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Ion channels in plasma membrane of filamentous fungi - methodology and advances in Mucoromycota
No full textThe activity of ion channels, driving exchange across the plasma membrane and electrical signaling, is an essential part of cell physiology. Although far better understood in animals, plants and even bacteria, there are a handful of ion channels or plasma membrane ion currents that have been characterized in filamentous fungi. There is evidence, albeit fragmentary, of their expected role in mycelial growth and regeneration, small organic acid exchange, and cellular and long-range signaling. This, still limited, knowledge is mainly the result of two main directions of fungal research: cloning of ion channels homologous to known types in other kingdoms, followed by characterization by expression in heterologous systems (1), and functional characterization in native membrane in specific experimental preparations, such as cytoplasmic droplets from sporangiophores of Phycomyces blakesleeanus, which we use (2). In addition, classical extracellular flux measurements with ion-selective electrodes (3) as well as the still underutilized imaging techniques and the registration of gross potential changes in mycelia also promise progress in this field. Recently, we have developed a femtosecond laser-based nanosurgery method for cell wall removal, which allowed us to obtain protoplasts with plasma membrane suitable for patch-clamp registration using P. blakesleeanus mycelia hyphae (4). Here we present our findings from both sporangiophore and hyphal membranes. Most of the identified ionic currents carry various anions, while none is potassium-based. ORIC is an ATP-dependent, outwardly rectifying anionic current from rapidly growing aerial sporangiophores that is osmotically dependent (5). ORAC, another sporangiophore anionic current, is blocked by malate and depolarization-activated for long periods of time. Both currents appear to be characteristic of sporangiophores. In hyphae, several types of ionic currents preferentially conduct glutamate rather than chloride, and only a small fraction can be attributed to calcium channels.International Fungal Biology Conference: from Molecules to Communities: IFBC-2025; 2025 Sep 27-30; Chania, Crete, Greece. International Fungal Biology Conference; 2025. p. 93
Multiphoton Live-Cell Metabolic Imaging and Femtosecond Laser Nanosurgery of Filamentous Fungi
No full textFilamentous fungi are of enormous ecological, medical and biotechnological importance. Studying their physiology and dynamics in vivo is crucial for understanding their interaction with the environment as well as for developing new therapeutic strategies for disease-causing pathogens. Nonlinear laser microscopy modalities, two-photon excitation fluorescence (TPEF) and third harmonic generation (THG) microscopy, were employed to image structural and functional changes in the cellular (lipid and energy) metabolism of model filamentous fungus Phycomyces blakesleeanus, at the single-cell level. Label-free THG method was used for the first time to image lipid droplets (LDs) in live filamentous fungi that predominantly have LDs smaller than 1 μm [1]. Under various environmental conditions, THG was able to detect changes in size, number and location of the LDs in tiny fungal cells. On the other hand, TPEF enabled the monitoring of morphological changes of mitochondria in real time under the same conditions as THG imaging. To study fungal physiology, the first femtosecond IR laser nanosurgery was performed on the cell wall of filamentous fungi, enabling electrophysiological measurements on the protoplasts released from the hyphae [2]. The high precision of fs laser ablation allowed us to cut a small portion of the cell wall without damaging the cell membrane only a few micrometers away. A reproducible and highly precise (diffraction-limited, submicron resolution) method for obtaining viable protoplasts was developed. Protoplast release from the nanosurgery-generated incisions in the cell wall was achieved from different regions of the hyphae. The introduction of this method opened the way to investigate the physiology of ion channels directly on the cell membrane of filamentous fungi, which is otherwise inaccessible due to the presence of a rigid chitinous cell wall. Figure 1. (a) Label-free THG image of lipid droplets in live P. blakesleeanus hypha (b) TPEF image of tubular mitochondria stained with 5 μM Rhodamine123 in live P. blakesleeanus hypha (c) In vivo femtosecond laser nanosurgery of chitinous cell wall of filamentous fungus P. blakesleeanus.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. 42
