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Povzetek: Plant genetic engineering represents an important aspect of modern agriculture, and new geneticallymodified (GM) crop varieties are entering the market on a regular basis. This necessitates thedevelopment of high throughput multi-target analytical methods to detect and quantify theirpresence for regulatory compliance. In this study, we present a multiplex dPCR method for discrimi-native quantification of 19 GM soybean events and the lectin (Le1) endogene on a nanowell plate-based all-in-one dPCR system. The method consists of four 5-plex assays, taking advantage of theplatform’s multiple fluorescence detection channels. The assays complied with the minimum perfor-mance requirements in terms of specificity, trueness, precision, sensitivity and dynamic range,making them suitable for use in routine detection and quantification of GM crops. This methodrepresents the most comprehensive multi-target GM soybean quantification approach to date with-out the need for prior screening and features a simplified workflow, making it suitable for widespreadadoption. Our study sets a precedent for rapid and straightforward development of multiplex dPCRGM crop quantification assays to address the evolving demands of regulatory monitoring.
Ključne besede: genetically modified (GM) crops, transgenic, soybean, Glycine max (L.) merr, quantification, digital PCR (dPCR), multiplex, multi-target
Objavljeno v DiRROS: 19.03.2026; Ogledov: 252; Prenosov: 208
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Povzetek: The European Union (EU) imposes strict regulations on the presence of genetically modified (GM) material in food and feed, requiring thorough testing of samples for various GM lines. Although traditional quantitative real-time PCR (qPCR) methods are sensitive and robust, they are not cost-effective for managing large numbers of GM events due to their limited multiplexing capabilities. Conversely, digital PCR (dPCR) is capable of robust quantitative multiplexing in addition to other benefits such as absolute quantification and better tolerance of PCR inhibitors. In this context, we present a protocol for multiplex quantification of 19 GM soybean lines using dPCR as an improvement over the currently used simplex qPCR approach. This method enables simple and robust quantification of common GM soybean lines with a relatively low number of reactions.
Ključne besede: genetically modified soybean, GMO, multiplexing, food and feed, food regulation, digital PCR (dPCR)
Objavljeno v DiRROS: 26.11.2025; Ogledov: 1031; Prenosov: 179
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Povzetek: Cell therapy product (CTP) developers face the significant challenge of developing appropriate potency tests for their CTPs. A review of the known potency tests used for the 31 United States Food and Drug Administration-approved CTPs (US FDA) can guide developers in designing effective potency tests for future CTPs. Data on these tests were primarily collected from publicly available regulatory documentation on the US FDA website (90%) as well as other sources (literature, company communications, etc.). Based on these data, an estimated 104 total potency tests have been used for the 31 CTPs. Of these, 33 are redacted (32%), leaving 71 non-redacted potency tests. On average, each CTP has 3.4 potency tests (standard deviation 2.0). The 71 non-redacted potency tests were categorized into 5 bins: “Viability and count” (37 tests, 52%), “Expression” (19 tests, 27%), “Bioassays” (7 tests, 7%), “Genetic modification” (6 tests, 9%) and “Histology” (2 tests, 3%). Measurements of gene or protein expression were used by 20 of the 31 CTPs (65%), and 19 CTPs (61%) used measurements of cell viability or cell count as a potency test. “Viability and count” and “Expression” are the two tests that have most often been used together for the same product, occurring for 16 CTPs (52%). It is unclear if bioassays are commonly used as potency tests since only 7 of 31 CTPs (23%) reported bioassays as potency tests. However, due to redactions, as many 24 (77%) CTPs could potentially have a bioassay as a potency test. Additionally, 26 of the 31 CTPs (84%) cite physicochemical assays (non-bioassays) as a potency test. This analysis of potency tests for approved CTPs provides valuable insights for developing potency tests for new CTPs.
Ključne besede: cell therapy product, potency, potency test, regenerative medicine
Objavljeno v DiRROS: 10.04.2025; Ogledov: 741; Prenosov: 574
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Povzetek: Innovative technological solutions are needed for water decontamination to combat the diverse pollutants present in water systems, as no single optimal decontamination technique is appropriate for all circumstances. Vacuum-ultraviolet (V-UV) radiation is a source of energetic photons that break molecular bonds, producing a plethora of chemically reactive agents, most notably OH● radicals, which can cause the degradation of harmful pollutants. Low-pressure gaseous plasma is a good source of V-UV radiation; however, its application to liquid water poses challenges. We constructed an inductively coupled radiofrequency plasma to produce high-intensity V-UV radiation, which was applied to contaminated water via a V-UV-transparent window. Plasma was sustained in hydrogen, as it produces the highest V-UV intensity among all gases at selected discharge parameters. Bacteriophage MS2 was used as an indicator of microbial decontamination efficiency. Reactive oxygen and nitrogen species were measured at various treatment setups to quantify their effect on MS2 inactivation and elucidate the primary inactivation factors. At optimal conditions, the concentration of active virus dropped by 9 log10 PFU/mL in 60 s. The optimal experimental setup was then used to treat bacteria E. coli, S. aureus, antibiotic tetracycline, and synthetic dye methylene blue as representatives of other types of pollutants, all of which were effectively removed/degraded within 10 min of treatment. A comparison of energy efficiency (EEO) to other disinfection setups was made for bacteriophage inactivation. With a low EEO value, we showcase the potential of this technique for further work in this field.
Ključne besede: water treatment, radical
Objavljeno v DiRROS: 07.02.2025; Ogledov: 1134; Prenosov: 549
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Povzetek: During the COVID-19 pandemic, face masks were the first line of defense against the spread of infection. However, infectious viruses may remain on medical textiles, potentially serving as an additional source of infection. Due to their chemical inertness, many textiles cannot be enhanced with antiviral functionalities. Through treatment with low-pressure gaseous plasma, we have activated the surface of a medical-grade melt-blown, non-woven polypropylene textile so that it can absorb sodium dodecyl sulfate, an antimicrobial surfactant. Within two hours of contact time, the functionalized textile has been able to inactivate over 7 log10 PFU mL−1 of bacteriophage phi6, a surrogate of enveloped viruses such as SARS-CoV-2, and it has retained its antiviral properties for over 100 days. The functionalized material has not disrupted facial mask filtration efficiency or breathability. In addition, the in vitro biocompatibility testing in accordance with ISO 10993-5 for testing of medical devices has demonstrated that the selected formulation causes no adverse effects on the mouse fibroblast cell line L-929. With the treatment processes that have been completed within seconds, the method seems to have great potential to produce antiviral textiles against future outbreaks.
Ključne besede: surgical face masks, plasma functionalization, antiviral materials, virus filtration, breathability
Objavljeno v DiRROS: 07.10.2024; Ogledov: 1366; Prenosov: 947
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