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Macroinvertebrate assemblages reveal ecological status of Belgrade suburban reservoirs
No full textReservoirs in urban and suburban areas are particularly vulnerable to anthropogenic impacts such as pollution from wastewater discharges, sedimentation, habitat modification, and recreational activities. These influences can alter biological assemblages, thereby affecting the overall ecological status of these aquatic systems. In this study, the ecological potential of three suburban reservoirs in Belgrade: Bela Reka,
Pariguz, and Duboki Potok is assessed based on macroinvertebrate assemblages. Macroinvertebrates are widely recognised bioindicators, as they are sensitive to changes in water quality, habitat, and pollution levels. Five biological indices were used to assess their ecological potential: Total Number of Taxa, Diversity Index (H'), Saprobic Index (SI), Biological Monitoring Working Party (BMWP), and Average Score Per Taxon (ASPT). These indices were calculated using ASTERICS software and interpreted according to Serbian national legislation. Sampling was carried out in August 2025 using a hand net with a mesh size of 500 μm, following a multi-habitat protocol (AQEM). A total of 33 taxa were recorded, with communities dominated by β-mesosaprobic species, indicating moderate organic pollution. Bela Reka had the highest taxonomic richness (23 taxa) and diversity (H' = 2.62), followed by Duboki Potok (21 taxa, H' = 2.47) and Pariguz (11
taxa, H' = 2.16). Diptera was the most diverse group at all sites and was most abundant at Bela Reka and Pariguz, while Gastropoda and Odonata dominated at Duboki Potok. SI values were similar across sites: Duboki Potok (2.34), Bela Reka (2.27), and Pariguz (2.25). BMWP scores ranged from 35 (Pariguz) to 76 (Bela Reka), with ASPT values between 4.75 (Bela Reka) and 5.0 (Pariguz). During this research, the ecological status varied among the reservoirs: Duboki Potok was classified as class II (good), Bela Reka as class III (moderate), and Pariguz as class V (very poor).Bosak S, Gračan R, Korać P, editors. Book of Abstracts: 15th Croatian Biological Congress: with international participation; 2025 Nov 20-23; Zagreb, Croatia. Zagreb: Croatian Biological Society; 2025. p. 261
Irrigation systems as reservoirs of diverse and pathogenic Pseudomonas syringae strains endangering crop health
No full textPseudomonas syringae (Psy) is a widely distributed bacterial species complex primarily recognized as a foliar pathogen but also inhabits diverse environments, including water habitats, where strains closely related to agricultural pathogens have been identified. The connection between Psy-caused epidemics and its potential presence in nearby irrigation systems remains underexplored. This study comprehensively examined the Psy complex in the Danube-Tisa-Danube Hydrosystem (DTD) in Serbia, assessing its abundance, phylogenetic diversity, and pathogenic potential. To reduce the reliance on the time-consuming steps of isolation and identification, we developed novel high-specific primers and probes for precise detection of strains belonging to phylogroup 2 within Psy complex. Our results demonstrated that dPCR, coupled with highly specific and sensitive primers, outperformed both traditional plating and qPCR in detecting the Psy complex and phylogroup 2 in irrigation waters, making Psy diagnostics more effective. Phylogenetic analysis indicated high strain diversity within the DTD, identifying phylogroups 1, 2, 7, 12, and 13 and haplotypes linked to strains previously encountered in epidemics on sugar beet in Serbia. Notably, 66.67% of the isolates from the DTD were capable of inducing disease. Phylogroup 2 isolates displayed a broad host range, suggesting that the dissemination of Psy from DTD through irrigation, poses a substantial threat to crop health and agricultural productivity
Cutibacterium acnes growth and biofilm formation is inhibited by flavonoids
