1211. RNA-dependent RNA polymerase 1 in potato (Solanum tuberosum) and its relationship to other plant RNA-dependent RNA polymerasesLydia J. R. Hunter, Samuel F. Brockington, Alex M. Murphy, Adrienne E. Pate, Kristina Gruden, Stuart A. MacFarlane, Peter Palukaitis, John P. Carr, 2016, original scientific article Abstract: Cellular RNA-dependent RNA polymerases (RDRs) catalyze synthesis of double-stranded RNAs that can serve to initiate or amplify RNA silencing. Arabidopsis thaliana has six RDR genes; RDRs 1, 2 and 6 have roles in anti-viral RNA silencing. RDR6 is constitutively expressed but RDR1 expression is elevated following plant treatment with defensive phytohormones. RDR1 also contributes to basal virus resistance. RDR1 has been studied in several species including A. thaliana, tobacco (Nicotiana tabacum), N. benthamiana, N. attenuata and tomato (Solanum lycopersicum) but not to our knowledge in potato (S. tuberosum). StRDR1 was identified and shown to be salicylic acid-responsive. StRDR1 transcript accumulation decreased in transgenic potato plants constitutively expressing a hairpin construct and these plants were challenged with three viruses: potato virus Y, potato virus X, and tobacco mosaic virus. Suppression of StRDR1 gene expression did not increase the susceptibility of potato to these viruses. Phylogenetic analysis of RDR genes present in potato and in a range of other plant species identified a new RDR gene family, not present in potato and found only in Rosids (but apparently lost in the Rosid A. thaliana) for which we propose the name RDR7.
Keywords: RNA polymerases, RNA, potato
Published in DiRROS: 25.07.2024; Views: 463; Downloads: 260
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1212. Primary metabolism, phenylpropanoids and antioxidant pathways are regulated in potato as a response to Potato virus Y infectionPolona Kogovšek, Maruša Pompe Novak, Marko Petek, Lena Fragner, Wolfram Weckwerth, Kristina Gruden, 2016, original scientific article Abstract: Potato production is one of the most important agricultural sectors, and it is challenged by various detrimental factors, including virus infections. To control losses in potato production, knowledge about the virus—plant interactions is crucial. Here, we investigated the molecular processes in potato plants as a result of Potato virus Y (PVY) infection, the most economically important potato viral pathogen. We performed an integrative study that links changes in the metabolome and gene expression in potato leaves inoculated with the mild PVYN and aggressive PVYNTN isolates, for different times through disease development. At the beginning of infection (1 day post-inoculation), virus-infected plants showed an initial decrease in the concentrations of metabolites connected to sugar and amino-acid metabolism, the TCA cycle, the GABA shunt, ROS scavangers, and phenylpropanoids, relative to the control plants. A pronounced increase in those metabolites was detected at the start of the strong viral multiplication in infected leaves. The alterations in these metabolic pathways were also seen at the gene expression level, as analysed by quantitative PCR. In addition, the systemic response in the metabolome to PVY infection was analysed. Systemic leaves showed a less-pronounced response with fewer metabolites altered, while phenylpropanoid-associated metabolites were strongly accumulated. There was a more rapid onset of accumulation of ROS scavengers in leaves inoculated with PVYN than those inoculated with PVYNTN. This appears to be related to the lower damage observed for leaves of potato infected with the milder PVYN strain, and at least partially explains the differences between the phenotypes observed.
Published in DiRROS: 25.07.2024; Views: 346; Downloads: 279
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1213. Improved protective effect of umbilical cord stem cell transplantation on cisplatin-induced kidney injury in mice pretreated with antithymocyte globulinŽeljka Večerić-Haler, Andreja Erman, Anton Cerar, Helena Motaln, Katja Kološa, Tamara Lah Turnšek, Snežna Sodin-Šemrl, Katja Lakota, Katjuša Mrak Poljšak, Špela Škrajnar, Simona Kranjc Brezar, Miha Arnol, Martina Perše, 2016, original scientific article Abstract: Mesenchymal stem cells (MSCs) are recognised as a promising tool to improve renal recovery in experimental models of cisplatin-induced acute kidney injury. However, these preclinical studies were performed on severely immunodeficient animals. Here, we investigated whether human umbilical cord derived MSC treatment could equally ameliorate acute kidney injury induced by cisplatin and prolong survival in mice with a normal immune system and those with a suppressed immune system by polyclonal antithymocyte globulin (ATG). We demonstrated that ATG pretreatment, when followed by MSC transplantation, significantly improved injured renal function parameters, as evidenced by decreased blood urea nitrogen and serum creatinine concentration, as well as improved renal morphology. This tissue restoration was also supported by increased survival of mice. The beneficial effects of ATG were associated with reduced level of inflammatory protein serum amyloid A3 and induced antioxidative expression of superoxide dismutase-1 (SOD-1), glutathione peroxidase (GPx), and hem oxygenase-1 (HO-1). Infused MSCs became localised predominantly in peritubular areas and acted to reduce renal cell death. In conclusion, these results show that ATG diminished in situ inflammation and oxidative stress associated with cisplatin-induced acute kidney injury, the effects that may provide more favourable microenvironment for MSC action, with consequential synergistic improvements in renal injury and animal survival as compared to MSC treatment alone.
