcomes evaluated for C1-C3 (n = 258) were prostate-specific antigen (PSA) response, time to PSA progression, and OS. Significance of continuous biomarkers (CPS; TMB; GEP) was prespecified at 0.05 for one-sided P values from logistic (ORR; DCR; PSA response) and Cox proportional hazard regression (rPFS; OS; PSA progression) adjusted for Eastern Cooperative Oncology Group performance status. Binary biomarkers (MSI) were analyzed using Fisher’s exact test (ORR; DCR; PSA response). Clinical data cutoff date: Jun 24, 2019. Results: Median TMB was 53.0 (interquartile range [IQR], 40.5 to 78.0), median CPS was 1 (IQR, 0 to 5), and median GEP was –0.64 (IQR, –0.88 to –0.46); 6 pts (2.3%) had MSI-high tumors. In C1-C3, TMB was associated with PSA response (one-sided nominal P = 0.0016) and time to PSA progression (one-sided nominal P = 0.00092). In C1-C3, PD-L1 CPS was associated with PSA response (one-sided nominal P = 0.046) and time to PSA progression (one-sided nominal P = 0.021). In C1-C3, GEP was not significantly associated with response. In C1-C3, MSI was associated with PSA response (one-sided nominal P = 0.019). Conclusions: In this biomarker analysis from KEYNOTE-199 C1-C3, TMB and PD-L1 CPS were associated with better PSA response; however, small pt numbers limit definitive conclusions on ORR, DCR, and OS. Further evaluation of molecular biomarkers in pts with mCRPC treated with pembrolizumab is warranted. Clinical trial information: NCT02787005 .
36
Takahashi, Naoki, Yutaka Matano, Akihiko Tsujibata, Kozue Murayama, Shoichi Miyazawa, TOMOHIRO MATSUSHIMA, Satoshi Shimizu, et al. "Comparison of immune-related gene expression between primary and metastatic site in advanced gastric cancer patients with peritoneal dissemination." Journal of Clinical Oncology 38, no. 4_suppl (February 1, 2020): 423. http://dx.doi.org/10.1200/jco.2020.38.4_suppl.423.
Full textAbstract:
423 Background: Immune checkpoints inhibitor (ICI) is effective and approved in some solid tumors including advanced gastric cancer (GC). Peritoneal dissemination is known as a poor prognostic factor and was reported to be associated with the resistance to ICI according to the previous reports. The aim of our study is to compare the immune-related gene expression between primary site and peritoneal lesion in advanced GC patients. Methods: Among advanced GC patients, we selected those who underwent surgical resection for both primary and peritoneal lesions simultaneously. Formalin-fixed paraffin-embedded (FFPE) tumor tissues of primary and peritoneal lesions were prepared and RNA was extracted by Maxwell RSC RNA FFPE kit (Promega). Immune-related gene expression was evaluated by using nCounter Max Analysis System (NanoString). We used nCounter PanCancer Immune Profiling Panel Kit which includes 770 immune-related genes. Results: Immune-related gene expression was evaluated by using twenty-four FFPE tumor tissues in twelve GC patients. Scatter plot and hierarchical clustering analyses showed that the pattern of immune-related gene expression was not much different between primary and peritoneal lesions beyond the individual differences. Regarding the T cell function, high expression of Immune-related genes was widely detected in patients with EB virus-positive (n = 2) and HER2-positive (n = 1). Gene expressions such as CD70, FAS, MAF and IL-3 were higher in peritoneal lesion compared with primary lesion (p < 0.05). Whereas, expressions of F2RL1 and IL-11 were lower in peritoneal lesion compared with primary lesion (p < 0.05). Conclusions: Our study indicated that there was not much difference of Immune-related gene expression between primary and peritoneal lesion in advanced GC patients. Positive status of EB virus and HER2 may be associated with high expression of immune-related genes. Further analysis to evaluate immune-related gene expression between primary and metastatic site may contribute the further understanding of cancer immunity in advanced GC.
37
Ulianova, Elena P., Vasilij V. Tokmakov, Iuliana S. Shatova, Aleksandr B. Sagakyants, Anna S. Goncharova, Ekaterina V. Zaikina, Elena N. Chernikova, et al. "Evaluation of prognostic significance of microRNA in tumors of luminal, primary operable breast cancer without Her2 neu overexpression in postmenopausal women." Journal of Clinical Oncology 38, no. 15_suppl (May 20, 2020): e12558-e12558. http://dx.doi.org/10.1200/jco.2020.38.15_suppl.e12558.
Full textAbstract:
e12558 Background: Breast cancer (BC) is associated with miRNAs, the qualitative and quantitative analysis of which shows significant differences between molecular subtypes, as well as between miRNAs circulating in the bloodstream and detected in tumor tissue. Since there is no clear characteristics for markers of luminal B primary operable BC without Her 2 neu overexpression, our purpose was an evaluation of prognostic significance of miRNA-21, 221, 20a, 200a, 196a in such postmenopausal patients in order to create a diagnostic panel. Methods: The study included 120 samples of healthy tissues of patients with luminal A BC (group 1, mean age 76.16±3.14 years) and tumor tissues of patients with luminal B BC (group 2, mean age 69.5±3.2 years); invasive ductal carcinoma in all patients. The expression status was analyzed using 5 mioRNA markers associated, according to the literature, with various molecular subtypes of BC (miRNA-21, 221, 20a, 200a, 196a). The Thermo Scientific PureLink RNA Mini Kit was used for the total RNA isolation. The reverse transcription reaction was performed using the ImProm-II Reverse Transcriptase kit (Promega). The Livak/2 method was used to assess the relative expression of miRNAs. Results: The average relative expression was: miRNA-21 in group 1 – 0.2±0.01, group 2 – 1.5±0.05; miRNA- 221 in group 1 – 0.19±0.02, group 2 – 0.43±0.04; miRNA -20a in group 1 – 0.12±0.01, group 2 – 0.13±0.01, miRNA -200a in group 1 – 0.22±0.02, group 2 – 0.96±0.06; miRNA- 196a in group 1 – 1.06±0.02, group 2 – 1.56±0.04. The differences between the groups were statistically significant (p = 0.0001); the difference was statistically insignificant only for miRNA -20a (p = 0.159955). Conclusions: Results of the study allow considering the overexpression of miR-21, 221, 200a and 196a and the relatively low expression of miR-20a as diagnostic markers for a diagnostic panel for luminal B BC without overexpression of HER2/neu.
