1. Nanosecond electric pulses are equally effective in electrochemotherapy with cisplatin as microsecond pulsesAngelika Vižintin, Stefan Marković, Janez Ščančar, Jerneja Kladnik, Iztok Turel, Damijan Miklavčič, 2022, original scientific article Keywords: electroporation, electrochemotherapy, nanosecond pulses, cisplatin
Published in DiRROS: 25.07.2024; Views: 168; Downloads: 124
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2. Loop-mediated isothermal amplification : rapid molecular detection of virulence genes associated with avian pathogenic Escherichia coli in poultryPolona Kogovšek, Jerneja Ambrožič, Alenka Dovč, Tanja Dreo, Hristo Hristov, Uroš Krapež, Maja Ravnikar, Brigita Slavec, Marjetka Lotrič, Jana Žel, Olga Zorman-Rojs, 2018, original scientific article Abstract: Infections with pathogenic Escherichia colican lead to different animal- and human-associated diseases. E. coliinfections are common in intensive poultry farming, and important economic losses can be expected during infections with avian pathogenic E. coli(APEC) strains followed by colibacillosis. Loop-mediated isothermal amplification (LAMP) assays were developed for rapid detection of 3 APEC-associated virulence genes: sitA, traT, and ompT. All 3 LAMP assays are shown to be specific, repeatable, and reproducible. High sensitivities of the assays are shown, where as few as 1,000 bacterial cells/mL can be detected in different matrices. On-site applicability of this LAMP method is demonstrated through testing of different sample types, from animal swabs and tissues, and from environmental samples collected from 6 commercial poultry farms. All 3 virulence genes were detected at high rates (above 85%) in samples from layer and broiler chickens with clinical signs and, interestingly, high prevalence of those genes was detected also in samples collected from clinically healthy broiler flock (above 75%) while lower prevalence was observed in remaining 3 clinically healthy chicken flocks (less than 75%). Importantly, these virulence genes were detected in almost all of the air samples from 11 randomly selected poultry houses, indicating air as an important route of E. colispread. Three LAMP assays that target APEC-associated virulence genes are shown to be sensitive and robust and are therefore applicable for rapid on-site testing of various sample types, from animal swabs to air. This on-site LAMP testing protocol offers rapid diagnostics, with results obtained in <35 min, and it can be applied to other important microorganisms to allow the required prompt measures to be taken.
Keywords: APEC virulence genes, isothermal amplification LAMP, on-site detection, colibacillosis
Published in DiRROS: 24.07.2024; Views: 188; Downloads: 78
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3. Morphological features of breast cancer circulating tumor cells in blood after physical and biological type of isolationTanja Jesenko, Živa Pišljar, Cvetka Grašič-Kuhar, Maja Čemažar, Urška Matkovič, Simona Miceska, Jerneja Varl, Anamarija Kuhar, Veronika Kloboves-Prevodnik, 2021, original scientific article Abstract: Background. Circulating tumor cells (CTCs) have become an important biomarker in breast cancer. Different iso-lation tech-niques based on their biological or physical features were established. Currently, the most widely used methods for visualization after their separation are based on immunofluorescent staining, which does not provide the information on the morphology.Materials and methods. The aim of this study was to evaluate how two different separation techniques affect cell morphology and to analyse cell morphology with techniques used in routine cytopathological laboratory. A direct side-by-side comparison of physical (Parsortix%) and biological (MACS%) separation technique was performed.Results. In the preclinical setting, both isolation techniques retained the viability and antigenic characteristics of MCF7 breast cancer cells. Some signs of degeneration such as cell swelling, cytoplasmic blebs, villous projections and vacuolization were observed. In metastatic breast cancer patient cohort, morphological features of isolated CTCs were dependent on the separation technique. After physical separation, CTCs with preserved cell morphology were detected. After biological separation the majority of the isolated CTCs were so degenerated that their identity was difficult to confirm.Conclusions. Taken together, physical separation is a suitable technique for detection of CTCs with preserved cell morphology for the use in a routine cytopathological laboratory.
