@article{MTMT:35294236, title = {Size and fluorescence calibrated imaging flow cytometry: From arbitrary to standard units}, url = {https://m2.mtmt.hu/api/publication/35294236}, author = {Woud, W.W. and Pugsley, H.R. and Bettin, B.A. and Varga, Zoltán and van, der Pol E.}, doi = {10.1002/cyto.a.24895}, journal-iso = {CYTOM PART A}, journal = {CYTOMETRY PART A}, unique-id = {35294236}, issn = {1552-4922}, abstract = {Imaging flow cytometry (IFCM) is a technique that can detect, size, and phenotype extracellular vesicles (EVs) at high throughput (thousands/minute) in complex biofluids without prior EV isolation. However, the generated signals are expressed in arbitrary units, which hinders data interpretation and comparison of measurement results between instruments and institutes. While fluorescence calibration can be readily achieved, calibration of side scatter (SSC) signals presents an ongoing challenge for IFCM. Here, we present an approach to relate the SSC signals to particle size for IFCM, and perform a comparability study between three different IFCMs using a plasma EV test sample (PEVTES). SSC signals for different sizes of polystyrene (PS) and hollow organosilica beads (HOBs) were acquired with a 405 nm 120 mW laser without a notch filter before detection. Mie theory was applied to relate scatter signals to particle size. Fluorescence calibration was accomplished with 2 μm phycoerythrin (PE) and allophycocyanin (APC) MESF beads. Size and fluorescence calibration was performed for three IFCMs in two laboratories. CD235a-PE and CD61-APC stained PEVTES were used as EV-containing samples. EV concentrations were compared between instruments within a size range of 100–1000 nm and a fluorescence intensity range of 3–10,000 MESF. 81 nm PS beads could be readily discerned from background based on their SSC signals. Fitting of the obtained PS bead SSC signals with Mie theory resulted in a coefficient of determination >0.99 between theory and data for all three IFCMs. 216 nm HOBs were detected with all instruments, and confirmed the sensitivity to detect EVs by SSC. The lower limit of detection regarding EV-size for this study was determined to be ~100 nm for all instruments. Size and fluorescence calibration of IFCM data increased cross-instrument data comparability with the coefficient of variation decreasing from 33% to 21%. Here we demonstrate – for the first time – scatter calibration of an IFCM using the 405 nm laser. The quality of the scatter-to-diameter relation and scatter sensitivity of the IFCMs are similar to the most sensitive commercially available flow cytometers. This development will support the reliability of EV research with IFCM by providing robust standardization and reproducibility, which are pre-requisites for understanding the biological significance of EVs. © 2024 International Society for Advancement of Cytometry.}, keywords = {STANDARDIZATION; reproducibility; Extracellular vesicles; Imaging flow cytometry; light scatter calibration}, year = {2024}, eissn = {1552-4930}, orcid-numbers = {Varga, Zoltán/0000-0002-5741-2669} } @article{MTMT:35134234, title = {Comparing novel small-angle x-ray scattering approaches for absolute size and number concentration measurements of spherical SiO2 particles to established methods}, url = {https://m2.mtmt.hu/api/publication/35134234}, author = {Schürmann, Robin and Gaál, Anikó and Sikora, Aneta and Ojeda, David and Bartczak, Dorota and Goenaga-Infante, Heidi and Korpelainen, Virpi and Sauvet, Bruno and Deumer, Jerome and Varga, Zoltán and Gollwitzer, Christian}, doi = {10.1088/1361-6528/ad568b}, journal-iso = {NANOTECHNOLOGY}, journal = {NANOTECHNOLOGY}, volume = {35}, unique-id = {35134234}, issn = {0957-4484}, abstract = {Biomedical analytical applications, as well as the industrial production of high-quality