38
Simarro Fagundes, N., V. Alice Michalczechen Lacerda, E. Siqueira Caixeta, G. Marinheiro Machado, E. de Oliveira Melo, R. Rumpf, M. Alves Nunes Dode, and M. Machaim Franco. "251 METHYLATION PATTERN OF A DIFFERENTIALLY METHYLATED REGION LOCATED IN THE LAST EXON OF THE Igf2 GENE IN OOCYTES FROM NELLORE COWS." Reproduction, Fertility and Development 22, no. 1 (2010): 283. http://dx.doi.org/10.1071/rdv22n1ab251.
Full textAbstract:
Assisted reproductive technologies are essential in modern cattle breeding. The relevance of in vitro embryo production (IVP) has been increasing around the world, particularly in Brazil. Acknowledging the positive effect of IVP regarding the utilization of female reproductive potential, its productivity can be improved. Methylation pattern of DNA in oocytes is a key factor for the improvement of IVP efficiency because it is related to oocyte competence. The objective of this study was to evaluate the methylation pattern of a differentially methylated region (DMR) located in the exon 10 of the imprint gene IGF2 in immature and in vitro-maturated oocytes from different follicle sizes. Small follicles (1-3 mm in diameter) and large follicles (≥8.1 mm in diameter) were dissected from slaughterhouse ovaries, and the COC were removed. Immature and in vitro-maturated oocytes, presenting the first polar body, were denuded and frozen in PBS at -80°C until DNA extraction. After extraction, DNA was treated with sodium bisulfite using EZ DNA Methylation™ Kit (Zymo Research, Orange, CA, USA). Bisulfite-treated DNA was amplified in a 2-round PCR strategy (nested-PCR), the amplicons purified with Geneclean® III Kit (MP Biomedicals, Irvine, CA, USA), cloned using pGEM®-T Easy Vector System (Promega, Madison, WI, USA), and sequenced. The number of follicles and in vitro maturation data were analyzed with chi-square test, and the methylation status of 27 CpG islands was analyzed with Student (normal distribution) or Mann-Whitney tests in the Prophet software (BBN Systems and Technologies, Cambridge, MA, USA). Cumulus-oocyte complexes from ≥8.1-mm follicles presented greater percentages of viable oocytes (40.5%) and nuclear maturation (60.6%) (P ≤ 0.05). Immature oocytes from 1- to 3-mm follicles were less methylated (33.33%) than those from ≥8.1-mm follicles (83.69%). After in vitro maturation, oocytes from ≥8.1-mm follicles were less methylated (18.51%) than the immature oocytes from ≥8.1-mm follicles (83.69%) and 1- to 3-mm maturated oocytes (49.62%). The less methylated pattern in the oocytes from ≥8.1-mm maturated follicles can be associated with greater competency of those oocytes, because this was the expected pattern. The lower level of methylation in the immature oocytes group (1- to 3-mm follicles) can be associated with the incomplete establishment of imprinting reprogramming pattern. Finally, we concluded that the methylation pattern of DMR, in the last exon of the IGF2 gene, can be used as a molecular marker for epigenetic reprogramming status in oocytes, helping the development of new IVP protocols. Supported by Embrapa Genetic Resources and Biotechnology and CAPES.
39
Vasseur, Damien, Cécile Jovelet, Nathalie Cozic, Julien Mazieres, Fabrice Barlesi, Jaafar Bennouna, Radj Gervais, et al. "Minimal residual disease (MRD) in patients with resected stage I NSCLC: Results of the prospective adjuvant IFCT-0703 trial." Journal of Clinical Oncology 39, no. 15_suppl (May 20, 2021): 8526. http://dx.doi.org/10.1200/jco.2021.39.15_suppl.8526.
Full textAbstract:
8526 Background: MRD aims to detect circulating biomarkers of micrometastatic disease and ultimately predict recurrences. The IFCT-0703 randomized phase II trial failed to show a benefit of 6 months adjuvant pazopanib (P) vs. placebo after resection of stage I NSCLC (7th TNM edition). The outcome of pts based on their MRD status has been evaluated. Methods: Blood samples were collected in EDTA tubes (Becton Dickinson Company) after surgery (T0), after 3 months (T3) of P or placebo and at the end of treatment (T6). Plasmas were obtained after double centrifugation of total blood. Total nucleic acid was extracted using the Maxwell RSC LV plasma kit (Promega) according to the manufacturer’s protocol. Samples were quantified using the QuBit dsDNA HS Assay kit on a QuBit 3.0 flurometer (Thermo Fisher Scientific). Molecular analysis was performed by next generation sequencing using the Oncomine Lung cfDNA Assay (ThermoFisher Scientific). Two MRD definitions were tested : 1) high level of DNA in the blood or 2) any mutation detected by the standard bioinformatic pipeline was considered present, whatever the allelic fraction. Results: 143 pts were randomized in 29 centers between March 2009 and August 2012, 71 and 72 in the placebo and P arms respectively. Among the 119 pts with evaluable T0 samples, 27 pts recurred and 14 died. Median DNA concentration ([DNA]) was 6.6 ng/ml and an increase of [DNA] of 10 ng/ml was found prognostic of poor DFS and OS, HR=1.4, 95%CI [1.14-1.72], p=0.0016 and HR=1.62, 95%CI [1.15-2.30], p=0.0057 respectively. In 81 pts with available T0-T6 samples, [DNA] variation had no different impact on DFS and OS, in the P arm and the placebo arm. ctDNA mutations (ctDNA+) were detected in 31/119 pts. ctDNA+ were more frequent in samples with high DNA quantity (p=0.0002). Genes mutated at T0 were TP53 in 16, NRAS in 6, MAP2K1 in 2, KRAS in 1, EGFR in 5, BRAF in 1, ALK in 2. 29 pts had 1 mutation, 2 had 2 mutations. DFS and OS were similar between pts with or without ctDNA+ : HR= 1.038 (95%CI 0.438-2.456, p=0.93) and 1.193 (95% CI 0.367-3.882, p=0.77) respectively. Among 27 pts with ctDNA+ at T0 and available sample at T6, 23 had no more mutations at T6. Two pts had a ctDNA+ only at T6 (not at T3), one of them had a recurrence at 7 months. Conclusions: Post-operative ctDNA mutations are found in 26.0% of the pts but their positivity had no impact on DFS or OS. In contrast, DFS and OS were poorer in pts with increased plasma DNA concentration. ctDNA mutations status do not recapitulate the complexity of MRD characterization. NGS will be performed on matched tissues in order to refine MRD definition. Clinical trial information: NCT00775307.