De Novo Transcriptome Assembly of Nepeta ernesti-mayeri Diklic & V.Nikolic leaf trichomes
No full textAssembled transcriptomes from paired-end RNA sequencing of Nepeta ernesti-mayeri Diklic & V.Nikolic leaf trichomes using DNBSEQ, in FASTA format. The dataset comprises 54162 assembled transcripts in a FASTA file. N. ernesti-mayeri transcriptome data are a part of phyloNEPETOME data platform
Biological and Computational Study of the Dual Antimicrobial and Anticancer Potential of 3,4-Dihydropyrimidin-2(1H)-ones and Thiones
No full textSixteen 3,4-dihydropyrimidin-2(1H)-ones and -thiones were synthesized and evaluated for antimicrobial activity against both resistant and nonresistant bacterial strains, as well as four fungal species. Antibacterial testing revealed MIC/MBC values ranging from 0.25 to 8.0 mg/mL, with compound 20 demonstrating the most potent activity (MIC: 0.42–2.0 mg/mL). Notably, compound 16 exhibited consistent efficacy against both resistant and nonresistant strains. Antifungal activity was assessed against Aspergillus fumigatus, Aspergillus niger, Penicillium funiculosum, and Candida albicans, with compound 20 again emerging as the most effective (MIC: 0.75–2.0 mg/mL). Compounds 16 and 25 also showed antibiofilm activity, with compound 16 inhibiting biofilm formation by 70.22%. Preliminary cytotoxicity screening against cancer cell lines indicated that several compounds exhibited moderate anticancer activity. ADMET (absorption, distribution, metabolism, excretion, and toxicity) analysis suggested favorable pharmacokinetic profiles, with no violations of Lipinski’s Rule of Five. Molecular docking studies suggest that the antibacterial mechanism may involve inhibition of the MurB enzyme, while antifungal activity is likely mediated through inhibition of cytochrome P450 (CYP) enzymes. These findings underscore the potential of 3,4-dihydropyrimidin-2(1H)-(thi)ones as promising lead compounds for the development of novel antibacterial and antifungal agents
Theodoxus transversalis (C. Pfeiffer, 1828) (Gastropoda: Neritidae) in the Western Balkans: an endangered freshwater snail bouncing back?
No full textPopulations of Theodoxus transversalis, striped nerite, have declined in recent decades, arguably due to negative
anthropogenic impacts on their habitats. The species is classified as endangered by the International Union for
Conservation of Nature, and there are generally few reports of its recent occurrence. This is especially true for the
Western Balkans and Serbia, where data on the distribution of T. transversalis are more than a decade old. Herein, we
present the latest data based on an extensive field study of this species conducted in 2023 and 2024. The Velika Morava-
Južna Morava-Nišava catchment and ecoregion (ER) 5 were confirmed as the most important refugium for this
rare snail in the investigated region. Of particular interest are the findings of an abundant population from the Drina
River (where the species has not been reported for 15 years), from the lower Serbian section of the Danube (reports
from the Danube are 30 years old), and the discovery of a new population from the Zapadna Morava River 100 km
upstream from the nearest known population. Our results show considerable progress in the restoration of the former
range of T. transversalis in the Western Balkans, although stable populations still appear to be localized. There is also a
possibility that this species is spreading into new areas, which is a possibility for the Zapadna Morava River catchment.
