1. Extracellular vesicle-bound DNA in urine is indicative of kidney allograft injuryIvana Sedej, Maja Štalekar, Magda Tušek-Žnidarič, Katja Goričar, Nika Kojc, Polona Kogovšek, Vita Dolžan, Miha Arnol, Metka Lenassi, 2022, izvirni znanstveni članek Povzetek: Extracellular vesicle-bound DNA (evDNA) is an understudied extracellular vesicle (EV) cargo, particularly in cancer-unrelated research. Although evDNA has been detected in urine, little is known about its characteristics, localization, and biomarker potential for kidney pathologies. To address this, we enriched EVs from urine of well-characterized kidney transplant recipients undergoing allograft biopsy, characterized their evDNA and its association to allograft injury. The SEC-based method enriched pure EVs from urine of kidney transplant recipients, regardless of the allograft injury. Urinary evDNA represented up to 29.2 ± 8% (mean ± SD) of cell-free DNA (cfDNA) and correlated with cfDNA in several characteristics but was less fragmented (P < 0.001). Importantly, using DNase treatment and immunogold labelling TEM, we demonstrated that evDNA was bound to the surface of urinary EVs. Normalised evDNA yield (P = 0.042) and evDNA copy number (P = 0.027) significantly differed between patients with normal histology, rejection injury and non-rejection injury, the later groups having significantly larger uEVs (mean diameter, P = 0.045) and more DNA bound per uEV. ddDNA is detectable in uEV samples of kidney allograft recipients, but its quantity is highly variable. In a proof-of-principle study, several evDNA characteristics correlated with clinical and histological parameters (P = 0.040), supporting that the potential of evDNA as a biomarker for kidney allograft injury should be further investigated. Ključne besede: DNA, donor-derived DNA, extracellular vesicles Objavljeno v DiRROS: 26.02.2025; Ogledov: 303; Prenosov: 199
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2. Incorporation of recombinant proteins into extracellular vesicles by Lactococcus cremorisTina Vida Plavec, Kristina Žagar, Giulia Della Pelle, Špela Zupančič, Robert Vidmar, Aleš Berlec, 2025, izvirni znanstveni članek Povzetek: Extracellular vesicles (EVs) are nanosized lipid bilayer particles released by various cellular organisms that carry an array of bioactive molecules. EVs have diagnostic potential, as they play a role in intercellular interspecies communication, and could be applied in drug delivery. In contrast to mammalian cell-derived EVs, the study of EVs from bacteria, particularly Gram-positive bacteria, received less research attention. This study aimed to investigate the production of EVs by lactic acid bacterium Lactococcus cremoris NZ9000 and to examine the impact of recombinant protein expression on their formation and protein content. Four different recombinant proteins were expressed in L. cremoris NZ9000, in different forms of expression and combinations, and the produced EVs were isolated using the standard ultracentrifugation method. The presence of vesicular structures (50–200 nm) in the samples was confirmed by transmission electron microscopy and by flow cytometry using membrane-specific stain. Mass spectrometry analyses confirmed the presence of recombinant proteins in the EVs fraction, with amounts ranging from 13.17 to 100%, highlighting their significant incorporation into the vesicles, together with intrinsic L. cremoris NZ9000 proteins that were either more abundant in the cytoplasm (ribosomal proteins, metabolic enzymes) or present in the membrane. The presence of the most abundant lactococcal proteins in EVs fraction suggests that protein cargo-loading of EVs in L. cremoris NZ9000 is not regulated. However, our data suggests that L. cremoris NZ9000 genetically engineered to express recombinant proteins can produce EVs containing these proteins in scalable manner. As L. cremoris NZ9000 is considered safe bacterium, EVs from L. cremoris NZ9000 could have several advantages over EVs from other bacteria, implying possible biotechnological applications, e.g. in therapeutic protein delivery. Ključne besede: Lactococcus cremoris, extracellular vesicles, recombinant proteins, delivery vehicle Objavljeno v DiRROS: 16.01.2025; Ogledov: 388; Prenosov: 155
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3. The effect of different surfactants and polyelectrolytes on nano-vesiculation of artificial and cellular membranesUrška Zagorc, Darja Božič, Vesna Arrigler, Žiga Medoš, Matej Hočevar, Anna Romolo, Veronika Kralj-Iglič, Ksenija Kogej, 2024, izvirni znanstveni članek Ključne besede: extracellular vesicles, liposomes, surfactants, polyelectrolytes, static light scattering, dinamic light scattering, microcalorimetry, drug delivery, cell-to-cell communication Objavljeno v DiRROS: 24.12.2024; Ogledov: 417; Prenosov: 193
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4. Assessment of extracellular particles directly in diluted plasma and blood by interferometric light microscopy : a study of 613 human and 163 canine samplesBoštjan Korenjak, Armando Tratenšek, Matevž Arko, Anna Romolo, Matej Hočevar, Matic Kisovec, Maxence Berry, Apolonija Bedina Zavec, David Drobne, Tomaž Vovk, Aleš Iglič, Alenka Nemec Svete, Vladimira Erjavec, Veronika Kralj-Iglič, 2024, izvirni znanstveni članek Ključne besede: extracellular particles, extracellular vesicles, exosomes, membrane vesiculation, platelets, blood products, platelet-rich plasma, liquid biopsy, membrane enclosed, drug delivery Objavljeno v DiRROS: 20.12.2024; Ogledov: 400; Prenosov: 210
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5. Short term effect of plant hybridosomes on growth of phaeodactylum tricornutum cultureAnna Romolo, Matej Hočevar, Aleš Iglič, Tjaša Griessler Bulc, Veronika Kralj-Iglič, 2023, objavljeni znanstveni prispevek na konferenci Ključne besede: Phaeodactylum tricornutum, hybridosomes, liposomes, nanoalgosomes, extracellular vesicles, small cellular particles, microalgae Objavljeno v DiRROS: 21.03.2024; Ogledov: 723; Prenosov: 384
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6. Small cellular particles from European spruce needle homogenateMarko Jeran, Anna Romolo, Vesna Spasovski, Matej Hočevar, Urban Novak, Roman Štukelj, Vid Šuštar, Matic Kisovec, Apolonija Bedina Zavec, Ksenija Kogej, Aleš Iglič, Polonca Trebše, Veronika Kralj-Iglič, 2023, izvirni znanstveni članek Ključne besede: Picea abies, spruce needles, extracellular particles, extracellular vesicles, small cellular particles, interferometric light microscopy, exosomes, cell-to-cell communication, drug delivery Objavljeno v DiRROS: 07.02.2024; Ogledov: 889; Prenosov: 502
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7. Autologous platelet and extracellular vesicle-rich plasma as therapeutic fluid : a reviewKaja Troha, Domen Vozel, Matevž Arko, Apolonija Bedina Zavec, Drago Dolinar, Matej Hočevar, Zala Jan, Matic Kisovec, Boštjan Kocjančič, Ljubiša Pađen, Manca Pajnič, Samo Penič, Anna Romolo, Neža Repar, Vesna Spasovski, Nejc Steiner, Vid Šuštar, Aleš Iglič, Damjana Drobne, Ksenija Kogej, Saba Battelino, Veronika Kralj-Iglič, 2023, pregledni znanstveni članek Ključne besede: platelet-rich plasma, platelets, centrifugation, extracellular vesicles, small cellular particles, wound healing, regeneration, fresh-frozen plasma, intercellular signaling peptides and proteins Objavljeno v DiRROS: 26.01.2024; Ogledov: 1000; Prenosov: 565
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