40
Barzi, Afsaneh, Mihaela Campan, Jonas Petterson, Lan Du, Tiffany Long, Louis Dubeau, Heinz-Josef Lenz, and Pamela Ward. "Assessment of microsatellite instability (MSI) in cell free DNA (cfDNA) of colorectal cancers (CRC) patients (pts)." Journal of Clinical Oncology 36, no. 4_suppl (February 1, 2018): 672. http://dx.doi.org/10.1200/jco.2018.36.4_suppl.672.
Full textAbstract:
672 Background: While there is accumulating evidence that tumor specific mutations can be accurately detected in cfDNA, the determination of MSI status in cfDNA has not been investigated, perhaps because the complexity of detecting tumor instability embedded within stutter amplification patterns generated from normal cfDNA was considered too challenging to allow the detection of altered microsatellite profiles. We explored the possibility of detecting MSI in cfDNA. Methods: Circulating cfDNA was isolated from Streck tubes from 13 patients with CRC; 6 pts had serial collections. Isolation and testing of cfDNA for MSI was performed using kits purchased from Promega. Microsatellite alterations were scored by comparing the amplicon stutter profiles of 5 mononucleotide markers generated from cfDNA to those present in genomic DNA extracted from buffy coats from the same pts. MSI was diagnosed based on a shift in the size of a predominant stutter peak together with a commensurate shift in the stutter profile resulting in the appearance of at least one new stutter in tumor-derived DNA compared to matched germline DNA. MSI was scored high when 30% or more of the markers showed MSI or low if present in less than 30% of the biomarkers examined. Results: No MSI was detected in 3 pts with microsatellite stable tumors and 2 Lynch syndrome pts with no evidence of disease. Five of six pts with MSI high tumors (sporadic and Lynch syndrome) had a detectable MSI high biomarker profile in cfDNA at baseline. Four pts with positive MSI in cfDNA started immunotherapy and had a decline in the relative amounts of tumor-derived microsatellite alleles in their subsequent specimens. The pt with MSI high in the primary tumor and no MSI in cfDNA had rapid progression on immunotherapy. Two other pts with MSI high who responded deeply to check point inhibitors had no evidence of MSI in cfDNA at the time of their specimen collection. In each case with MSI in cfDNA, the electrophoretic profile matched that in the respective primary tumor. Conclusions: Detection of MSI in cfDNA is feasible and may be reflective of burden of the disease. A prospective study to establish the validity of cfDNA in the diagnosis and follow up care in pts with metastatic CRC is planned.
41
Scherrer Júnior, Gerson, Maria Isabel Barbosa Rodrigues, Kleyton Góes Passos, Odete Teresinha Portela, Angélica Castilho Alonso, and Angélica Gonçalves Silva Belasco. "Fatores associados à dependência de idosos residentes em instituições públicas." Revista Remecs - Revista Multidisciplinar de Estudos Científicos em Saúde 4, no. 6 (June 26, 2019): 3. http://dx.doi.org/10.24281/rremecs2526-2874.2019.4.6.3-11.
Full textAbstract:
Comparar o grau de dependência e correlacionar a dependência com as variáveis sociodemográficas, cognição e estado nutricional de idosos residentes em instituições de longa permanência pública de São Paulo. Estudo transversal e analítico, com 154 idosos, moradores de instituições, inseridas no seio da comunidade, que acolhem idosos em situação de vulnerabilidade social. Os dados foram coletados em agosto de 2016. As informações sociais, culturais, epidemiológicas e de estilo de vida foram transcritas dos prontuários, as atividades básicas da vida diária avaliadas pelo Índice de Katz, o estado mental pelo mini exame do estado mental, condição nutricional pelo mini avaliação nutricional. O aumento da idade, maior quantidade de doenças e tempo de moradia, má condição cognitiva e nutricional aumenta o grau de dependência dos idosos institucionalizados. Os achados recomendam às instituições a oferta de um ambiente que promova a cognição, o físico e estilo de vida saudável.Descritores: Instituição de Longa Permanência para Idosos, Envelhecimento, Atividades Cotidianas. Factors associated with the dependence of elderly residents in public institutionsAbstract: To compare the degree of dependence and correlate dependence with the sociodemographic variables, cognition and nutritional status of elderly people living in long - term public institutions in. A cross-sectional and analytical study, with 154 elderly people, residents of institutions within the community, who welcome elderly people in situations of social vulnerability. The data were collected in August 2016. The social, cultural, epidemiological and lifestyle information was transcribed from the charts, the basic activities of daily living evaluated by the Katz Index, the mental state by the mini-mental state examination, nutritional condition by the mini nutritional assessment. Increased age, higher amount of illnesses and length of stay, poor cognitive and nutritional status increase the degree of dependence of the institutionalized elderly. The findings recommend that institutions offer an environment that promotes cognition, physical and healthy lifestyle.Descriptors: Homes for the Aged, Aging, Activities of Daily Living.Factores asociados a la dependencia de ancianos residentes en instituiciones públicasResumen: Comparar o determinar la dependencia y la correlación con la dependencia como variables sociodemográficas, conocimiento y estado nutricional de los residentes en instituciones de larga duración pública de São Paulo. Estudio transversal y analítico, con 154 ancianos, habitantes de instituciones, insertas en el seno de la comunidad, que acogen a ancianos en situación de vulnerabilidad social. Los datos fueron recolectados en agosto de 2016. Las informaciones sociales, culturales, epidemiológicas y de estilo de vida fueron transcritas de los archivos, actividades básicas de la vida cotidiana evaluadas por el índice de Katz, el estado mental por el mini examen del estado mental. El aumento de la edad, mayor cantidad de enfermedades y tiempo de morada, mala condición cognitiva y nutricional aumenta el grado de dependencia de los ancianos institucionalizados. Los hallazgos recomiendan a las instituciones la oferta de un ambiente que promueve la cognición, el físico y el estilo de vida saludable.Descriptores: Hogares para Ancianos, Envejecimiento, Actividades Cotidianas.
42
Perecin, F., S. C. Méo, W. Yamazaki, C. R. Ferreira, F. H. Biase, G. K. F. Merighe, F. V. Meirelles, and J. M. Garcia. "187 IMPRINTED GENE EXPRESSION IN IN VIVO-AND IN VITRO-PRODUCED BOVINE FETUSES AND PLACENTAS." Reproduction, Fertility and Development 20, no. 1 (2008): 173. http://dx.doi.org/10.1071/rdv20n1ab187.
