1651. Digital PCR for the characterization of reference materialsMegan H. Cleveland, Hua-Jun He, Mojca Milavec, Young-Kyung Bae, Peter M. Vallone, Jim F. Huggett, 2024, review article Abstract: Well-characterized reference materials support harmonization and accuracy when conducting nucleic acid-based tests (such as qPCR); digital PCR (dPCR) can measure the absolute concentration of a specific nucleic acid sequence in a background of non-target sequences, making it ideal for the characterization of nucleic acid-based reference materials. National Metrology Institutes are increasingly using dPCR to characterize and certify their reference materials, as it offers several advantages over indirect methods, such as UV-spectroscopy. While dPCR is gaining widespread adoption, it requires optimization and has certain limitations and considerations that users should be aware of when characterizing reference materials. This review highlights the technical considerations of dPCR, as well as its role when developing and characterizing nucleic acid-based reference materials.
Keywords: digital PCR, dPCR, reference materials, UV-spectroscopy
Published in DiRROS: 03.06.2024; Views: 511; Downloads: 188
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1652. International Conference Religious Conversions and Atheization in 20th Century Central and Eastern Europe : Koper, 22-24 April 20242024, other monographs and other completed works Abstract: Using a broad-based understanding of religious conversions which includes atheization and religious revival, the conference aims to address the issues related to religious transformation in Central and Eastern Europe in the 20th century and different types of religious agency that transcend established norms, legal frameworks and cultural expectations. The speakers will go beyond the usual concept of religious conversion as an exclusively individual, voluntary spiritual phenomenon and will examine conversions that embody the myriad motives, forms, and socio-political particularities which condition the changes of worldviews as a process that frequently encompasses obstacles, pragmatism and profound life alterations. The conference will contribute to wide-ranging but – especially in Central and Eastern Europe – largely unexplored topics from a more nuanced understanding of the mechanisms that drive religious change and religion-state relations.
Taking into account new research perspectives, scholars from the fields of history, anthropology, study of religions, ethnology, sociology, theology and political science will provide new insights into the following topics:
- religious conversions as mechanisms for personal gain, coercive tactics and the (gradual) disestablishment of religious institutions;
- state-imposed atheization processes;
- religious upsurges of (non-)dominant religions, relations between religion and state, and the (re)shaping of religious policies in practise.
Published in DiRROS: 03.06.2024; Views: 374; Downloads: 284
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1653. Distribution of toxigenic cyanobacteria in Alpine lakes and rivers as revealed by molecular screeningMaša Jablonska, Leonardo Cerasino, Adriano Boscaini, Camilla Capelli, Claudia Greco, Aleksandra Krivograd-Klemenčič, Ute Mischke, Nico Salmaso, Rainer Kurmayer, 2024, original scientific article Abstract: The increasing frequency of cyanobacteria blooms in waterbodies caused by ecosystem eutrophication could endanger human health. This risk can be mitigated by effective monitoring incorporating molecular methods. To date, most molecular studies on toxigenic cyanobacteria have been limited to microcystins (MCs), disregarding other cyanotoxins, to freshwater planktic habitats while ignoring benthic habitats, and to limited geographic areas (usually one or a few specific waterbodies). In this study, we used PCR-based methods including PCR product sequencing and chemical-analytical methods (LC-MS/MS) to screen many plankton (n = 123) and biofilm samples (n = 113) originating from 29 Alpine lakes and 18 rivers for their cyanotoxin production potential. Both mcyE (indicating MC synthesis) and anaC (indicating anatoxin (ATX) synthesis) gene fragments were able to qualitatively predict MC or ATX occurrence. The abundance of mcyE gene fragments was significantly related to MC concentrations in plankton samples (R2 = 0.61). mcyE gene fragments indicative of MC synthesis were most abundant in planktic samples (65 %) and were assigned to the genera Planktothrix and Microcystis. However, mcyE rarely occurred in biofilms of lakes and rivers, i.e., 4 % and 5 %, respectively, and were assigned to Microcystis, Planktothrix, and Nostoc. In contrast, anaC gene fragments occurred frequently in planktic samples (14 % assigned to Tychonema, Phormidium (Microcoleus), and Oscillatoria), but also in biofilms of lakes (49 %) and rivers (18 %) and were assigned to the genera Phormidium, Oscillatoria, and Nostocales. The cyrJ gene fragment indicating cylindrospermopsin synthesis occurred only once in plankton (assigned to Dolichospermum), while saxitoxin synthesis potential was not detected. For plankton samples, monomictic and less eutrophic conditions were positively related to mcyE/MC occurrence frequency, while oligomictic conditions were related to anaC/ATX frequency. The anaC/ATX frequency in biofilm was related to the lake habitats generally showing higher biodiversity as revealed from metabarcoding in a parallel study.