While it appears that T. transversalis is more resilient to negative anthropogenic impacts than previously described
in the available literature, a more detailed ecological and genetic study should shed more light on the presence and
prospects of the species in the region
Identifying Lipidomic Signatures and Microbial Metabolites in Patients with Metabolic Dysfunction-Associated Steatotic Liver Disease from Serbia: A Metabolomics-Based Approach to Liver Dysfunction
No full textMetabolic dysfunction-associated steatotic liver disease (MASLD) is the most prevalent chronic liver disease in the Western world, and is rapidly increasing
worldwide. Serbia, along with other Balkan countries, faces particularly high rates of MASLD, yet remaining underrepresented in large-scale European
research initiatives, including the MASLD registry. The absence of structured clinical and molecular datasets from this region emphasize the need for
locally conducted, population-specific studies. This work presents preliminary results from the first comprehensive multi-omics study of MASLD in the
population from Serbia, aiming to identify a lipidomic signature of liver dysfunction and to explore microbial metabolites potentially involved in disease
pathophysiology, using both untargeted and targeted metabolomics approaches.
Plasma and fecal samples were collected from 122 participants, including MASLD patients and sex-, age-, and BMI-matched healthy controls. Lipids from
plasma, as well as semi-polar and polar metabolites, were extracted using a monophasic extraction solution (MeOH:MTBE:IPA (20:15:15)) containing
internal standards. The samples were analyzed using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass
spectrometry (UHPLC-QTOF-MS, Agilent Technologies, Santa Clara, CA, USA). The analysis was performed on an ACQUITY UPLC BEH C18 column
(2.1 mm × 100 mm, particle size 1.7 µm; Waters, Milford, USA). Data were processed and analyzed using MZmine 3 software, with lipid and metabolite
identification based on an in-house library.
A total of 1751 lipids and 1315 semi-polar/polar metabolites were detected in plasma samples, with 214 lipids and 132 semi-polar/polar metabolites identified.
Principal component analysis did not reveal significant separation between MASLD patients and healthy controls. At the lipid class level, however, distinct
clustering patterns emerged, pinpointing alterations in lipid metabolism. Ongoing analyses aim to further refine these observations and investigate
associations with clinical parameters, with the goal of uncovering meaningful patterns that could contribute to the discovery of disease biomarkers and the
elucidation of pathophysiological mechanisms.Oral and Posterts Abstracts: 21st Annual Conference of the Metabolomics Society: Metabolomics2025; 2025 Jun 22-26; Prague, Czech Republic. Metabolomcis Society; 2025. p. 275
Is there a limit to the adaptive potential of chromosomal inversions with regard to global warming?
No full textGlobal warming is a fact, and organisms have the following options to avoid extinction: to acclimate (phenotypic plasticity), to adapt through genetic changes or to migrate to more favorable regions. Useful genetic markers to study genetic adaptations are chromosomal inversions. In this context, Drosophila subobscura is an excellent model species, because it presents inversions in its whole karyotype. Also, it has been possible to define 'cold', 'warm' and 'non-thermal adapted inversions, both in the Palearctic and American colonized regions. The aims of this research were to study the differentiation of this polymorphism in two populations from Jastrebac Mt. (Serbia) which differed in habitat conditions (beech and oak forests) sampled in 2023. Also, it was possible to compare changes of this polymorphism over time, comparing both populations with those from 1990, 1993, 1994 and 2023. Furthermore, the CTI (Chromosomal Thermal Index), which measured the thermal adaptation of the whole karyotype was computed and compared among all populations. The possible relation between climatic variables (mean, minimum and maximum temperatures, humidity and rainfall) and chromosomal inversions was also studied.Abstract Book: Congress of the European Sociey for Evolutionary Biology; 2025 Aug 17-22; Barcelona, Spain. European Society for Evolutionary Biology; 2025. p. 1604
Physico-Chemical Properties and Push-Out Bond Strength to Root Dentine of Calcium Silicate-Based Sealers
No full textThe calcium silicate-based sealers currently available on the market have different
compositions and formulations, which is why their physical and chemical properties may
vary. (1) The aim of the study was to measure the physico-chemical properties of calcium
silicate-based sealers and their push-out bond strength to root dentine, comparing two
push-out testing protocols. (2) Standardized specimens of EndoSequence BC, BioRoot RCS,
MTA Fillapex, and AH Plus (control) were subjected to pH measurements over 28 days.