Keywords: mesenchymal stem cells, nephrotoxicity
Published in DiRROS: 25.07.2024; Views: 348; Downloads: 275
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1214. Use of hydrodynamic cavitation in (waste)water treatmentMatevž Dular, Tjaša Griessler Bulc, Ion Gutiérrez-Aguirre, Ester Heath, Tina Kosjek, Aleksandra Krivograd-Klemenčič, Martina Oder, Martin Petkovšek, Nejc Rački, Maja Ravnikar, Andrej Šarc, Brane Širok, Mojca Zupanc, Miha Žitnik, Boris Kompare, 2016, original scientific article Abstract: The use of acoustic cavitation for water and wastewater treatment (cleaning) is a well known procedure. Yet, the use of hydrodynamic cavitation as a sole technique or in combination with other techniques such as ultrasound has only recently been suggested and employed.
In the first part of this paper a general overview of techniques that employ hydrodynamic cavitation for cleaning of water and wastewater is presented.
In the second part of the paper the focus is on our own most recent work using hydrodynamic cavitation for removal of pharmaceuticals (clofibric acid, ibuprofen, ketoprofen, naproxen, diclofenac, carbamazepine), toxic cyanobacteria (Microcystis aeruginosa), green microalgae (Chlorella vulgaris), bacteria (Legionella pneumophila) and viruses (Rotavirus) from water and wastewater.
As will be shown, hydrodynamic cavitation, like acoustic, can manifest itself in many different forms each having its own distinctive properties and mechanisms. This was until now neglected, which eventually led to poor performance of the technique. We will show that a different type of hydrodynamic cavitation (different removal mechanism) is required for successful removal of different pollutants.
The path to use hydrodynamic cavitation as a routine water cleaning method is still long, but recent results have already shown great potential for optimisation, which could lead to a low energy tool for water and wastewater cleaning.
Keywords: wastewater, cleaning, hydrodynamic cavitation, pharmaceuticals, cyanobacteria, microalgae, viruses, Legionella bacteria
Published in DiRROS: 25.07.2024; Views: 314; Downloads: 173
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1215. Assessment of the real-time PCR and different digital PCR platforms for DNA quantificationJernej Pavšič, Jana Žel, Mojca Milavec, 2016, original scientific article Abstract: Digital PCR (dPCR) is beginning to supersede real-time PCR (qPCR) for quantification of nucleic acids in many different applications. Several analytical properties of the two most commonly used dPCR platforms, namely the QX100 system (Bio-Rad) and the 12.765 array of the Biomark system (Fluidigm), have already been evaluated and compared with those of qPCR. However, to the best of our knowledge, direct comparison between the three of these platforms using the same DNA material has not been done, and the 37 K array on the Biomark system has also not been evaluated in terms of linearity, analytical sensitivity and limit of quantification. Here, a first assessment of qPCR, the QX100 system and both arrays of the Biomark system was performed with plasmid and genomic DNA from human cytomegalovirus. With use of PCR components that alter the efficiency of qPCR, each dPCR platform demonstrated consistent copy-number estimations, which indicates the high resilience of dPCR. Two approaches, one considering the total reaction volume and the other considering the effective reaction size, were used to assess linearity, analytical sensitivity and variability. When the total reaction volume was considered, the best performance was observed with qPCR, followed by the QX100 system and the Biomark system. In contrast, when the effective reaction size was considered, all three platforms showed almost equal limits of detection and variability. Although dPCR might not always be more appropriate than qPCR for quantification of low copy numbers, dPCR is a suitable method for robust and reproducible quantification of viral DNA, and a promising technology for the higher-order reference measurement method.
Keywords: real-time PCR, molecular diagnostics, human cytomegalovirus, DNA quantification, digital PCR
Published in DiRROS: 25.07.2024; Views: 375; Downloads: 218
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1216. Digital PCR for direct quantification of viruses without DNA extractionJernej Pavšič, Jana Žel, Mojca Milavec, 2016, original scientific article Abstract: DNA extraction before amplification is considered an essential step for quantification of viral DNA using real-time PCR (qPCR). However, this can directly affect the final measurements due to variable DNA yields and removal of inhibitors, which leads to increased inter-laboratory variability of qPCR measurements and reduced agreement on viral loads. Digital PCR (dPCR) might be an advantageous methodology for the measurement of virus concentrations, as it does not depend on any calibration material and it has higher tolerance to inhibitors. DNA quantification without an extraction step (i.e. direct quantification) was performed here using dPCR and two different human cytomegalovirus whole-virus materials. Two dPCR platforms were used for this direct quantification of the viral DNA, and these were compared with quantification of the extracted viral DNA in terms of yield and variability. Direct quantification of both whole-virus materials present in simple matrices like cell lysate or Tris-HCl buffer provided repeatable measurements of virus concentrations that were probably in closer agreement with the actual viral load than when estimated through quantification of the extracted DNA. Direct dPCR quantification of other viruses, reference materials and clinically relevant matrices is now needed to show the full versatility of this very promising and cost-efficient development in virus quantification.