nano- and sub-micrometre particles, require accurate methods to quantify the absolute number concentration of particles. In this context, small-angle x-ray scattering (SAXS) is a powerful tool to determine the particle size and concentration traceable to the Syst & egrave;me international d'unit & eacute;s (SI). Therefore, absolute measurements of the scattering cross-section must be performed, which require precise knowledge of all experimental parameters, such as the electron density of solvent and particles, whereas the latter is often unknown. Within the present study, novel SAXS-based approaches to determine the size distribution, density and number concentrations of sub-micron spherical silica particles with narrow size distributions and mean diameters between 160 nm and 430 nm are presented. For the first-time traceable density and number concentration measurements of silica particles are presented and current challenges in SAXS measurements such as beam-smearing, poorly known electron densities and moderately polydisperse samples are addressed. In addition, and for comparison purpose, atomic force microscopy has been used for traceable measurements of the size distribution and single particle inductively coupled plasma mass spectrometry with the dynamic mass flow approach for the accurate quantification of the number concentrations of silica particles. The possibilities and limitations of the current approaches are critically discussed in this study.}, keywords = {NANOPARTICLES; SILICA PARTICLES; small-angle X-ray scattering; Number concentration; Materials Science, Multidisciplinary; Nanoscience & Nanotechnology; Metrology; spICPMS}, year = {2024}, eissn = {1361-6528}, orcid-numbers = {Gaál, Anikó/0000-0003-4064-1825; Varga, Zoltán/0000-0002-5741-2669} } @article{MTMT:34981962, title = {Optimizing lipopeptide bioactivity: The impact of non-ionic surfactant dressing}, url = {https://m2.mtmt.hu/api/publication/34981962}, author = {Ábrahám, Ágnes and Gyulai, Gergő and Mihály, Judith and Horváth, Andrea and Dobay, Orsolya and Varga, Zoltán and Kiss, Éva and Horváti, Kata}, doi = {10.1016/j.jpha.2024.101020}, journal-iso = {J PHARM ANAL}, journal = {Journal of Pharmaceutical Analysis}, unique-id = {34981962}, issn = {2095-1779}, year = {2024}, eissn = {2214-0883}, orcid-numbers = {Ábrahám, Ágnes/0000-0003-2586-8650; Gyulai, Gergő/0000-0002-1352-2014; Horváth, Andrea/0000-0003-4208-3417; Dobay, Orsolya/0000-0001-7094-2288; Varga, Zoltán/0000-0002-5741-2669; Kiss, Éva/0000-0002-4757-4437; Horváti, Kata/0000-0003-4092-6011} } @article{MTMT:34966111, title = {Interfacial Behavior of Biodegradable Poly(lactic- co -glycolic acid)-Pluronic F127 Nanoparticles and Its Impact on Pickering Emulsion Stability}, url = {https://m2.mtmt.hu/api/publication/34966111}, author = {Fülöp, Dániel and Varga, Zoltán and Kiss, Éva and Gyulai, Gergő}, doi = {10.1021/acs.langmuir.4c00147}, journal-iso = {LANGMUIR}, journal = {LANGMUIR}, volume = {40}, unique-id = {34966111}, issn = {0743-7463}, abstract = {Biodegradable nanoparticle-based emulsions exhibit immense potential in various applications, particularly in the pharmaceutical, cosmetic, and food industries. This study delves into the intricate interfacial behavior of Pluronic F127 modified poly(lactic-co-glycolic acid) (PLGA-F127) nanoparticles, a crucial determinant of their ability to stabilize Pickering emulsions. Employing a combination of Langmuir balance, surface tension, and diffusion coefficient measurements, we investigate the interfacial dynamics of PLGA-F127 nanoparticles under varying temperature and ionic strength conditions. Theoretical calculations are employed to elucidate the underlying mechanisms governing these phenomena. Our findings reveal a profound influence of temperature-dependent Pluronic layer behavior and electrostatic and steric interactions on the interfacial dynamics. Nonlinear changes in surface tension are observed, reflecting the interplay of these factors. Particle aggregation is found to be prevalent at elevated temperatures and ionic strengths, compromising the stability and emulsification efficiency of the formed emulsions. This work provides insights into the rational design of stable and efficient biodegradable nanoparticle-based Pickering emulsions, broadening their potential applications in various fields.