Radiopacity was measured using a CCD sensor, and flexural strength was assessed using a
three-point bending setup. Push-out bond strength was measured in coronal, middle, and
apical sections of 40 single-root teeth (conventional method), and cylindrical cavities were
prepared for all sealers on the same root dentine disks in 11 third molars (disk method).
(3) EndoSequence BC exhibited a higher pH than MTA Fillapex and the highest radiopacity
(p < 0.05). The highest flexural and push-out bond strengths were found for AH Plus.
The push-out bond strength of EndoSequence BC and BioRoot RCS was higher than MTA
Fillapex (p < 0.05). The conventional and disk methods exhibited similar push-out bond
strength results, but the data were more homogeneously distributed in the disk method.
(4) All calcium silicate-based sealers exhibited a higher pH than AH Plus. MTA Fillapex
did not meet the ISO standard. Calcium silicate-based sealers showed weaker performance
in terms of physical properties compared to AH Plus
A Preliminary Insight into Under-Researched Plants from the Asteraceae Family in the Balkan Peninsula: Bioactive Compound Diversity and Antioxidant Potential
No full textNatural resources rich in polyphenols from plants belonging to the Asteraceae family remain largely unexplored. The main goal of this study was to characterize under-studied Asteraceae plants in terms of different bioactive compounds, antioxidant potential, and chemical profile. Twenty-three samples from 19 plant species were analyzed using conventional solid/liquid extraction, and the contents of total phenolics (TP), flavonoids (TF), flavonols (FL), hydroxycinnamic acids (HCA) and condensed tannins (CT), as well as extraction yield were determined. Antioxidant activity was assessed using DPPH, ABTS and FRAP assays, and five plant samples were subjected to LC-MS analysis. Extraction yields ranged from 0.57% to 1.74%. Solidago virgaurea had the highest TP and FL contents, while Tanacetum vulgare showed the highest TF and HCA levels. The highest CT content was found in the roots of Helianthus tuberosus. Asteraceae species such as S. virgaurea, Tussilago farfara, Cota tinctoria, T. vulgare, and Inula ensifolia demonstrated the greatest antioxidant potential, with about 130 different identified compounds. Given the promising chemical richness of these under-researched species, future studies should focus on enhancing extraction of bioactive compounds using novel techniques and incorporating extracts as natural, non-synthetic preservatives in various products to improve their nutritional and biological properties
Identification of microplastics in the lake water of the "Kraljevac" Special Nature Reserve
No full textMikroplastika predstavlja čvrste čestice manje od pet milimetara. Površinske vode smatraju
se koridorima disperzije (mikro)plastičnog zagađenja. U ovom radu, ispitivali smo prisustvo
mikroplastike u uzorcima jezerske vode Specijalnog rezervata prirode "Kraljevac". Ukupno
je detektovano 23 čestice mikroplastike u 1L jezerske vode. Fibrili su bili zastupljeni sa
78,26 %, a fragmenti sa 21,74%. Neadekvatno upravljanje otpadom možemo dovesti u vezu
sa pojavom mikroplastike u jezeru Kraljevac.Microplastics are solid particles smaller than five millimeters. Surface waters are regarded
distribution corridors for (micro)plastic pollution. In this study, we investigated the presence of microplastics in water samples from the Kraljevac Special Nature Reserve. A
total of 23 microplastic particles were detected in 1 liter of lake water. The proportion of
fibrils was 78.26 % and that of fragments 21.74 %. Inadequate waste management can be
linked to the occurrence of microplastic particles in Lake Kraljevac.Đukić A, editor. Conference proceedings: 54th Annual Conference of the Serbian Water Pollution Control Society: Water 2025 ; 2025 Jun 16-18; Kopaonik, Serbia. Belgrade: Serbian Water Pollution Control Society; 2025. p. 177-84