Keywords: circulating tumor cells, breast cancer, morphology
Published in DiRROS: 22.07.2024; Views: 223; Downloads: 186
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4. Biodiversity offset mechanisms and compensation for loss from exceptional to popular : rediscovering environmental lawJerneja Penca, 2024, original scientific article Abstract: The use of compensatory mechanisms for biodiversity conservation, also known as biodiversity offsets, has increased significantly in recent decades. The Kunming Montreal Global Biodiversity Framework mentions them as an innovative scheme in support of substantially and progressively increasing the level of financial resources for biodiversity conservation. This article traces the origin of compensatory mechanisms in international environmental law and their development in transnational biodiversity governance. The article points to the shifts in the application of the biodiversity offsets: from the context of wetlands to other habitats and ecosystems; from its use in intergovernmental conventions to an increasing number of transnational (business) networks; and from an instrument of last resort to a source of additional funding for biodiversity conservation. In the evolution, compensatory mechanisms have been decoupled from their original purpose as an exceptional mitigation measure and a strong focus of environmental law on the preventive function. The increased rhetoric of commitment to no net loss, net gain, restoration, and the mitigation hierarchy has not been matched by an improved status of wetlands and other ecosystems. The processes within the biodiversity conventions (Ramsar and CBD) have accepted an ongoing destruction of nature and limited the role of environmental law to minimizing harmful impacts on nature and consolidating the decline, rather than shaping socio-ecological outcomes. An ambiguous position about the spread of compensatory mechanisms has been part and parcel of this; biodiversity conventions have neither endorsed nor distanced themselves from the application, promotion, and justification of compensatory mechanisms. To maintain the integrity of environmental law, the rules that prevent biodiversity loss need to be emphasised and enforced.
Keywords: biodiversity offsets, compensatory mechanisms, biodiversity governance, environmental law, preventive principles
Published in DiRROS: 17.07.2024; Views: 665; Downloads: 223
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5. Evaluation of the training program for p16/ Ki-67 dual immunocytochemical staining interpretation for laboratory staff without experience in cervical cytology and immunocytochemistryVeronika Kloboves-Prevodnik, Živa Pohar-Marinšek, Janja Zalar, Hermina Rozina, Nika Kotnik, Tine Jerman, Jerneja Varl, Urška Ivanuš, 2020, original scientific article Abstract: ackground p16/Ki-67 dual immunocytochemical staining (DS) is considered easy to interpret if evaluators are properly trained, however, there is no consensus on what constitutes proper training. In the present study we evaluated a protocol for teaching DS evaluation on students inexperienced in cervical cytology. Methods Initial training on 40 DS conventional smears was provided by a senior cytotechnologist experienced in such evaluation. Afterwards, two students evaluated 118 cases. Additional training consisted mainly of discussing discrepant cases from the first evaluation and was followed by evaluation of new 383 cases. Agreement and accuracy of students' results were compared among the participants and to the results of the reference after both evaluations. We also noted time needed for evaluation of one slide as well as intra-observer variability of the teacher's results. Results At the end of the study, agreement between students and reference was higher compared to those after initial training (overall percent agreement [OPA] 81.4% for each student, kappa 0.512 and 0.527 vs. OPA 78.3% and 87.2%, kappa 0.556 and 0.713, respectively). However, accuracy results differed between the two students. After initial training sensitivity was 4.3% points and 2.9% points higher, respectively compared to the reference, while specificity was 30.6% points and 24.4% points lower, respectively, compared to the reference. At the end of the study, the sensitivity reached by one student was the same as that of the reference, while it was 2.6% points lower for the other student. There was a statistically significant difference in specificity between one student and the reference and also between students (16.7 and 15.1% points). Towards the end of the study, one student needed 5.2 min for evaluating one slide while the other needed 8.2 min. The intra-observer variability of the senior cytotechnologist was in the range of "very good" in both arms of the study. Conclusions In teaching DS evaluation, the students' progress has to be monitored using several criteria like agreement, accuracy and time needed for evaluating one slide. The monitoring process has to continue for a while after students reach satisfactory results in order to assure a continuous good performance. Monitoring of teacher's performance is also advisable.