}, year = {2024}, eissn = {1520-5827}, pages = {12353-12367}, orcid-numbers = {Varga, Zoltán/0000-0002-5741-2669; Kiss, Éva/0000-0002-4757-4437; Gyulai, Gergő/0000-0002-1352-2014} } @article{MTMT:34875002, title = {Extracellular vesicles promote migration despite BRAF inhibitor treatment in malignant melanoma cells.}, url = {https://m2.mtmt.hu/api/publication/34875002}, author = {Németh, Afrodité and Bányai, Gréta Lilla and Dobos, Nikolett K and Kós, Tamás and Gaál, Anikó and Varga, Zoltán and Buzás, Edit Irén and Khamari, Delaram and Dank, Magdolna and Takács, István and Szász, Attila Marcell and Garay, Tamás}, doi = {10.1186/s12964-024-01660-4}, journal-iso = {CELL COMM SIGN}, journal = {CELL COMMUNICATION AND SIGNALING}, volume = {22}, unique-id = {34875002}, issn = {1478-811X}, abstract = {Extracellular vesicles (EVs) constitute a vital component of intercellular communication, exerting significant influence on metastasis formation and drug resistance mechanisms. Malignant melanoma (MM) is one of the deadliest forms of skin cancers, because of its high metastatic potential and often acquired resistance to oncotherapies. The prevalence of BRAF mutations in MM underscores the importance of BRAF-targeted therapies, such as vemurafenib and dabrafenib, alone or in combination with the MEK inhibitor, trametinib. This study aimed to elucidate the involvement of EVs in MM progression and ascertain whether EV-mediated metastasis promotion persists during single agent BRAF (vemurafenib, dabrafenib), or MEK (trametinib) and combined BRAF/MEK (dabrafenib/trametinib) inhibition.Using five pairs of syngeneic melanoma cell lines, we assessed the impact of EVs - isolated from their respective supernatants - on melanoma cell proliferation and migration. Cell viability and spheroid growth assays were employed to evaluate proliferation, while migration was analyzed through mean squared displacement (MSD) and total traveled distance (TTD) measurements derived from video microscopy and single-cell tracking.Our results indicate that while EV treatments had remarkable promoting effect on cell migration, they exerted only a modest effect on cell proliferation and spheroid growth. Notably, EVs demonstrated the ability to mitigate the inhibitory effects of BRAF inhibitors, albeit they were ineffective against a MEK inhibitor and the combination of BRAF/MEK inhibitors. In summary, our findings contribute to the understanding of the intricate role played by EVs in tumor progression, metastasis, and drug resistance in MM.}, keywords = {cell migration; Melanoma; Extracellular vesicles; Vemurafenib; Trametinib; dabrafenib; Single cell tracking}, year = {2024}, eissn = {1478-811X}, orcid-numbers = {Gaál, Anikó/0000-0003-4064-1825; Varga, Zoltán/0000-0002-5741-2669; Buzás, Edit Irén/0000-0002-3744-206X; Dank, Magdolna/0000-0002-4442-8733; Takács, István/0000-0002-7810-4833; Szász, Attila Marcell/0000-0003-2739-4196; Garay, Tamás/0000-0003-0329-9207} } @article{MTMT:34849850, title = {Limited miscibility in hydrated DPPC – Lyso-PPC systems}, url = {https://m2.mtmt.hu/api/publication/34849850}, author = {Bóta, Attila and Wacha, András Ferenc and Trif, László and Varga, Zoltán and Mihály, Judith}, doi = {10.1016/j.molliq.2024.124960}, journal-iso = {J MOL LIQ}, journal = {JOURNAL OF MOLECULAR