Full textAbstract:
Some gestational alterations associated with bovine somatic cell nuclear transfer (SCNT) are presumably consequences of abnormal imprinted gene expression. This work aimed to evaluate the expression patterns of imprinted genes IGF2 and IGF2R in bovine fetuses and chorioallantoic membranes derived from in vivo- and in vitro-produced embryos. Fetuses were produced by AI (in vivo group, n = 3), IVF (n = 3), parthenogenesis (n = 3), or SCNT (n = 2). Cows with positive pregnancy diagnosis after ultrasonographic examination were slaughtered between Days 33 and 36 of gestation. The reproductive tract was transported on ice to the laboratory, where fetuses and chorioallantoic fragments were collected and stored in liquid nitrogen. Total RNA extraction was performed using TRIzol, according to manufacturer's instructions, and the reverse transcription reaction was carried out with 1 µg of total RNA, 6.75 µm oligo pd(T)12–18, and 50 U of reverse transcriptase (Improm-II, Promega, Madison, WI, USA). The relative quantification of IGF2 and IGF2R transcripts was done using real-time PCR with SYBR Green dye. The average efficiency of PCR amplifications was estimated for each gene using a linear regression on the logarithm of fluorescence per cycle (Ramakers et al. 2003 Neurosci. Lett. 339, 62–66), and the expression ratios were calculated according to the method described previously by Livak and Schmittgen (2001 Methods 25, 402–408). To verify statistical differences, a pair-wise fixed reallocation randomization test (Pfaffl et al. 2002 Nucl. Acids Res. 30, e36) was used. All expression ratios were normalized by glyceraldehyde 3-phosphate dehydrogenase expression and calibrated by the in vivo group (expression assumed as 1.00 for all genes and tissues). The analysis of relative differences on transcript levels of imprinted genes in fetuses revealed IGF2 down-regulation (P < 0.05) in the SCNT (0.19) and parthenogenetic (0.02) groups when compared to the in vivo group and IVF fetuses (2.02). In chorioallantois, IGF2 was down-regulated (P < 0.001) in parthenotes (0.001) when compared to the in vivo, IVF (3.13), and SCNT (0.98) groups. IGF2R was down-regulated (P < 0.001) in SCNT chorioallantois (0.25) when compared to the in vivo group. Low expression of IGF2 in parthenogenetic fetuses and chorioallantois confirms its imprinted status in bovine. Alterations in the relative frequency of IGF2 and IGF2R transcripts were observed in bovine SCNT-derived fetuses and chorioallantoic membranes, respectively, supporting the hypothesis that abnormalities in the expression of imprinted genes are causes for the low efficiency of SCNT procedures in this species. Such alterations suggest modifications in DNA methylation patterns at IGF2 and IGF2R imprinting centers.
43
Michalczechen-Lacerda, V. A., F. C. Rodrigues, R. V. de Sousa, R. Rumpf, and M. M. Franco. "53 IMPACTS OF USING PROCAINE AS A DNA-DEMETHYLATING AGENT IN IN VITRO CULTURE OF BOVINE CELLS." Reproduction, Fertility and Development 23, no. 1 (2011): 132. http://dx.doi.org/10.1071/rdv23n1ab53.
Full textAbstract:
Euchromatin and heterochromatin organisation define the specificity of each cell type. This structure is controlled by epigenetic modifications and the DNA methylation is one of the best known for inducing transcriptional repression. Recently, procaine was uncovered as a DNA-demethylating agent, but there are few reports about its dynamic epigenetic action on somatic cells. Mono-allelic expression of imprinted genes is controlled by DNA methylation and inherited to somatic tissues of a sex-specific manner. The aim was to investigate the effects of using procaine, a DNA-demethylating agent, in in vitro culture of bovine (Bos taurus indicus) fibroblast for 72 h (passage 4). We have evaluated cell viability, chromosome integrity, and DNA methylation patterns. To evaluate cell viability, we have used trypan blue 0.4%. To evaluate chromosome integrity, we have used conventional cytogenetic analysis. To investigate DNA methylation patterns, we have analysed 2 differentially methylated regions (DMR) located into the exon 10 of IGF2 and exon 1 of XIST imprinted genes, using the bisulfite sequencing method (EZ DNA methylation kit, Zymo Research, Orange, CA, USA). After bisulfite treatment and nested-PCR, the amplicons were separated in agarose gel electrophoresis, purified with GenClean III kit (MP Biomedicals, Irvine, CA, USA), cloned in a pGEM-T easy vector system (Promega, Madison, WI), and sequenced. The DNA sequences were analysed using the BiQ Analyzer v. 2.0 (2008) software. The cell viability data were analysed using ANOVA and Tukey or Kruskal-Wallis and Mann-Whitney tests, and the methylation status were analysed using Student’s t-test or Mann-Whitney tests in the Prophet software (BBN Systems and Technologies). Cell culture using 0.1 mM or 0.5 mM of procaine were viable and the number of cells with intact membrane was higher than the control and 2.0 mM of procaine groups (P ≤ 0.05). The total number of cells was lower in the group with 2.0 mM of procaine (P ≤ 0.01). Cytogenetic analysis showed no differences among the groups, with no chromosome abnormalities detected. The methylation pattern was not different for both DMR evaluated among the groups. We have observed that there was a beneficial effect to the cells that have received supplementation with 0.1 mM or 0.5 mM of procaine, because there was an increase in the number of viable cells without chromosomal abnormalities. We cannot ignore that a global DNA demethylation may have occurred, which was not detected in the specific analysed regions. The results obtained here may contribute to improving the efficiency of animal cloning, transgenic animal production, and the knowledge about stem cells. Supported by Embrapa Genetic Resources and Biotechnology and CAPES.
44
Denis, Marc G., Audrey Vallee, Christine Sagan, Guillaume Herbreteau, Sandrine Theoleyre, Jaya Rajamani, Michael Lee, and Ellen Ordinario. "Detection of ALK and ROS1 fusion transcripts in FFPE samples of non-small cell lung cancer patients using a novel RT-PCR based assay and targeted RNA sequencing." Journal of Clinical Oncology 38, no. 15_suppl (May 20, 2020): e21695-e21695. http://dx.doi.org/10.1200/jco.2020.38.15_suppl.e21695.