Keywords: cyanotoxins, planktic and benthic habitats, water quality monitoring, early warning, toxigenic cyanobacteria, European Alps
Published in DiRROS: 03.06.2024; Views: 511; Downloads: 316
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1654. The need for innovations to secure the future of artisanal mussel farming in the coastal sea of the Gulf of Trieste (Slovenia)Andreja Ramšak, Tine Bizjak, Uroš Robič, Manca Kovač Viršek, 2024, original scientific article Abstract: Mariculture along the Slovene coastline is mainly mussel culture operated as a family business; in one case, it is combined with sea bass farming. It started in the early 70 s with cultivation of Mediterranean mussels, while today, the two largest producers incorporate the cultivation of Venus clams and oysters as species with higher economic value on the market. Currently, all mussels and clams produced in Slovenia are sold in the Slovenian, Italian, Croatian and French markets. The production of Mediterranean mussels (Mytilus galloprovincialis) has increased steadily, with the main obstacles being a limited area for cultivation, occasional harmful algal blooms, predation by gilthead seabream and flatworms. In addition, more recent summer heatwaves negatively affected mussel production when seawater reached high temperatures at the thermal limits unsuitable for mussel growth. This study aimed to collect first-hand information about the current mussel production technology, processes and needs to identify opportunities for innovations that could benefit the entire sector (increased knowledge, production yield, reduced workload and effort with administrative issues). The study was based on a semi-structured questionnaire with the owners of mussel farms. Analysis of the questionnaire was supplemented with current knowledge from the field and provided comprehensive insight into a small sector of mussel farming along the Slovenian coast and its possible development in the future.
Keywords: mussel farming, Mediterranean mussel, aquaculture, socio-economic impact, Gulf of Trieste, Slovenia
Published in DiRROS: 03.06.2024; Views: 419; Downloads: 342
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1655. Exploring BPA alternatives : environmental levels and toxicity reviewOndrej Adamovsky, Ksenia J Groh, Anna Białk-Bielińska, Beate I. Escher, R. Beaudouin, Liadys Mora Lagares, K. E. Tollefsen, Alja Štern, Tina Eleršek, Marjan Vračko, Bojana Žegura, 2024, review article Abstract: Bisphenol A alternatives are manufactured as potentially less harmful substitutes of bisphenol A (BPA) that offer similar functionality. These alternatives are already in the market, entering the environment and thus raising ecological concerns. However, it can be expected that levels of BPA alternatives will dominate in the future, they are limited information on their environmental safety. The EU PARC project highlights BPA alternatives as priority chemicals and consolidates information on BPA alternatives, with a focus on environmental relevance and on the identification of the research gaps. The review highlighted aspects and future perspectives. In brief, an extension of environmental monitoring is crucial, extending it to cover BPA alternatives to track their levels and facilitate the timely implementation of mitigation measures. The biological activity has been studied for BPA alternatives, but in a non-systematic way and prioritized a limited number of chemicals. For several BPA alternatives, the data has already provided substantial evidence regarding their potential harm to the environment. We stress the importance of conducting more comprehensive assessments that go beyond the traditional reproductive studies and focus on overlooked relevant endpoints. Future research should also consider mixture effects, realistic environmental concentrations, and the long-term consequences on biota and ecosystems.
Keywords: BPA alternatives, biological activity, in silico, invertebrates, vertebrates, toxicity
Published in DiRROS: 03.06.2024; Views: 445; Downloads: 363
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1656. Protein G-quadruplex interactions and their effects on phase transitions and protein aggregationBikash Ranjan Sahoo, Vojč Kocman, Nathan Clark, Nikhil Myers, Xiexiong Deng, Ee L. Wong, Harry J. Yang, Anita Kotar, Bryan B. Guzman, Daniel Dominguez, Janez Plavec, James C. A. Bardwell, 2024, original scientific article Published in DiRROS: 03.06.2024; Views: 401; Downloads: 428
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1657. Dynamic susceptibility contrast enhanced (DSC) MRI perfusion and plasma cytokine levels in patients after tonic-clonic seizuresTatjana Filipovič, Katarina Šurlan Popović, Alojz Ihan, David B. Vodušek, 2017, original scientific article Abstract: Background Inflammatory events in brain parenchyma and glial tissue are involved in epileptogenesis. Blood concentration of cytokines is shown to be elevated after tonic-clonic seizures. As a result of inflammation, blood-brain barrier leakage occurs. This can be documented by imaging techniques, such is dynamic susceptibility contrast enhanced (DSC) MRI perfusion. Our aim was to check for postictal brain inflammation by studying DSC MRI perfusion and plasma level of cytokines. We looked for correlations between number and type of introducing seizures, postictal plasma level of cytokines and parameters of DSC MRI perfusion. Furthermore, we looked for correlation of those parameters and course of the disease over one year follow up. Patients and methods We prospectively enrolled 30 patients, 8%24 hours after single or repeated tonic-clonic seizures. Results 25 of them had normal perfusion parameters, while 5 had hyperperfusion. Patients with hyperperfusion were tested again, 3 months later. Two of 5 had hyperperfusion also on control measurements. Number of index seizures negatively correlated with concentration of proinflammatory cytokines IL-10, IFN-[gamma] and TNF-[alpha] in a whole cohort. In patients with hyperperfusion, there were significantly lower concentrations of antiinflammatory cytokine IL-4 and higher concentrations of proinflammatory TNF-[alpha]. Conclusions Long lasting blood- brain barrier disruption may be crucial for epileptogenesis in selected patients.