No full textCutibacterium acnes is the cause of inflammatory acne and implanted devices associated infections. This study investigated flavonoids belonging to the 4 classes: flavanones (hesperidin, neohesperidin, naringenin, naringin, and eriodictyol), flavones (diosmetin, diosmin), isoflavone (genistein), and flavonols (kaempferol, galangin) as inhibitors of C. acnes growth in planktonic and biofilm form and assessed its potential to impact the integrity of cell membrane. Flavonoids could inhibit C. acnes growth, minimal inhibitory concentration (MIC) was in range 50–200 mg/L, with the lowest MIC (50 mg/L) for neohesperidin and eriodictyol. Minimal biofilm inhibitory concentrations were in range 25–200 mg/L, with the lowest values and highest antibiofilm potential recorded for eriodictyol, genistein, and kaempferol. Impact on cell membrane was significant in the presence of almost all examined flavonoids, as determined by crystal violet uptake assay. Flavonoids, especially eriodictyol, represent promising underexplored agents for combating infections and processes linked with the presence of C. acnes
Valorization of Waste from Bell Pepper: Chemical Composition and Bioactive Potential
No full textAgri-food waste, especially in the fruit and vegetable sector, presents environmental and economic challenges. The bell pepper industry generates significant waste, offering potential for resource recovery within a circular economy. This study analyzed the nutritional and biochemical composition of waste from green, orange, and red bell peppers (Capsicum annuum L.) and evaluated the bioactive potential of hydroethanolic extracts from fruits, stalks, and seeds. Carbohydrates were the main macronutrients, followed by proteins and ashes. Fructose was dominant in orange and red peppers, while glucose prevailed in green peppers. Waste samples contained organic acids, fatty acids, and phenolic compounds, including oxalic and malic acids, polyunsaturated fatty acids, 22 bioactive phenolics, and 34 volatile compounds. Hydroethanolic extracts demonstrated strong antioxidant activity, with green bell pepper waste showing the highest levels. The extracts also exhibited antibacterial effects against Yersinia enterocolitica and Bacillus cereus and antifungal activity against Aspergillus brasiliensis. These findings highlight the potential of bell pepper waste as a rich source of bioactive compounds with applications in food, cosmetics, and pharmaceuticals. Utilizing these by-products promotes sustainability, supports the circular economy, and addresses global waste challenges
Cutibacterium acnes growth and biofilm formation is inhibited by flavonoids
No full textCutibacterium acnes is the cause of inflammatory acne and implanted devices associated infections. This study investigated flavonoids belonging to the 4 classes: flavanones (hesperidin, neohesperidin, naringenin, naringin, and eriodictyol), flavones (diosmetin, diosmin), isoflavone (genistein), and flavonols (kaempferol, galangin) as inhibitors of C. acnes growth in planktonic and biofilm form and assessed its potential to impact the integrity of cell membrane. Flavonoids could inhibit C. acnes growth, minimal inhibitory concentration (MIC) was in range 50–200 mg/L, with the lowest MIC (50 mg/L) for neohesperidin and eriodictyol. Minimal biofilm inhibitory concentrations were in range 25–200 mg/L, with the lowest values and highest antibiofilm potential recorded for eriodictyol, genistein, and kaempferol. Impact on cell membrane was significant in the presence of almost all examined flavonoids, as determined by crystal violet uptake assay. Flavonoids, especially eriodictyol, represent promising underexplored agents for combating infections and processes linked with the presence of C. acnes.This is an Accepted Manuscript version of the following article, accepted for publication in Natural Product Research. Ivano M, Đorđevski N, Dabić J, Stojković D. Cutibacterium acnes growth and biofilm formation is inhibited by flavonoids. Nat Prod Res. 2025. It is deposited under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License [http://creativecommons.org/licenses/by-nc-nd/4.0/], which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any wa
Sepsis prevents the development of experimental type 1 diabetes
No full textIntroduction: While both sepsis and autoimmunity are characterized by
dysregulated immune responses, their mutual influence remains only
partially understood.
Methods: In this study, we investigated how sepsis affects the development of
type 1 diabetes (T1D), an autoimmune disease characterized by the destruction of
pancreatic b-cells. Specifically, we examined the impact of polymicrobial sepsis
on the progression of T1D induced by multiple low doses of streptozotocin
(MLDS). C57BL/6 mice were subjected to cecal ligation and puncture (CLP) to
induce sepsis, and T1D was subsequently induced using the MLDS protocol after
the mice had fully recovered from the acute phase of sepsis.