Full textAbstract:
e21695 Background: Detection of genomic rearrangements like ALK and ROS1 fusions are mandatory in non-small cell lung cancer (NSCLC) as those alterations can be targeted by an increasing number of drugs. Fluorescence in-situ hybridization (FISH) has been so far the gold standard but multiplexed technologies would allow analysis of many potential targets simultaneously. We have evaluated a novel RT-PCR based assay and a targeted RNA sequencing solution for the detection of ALK and ROS1 rearrangement in NSCLC patients. Methods: 41 patients with late stage NSCLC were included in the study. ALK and ROS1 status were screened on FFPE tissue sections using immunohistochemistry and confirmed with FISH. Total nucleic acids were extracted from tissue sections using the Maxwell RSC RNA FFPE kit (Promega). Detection of rearrangements was performed by RT-PCR using the prototype ALK/RET/ROS1 Fusion Panel assay (Roche), which detects 7 ALK fusion variants, 13 ROS1 variants and 6 RET variants. For RNA sequencing, the libraries were prepared with the QIAseq Targeted RNAscan Lung Cancer Panel (Qiagen). Sequencing was performed on a MiSeq instrument (Illumina). Results: We tested 16 ALK FISH+ tumors, 4 ROS1 FISH+ tumors, and 21 samples that were either IHC- or IHC+/FISH-. The RT-PCR assay detected 15 of the 16 ALK FISH+ tumors, and 3 of the 4 ROS1 FISH+ tumors. In the 21 IHC- or IHC+/FISH- samples, 1 additional ROS1 fusion and 1 RET fusion were detected. The remaining 19 assays samples were negative. 2 fusions were missed by RT-PCR because the assay design does not cover these rearrangements (KLC1-ALK and CTNND1-ROS1). With targeted RNA sequencing, we were able to detect 14 of the 16 ALK FISH+ tumors and all 4 of the ROS1 FISH+ tumors. We also identified 1 additional ROS1 fusion and 1 RET fusion in the 21 IHC- or IHC+/FISH- samples. 17 patients were negative and 2 tests were not contributive. Overall, using FISH as a reference, the sensitivity of both assays was 90% (18/20). Conclusions: Using limited amount of biological material, both RT-PCR and targeted RNA sequencing were able in one assay to detect the overwhelming majority of ALK and ROS1 fusions identified with FISH. These approaches also allowed us to detect gene rearrangements that were missed by the initial IHC/FISH screening. The RT-PCR assay requires less RNA (as little as 3 ng) and has a much faster turn-around time. The RNA sequencing approach allowed us to identify the fusion partners, but it requires much more RNA and is rather difficult to implement in routine practice.
45
Alonso Alonso, Ruth, Rufino Mondejar García, Jose Pedro Vaqué Díez, Nuria García Díaz, Cristina Pérez Menéndez, Francis Burrows, Catherine Scholz, Antonio Gualberto, and Miguel A. Piris. "Identification of Tipifarnib Sensitivity Biomarkers in T-Cell Tumor Cell Lines." Blood 132, Supplement 1 (November 29, 2018): 2851. http://dx.doi.org/10.1182/blood-2018-99-118291.
Full textAbstract:
Abstract Background T-cell leukemia and lymphoma (TCL) is a very aggressive and heterogeneous group of hematological malignancies with poor prognosis and inadequate response to current therapies. The standard first-line treatments have resulted in unsatisfactory patient outcomes and unfortunately, targeted treatments are still at a preliminary phase. The RAS/MAPK pathway is crucial for TCR signaling of T-cells and it is deregulated in TCL. We aimed to determine the therapeutic value of farnesyl transferase inhibitor (FTIs) tipifarnib in TCL cell lines. Effects on cell viability, apoptosis, cell cycle and gene expression were evaluated. Tipifarnib blocks the localization of some farnesylated proteins, including some RAS family members, to cellular membranes, thereby inhibiting their activation. Tipifarnib is a potent and specific FTI with prominent anti-proliferative effects. Methods We tested tipifarnib in 18 T-cell leukemia and 4 T-cell lymphoma cell lines for in vitro sensitivity and for biomarker discovery, both genomic and immunohistochemical. We selected cell lines with available genomic data from COSMIC, CCLE or generated by our group. The MAPK, NFAT, NFKB and JAK/STAT pathways were tested by immunohistochemistry analysis under basal conditions. The range of drug concentrations to perform IC50 analysis was established between 0-10,000 nM (ten points), using CellTiter-Glo® Luminescent Cell Viability Assay kit (Promega, Madison, WI, USA) following manufacturer´s instructions at 0h, 48h and 96h. All experiments were done in sextuplet and all numerical data were expressed as the average of the values ± the standard error of the mean. IC50 analyses were performed with GraphPad Prism v5. Clinically-relevant drug sensitivity was defined as IC50 <100nM at 96h (defined as sensitive). RNAseq was performed in 4 sensitive cell lines with tipifarnib and DMSO (vehicle) in three biological replicates at 72h. Differential gene expression (DE) analysis was carried out under these two conditions. Pathway analysis was done using Gene Set Enrichment Analysis (GSEA). Induction of apoptosis and cell viability were tested by flow cytometry in sensitive cell lines. Results 59.1% (n=13) of cell lines were sensitive to tipifarnib at concentrations which are readily achievable in the clinic. RAS, RAS-guanine nucleotide exchange factors (GEFs) and RAS-GTPase activating proteins (GAPs) genes were mutated in 45.5%, 50% and 27.3%, of cell lines respectively, but their mutational status was not associated with drug sensitivity. We found TP53 and DNMT3 mutated in 80% and 33.3% in sensitive cell lines respectively, and in 63.6% and 0% in resistant cell lines. The mutational state of NOTCH1 was associated with tipifarnib sensitivity; 66.7% in sensitive cell lines versus 9.1% in resistant cell lines (p=0.005). ERK activation (MAPK pathway) was significantly associated with drug sensitivity (p=0.046). Conversely, RelB (NFκB pathway) was associated with drug resistance (p=0.007). GSEA analysis showed that tipifarnib downregulated cell cycle, protein localization to membranes, metabolism, and ribosome and mitochondrial activity. The effect of tipifarnib on cell cycle progression and apoptosis was validated by flow cytometry assays. Tipifarnib decreased cellular viability, increased apoptosis and blocked cell cycle progression in G1 phase. GSEA of molecular pathways regulated by tipifarnib identified downregulation of MYC targets and mTOR and MAPK pathways. MYC is a target of NOTCH1, mutated in sensitive cases and associated with tipifarnib sensitivity. Conclusions In vitro response to tipifarnib in T-cell lymphoma and leukemia cell lines can be predicted using several biomarkers. The mutational NOTCH1 status, together with p-ERK (MAPK activation) were associated with increased sensitivity while RelB expression was associated with resistance. Disclosures Gualberto: Kura Oncology: Employment, Equity Ownership. Piris:Takeda: Honoraria; Janssen: Honoraria; Gilead: Honoraria; Kura: Honoraria.
