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Fast and accurate multiplex identification and quantification of seven genetically modified soybean lines using six-color digital PCR
Alexandra Bogožalec Košir, Sabine Muller, Jana Žel, Mojca Milavec, Allison C. Mallory, David Dobnik, 2023, izvirni znanstveni članek

Povzetek: The proliferation of genetically modified organisms (GMOs) presents challenges to GMO testing laboratories and policymakers. Traditional methods, like quantitative real-time PCR (qPCR), face limitations in quantifying the increasing number of GMOs in a single sample. Digital PCR (dPCR), specifically multiplexing, offers a solution by enabling simultaneous quantification of multiple GMO targets. This study explores the use of the Naica six-color Crystal dPCR platform for quantifying five GM soybean lines within a single six-plex assay. Two four-color assays were also developed for added flexibility. These assays demonstrated high specificity, sensitivity (limit of detection or LOD < 25 copies per reaction) and precision (bias to an estimated copy number concentration <15%). Additionally, two approaches for the optimization of data analysis were implemented. By applying a limit-of-blank (LOB) correction, the limit of quantification (LOQ) and LOD could be more precisely determined. Pooling of reactions additionally lowered the LOD, with a two- to eight-fold increase in sensitivity. Real-life samples from routine testing were used to confirm the assays’ applicability for quantifying GM soybean lines in complex samples. This study showcases the potential of the six-color Crystal dPCR platform to revolutionize GMO testing, facilitating comprehensive analysis of GMOs in complex samples.
Ključne besede: digital PCR, dPCR, quantification, multiplexing, genetically modified organisms, 6-color system, virus diagnostics, virology
Objavljeno v DiRROS: 29.03.2024; Ogledov: 201; Prenosov: 80
.pdf Celotno besedilo (1,83 MB)
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13.
Inhibition of cathepsin X enzyme influences the immune response of THP-1 cells and dendritic cells infected with Helicobacter pylori
Miha Skvarč, David Štubljar, Andreja Nataša Kopitar, Samo Jeverica, Bojan Tepeš, Janko Kos, Alojz Ihan, 2013, izvirni znanstveni članek

Povzetek: Background. The immune response to Helicobacter pylori importantly determines the outcome of infection as well as the success of eradication therapy. We demonstrate the role of a cysteine protease cathepsin X in the immune response to H. pylori infection. Materials and methods. We analysed how the inhibition of cathepsin X influenced the immune response in experiments when THP-1 cells or dendritic cells isolated from patients were stimulated with 48 strains of H. pylori isolated from gastric biopsy samples of patients which had problems with the eradication of bacteria. Results. The experiments, performed with the help of a flow cytometer, showed that the expression of Toll-like receptors (TLRs), especially TLR-4 molecules, on the membranes of THP-1 cells or dendritic cells was higher when we stimulated cells with H. pylori together with inhibitor of cathepsin X 2F12 compared to THP-1 cells or dendritic cells stimulated with H. pylori only, and also in comparison with negative control samples. We also demonstrated that when we inhibited the action of cathepsin X in THP-1 cells, the concentrations of pro-inflammatory cytokines were lower than when THP-1 cell were stimulated with H. pylori only. Conclusions. We demonstrated that inhibition of cathepsin X influences the internalization of TLR-2 and TLR-4. TLR-2 and TLR-4 redistribution to intra-cytoplasmic compartments is hampered if cathepsin X is blocked. The beginning of a successful immune response against H. pylori in the case of cathepsin X inhibition is delayed.
Ključne besede: cathepsin X, macrophage, dendritic cells
Objavljeno v DiRROS: 22.03.2024; Ogledov: 131; Prenosov: 60
.pdf Celotno besedilo (564,38 KB)
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The automorphism group of the zero-divisor digraph of matrices over an antiring
David Dolžan, Gabriel Verret, 2024, izvirni znanstveni članek

Povzetek: We determine the automorphism group of the zero-divisor digraph of the semiring of matrices over an antinegative commutative semiring with a finite number of zero-divisors.
Ključne besede: automorphism group of a graph, zero-divisor graph, semirings
Objavljeno v DiRROS: 16.02.2024; Ogledov: 199; Prenosov: 72
.pdf Celotno besedilo (268,74 KB)
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17.
Effect of demineralization and ball milling treatments on the properties of Arundo donax and olive stone-derived biochar
Mariem Zouari, Laetitia Sarah Jennifer Marrot, David Brian DeVallance, 2024, izvirni znanstveni članek

Povzetek: The structural and physio-chemical properties of biochar are crucial to determining biochar’s quality and the adequate application. Specifically, the large porosity of biochar has been known as a favorable feature, especially for environmental remediation. In this regard, physical and chemical modifications have been used to improve biochar’s porosity which requires high-energy consumption and involves chemical agents. The objective of this study was to prepare biochar with developed porosity using mild treatments. Arundo donax and olive stone were demineralized by a water-washing method. Treated and non-treated biomasses were pyrolyzed, and part of the derived samples was subjected to wet ball milling. Samples were characterized with proximate, Fourier transform infrared, particle size, and physisorption analyses. The effect of demineralization depended on the biomass type, as ash reduction only influenced Arundo donax-derived biochar, which was attributed to the difference in initial ash content that was relatively low for olive stone. The carbonization yield decreased by 46% for the Arundo donax biomass after demineralization. Moreover, demineralization expanded the surface area and total pore volume of the Arundo donax biochar. The ball milling was effective in producing micro-sized biochar particles with a mean size ranging between 30 ± 2 µm and 42 ± 2 µm and between 13 ± 1 µm and 22 ± 2 µm for Arundo donax and olive stone without and with demineralization, respectively. Ball milling increased the surface area of non-demineralized Arundo donax by 47% and demineralized Arundo donax by 124%. Additionally, ball milling increased the surface area of non-demineralized olive stone by 65% and demineralized olive stone by 62%.
Ključne besede: biomass, carbonisation, porosity
Objavljeno v DiRROS: 13.02.2024; Ogledov: 184; Prenosov: 102
.pdf Celotno besedilo (2,97 MB)
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18.
A web-application that extends functionality of medical device for tumor treatment by means of electrochemotherapy
Ivan Pavlović, Peter Kramar, Selma Čorović, David Cukjati, Damijan Miklavčič, 2004, izvirni znanstveni članek

