1. Amphiphilic coumarin-based probes for live-cell STED nanoscopy of plasma membraneHana Kokot, Boštjan Kokot, Anja Pišlar, Hana Esih, Alen Gabrič, Dunja Urbančič, Rojbin El, Iztok Urbančič, Stane Pajk, 2024, izvirni znanstveni članek Povzetek: Plasma membranes are vital biological structures, serving as protective barriers and participating in various cellular processes. In the field of super-resolution optical microscopy, stimulated emission depletion (STED) nanoscopy has emerged as a powerful method for investigating plasma membrane-related phenomena. However, many applications of STED microscopy are critically restricted by the limited availability of suitable fluorescent probes. This paper reports on the development of two amphiphilic membrane probes, SHE-2H and SHE-2N, specially designed for STED nanoscopy. SHE-2N, in particular, demonstrates quick and stable plasma membrane labelling with negligible intracellular redistribution. Both probes exhibit outstanding photostability and resolution improvement in STED nanoscopy, and are also suited for two-photon excitation microscopy. Furthermore, microscopy experiments and cytotoxicity tests revealed no noticeable cytotoxicity of probe SHE-2N at concentration used for fluorescence imaging. Spectral analysis and fluorescence lifetime measurements conducted on probe SHE-2N using giant unilamellar vesicles, revealed that emission spectra and fluorescence lifetimes exhibited minimal sensitivity to lipid composition variations. These novel probes significantly augment the arsenal of tools available for high-resolution plasma membrane research, enabling a more profound exploration of cellular processes and dynamics
Objavljeno v DiRROS: 24.10.2025; Ogledov: 292; Prenosov: 106
 Celotno besedilo (4,33 MB) |
2. Inducirane pluripotentne matične celice pri odkrivanju in vrednotenju zdravilnih učinkovinJaka Rotman Primec, Zala Krajšek, Dunja Urbančič, 2025, pregledni znanstveni članek Povzetek: Inducirane pluripotentne matične celice so v zadnjih letih postale pomembno orodje za odkrivanje zdravilnih učinkovin, saj ponujajo bistvene prednosti pred tradicionalnimi modeli, kot so primarne celice, rakave celične linije in živalski modeli. Pridobimo jih z reprogramiranjem somatskih celic v pluripotentno stanje, kar omogoča njihovo diferenciacijo v različne tipe celic ob ohranjanju dedne informacije posameznika. Pri uporabi induciranih pluripotentnih matičnih celic za odkrivanje zdravilnih učinkovin je na voljo več modelov, vključno z dvodimenzionalnimi kulturami, enostavnejšimi tridimenzionalnimi modeli in naprednejšimi modeli – organi na čipu. Tovrstni modeli omogočajo natančnejšo oceno toksičnosti in učinkovitosti zdravilnih učinkovin, njihovih medsebojnih interakcij ter proučevanje mehanizmov bolezni. Izzivi, s katerimi se soočamo ob uporabi induciranih pluripotentnih matičnih celic, so: njihova nezadostna zrelost, variabilnost med pridobljenimi celičnimi linijami in visoki stroški, kar trenutno omejuje njihovo širšo uporabo v farmaciji. Kljub tem omejitvam imajo inducirane pluripotentne matične celice velik potencial za razvoj personaliziranih oblik zdravljenja in odkrivanje novih terapevtskih pristopov.
Ključne besede: celični modeli in vitro, inducirane pluripotentne matične celice, odkrivanje novih zdravilnih učinkovin, predklinične študije
Objavljeno v DiRROS: 08.07.2025; Ogledov: 514; Prenosov: 182
Celotno besedilo (1,99 MB) |
3. Robust saliva-based RNA extraction-free one-step nucleic acid amplification test for mass SARS-CoV-2 monitoringEva Rajh, Tina Šket, Arne Praznik, Petra Sušjan, Alenka Šmid, Dunja Urbančič, Irena Mlinarič-Raščan, Polona Kogovšek, Tina Demšar, Mojca Milavec, Katarina Prosenc, Žiga Jensterle, Mihaela Zidarn, Viktorija Tomič, Gabriele Turel, Tatjana Lejko-Zupanc, Roman Jerala, Mojca Benčina, 2021, izvirni znanstveni članek Povzetek: Early diagnosis with rapid detection of the virus plays a key role in preventing the spread of infection and in treating patients effectively. In order to address the need for a straightforward detection of SARS-CoV-2 infection and assessment of viral spread, we developed rapid, sensitive, extraction-free one-step reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and reverse transcription loop-mediated isothermal amplification (RT-LAMP) tests for detecting SARS-CoV-2 in saliva. We analyzed over 700 matched pairs of saliva and nasopharyngeal swab (NSB) specimens from asymptomatic and symptomatic individuals. Saliva, as either an oral cavity swab or passive drool, was collected in an RNA stabilization buffer. The stabilized saliva specimens were heat-treated and directly analyzed without RNA extraction. The diagnostic sensitivity of saliva-based RT-qPCR was at least 95% in individuals with subclinical infection and outperformed RT-LAMP, which had at least 70% sensitivity when compared to NSBs analyzed with a clinical RT-qPCR test. The diagnostic sensitivity for passive drool saliva was higher than that of oral cavity swab specimens (95% and 87%, respectively). A rapid, sensitive one-step extraction-free RT-qPCR test for detecting SARS-CoV-2 in passive drool saliva is operationally simple and can be easily implemented using existing testing sites, thus allowing high-throughput, rapid, and repeated testing of large populations. Furthermore, saliva testing is adequate to detect individuals in an asymptomatic screening program and can help improve voluntary screening compliance for those individuals averse to various forms of nasal collections.
Ključne besede: SARS-CoV-2, COVID-19, COVID-19 serological testing, real-time polymerase chain reaction, saliva, oral cavity swab, passive drool, pooling
Objavljeno v DiRROS: 09.11.2021; Ogledov: 2341; Prenosov: 1009
 Povezava na celotno besedilo |