46
Barrera, A. D., E. V. García, M. Hamdi, M. J. Sánchez-Calabuig, D. Rizos, and A. Gutiérrez-Adán. "80 EFFECT OF BOVINE OVIDUCTAL FLUID ON DNA METHYLATION OF BOVINE BLASTOCYSTS PRODUCED IN VITRO." Reproduction, Fertility and Development 29, no. 1 (2017): 147. http://dx.doi.org/10.1071/rdv29n1ab80.
Full textAbstract:
During the transit through the oviduct, the early embryo undergoes an epigenetic reprogramming of its genome, which induces changes in DNA methylation pattern. Given that epigenetic modifications are susceptible to environmental influence, the oviducal milieu may affects DNA methylation marks in the developing embryo. The aim of this study was to evaluate whether bovine oviducal fluid (OF) exerts an effect on methylation status of genomic regions at different time points of embryo development. In vitro-produced zygotes were cultured in SOF + 3 mg mL−1 BSA (control, C) or in SOF + 1.25% OF at 3 different time points: until 98 h post-insemination (hpi) (OF1–16: 1–16 cell), 52 hpi (OF1–8: 1–8 cell), or from 52 until 98 hpi (OF8–16: 8–16 cell). The OF used was acquired from Embryocloud (Murcia, Spain) from cow oviducts at the early luteal phase (Day 1–4). After, embryo culture took place in control medium up to Day 8. For all the groups, the speed of development was considered, and normal developing embryos that reached ≥6 cells at 52 hpi and ≥16 cells at 98 hpi were selected and separately cultured from slow developing embryos. Cleavage (52 hpi) and blastocyst yield (Day 7–8) were analysed by ANOVA (8 replicates). Expanding blastocysts (Day 7–8) from the normal developing groups were collected for bisulfite sequencing analysis. The DNA bisulfite conversion was performed with a MethylEdge Bisulfite Conversion System kit (Promega, WI, USA) in groups of 20 blastocysts obtained from 5 replicates. Methylation status was analysed on regions localised in 4 developmental important genes (MTERF2, ABCA7, OLFM1, and GMDS) and within 2 LINE L1 elements located on chromosomes 9 (L9) and 29 (L29). Methylation percentages (10 sequenced clones/group) were compared using statistical z-test. No significant differences were found on cleavage rate (C: 89.7 ± 1.0, OF1–16: 84.9 ± 1.7; OF1–8: 85.4 ± 1.9; OF8–16: 89.1 ± 1.9%) and blastocyst yield between normal developing embryos (C: 36.8 ± 5.3; OF1–16: 34.7 ± 3.7; OF1–8: 41.0 ± 3.8; OF8–16: 43.9 ± 5.1%). Blastocysts derived from all OF groups showed the CpG region of MTERF2 hypomethylated compared with C group (20.0, 26.2, and 32.9% v. 56.2%, respectively; P < 0.001). The CpG sequence of ABCA7 exhibited significant hypomethylation in embryos from OF1–16 group compared with OF1–8, OF8–16, and C groups (31.1 v. 56.8, 57.9, and 65.8%, respectively; P < 0.001). Although the methylation of the CpG region within OLFM1 did not differ between OF1–16 and C groups (24.1 v. 19.4%, respectively), embryos from OF1–8 group showed a highly methylated region (47.1%) compared with OF1–16 and C groups (P < 0.001). The CpG sequence on L9 showed a high methylation level in blastocysts derived from OF1–16 group compared with OF8–16 and C groups (36.4 v. 14.5 and 20.0%, respectively; P < 0.05). There were no differences in methylation marks between groups examined for CpG regions of GMDS and L29. These results indicated that embryos exhibit a temporal sensitivity to OF at early embryonic stages, which is reflected by DNA methylation changes of specific genes at blastocyst stage. This is the first report describing that OF could modify specific epigenetic marks of the bovine embryonic genome.
47
Pérez, Cristina, Julia Gonzalez-Rincon, Carmen Almaraz, Soraya Curiel, Nuria Garcia, Helena Pisonero, Sagrario Gomez, et al. "A Role of JAK/STAT Pathway in Cutaneous T-Cell Lymphomas: Exploring Its Effects for Targeted Therapy." Blood 124, no. 21 (December 6, 2014): 4498. http://dx.doi.org/10.1182/blood.v124.21.4498.4498.
