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Proteolytically activated CRAC effectors through designed intramolecular inhibition
Vid Jazbec, Roman Jerala, Mojca Benčina, 2022, izvirni znanstveni članek

Ključne besede: STIM1, Orai, TEV protease, PPV protease, calcium signaling, coiled-coil peptides
Objavljeno v DiRROS: 20.07.2022; Ogledov: 651; Prenosov: 399
.pdf Celotno besedilo (5,62 MB)
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Robust saliva-based RNA extraction-free one-step nucleic acid amplification test for mass SARS-CoV-2 monitoring
Eva 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: 1034; Prenosov: 532
URL Povezava na celotno besedilo

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Decrease in cellular nanovesicles concentration in blood of athletes more than 15 hours after marathon
Zala Jan, Mitja Drab, Damjana Drobne, Apolonija Bedina Zavec, Mojca Benčina, Barbara Drašler, Matej Hočevar, Judita Lea Krek, Ljubiša Pađen, Manca Pajnič, Neža Repar, Boštjan Šimunič, Roman Štukelj, Veronika Kralj-Iglič, 2021, izvirni znanstveni članek

Povzetek: Introduction: Cellular nanovesicles (CNVs), that are shed from cells, have been recognized as promising indicators of health status. We analyzed the effect of long-distance running on concentration of CNVs, along with some standard blood parameters, in 27 athletes two days before and > 15 hours after physical effort. Methods: CNVs were isolated by repetitive centrifugation and washing of samples, and assessed by flow cytometry. Cholinesterase (ChE) and glutathione S-transferase (GST) activity were measured spectrophotometrically. Interleukin 6 (IL-6) and tumor necrosis factor-% (TNF-%) concentrations were measured using enzyme-linked immunosorbent assay (ELISA). C-reactive protein (CRP) was measured with immunoturbidimetric determination and lipidogram parameters were measured by enzymatic colorimetric assay. Flow cytometry was used for blood cell count and mean platelet volume (MPV) measurement. Results: More than 15 hours after physical effort a decrease was found in CNVs% concentration in isolates from blood (46%; p< 0.05), in ChE activity in whole blood (47%; p< 0.001), in plasma (34%; p< 0.01), and in erythrocyte suspension (54%; p< 0.001), as well as in GST activity in erythrocyte suspension (16%; p< 0.01) and in IL-6 concentration in plasma (63%; p< 0.05). We found no change in GST activity in plasma and in TNF-% concentration in plasma. Correlations (> 0.8; p< 0.001) between CNVs% concentration and ChE activity, and GST activity, respectively, in erythrocyte suspension were found. Conclusion: We found that > 15 hours post-physical effort, CNVs% concentration was below the initial value, concomitant with other measured parameters: ChE and GST activity as well as IL-6 concentration, indicating a favorable effect of physical effort on health status. CNVs% concentration and ChE activity in isolates from peripheral blood proved to have potential as indicators of the response of the human body to inflammation after physical effort. Physical activity should be considered as an important factor in preparation of subjects for blood sampling in procedures focusing on CNV-containing diagnostic and therapeutic compounds.
Ključne besede: membrane vesiculation, physical effort, blood samples, inflammation process, cellular nanovesicles, marathon
Objavljeno v DiRROS: 25.01.2021; Ogledov: 1243; Prenosov: 1015
.pdf Celotno besedilo (1,88 MB)
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Engineering and rewiring of a calcium-dependent signaling pathway
Maja Meško, Tina Lebar, Petra Dekleva, Roman Jerala, Mojca Benčina, 2020, izvirni znanstveni članek

Objavljeno v DiRROS: 25.11.2020; Ogledov: 1406; Prenosov: 894
.pdf Celotno besedilo (3,76 MB)
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