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22. 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, original scientific article Abstract: 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.
Keywords: SARS-CoV-2, COVID-19, COVID-19 serological testing, real-time polymerase chain reaction, saliva, oral cavity swab, passive drool, pooling
Published in DiRROS: 09.11.2021; Views: 1087; Downloads: 546
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23. Treatment outcome clustering patterns correspond to discrete asthma phenotypes in childrenIvana Banić, Mario Lovrić, Gerald Cuder, Roman Kern, Matija Rijavec, Peter Korošec, Mirjana Kljajić-Turkalj, 2021, original scientific article Abstract: Despite widely and regularly used therapy asthma in children is not fully controlled. Recognizing the complexity of asthma phenotypes and endotypes imposed the concept of precision medicine in asthma treatment. By applying machine learning algorithms assessed with respect to their accuracy in predicting treatment outcome, we have successfully identified 4 distinct clusters in a pediatric asthma cohort with specific treatment outcome patterns according to changes in lung function (FEV1 and MEF50), airway inflammation (FENO) and disease control likely affected by discrete phenotypes at initial disease presentation, differing in the type and level of inflammation, age of onset, comorbidities, certain genetic and other physiologic traits. The smallest and the largest of the 4 clusters- 1 (N = 58) and 3 (N = 138) had better treatment outcomes compared to clusters 2 and 4 and were characterized by more prominent atopic markers and a predominant allelic (A allele) effect for rs37973 in the GLCCI1 gene previously associated with positive treatment outcomes in asthmatics. These patients also had a relatively later onset of disease (6 + yrs). Clusters 2 (N = 87) and 4 (N = 64) had poorer treatment success, but varied in the type of inflammation (predominantly neutrophilic for cluster 4 and likely mixed-type for cluster 2), comorbidities (obesity for cluster 2), level of systemic inflammation (highest hsCRP for cluster 2) and platelet count (lowest for cluster 4). The results of this study emphasize the issues in asthma management due to the overgeneralized approach to the disease, not taking into account specific disease phenotypes.
Keywords: asthma, allergy and immunology, pediatrics, machine learning, treatment outcome, phenotypes, childhood asthma, clustering
Published in DiRROS: 16.08.2021; Views: 938; Downloads: 655
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24. Somatic mutations and the risk of undifferentiated autoinflammatory disease in MDS : an under-recognized but prognostically important complicationAbdulla Watad, Mark Kačar, Nicola Luigi Bragazzi, Qiao Zhou, Miriam Jassam, Jan Taylor, Eve Roman, Alexandra Smith, Richard A. Jones, Howard Amital, 2021, original scientific article Abstract: Objectives: We theorized that myelodysplastic syndrome (MDS) with somatic mutations and karyotype abnormalities are associated with autoinflammation, and that the presence of autoinflammatory disease affected prognosis in MDS. Methods: One hundred thirty-four MDS patients were assessed for the prevalence of autoinflammatory complications and its link with karyotypes and somaticmutation status. Autoinflammatory complications were described either as well-defined autoinflammatory diseases (AD) or undifferentiated "autoinflammatory disease" (UAD) (defined as CRP over 10.0 mg/L on five consecutive occasions, taken at separate times and not explained by infection). Several patient characteristics including demographic, clinical, laboratory, cytogenetics charts, and outcomes, were compared between different groups. Results: Sixty-two (46.3%) patients had an autoinflammatory complication manifesting as arthralgia (43.5% vs. 23.6%, p = 0.0146), arthritis (30.6% vs. 15.3%, p = 0.0340), skin rash (27.4% vs. 12.5%, p = 0.0301), pleuritis (14.5% vs. 4.2%, p = 0.0371) and unexplained fever (27.4% vs. 0%, p < 0.0001). AD were found in 7.4% of MDS patients (with polymyalgia rheumatic being the most frequently one). Classical autoimmune diseases were found only in 4 MDS patients (3.0%). Transcription factor pathway mutations (RUNX1, BCOR, WTI, TP53) (OR 2.20 [95%CI 1.02-4.75], p = 0.0451) and abnormal karyotypes (OR 2.76 [95%CI 1.22-6.26], p = 0.0153) were associated with autoinflammatory complications. Acute leukaemic transformation was more frequent in MDS patients with autoinflammatory features than those without (27.4% vs. 9.7%, p = 0.0080). Conclusions: Autoinflammatory complications are common inMDS. Somatic mutations of transcription factor pathways and abnormal karyotypes are associated with greater risk of autoinflammatory complications, which are themselves linked to malignant transformation and a worse prognosis.
Keywords: myelodysplastic syndromes - genetics, autoinflammation, undifferentiated autoinflammatory disease, molecular characterization, somatic mutations
Published in DiRROS: 31.03.2021; Views: 1181; Downloads: 801
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25. Triangular in vivo self-assembling coiled-coil protein origamiSabina Božič Abram, Helena Gradišar, Jana Aupič, Adam Round, Roman Jerala, 2021, original scientific article Keywords: coiled coil, enzyme clustering, biosynthesis, resveratrol, mevalonate
Published in DiRROS: 22.03.2021; Views: 1463; Downloads: 911
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26. Decrease in cellular nanovesicles concentration in blood of athletes more than 15 hours after marathonZala 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, original scientific article Abstract: 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.
Keywords: membrane vesiculation, physical effort, blood samples, inflammation process, cellular nanovesicles, marathon
Published in DiRROS: 25.01.2021; Views: 1305; Downloads: 1614
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27. Platformne storitveAna Robnik, Andrej Ciglič, Aleš Balanč, Dušan Gabrijelčič, Jan Porekar, Arso Savanović, Polona Škraba, Roman Kužnar, Ignac Zupan, 2015, published scientific conference contribution Published in DiRROS: 08.12.2020; Views: 1298; Downloads: 391
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29. Spremljanje vektorjev borove ogorčice (Monochamus spp.) in druge entomofavne v Sloveniji od leta 2007 do leta 2013Maja Jurc, Roman Pavlin, Gregor Meterc, Danijel Borkovič, 2013, professional article Keywords: zdravje gozdov, gozdovi, varstvo gozdov, borova ogoričica, Monochamus spp., entomofavna
Published in DiRROS: 29.07.2020; Views: 1536; Downloads: 845
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30. Pinijev sprevodni prelec (Thaumetopoea pityocampa) v SlovenijiMaja Jurc, Tine Hauptman, Roman Pavlin, Danijel Borkovič, Zoran Zavratnik, Vida Papler-Lampe, 2019, professional article Keywords: gozdovi, Varstvo gozdov, Pinus spp., podnebne spremembe, širjenje areala vrste, monitoring
Published in DiRROS: 28.07.2020; Views: 1561; Downloads: 884
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