Results and discussion: Although CLP triggered transient hypoglycemia, it did
not impair the structure or function of the endocrine pancreas, and the mice
were normoglycemic at the time of T1D induction. Notably, CLP limited immune
cell infiltration into the pancreas following MLDS treatment, thereby preventing
the onset of T1D and the development of hyperglycemia. CD4+ T cells are
important for initiating the autoimmune attack on pancreatic islets by activating
CD8+ T cells and macrophages. Thus, it seems plausible that the protective effect
observed in CLP-exposed mice was due to the reduction in CD4+ T cells, but not
CD8+ T cells, in the pancreatic lymph nodes. Additionally, CD4+ T cells and
regulatory T cells in the spleens of CLP-treated mice exhibited elevated
expression of inhibitory/exhaustion markers. These findings suggest that
sepsis-induced alterations in the CD4+ T cell compartment within pancreasassociated lymphoid organs confer protection against MLDS-induced T1
Innovative Wound Healing Utilizing Bioactive Fabrics Functionalized with Tormentillae rhizoma Extract: An In Vivo Study on Wistar Albino Rats
No full textThis paper presents an innovative protocol for fabric functionalization using Tormentillae rhizoma extract, the chemical composition of which was proved via LC/MS analysis. The extract demonstrated antioxidant activity > 99%, and antibacterial efficacy against E. coli and S. aureus > 99%. Cotton, wool, polyamide, and cellulose acetate were functionalized with the prepared extract, all showing > 90% antioxidant activity. Functionalized cotton, wool, and polyamide exhibited > 99% antibacterial activity against both bacteria. Based on these findings and the fabrics’ ability to release bioactive compounds, functionalized cotton and polyamide fabrics having excellent bioactivity but a lower ability to release bioactive compounds can serve as protective fabrics for people with sensitive skin prone to wounds, and various products for hospitals. Functionalized wool was identified as the most suitable wound dressing for in vivo preclinical investigation on Wistar albino rats. The obtained results showcased a wound-healing rate of 95.54%, and hydroxyproline content of 8.08 µg/mg dry tissue for rats treated with functionalized wool. Compared to negative, positive, and a group of rats treated with non-functionalized wool, those treated with functionalized wool demonstrated elevated values of tissue redox state parameters, superoxide dismutase (SOD) and catalase (CAT), and a notable reduction in thiobarbituric acid reactive substances (TBARS) value. Analysis of the blood samples of rats treated with functionalized wool indicated increased levels of antioxidant defense system parameters (SOD and CAT) and decreased pro-oxidative markers superoxide (O2−) and TBARS. Further clinical trials are needed to validate these findings
Generation of Insulin-Producing Alpha TC1- 6 Cells Using EpiCRISPR System for Targeted DNA Methylation
No full textDiabetes lacks concrete curative strategies due to diverse aetiologies and, therefore, represents the perfect candidate for cell replacement therapy, since it is caused by either an absolute (type 1 diabetes) or relative (type 2 diabetes) defect in the insulin-producing beta cells of the pancreas. Pancreatic alpha cells are a promising source for transdifferentiation into insulin-producing cells as they share a common developmental origin with beta cells and exhibit a certain degree of cellular plasticity. Furthermore, impairment of glucagon signaling in diabetes leads to a marked increase in alpha cell mass, raising the possibility that such alpha cell hyperplasia provides an increased supply of alpha cells for their transdifferentiation into new beta cells.
In this protocol, we used the modular epigenetic CRISPR/dCas9 toolbox for targeted DNA methylation (EpiCRISPR) and silencing of the Arx gene (Aristaless Related Homeobox, Arx), which is essential for the maintenance of alpha cell identity. Methylation-based silencing of Arx initiates the reprogramming of pancreatic alpha cells into insulin-producing cells. As a key novelty, this protocol provides a direct route for epigenetically induced transdifferentiation of mouse pancreatic alpha TC1-6 cells into insulin-producing cells and thereby confirms a proof of concept of reversible cellular epigenetic reprogramming in vitro. In addition, this streamlined workflow addresses the inherent challenges of transfecting clustered alpha TC1-6 cells by optimizing their dissociation into single-cell suspensions, thereby improving uptake and reproducibility.
In summary, this approach for cell transdifferentiation involves precise epigenetic editing of a lineage-specific marker gene, thereby enabling direct lineage conversion in a safe and versatile strategy to generate insulin-producing cells by epigenetic reprogramming. In contrast to approaches that rely on viral vectors or permanent genome editing, this method reduces the risk of off-target effects and immunogenic responses while ensuring reproducibility. The combination of efficiency and precision makes it a valuable tool to advance regenerative approaches for diabetes therapy and to explore the epigenetic regulation of cell identity
Performance Evaluation of Galvanometric Mirrors for Fast Scanning in Multimodal Laser Microscopy
No full textIn the development of a versatile nonlinear microscope optimized for multimodal imaging of biological samples, we implemented a custom-built setup combining two-photon excitation fluorescence, second harmonic generation and up-conversion imaging modalities. While the core system has been previously validated and applied to diverse samples ranging from starch granules to fungal hyphae, further improvements in imaging precision motivated a more detailed investigation into the role of galvanometric scanner performance.