Povzetek: Electrochemotherapy (ECT) is a novel method for efficient tumor treatment in clinical environment. It combines local drug delivery and application of shorthigh voltage pulses, which permeabilize the plasma membrane by electroporation. Drug can enter only the cells with permeabilzed membrane. Recently, medical device CliniporatorTM for controlled electroporation was developed. Here, we present a web-application that extends the functionality of this medical device. The aim of the application is to collect, store and toallow the analysis of every ECT application using this medical device. The application helps transferring data collected by devčce during the electroporation process to the central database, and enables filling of medical records through the web forms. The application is based on technologies ASP, HTML, Flash, JavaScript, XML and others. The application main advantages are easy and rapid data access, scalability and independence of client computer operating system as well as easy application debugging and upgrading.
Ključne besede: neoplasms- drug therapy, drug delivery systems, elektroporation instrumentation
Objavljeno v DiRROS: 07.02.2024; Ogledov: 215; Prenosov: 54
.pdf Celotno besedilo (133,39 KB)

19.
Extending BIM for air quality monitoring
Michael Nicolas Mrissa, Jan Vcelak, László Hajdu, Balázs Dávid, Miklós Ferenz Krész, Jakub Michal Sandak, Anna Malgorzata Sandak, Rok Kanduti, Monika Varkonji, Anja Jutraž, Katja Malovrh Rebec, 2020, objavljeni znanstveni prispevek na konferenci

Povzetek: As we spend more than 90% of our time inside buildings, indoor environmental quality is a major concern for healthy living. Recent studies show that almost 80% of people in European countries and the United States suffer from SBS (Sick Building Syndrome), which affects physical health, productivity and psychological well-being. In this context, environmental quality monitoring provides stakeholders with crucial information about indoor living conditions, thus facilitating building management along its lifecycle, from design, construction and commissioning to usage, maintenance and end-of-life. However, currently available modelling tools for building management remain limited to static models and lack integration capacities to efficiently exploit environmental quality monitoring data. In order to overcome these limitations, we designed and implemented a generic software architecture that relies on accessible Building Information Model (BIM) attributes to add a dynamic layer that integrates environmental quality data coming from deployed sensors. Merging sensor data with BIM allows creation of a digital twin for the monitored building where live information about environmental quality enables evaluation through numerical simulation. Our solution allows accessing and displaying live sensor data, thus providing advanced functionality to the end-user and other systems in the building. In order to preserve genericity and separation of concerns, our solution stores sensor data in a separate database available through an application programming interface (API), which decouples BIM models from sensor data. Our proof-of-concept experiments were conducted with a cultural heritage building located in Bled, Slovenia. We demonstrated that it is possible to display live information regarding environmental quality (temperature, relative humidity, CO2, particle matter, light) using Revit as an example, thus enabling end-users to follow the conditions of their living environment and take appropriate measures to improve its quality
Ključne besede: Building Information Model, internet of things, environmental quality monitoring, healthy living
Objavljeno v DiRROS: 19.01.2024; Ogledov: 245; Prenosov: 105
.pdf Celotno besedilo (18,89 MB)
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20.
Efficacy of trapping protocols for Agrilus jewel beetles : a multi-country assessment
Giacomo Santoiemma, David Williams, Everett G. Booth, Giacomo Cavaletto, James Connell, Gianfranco Curletti, Maarten De Groot, Sarah M. Devine, Abigail Enston, Joseph A. Francese, Eva Groznik, Tine Hauptman, 2024, izvirni znanstveni članek

Povzetek: The genus Agrilus is one of the most diverse insect genera worldwide. The larval feeding activity causes extensive damage in both forests and orchards. In addition, more than 30 species have been introduced outside their native range so far, including the emerald ash borer Agrilus planipennis Fairmaire. Thus, the availability of efficient trapping protocols for early detection of Agrilus species at entry points is of utmost importance. In this study we tested whether trapping protocols developed for surveillance of A. planipennis in North America were also effective for other Agrilus species. In particular, through a multi-country assessment we compared the efficacy of detecting Agrilus species on: (i) green glue-coated prism traps vs. green Fluon-coated multi-funnel traps when baited with the green leaf volatile (Z)-3-hexenol or left unbaited; and (ii) green multi-panel traps vs. green multi-panel traps baited with dead adult Agrilus beetles (decoys). A total of 23,481 individuals from 45 Agrilus species were caught. Trap design significantly affected both species richness and abundance of Agrilus species in several of the countries where the trapping experiments were carried out, and green prism traps outperformed green multi-funnel traps in most cases. On the contrary, the addition of a (Z)-3-hexenol lure or dead adult beetle decoys on to traps did not improve trap catches. Our study highlights that reliable trap models to survey Agrilus species are already available, but also that there is the clear need to further investigate chemical ecology of Agrilus species to develop semiochemical lures that can improve detection efficacy.
Ključne besede: biosecurity, early detection, forest pests, lures, traps, visual stimuli, Agrilus, beetles
Objavljeno v DiRROS: 19.01.2024; Ogledov: 282; Prenosov: 214
.pdf Celotno besedilo (3,11 MB)
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