LIQUIDS}, volume = {404}, unique-id = {34849850}, issn = {0167-7322}, year = {2024}, eissn = {1873-3166}, pages = {124960-124969}, orcid-numbers = {Wacha, András Ferenc/0000-0002-9609-0893; Trif, László/0000-0002-3960-1829; Varga, Zoltán/0000-0002-5741-2669} } @article{MTMT:34848253, title = {Determination of the Nanoscale Silica Mass Fraction by AF4/ICP-MS with Isotope Dilution Analysis Using 29 Si-Enriched Silica Nanoparticles}, url = {https://m2.mtmt.hu/api/publication/34848253}, author = {Bartczak, Dorota and Cuello-Nuñez, Susana and Pálmai, Marcell and Hill, Sarah and Petrov, Panayot and Varga, Zoltán and Szalay, Roland and Goenaga-Infante, Heidi}, doi = {10.1021/acs.analchem.4c00021}, journal-iso = {ANAL CHEM}, journal = {ANALYTICAL CHEMISTRY}, unique-id = {34848253}, issn = {0003-2700}, year = {2024}, eissn = {1520-6882}, orcid-numbers = {Hill, Sarah/0000-0002-7480-0655; Varga, Zoltán/0000-0002-5741-2669; Szalay, Roland/0000-0003-3535-5365} } @article{MTMT:34819821, title = {In situ captured antibacterial action of membrane-incising peptide lamellae}, url = {https://m2.mtmt.hu/api/publication/34819821}, author = {El Battioui, Kamal and Chakraborty, Sohini and Wacha, András Ferenc and Molnár, Dániel and Quemé Peña, Mayra and Szigyártó, Imola Csilla and Szabó, Csenge Lilla and Bodor, Andrea and Horváti, Kata and Gyulai, Gergő and Bősze, Szilvia and Mihály, Judith and Jezsó, Bálint and Románszki, Loránd and Tóth, Judit and Varga, Zoltán and Mándity, István and Juhász, Tünde and Beke-Somfai, Tamás}, doi = {10.1038/s41467-024-47708-4}, journal-iso = {NAT COMMUN}, journal = {NATURE COMMUNICATIONS}, volume = {15}, unique-id = {34819821}, issn = {2041-1723}, abstract = {Developing unique mechanisms of action are essential to combat the growing issue of antimicrobial resistance. Supramolecular assemblies combining the improved biostability of non-natural compounds with the complex membrane-attacking mechanisms of natural peptides are promising alternatives to conventional antibiotics. However, for such compounds the direct visual insight on antibacterial action is still lacking. Here we employ a design strategy focusing on an inducible assembly mechanism and utilized electron microscopy (EM) to follow the formation of supramolecular structures of lysine-rich heterochiral β 3 -peptides, termed lamellin-2K and lamellin-3K, triggered by bacterial cell surface lipopolysaccharides. Combined molecular dynamics simulations, EM and bacterial assays confirmed that the phosphate-induced conformational change on these lamellins led to the formation of striped lamellae capable of incising the cell envelope of Gram-negative bacteria thereby exerting antibacterial activity. Our findings also provide a mechanistic link for membrane-targeting agents depicting the antibiotic mechanism derived from the in-situ formation of active supramolecules.}, year = {2024}, eissn = {2041-1723}, orcid-numbers = {Wacha, András Ferenc/0000-0002-9609-0893; Szabó, Csenge Lilla/0000-0002-6508-3439; Bodor, Andrea/0000-0002-7422-298X; Horváti, Kata/0000-0003-4092-6011; Gyulai, Gergő/0000-0002-1352-2014; Bősze, Szilvia/0000-0001-9555-699X; Jezsó, Bálint/0000-0002-1306-4797; Tóth, Judit/0000-0002-0965-046X; Varga, Zoltán/0000-0002-5741-2669; Mándity, István/0000-0003-2865-6143} } @article{MTMT:34766204, title = {Extracellular Vesicles of Patients on Peritoneal Dialysis Inhibit the TGF-β- and PDGF-B-Mediated Fibrotic Processes}, url = {https://m2.mtmt.hu/api/publication/34766204}, author = {Szebeni, Beáta and Veres-Székely, Apor and Pap, Domonkos and Bokrossy, Péter and Varga, Zoltán and Gaál, Anikó and Mihály, Judith and Pállinger, Éva and Takács, István Márton and Pajtók, Csenge and Bernáth, Mária and Reusz, György and Szabó, Attila and Vannay, Ádám}, doi = {10.3390/cells13070605}, journal-iso = {CELLS-BASEL}, journal = {CELLS}, volume = {13}, unique-id = {34766204}, abstract = {Among patients on peritoneal dialysis (PD), 50–80% will develop peritoneal fibrosis, and 0.5–4.4% will develop life-threatening encapsulating peritoneal sclerosis (EPS). Here, we investigated the role of extracellular vesicles (EVs) on the TGF-β- and PDGF-B-driven processes of peritoneal fibrosis. EVs were isolated from the peritoneal dialysis effluent (PDE) of children receiving continuous ambulatory PD. The impact of PDE-EVs on the epithelial–mesenchymal transition (EMT) and collagen production of the peritoneal mesothelial cells and fibroblasts were investigated in vitro and in vivo in the chlorhexidine digluconate (CG)-induced mice model of peritoneal fibrosis. PDE-EVs showed spherical morphology in the 100 nm size range, and their spectral features, CD63, and annexin positivity were characteristic of EVs. PDE-EVs penetrated into the peritoneal mesothelial cells and fibroblasts and reduced their PDE- or PDGF-B-induced proliferation. Furthermore, PDE-EVs inhibited the PDE- or TGF-β-induced EMT and collagen production of the investigated cell types. PDE-EVs contributed to the mesothelial layer integrity and decreased the submesothelial thickening of CG-treated mice. We demonstrated that PDE-EVs significantly inhibit the PDGF-B- or TGF-β-induced fibrotic processes in vitro and in vivo, suggesting that EVs may contribute to new therapeutic strategies to treat peritoneal fibrosis and other fibroproliferative diseases.}, year = {2024}, eissn = {2073-4409}, orcid-numbers = {Szebeni, Beáta/0000-0001-7577-9803; Veres-Székely, Apor/0000-0002-6830-8779; Pap, Domonkos/0000-0002-1718-1210; Varga, Zoltán/0000-0002-5741-2669; Gaál, Anikó/0000-0003-4064-1825; Pállinger, Éva/0000-0002-5789-0951; Reusz, György/0000-0003-0396-043X; Szabó, Attila/0000-0001-7321-9861; Vannay, Ádám/0000-0001-7412-4733} } @article{MTMT:34567532, title = {Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches}, url = {https://m2.mtmt.hu/api/publication/34567532}, author = {Welsh, Joshua A. and Goberdhan, Deborah C. I. and O'Driscoll, Lorraine and Buzás, Edit Irén and Blenkiron, Cherie and Bussolati, Benedetta and Cai, Houjian and Di Vizio, Dolores and Driedonks, Tom A. P. and Erdbrügger, Uta and Falcon‐Perez, Juan M. and Fu, Qing‐Ling and Hill, Andrew F. and Lenassi, Metka and Lim, Sai Kiang and Mahoney, Mỹ G. and Mohanty, Sujata and Möller, Andreas and Nieuwland, Rienk and Ochiya, Takahiro and Sahoo, Susmita and Torrecilhas, Ana C. and Zheng, Lei and Zijlstra, Andries and Abuelreich, Sarah and Bagabas, Reem and Bergese, Paolo and Bridges, Esther M. and Brucale, Marco and Burger, Dylan and Carney, Randy P. and Cocucci, Emanuele and Colombo, Federico and Crescitelli, Rossella and Hanser, Edveena and Harris, Adrian L. and Haughey, Norman J. and Hendrix, An and Ivanov, Alexander R. and Jovanovic‐Talisman, Tijana and Kruh‐Garcia, Nicole A. and Ku'ulei‐Lyn Faustino, Vroniqa and Kyburz, Diego and Lässer, Cecilia and Lennon, Kathleen M. and Lötvall, Jan and Maddox, Adam L. and Martens‐Uzunova, Elena S. and Mizenko, Rachel R. and Newman, Lauren A. and Ridolfi, Andrea and Rohde, Eva and Rojalin, Tatu and Rowland, Andrew and Saftics, Andras and Sandau, Ursula S. and Saugstad, Julie A. and Shekari, Faezeh and Swift, Simon and Ter‐Ovanesyan, Dmitry and Tosar, Juan P. and Useckaite, Zivile and Valle, Francesco and Varga, Zoltán and van der Pol, Edwin and van Herwijnen, Martijn J. C. and Wauben, Marca H. M. and Wehman, Ann M. and Williams, Sarah and Zendrini, Andrea and Zimmerman, Alan J. and Théry, Clotilde and Witwer, Kenneth W. and Haseeb, Zubair}, doi = {10.1002/jev2.12404}, journal-iso = {J EXTRACELLULAR VESICL}, journal = {JOURNAL OF EXTRACELLULAR VESICLES}, volume = {13}, unique-id = {34567532}, abstract = {Extracellular vesicles (EVs), through