In this work, we present a comparative characterization of two different pairs of galvanometric mirrors used in beam steering within our system. To evaluate how scanner dynamics – including step response behavior, dynamic artefacts such as overshoot, settling time and possible jitter – affect both beam position accuracy and image quality, we implemented several image acquisition modes operating at different scanning speeds. A custom-developed software platform allows full control of galvo driving signals and scanning trajectories, enabling precise and programmable scan patterns optimized for various imaging tasks. This flexibility enables a direct comparison of scanner performance under matched conditions and allows testing under regimes such as point-by-point acquisition. Preliminary measurements were conducted using photodetector and oscilloscope-based monitoring of scanner response to control signals as well as test imaging on simple samples. Results indicate notable differences in response profiles, which could be relevant for applications requiring rapid and precise scanning.Rabasović M, Ralević U, Lekić M, Krmpot A, editors. Abstracts of tutorial, keynote, invited lectures, progress reports and contributed papers: X International School and Conference on Photonics PHOTONICA 2025; 2025 Aug 25-29; Belgrade, Serbia. Belgrade: Institute of Physics; 2025. p. 95
Expression of GSK3β and S6K1 mRNA in triple negative breast cancer
No full textIntroduction. Breast cancer is the most common type of malignancy and the leading cancer-related cause of death in women world wide. Triple negative breast cancer (TNBC) is characterized by absence of ER, PR and HER-2 receptors and it accounts for 10-15% of all breast cancer cases. In comparison to other types of breast cancer, TNBC is more aggressive, has higher recurrence, metastasis and mortality rates. The only available therapy for TNBC is conventional chemotherapy, to which it readily develops resistance. Aberrant signaling in both PI3K/AKT/mTOR(PAM) pathway and Wnt/β-catenin pathway is proved to contribute to acquiring resistance to therapy. These signaling pathways are interconnected by glycogen synthase kinase-3β (GSK3β) which is a key kinase in regulating β-catenin degradation, while its own activity is regulated by kinases from PAM pathway: S6 kinase 1(S6K1) and AKT. GSK3β also suppresses the activity of PTEN, a PAM pathway suppressor. The inter relation between PAM and Wnt pathways provided by GSK3β could enable the tumor cells to simultaniously activate both pathways in order to survive. Therefore, GSK3β andS6K1 may be implicated in progression and development of drug resistance.
Material and method. Total RNA was extracted from FFPE samples of paired tumor and normal tissues of 119 patients diagnosed with TNBC. RNA was reverse transcribed and obtained cDNA was used for real-time PCR expression analysis ofGSK3β and S6K1 genes. Obtained results were analysed with clinical, pathohistological and immunohisto-chemistry data of Ki-67, EGFR, PD-L1 and androgen receptor (AR) proteins.
Result and discussion. The analysis of mRNA expression shows that it significantly changes in 45,4% patients for GSK3β and in25.2% for S6K1. In majority of those patients the mRNA level is decreased both for GSK3β (90.7%) and S6K1(56.7%). There is a positive correlation between expression of GSK3β and S6K1 mRNA (p<0.0001, r =0.3511). We further analysed association of GSK3β andS6K1 expression with pathohistological parameters as well as protein expression of Ki-67, EGFR, PD-L1 and AR, assessed by immunohistochemistry. The expression of S6K1 is negatively correlated with EGFR protein level(p = 0.004, r = -0.2618), while there is also association between change in S6K1 expression and low level of AR(p = 0.0238). The analysis in simultanious changes inGSK3β and S6K1 mRNA levels also shows association of decreased levels of both mRNA with low AR and high EGFR protein expression (p = 0.0083 and p = 0.0385respectively). We previously established that high expression of EGFR combined with low expression of both PD-L1 and AR represent a “high-risk” phenotype of TNBC. Conclusion. Our findings show that GSK3β and S6K1 mRNA levels are positively correlated and that the decrease inexpression of both genes could be considered as a contributing factor in “high-risk” profile of TNBC. Therefore we suggest EGFR+, PD-L1-, AR-, GSK3β-and S6K1- as up to date high risk profile for TNBC patients.EACR 2025 Congress: Innovative Cancer Science; 2025 Jun 16-19; Lisbon, Portugal. John Wiley and Sons; 2025. p. 114-5. (Molecular Oncology; Vol. 19; Suppl. 1)