Keywords: cervical cytology, cervical cancer, immunocytochemistry, accuracy
Published in DiRROS: 11.07.2024; Views: 252; Downloads: 186
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6. Clinical relevance of the borderline results of the Hybrid Capture 2 High-Risk HPV DNA assay with cervical samples collected in Specimen Transport MediumJerneja Varl, Urška Ivanuš, Živa Pohar-Marinšek, Tine Jerman, Anja Oštrbenk Valenčak, Mario Poljak, Veronika Kloboves-Prevodnik, 2019, original scientific article Keywords: Hybrid Capture 2, cervical cancer, HPV test, borderline results
Published in DiRROS: 05.07.2024; Views: 262; Downloads: 132
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7. Building competences for researchers working towards ocean sustainabilityJerneja Penca, Andrea Barbanti, Christopher Cvitanovic, Amel Hamza-Chaffai, Ahmed Elshazly, Jean-Baptiste Jouffray, Nezha Mejjad, Melita Mokos, 2024, original scientific article Abstract: The challenges of achieving just, equitable and sustainable ocean futures require a new type of transdisciplinary and action-oriented science that integrates across disciplines and knowledge systems. Scientists and researchers in academia, industry or government, who contribute to knowledge creation, innovation, and policy development for the ocean, must be empowered with a fresh set of competences. This paper maps the knowledge, skills, and attitudes required to enable such a shift. The proposed skillset serves as a foundation for the design and operationalisation of modern training for ocean sustainability and is envisaged to be used by researchers both individually and in teams. It also highlights the potential for career diversification beyond the traditional ’blue jobs’ legitimated by existing sectors. To ensure the short-term practical implementation of the competence framework, self-awareness and self-reflection are encouraged among learners and teachers, along with pragmatic actions to overcome barriers to transdisciplinarity. For long-term impact, system interventions will be necessary to improve organisations’ readiness to absorb and valorise researchers trained in this new framework. This will require re-training the current pedagogical workforce as well as reframing existing knowledge systems and incentives.
Keywords: sustainable blue economy, sustainability science, marine sciences, transformative research, interdisciplinary sciences
Published in DiRROS: 07.05.2024; Views: 311; Downloads: 399
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8. Safety and efficacy of IL-12 plasmid DNA transfection into pig skin : supportive data for human clinical trials on gene therapy and vaccinationUrša Lampreht Tratar, Tanja Jesenko, Maša Omerzel, Alenka Seliškar, Urban Stupan, Mihajlo Djokić, Jerneja Sredenšek, Blaž Trotovšek, Gregor Serša, Maja Čemažar, 2024, original scientific article Abstract: Gene electrotransfer (GET) of plasmids encoding interleukin 12 (IL-12) has already been used for the treatment of various types of tumors in human oncology and as an adjuvant in DNA vaccines. In recent years, we have developed a plasmid encoding human IL-12 (phIL12) that is currently in a phase I clinical study. The aim was to confirm the results of a non-clinical study in mice on pharmacokinetic characteristics and safety in a porcine model that better resembled human skin. The GET of phIL12 in the skin was performed on nine pigs using different concentrations of plasmid phIL12 and invasive (needle) or noninvasive (plate) types of electrodes. The results of our study demonstrate that the GET of phIL-12 with needle electrodes induced the highest expression of IL-12 at the protein level on day 7 after the procedure. The plasmid was distributed to all tested organs; however, its amount decreased over time and was at a minimum 28 days after GET. Based on plasmid copy number and expression results, together with blood analysis, we showed that IL-12 GET is safe in a porcine animal model. Furthermore, we demonstrated that pigs are a valuable model for human gene therapy safety studies.