Keywords: molecular diagnostics, direct quantification, viruses, virus reference materials, human cytomegalovirus, digital PCR
Published in DiRROS: 25.07.2024; Views: 337; Downloads: 262
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1217. Assessment of short-term effect of platelet-rich plasma treatment of tendinosis using texture analysis of ultrasound imagesKarlo Pintarić, Vladka Salapura, Žiga Snoj, Andrej Vovk, Mojca Božič, Jernej Vidmar, 2023, original scientific article Keywords: texture analysis, ultrasonography, tendinosis
Published in DiRROS: 25.07.2024; Views: 369; Downloads: 138
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1219. The effects of normobaric and hyperbaric oxygenation on MRI signal intensities in T1-weighted, T2-weighted and FLAIR images in human brainVida Velej, Ksenija Cankar, Jernej Vidmar, 2023, original scientific article Abstract: Dissolved oxygen has known paramagnetic effects in magnetic resonance imaging (MRI). The aim ofthis study was to compare the effects of normobaric oxygenation (NBO) and hyperbaric oxygenation (HBO) on hu-man brain MRI signal intensities.Patients and methods. Baseline brain MRI was performed in 17 healthy subjects (mean age 27.8 ± 3.2). MRI wasrepeated after exposure to the NBO and HBO at different time points (0 min, 25 min, 50 min). Signal intensities in T1-weighted, T2-weighted images and fluid attenuated inversion recovery (FLAIR) signal intensities of several intracranialstructures were compared between NBO and HBO.Results. Increased T1-weighted signal intensities were observed in white and deep grey brain matter, cerebrospinalfluid (CSF), venous blood and vitreous body after exposure to NBO as well as to HBO compared to baseline (Dunnett’stest, p < 0.05) without significant differences between both protocols. There was also no significant difference in T2-weighted signal intensities between NBO and HBO. FLAIR signal intensities were increased only in the vitreous bodyafter NBO and HBO and FLAIR signal of caudate nucleus was decreased after NBO (Dunnett’s test, p < 0.05). Thestatistically significant differences in FLAIR signal intensities were found between NBO and HBO (paired t-test, p < 0.05)in most observed brain structures (paired t-test, p < 0.05).Conclusions. Our results show that NBO and HBO alters signal intensities T1-weighted and FLAIR images of humanbrain. The differences between NBO and HBO are most pronounced in FLAIR imaging.
Keywords: hyperbaric oxygen, normobaric oxygen, magnetic resonance, human brain
Published in DiRROS: 25.07.2024; Views: 330; Downloads: 132
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1220. Local control and survival after stereotactic body radiation therapy of early-stage lung cancer patients in SloveniaKarmen Stanič, Jasna But-Hadžić, Jan Žagar, Martina Vrankar, 2023, original scientific article Abstract: Background. Stereotactic body radiation therapy (SBRT) precisely and non-invasively delivers ablative radiationdose to tumors in early-stage lung cancer patients who are not candidates for surgery or refuse it. The aim of researchwas to evaluate local control, overall survival (OS), local progression free survival (LPFS), distant metastases free survival(DMFS), disease free survival (DFS) and toxicity in early-stage lung cancer patients treated with SBRT in a single tertiarycancer centre.Patients and methods. We retrospectively evaluated medical records and radiation treatment plan parametersof 228 tumors irradiated in 206 early-stage lung cancer patients between 2016 and 2021 at the Institute of OncologyLjubljana.Results. After 25 months of median follow up, 68 of 206 (33%) patients died. Median OS was 46 months (CI 36 −56),1-year, 2-year and 3-year OS were 87%, 74% and 62% and 5-year OS was 31%. A total of 45 disease progressions havebeen identified in 41 patients. Local progress only was noticed in 5 (2%) patients, systemic progress in 32 (16%) andcombined systemic and local in 4 (2%) patients. Local control rate (LCR) at 1 year was 98%, at 2 and 3 years 96%and 95% at 5 years. The 1-, 2- and 3-year LPFS were 98%, 96% and 94%, respectively and 5-year LPFS was 82%. One,2-, 3- and 5-year DFS w ere 89%, 81%, 72% and 49%, respectively. Among 28 toxicities recorded only one was Grade4 (pneumonitis), all others were Grade 1 or 2. No differences in LCR, LPFS, DFS were found in univariate analysis com-paring patient, tumor, and treatment characteristics. For OS the only statistically significant difference was found inpatients with more than 3 comorbidities compared to those with less comorbidities.Conclusions. Early lung cancer treated with SBRT at single tertiary cancer centre showed that LCR, LPFS, DFS, DMFSand OS were comparable to published studies. Patients with many comorbidities had significantly worse overallsurvival compared to those with less comorbidities. No other significant differences by patient, tumor, or treatmentcharacteristics were found for DMFS, LPFS, and DFS. Toxicity data confirmed that treatment was well tolerated.
Keywords: stereotactic body radiotherapy, early-stage lung cancer, lung cancer
Published in DiRROS: 25.07.2024; Views: 338; Downloads: 175
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