Full textAbstract:
Abstract INTRODUCTION Development of targeted therapy in Cutaneous T-Cell Lymphoma (CTCL) patients still requires actionable mutated genes and deregulated pathways to be identified. We have recently published the mutational status of a number of human CTCL lesions, and found JAK/STAT signaling pathway to frequently harbor somatic mutations (Vaqué et al 2014). In this line of evidence, activating mutations in JAK kinases have been reported in human hematological malignancies (Kameda T1 2010) and may serve as indicators for targeted therapy. Therefore, we decided to analyze the mutational status of JAK/STAT pathway in a greater cohort of human CTCL patient samples and cell lines, by using massive parallel sequencing techniques, to shed light on its possible role in the pathogenesis and to uncover its potential as a new therapeutic target for this disease. To this end, we have studied the biological and molecular effects that targeted inhibition of the JAK/STAT signaling pathway, exerts in human CTCL cells, and explore its potential use as a targeted therapy. MATERIAL AND METHODS NGS: We searched for mutations in the catalytic domain of JAK and STAT genes in 39 CTCL patients using targeted NGS techniques: HaloPlex (Agilent) and sequencing in IonTorrent (LifeTech) and Illumina sequencers. Proliferation, apoptosis, cell cycle and DNA synthesis assays: CTCL cell lines were: My-La (MF), HH (MF) and HUT-78 (SS) (ECACC,Salisbury, UK). Effects in proliferation was assessed using CellTiter-Glo® Luminescent Cell Viability Assay (Promega, USA). To analyze the effects on cell survival, we evaluated early and delayed cell death provoked by FACS using FlowCellect Annexin Red Kit (EMD Millipore Corporation, USA). The distribution of cells among different phases of the cell cycle was evaluated by FACS using propidium iodide (PI, Sigma-Aldrich, USA) and Click-iT® EdU Alexa Fluor® 488 Flow Cytometry Assay Kit (Technologies-Thermo Fisher Scientific, USA) according to manufacturer’s instruction. Gene expression (GEP) studies: mRNA was extracted with Trizol and mRNA Array-based expression analysis was perfomed using a Whole Human Genome Agilent 4 × 44K v1 Oligonucleotide Microarray. RESULTS: Using our approach we found JAK/STAT pathway mutated in up to 24% of the CTCL samples. 3 mutations were found in JAK1, 5 in JAK3 and 1 in STAT5A genes. Interestingly, most of these mutations affected the tyrosine kinase domain of JAK kinases, a hotspot for activating mutations described in multiple types of human cancer. Thereafter, we decided to explore the biological effects of targeted JAK/STAT inhibition, using specific JAK inhibitors (JAKi) in preclinical CTCL models. To this end, CTCL cell lines were incubated at different time points with increasing concentrations of JAKi. These treatments inhibited JAK/STAT signaling in CTCL cells, as assessed by western-blot using P-STAT-1,-3 and -5 antibodies. Biologically, JAKi provoked a marked inhibition of cell proliferation by a mechanism impinging the control of DNA replication. This was accompanied with a modest increase in cell death. To better understand the molecular mechanisms controlled by aberrant JAK/STAT signaling, we also performed an array-based mRNA expression analysis (GEP) in HUT78 cell line (JAK1Y654F) treated at different time points (0, 0.5 and 3h) with JAKi (at a concentration of IC50). Our results showed a number of genes regulated by JAK/STAT signaling which activity has been described as T-cell activation, differentiation and proliferation (i.e. PAG1,TNF or EGR1).These genes may serve as potential indicators for therapy using JAK/STAT inhibitors or as surrogate markers for drug response. CONCLUSIONS Our results show frequent mutations affecting JAK/STAT downstream signaling in samples from patients with CTCL. Targeted inhibition of this pathway reveals an aberrant CTCL growing controlled by this signaling. Thus, these results provide evidence for the use of JAK/STAT inhibitors on the basis of a targeted massive sequencing analysis in specific cases of CTCL. Disclosures No relevant conflicts of interest to declare.
48
Gill, Harinder, Cheuk Him Man, Karen Lam, Raymond Liang, Edmond Ma, Yok Lam Kwong, and Anskar Y. H. Leung. "Acquired Fms-Like Tyrosine Kinase 3 Internal Tandem Duplication (FLT3 ITD) During Leukaemic Transformation From Underlying Myelodysplasia: Successful Rescue with Sorafenib." Blood 120, no. 21 (November 16, 2012): 4927. http://dx.doi.org/10.1182/blood.v120.21.4927.4927.
Full textAbstract:
Abstract Abstract 4927 Leukemic transformation from antecedent myelodysplastic syndromes (MDS) has a dismal prognosis and is often resistant to conventional intensive chemotherapy. The leukemic process may involve stepwise acquisition of genetic mutations and emergence of subclones with proliferative and survival advantages. Identifying molecular targets during leukemic progression may provide new treatment options for this group of patients. In this study, the mutational status of a gene encoding for FMS-like tyrosine kinase 3 (FLT3) at MDS and leukemic transformation as well as the clinical responseto FLT3 inhibitor sorafenib were evaluated. Eighty patients with MDS according to the World Health Organization 2008 classification were prospectively evaluated. Patients who subsequently developed secondary acute myeloid leukemia (sAML) were identified. Bone marrow (BM) samples at the diagnosis of MDS and sAML were collected and genomic DNA (gDNA) was extracted. Internal tandem duplication (ITD) of FLT3 at the juxtamembrane domain and tyrosin kinase 1 domain spanning exons 14 and 15were identified by polymerase chain reaction (PCR). To increase the sensitivity so that small MDS clones with FLT3-ITD could be detectable, an ITD-specific PCR was designed in which the PCR products from FLT3-ITD+ sAML samples were cloned into pGEMT-Easy vector (Promega, USA), amplified, extracted and sequenced by Sanger sequencing. PCR primers targeting specific ITD sequence were designed. Patients with FLT3-ITD+ sAML were treated with Sorafenib at 400mg twice daily. Bone marrow aspiration and biopsy was performed around 3 weeks following treatment. Between August 2009 to August 2012, 12 patients with sAML from antecedent MDS were identified of whom 5 were FLT3 ITD+ sAML. ITD-specific PCR performed on the respective MDS samples were negative in four and weakly positive in one patients. Four patients (all with refractory anaemia with excess blasts – 2 with normal karyotype at initial diagnosis) received sorafenib 400mg twice daily of whom two achieved complete remission (CR) and one CR with insufficient hematological recovery (CRi) three weeks after treatment. There was no overt dysplasia at morphological remission. Two of these patients respectively underwent subsequent allogeneic hematopoietic stem cell transplantation (HSCT) at 5 and 6 months after treatment (sorafenib was stopped before transplantation) and remained in remission for 10 and 12 months after initial CRi/CR. The third patient who was not eligible for HSCT received maintenance azacitidine concurrence with continuous sorafenib treatment and was 2 months in remission after CR. The fourth patient has just received sorafenib, awaiting bone marrow reassessment. Despite the small number of patients recruited, our observation showed that acquisition of FLT3 ITD is associated with leukemic progression in MDS and most importantly the use of FLT3 inhibitor effectively induced remission that can be further consolidated with allogeneic HSCT or demethylating agents. The effective clearance of myeloblasts with sorafenib also provided an opportunity whereby MDS clonal heterogeneity and evolution during leukemic transformation can be biologically evaluated. Disclosures: No relevant conflicts of interest to declare.
49
Xue, Kai, Juan Gu, Cory Mavis, Francisco Hernandez, Myron S. Czuczman, and Ye Guo. "Vorinostat, a Histone Deacetylase (HDAC) Inhibitor, Is Active in Chemotherapy Resistant B-Cell Non-Hodgkin Lymphoma and Enhances the Anti-Tumor Activity of Chemotherapy Agents." Blood 124, no. 21 (December 6, 2014): 5486. http://dx.doi.org/10.1182/blood.v124.21.5486.5486.
