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Abstract: Muscle tissues of Mullus barbatus and Merluccius merluccius were analyzed for the presence of selected trace elements (As, Hg, Cd, and Pb) to determine the value of the daily intake of trace elements from fish consumption and to assess the risk to human health. The mean concentrations of As in the muscle tissue of M. barbatus and M. merluccius for the entire period were 19.689 mg/kg wet weight (ww) and 8.356 mg/kg ww, Hg 0.497 mg/kg ww and 0.153 mg/kg ww, and Pb 0.031 mg/kg ww and 0.025 mg/kg ww, respectively. The concentrations of Cd in all fish sampled were below the detection limit (<0.02 mg/kg ww). The evaluation of the potential health risk assessments based on the target hazard quotient (THQ) and estimated daily intake (EDI) indicated that the intake of As in both fish species and Hg for M. barbatus could pose an appreciable risk to human health. The calculated hazard index (HI) was higher than 1 for both fish species. The continuous monitoring of trace elements’ concentrations in fish is strongly recommended, as the results demonstrate potential health risks due to the presence of As and Hg.
Keywords: trace elements, Mullus barbatus, Merluccius merluccius, human health risk assessment, Montenegro, Adriatic Sea, hydrobiology
Published in DiRROS: 12.07.2024; Views: 9; Downloads: 3
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Keywords: Campylobacter jejuni, luxS, intercellular signaling, transcriptome, proteome, metabolism, stress response
Published in DiRROS: 12.07.2024; Views: 21; Downloads: 7
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Abstract: Purpose: Glioblastoma (GBM) is the most common primary brain tumour and one of the deadliest cancers. In addition to late diagnosis and inadequate treatment, the extremely low survival rate is also due to the lack of appropriate therapeutic biomarkers and corresponding therapeutic agents. One of the potential therapeutic biomarkers is the intermediate filament vimentin, which is associated with epithelial-mesenchymal transition (EMT). The purpose of this study was to analyse the effect of the anti-vimentin nanobody Nb79 on cell invasion in vitro and in vivo. To further our understanding of the mechanism of action, we investigated the association between Nb79 and EMT in GBM and GBM stem cells by analysing the expression levels of key EMT-related proteins. Methods: The expression of vimentin in glioma tissues and cells was determined by RT-qPCR. An invasion assay was performed on differentiated glioblastoma cell line U-87 MG and stem cell line NCH421k in vitro as well as in vivo in zebrafish embryos. The effect of Nb79 on expression of EMT biomarkers beta-catenin, vimentin, ZEB-1 and ZO1 was determined by Western blot and immunocytochemistry. Results: Our study shows that vimentin is upregulated in glioblastoma tissue compared to lower grade glioma and non-tumour brain tissue. We demonstrated that treatment with Nb79 reduced glioblastoma cell invasion by up to 64% in vitro and up to 21% in vivo. In addition, we found that the tight junction protein ZO-1 had higher expression on the cell membrane, when treated with inhibitory anti-vimentin Nb79 compared to control. Conclusion: In conclusion, our results suggest that anti-vimentin nanobody Nb79 is a promising tool to target glioblastoma cell invasion.
Keywords: glioblastoma, vimentin, nanobody
Published in DiRROS: 12.07.2024; Views: 12; Downloads: 5
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Abstract: The fungus Phyllosticta citricarpa is a quarantine pathogen in the EU and is of high economic importance in many parts of the world where favourable climate conditions drive the development of citrus black spot (CBS) disease. Disease symptoms include necrotic lesions on leaves and fruits. Low disease pressure can reduce crop market-ability, while higher disease pressure can cause premature fruit drop, significantly increasing crop losses. The wind-dispersed spores of P. citricarpa are especially prob-lematic for rapid pathogen dispersal, but also provide an opportunity for early detec-tion of the disease spreading into a new area. In this study we have developed and validated a quantitative PCR (qPCR) assay based on the TEF1-α sequence. Specificity testing demonstrated that it is currently the only qPCR assay that does not cross- react with closely related Phyllosticta species. The assay is sensitive and can detect a single copy of the TEF1 gene in a reaction, it is highly repeatable and reproducible and can be used for testing of the sticky tapes from spore traps as well as citrus fruit sam-ples. High-throughput sequencing (HTS) of the DNA barcodes ITS1 and TEF1 was also explored for the detection and discrimination of P. citricarpa. The limit of detection of the HTS was 1000 spores on a daily spore trap tape. This study makes an important improvement to the diagnostics of the CBS and the methods developed can also be applied to improve the surveillance and early detection of the pathogen when linked to spore samplers in the field.
Keywords: detection, fungal spore sampling, internal transcribed region (ITS), translation elongation factor 1-α (TEF1)
Published in DiRROS: 12.07.2024; Views: 11; Downloads: 4
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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: 12; Downloads: 5
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