their complex cargo, can reflect the state of their cell of origin and change the functions and phenotypes of other cells. These features indicate strong biomarker and therapeutic potential and have generated broad interest, as evidenced by the steady year‐on‐year increase in the numbers of scientific publications about EVs. Important advances have been made in EV metrology and in understanding and applying EV biology. However, hurdles remain to realising the potential of EVs in domains ranging from basic biology to clinical applications due to challenges in EV nomenclature, separation from non‐vesicular extracellular particles, characterisation and functional studies. To address the challenges and opportunities in this rapidly evolving field, the International Society for Extracellular Vesicles (ISEV) updates its ‘Minimal Information for Studies of Extracellular Vesicles’, which was first published in 2014 and then in 2018 as MISEV2014 and MISEV2018, respectively. The goal of the current document, MISEV2023, is to provide researchers with an updated snapshot of available approaches and their advantages and limitations for production, separation and characterisation of EVs from multiple sources, including cell culture, body fluids and solid tissues. In addition to presenting the latest state of the art in basic principles of EV research, this document also covers advanced techniques and approaches that are currently expanding the boundaries of the field. MISEV2023 also includes new sections on EV release and uptake and a brief discussion of in vivo approaches to study EVs. Compiling feedback from ISEV expert task forces and more than 1000 researchers, this document conveys the current state of EV research to facilitate robust scientific discoveries and move the field forward even more rapidly.}, year = {2024}, eissn = {2001-3078}, orcid-numbers = {Welsh, Joshua A./0000-0002-1097-9756; Goberdhan, Deborah C. I./0000-0003-0645-6714; Buzás, Edit Irén/0000-0002-3744-206X; Bussolati, Benedetta/0000-0002-3663-5134; Cai, Houjian/0000-0003-4887-2652; Falcon‐Perez, Juan M./0000-0003-3133-0670; Hill, Andrew F./0000-0001-5581-2354; Lenassi, Metka/0000-0002-9488-6855; Mohanty, Sujata/0000-0002-0047-4914; Nieuwland, Rienk/0000-0002-5671-3400; Ochiya, Takahiro/0000-0002-0776-9918; Sahoo, Susmita/0000-0002-7279-1564; Torrecilhas, Ana C./0000-0001-5724-2199; Zheng, Lei/0000-0003-2576-8780; Zijlstra, Andries/0000-0001-8460-8803; Brucale, Marco/0000-0001-7244-4389; Carney, Randy P./0000-0001-8193-1664; Crescitelli, Rossella/0000-0002-1714-3169; Haughey, Norman J./0000-0001-5194-4122; Martens‐Uzunova, Elena S./0000-0002-5363-2525; Newman, Lauren A./0000-0003-3303-1666; Rohde, Eva/0000-0001-8692-886X; Sandau, Ursula S./0000-0002-3646-7089; Saugstad, Julie A./0000-0002-2996-9611; Shekari, Faezeh/0000-0001-6026-5412; Tosar, Juan P./0000-0002-2021-2479; Varga, Zoltán/0000-0002-5741-2669; Wauben, Marca H. M./0000-0003-0360-0311; Wehman, Ann M./0000-0001-9826-4132; Zimmerman, Alan J./0000-0001-6280-4790; Théry, Clotilde/0000-0001-8294-6884; Witwer, Kenneth W./0000-0003-1664-4233; Bodnár, Bernadett Réka/0000-0003-3347-9225; Bukva, Mátyás/0000-0002-5225-0285; Buzás, Edit Irén/0000-0002-3744-206X; Buzás, Krisztina/0000-0001-8933-2033; Dobra, Gabriella/0000-0002-2814-7720; Försönits, András/0000-0002-9298-8890; Ghosal, Sayam/0000-0001-6618-930X; Gyukity-Sebestyén, Edina/0000-0003-1383-6301; Koncz, Anna/0000-0003-2511-2394; Lőrincz, Márton Ákos/0000-0002-2819-5116; Németh, Krisztina/0000-0002-3825-2137; Oláh, Attila/0000-0003-4122-5639; Osteikoetxea, Xabier/0000-0003-3628-0174; Pálóczi, Krisztina/0000-0001-7065-3582; Stepanova, Ganna/0000-0002-8285-2762; Visnovitz, Tamás/0000-0002-7962-5083; Wiener, Zoltán/0000-0001-7056-4926; Harmati, Mária/0000-0002-4875-5723; Hegyesi, Hargita/0000-0002-8800-5169} }