Full textAbstract:
Abstract Pre-clinical models of chemotherapy resistance and clinical observations derived from the CORAL study suggest that primary refractory/relapsed B-Cell Non-Hodgkin Lymphoma (NHL) is a more aggressive and resistant lymphoma to current available treatments. Pre-clinically, we demonstrated that chemotherapy resistance is associated with a deregulation on the apoptotic machinery rendering lymphoma cells resistant to apoptotic stimuli. There is a dire need to develop agents capable to execute alternative pathways of cell death in an attempt to overcome chemotherapy resistance. Post-transcriptional histone modification plays an important role in regulating gene transcription, and is altered by histone acetyltransferases (HATs) and histone deacetylases (HDACs). HDACs regulate several key cellular functions, including: cell proliferation, cell cycle, apoptosis, angiogenesis, migration, antigen presentation, and/or immune regulation. Given their influence in multiple regulatory pathways, HDAC inhibition is an attractive strategy to evaluate its anti-tumor activity in cancer cells. To this end we studied the anti-tumor activity and mechanisms of action of suberoylanilide hydroxamic acid (SAHA, Vorinostat) in drug-resistant pre-clinical models. A panel of rituximab and chemotherapy resistant cell lines and primary tumor cells isolated from B-cell NHL patients (N=42: Denovo=27, Relapse/Rafractory=15) were exposed to escalating doses of vorinostat. Changes in cell viability were determined by Cell Titer Glo ® (Promega, Fitchburg, WI) luminescent assays. Changes in cell cycle were determined by flow cytometric analysis. Subsequently, protein lysates were isolated from vorinostat exposed cells and changes in members of apoptosis, cell cycle family proteins and the acetylation status of histone H3 were evaluated by Western blotting. In addition, cell lines were pre-exposed to vorinostat for 48 hrs and subsequently plated in 384 well plates and exposed to several chemotherapy agents (cisplatin, etoposide, or gemcitabine) changes in cell viability were determined by Cell Titer Glo and synergistic activity was evaluated using the Calcusm® software. Vorinostat induced dose-and time- dependent cell death in cell lines and in primary tumor cells. In addition, in vitro exposure of lymphoma cells to vorinostat resulted in an increase in p21, decrease cyclin dependent kinase2 (CDK2) and cyclin E; acetylation of histone H3 and was associated with a G1 cell cycle arrest. As expected, PARP cleavage was not observed in cell lines exposed to vorisnostat at IC50 doses, and Caspase inhibitor experiment further showed that blockage of caspase pathway cannot rescue cells and primary patient tumor samples from vorinostat induced cell death, suggesting that alternative cell death pathways were executed (i.e. irreversible cell cycle arrest). Moreover vorinostat was found to enhances the anti-tumor activity of chemotherapy agents. Our data suggests that vorinostat is active in drug-resistant pre-clinical models and that in cell lines with known defective apoptotic machinery, it can active alternative pathway of cell death. Given the multiple pathways affected by HDAC inhibition, vorisnostat can potentially be used to overcome acquired resistant to chemotherapy in aggressive B-cell lymphoma. (Research, in part, supported by a NIH grant R01 CA136907-01A1 awarded to Roswell Park Cancer Institute and The Eugene and Connie Corasanti Lymphoma Research Fund) Disclosures No relevant conflicts of interest to declare.
50
Kanuya, E., L. A. Clayton, R. A. Naidu, and A. V. Karasev. "First Report of Grapevine fleck virus in Idaho Grapevines." Plant Disease 96, no. 11 (November 2012): 1705. http://dx.doi.org/10.1094/pdis-06-12-0574-pdn.
Full textAbstract:
Idaho has a growing viticulture industry, with nearly 1,600 acres of wine grapes (Vitis vinifera L.). Production is largely concentrated in two locations, the Snake River valley, which includes Canyon County in the southwest, and the Clearwater River valley, primarily Nez Perce County in the northwest. Grapevine fleck virus (GFkV) belongs to the genus Maculavirus, family Tymoviridae, comprising positive-sense, single-stranded RNA viruses with ca. 7.6-kb genome (3). It is one of five non-mechanically transmitted viruses associated with the fleck disease complex and has been previously documented to occur in the neighboring state of Washington (2). Main sources of wine grape nursery material imported to Idaho reside in Washington or in California, and it is important to monitor virus status of the planting material brought to the state. However, no information was available on the occurrence and prevalence of GFkV in wine grapes in Idaho. During three growing seasons in 2009 through 2011, random grapevine samples were collected in 14 vineyards in Canyon, Elmore, Ada, and Nez Perce counties. A total of 434 samples were tested by one step RT-PCR using GFkV-specific primers, GFkVf: 5′-TGACCAGCCTGCTGTCTCTA-3′ and GFkVr: 5′-TGGACAGGGAGGTGTAGGAG-3′ designed to amplify a fragment of the GFkV capsid protein gene (1). Twenty-four samples tested positive for GFkV by RT-PCR and produced the expected 179-bp DNA fragment. These samples came from five vineyards sampled across all surveyed counties, and represented seven wine grape cultivars, including Pinot Noir, Cabernet Sauvignon, Syrah, Lemberger, Riesling, Chardonnay, Pinot Gris, and one unknown table grape cultivar. Twelve PCR products were cloned into the pGEM-T Easy plasmid vector (Promega), sequenced (numbered ID1 to 12, available upon request), and confirmed to represent fragments of the GFkV CP gene between positions 6,453 and 6,631 in the genome of GFkV isolate MT48 (GenBank Accession No. AJ309022.1). Eight of the Idaho GFkV sequences (ID2, ID3, ID7 to 11, and ID12) matched closely with other GFkV sequences from Washington State, Italy, India, and South America, showing 97 to 99% identity at the nucleotide level in pair-wise comparisons. Four GFkV sequences from Idaho (ID1 and ID4 to 6) showed only modest (90 to 92%) identity in pair-wise comparisons with GFkV sequences available in GenBank. Consequently, in phylogenetic reconstructions eight Idaho GFkV sequences clustered in the same lineage with the six GFkV sequences deposited in GenBank, and four other GFkV sequences were placed outside of this main clade. It is possible that this phylogeny of the Idaho GFkV reflects different sources of the virus-infected planting material brought to the state. In the absence of symptoms expressed in wine grape cultivars infected with GFkV, laboratory methods remain the only tool to detect the virus. To our knowledge, this is the first report of GFkV found in wine grapes in Idaho demonstrating its substantial presence in production areas. References: (1) G. Gambino and I. Gribaudo. Phytopathology 96:1223, 2006. (2) R. A. Naidu et al. Plant Dis. 94:784, 2010. (3) S. Sabanadzovic et al. J. Gen. Virol. 82:2009, 2001.