Keywords: interleukin 12, gene electrotransfer, immunotherapy
Published in DiRROS: 18.04.2024; Views: 437; Downloads: 193
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9. Elektronske cigarete, ogrevani tobačni izdelki, nikotinske vrečke in brezdimni tobačni izdelki : kaj o teh izdelkih s tobakom ali nikotinom povedo raziskave?Helena Koprivnikar, Jerneja Farkaš-Lainščak, 2023, review article Abstract: V Sloveniji je kajenje tobaka še vedno med najpomembnejšimi preprečljivimi dejavniki tveganja za smrt in na prvem mestu za izgubljena zdrava leta življenja. V Sloveniji in številnih razvitih državah beležimo zniževanje prevalence kajenja. Industrija je za ohranitev oziroma večanje prodaje in dobička dala na trg različne vrste izdelkov s tobakom ali nikotinom, kot so elektronske cigarete, ogrevani tobačni izdelki in nikotinske vrečke, ki so prisotni tudi v Sloveniji. Vse bolj priljubljeni v Sloveniji postajajo tudi brezdimni tobačni izdelki. V bibliografski podatkovni zbirki PubMed smo poiskali sistematične preglede in metaanalize o vplivih na zdravje pri uporabi teh izdelkov, poiskali poročila mednarodnih organizacij o njihovem vplivu na zdravje in okolje ter uporabili najnovejše dostopne podatke iz slovenskih nacionalnih raziskav za prikaz razširjenosti njihove uporabe v Sloveniji. Ogrevani tobačni izdelki, elektronske cigarete in nikotinske vrečke izpostavijo uporabnika znatnim ravnem zdravju škodljivih snovi. Dolgoročni učinki njihove uporabe na zdravje še niso ustrezno raziskani, saj so v uporabi še prekratek čas, dostopni podatki pa kažejo, da imajo škodljive učinke na zdravje, nekateri že kratkoročno, in da bodo verjetno privedli do pomembnih posledic za zdravje pri dolgoročni uporabi. O manjši škodljivosti teh izdelkov v primerjavi s kajenjem cigaret ni možno govoriti, saj je na voljo premalo kakovostnih raziskav, poznani pa so škodljivi učinki dolgoročne uporabe brezdimnih tobačnih izdelkov. Vsi omenjeni izdelki se pogosto promovirajo tudi kot učinkoviti pripomočki za opuščanje kajenja, a tudi na tem področju ni na voljo zadosti kakovostnih raziskav, da bi lahko sklepali zaključke o njihovi vlogi v opuščanju kajenja, zato se jih v te namene ne priporoča. Vsi omenjeni izdelki vsebujejo nikotin (izjema so elektronske cigarete brez nikotina) in uporabnika zasvojijo. Uporaba izdelkov z nikotinom med mladimi pa je povezana še z dodatnimi pomembnimi tveganji; poleg zasvojenosti, ki se razvije hitreje in močneje kot pri odraslih, so tu še škodljivi učinki na razvoj možganov, pri elektronskih cigaretah in brezdimnih tobačnih izdelkih pa tudi zvišano tveganje za kajenje cigaret v prihodnosti. Podatki za Slovenijo kažejo, da uporaba teh izdelkov med mladimi narašča s starostjo ter da med mladimi odstotki uporabnikov večinoma presegajo odstotek med populacijo, starejšo od 25 let. V zadnjih letih odstotki uporabnikov elektronskih cigaret in brezdimnih tobačnih izdelkov med mladostniki naraščajo izraziteje kot med odraslimi. Ključna je čimprejšnja strožja zakonodajna ureditev teh izdelkov, da preprečimo in zmanjšamo njihovo uporabo, predvsem med mladimi.
Keywords: tobak, nikotin, elektronski sistemi za dovajanje nikotina, tobačni izdelki, mladostniki
Published in DiRROS: 05.02.2024; Views: 556; Downloads: 116
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