Keywords: cytokines, blood-brain barrier, tonic-clonic seizures
Published in DiRROS: 03.06.2024; Views: 396; Downloads: 245
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1658. Expression of LOC285758, a potential long non-coding biomarker, is methylation- dependent and correlates with glioma malignancy gradeAlenka Matjašič, Mara Popović, Boštjan Matos, Damjan Glavač, 2017, original scientific article Abstract: Background. Identifying the early genetic drivers can help diagnose glioma tumours in their early stages, before becoming malignant. However, there is emerging evidence that disturbance of epigenetic mechanisms also con- tributes to cell's malignant transformation and cancer progression. Long non-coding RNAs are one of key epigenetic modulators of signalling pathways, since gene expression regulation is one of their canonical mechanisms. The aim of our study was to search new gliomagenesis-specific candidate lncRNAs involved in epigenetic regulation. Patients and methods. We used a microarray approach to detect expression profiles of epigenetically involved lncRNAs on a set of 12 glioma samples, and selected LOC285758 for further qPCR expression validation on 157 glioma samples of different subtypes. To establish if change in expression is a consequence of epigenetic alterations we determined methylation status of lncRNA's promoter using MS-HRM. Additionally, we used the MLPA analysis for de- termining the status of known glioma biomarkers and used them for association analyses. Results. In all glioma subtypes levels of LOC285758 were significantly higher in comparison to normal brain reference RNA, and expression was inversely associated with promoter methylation. Expression substantially differs between astrocytoma and oligodendroglioma, and is elevated in higher WHO grades, which also showed loss of methylation. Conclusions. Our study revealed that lncRNA LOC285758 changed expression in glioma is methylation-dependent and methylation correlates with WHO malignancy grade. Methylation is also distinctive between astrocytoma I-III and other glioma subtypes and may thus serve as an additional biomarker in glioma diagnosis.
Keywords: glioma, epigenetics, methylation
Published in DiRROS: 03.06.2024; Views: 416; Downloads: 282
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1660. Nature-inspired substituted 3-(imidazol-2-yl) morpholines targeting human topoisomerase IIα : dynophore-derived discoveryBarbara Herlah, Matej Janežič, Iza Ogris, Simona Golič Grdadolnik, Katja Kološa, Sonja Žabkar, Bojana Žegura, Andrej Perdih, 2024, original scientific article Abstract: The molecular nanomachine, human DNA topoisomerase IIα, plays a crucial role in replication, transcription, and recombination by catalyzing topological changes in the DNA, rendering it an optimal target for cancer chemotherapy. Current clinical topoisomerase II poisons often cause secondary tumors as side effects due to the accumulation of double-strand breaks in the DNA, spurring the development of catalytic inhibitors. Here, we used a dynamic pharmacophore approach to develop catalytic inhibitors targeting the ATP binding site of human DNA topoisomerase IIα. Our screening of a library of nature-inspired compounds led to the discovery of a class of 3-(imidazol-2-yl) morpholines as potent catalytic inhibitors that bind to the ATPase domain. Further experimental and computational studies identified hit compound 17, which exhibited selectivity against the human DNA topoisomerase IIα versus human protein kinases, cytotoxicity against several human cancer cells, and did not induce DNA double-strand breaks, making it distinct from clinical topoisomerase II poisons. This study integrates an innovative natural product-inspired chemistry and successful implementation of a molecular design strategy that incorporates a dynamic component of ligand-target molecular recognition, with comprehensive experimental characterization leading to hit compounds with potential impact on the development of more efficient chemotherapies.
Keywords: topoisomerase II, catalytic inhibitors, chemotherapy, DNA damage, cancer
Published in DiRROS: 03.06.2024; Views: 564; Downloads: 327
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