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The Balkan Region and the “Nano Gap”: An Underexplored Dimension of In Vitro Biotechnology for Woody Plants
No full textAlthough nanotechnology is increasingly applied in plant tissue culture in many parts of Europe, its use in the Balkans remains limited, revealing a regional gap with untapped potential for advancing in vitro propagation and preservation of woody plant species. Building upon a recently published regional review covering 2001–2024, which analyzed in vitro biotechnology progress in nine Balkan countries, this paper introduces the concept of a “nano gap”, referring to the limited connection between existing nanotechnology research potential and its use in in vitro woody plant biotechnology. In Serbia, Greece, Bulgaria, Croatia, and Albania, significant progress has been made in optimizing micropropagation and in vitro conservation strategies by introducing temporary immersion systems, synthetic seed technology, adapting genotype-specific sterilization and multiplication protocols, and modifying established cryopreservation methods for regional woody species. However, the integration of nanotechnology into these systems remains largely unexplored. To date, there are no published results or validated applications for nano-enhanced media or nanoscale delivery systems for micropropagation and in vitro conservation of woody species. The limited integration of nanotechnology may be due to insufficient funding, lack of specialized infrastructure, and limited interdisciplinary expertise. Nevertheless, many Balkan countries possess growing capacities in nano-applications within agriculture and environmental sciences and are ready to advance toward interdisciplinary research and innovation. By mapping both scientific readiness and structural barriers, this review provides a strategic framework for bridging the “nano gap” and offers a novel regional perspective with broader implications for European research policy, sustainable agriculture, biodiversity preservation, and green innovation
Pharmacological Challenge with D-Amphetamine reveals ADHD-related psychomotor activity in the A53T mouse model of Parkinson’s Disease
No full textThe Hualpha-Syn(A53T) transgenic mouse line G2-3 (JAX006823) is a model forstudying Parkinson’s disease (PD) and related synucleinopathies. Previous studieshave reported hyperlocomotion in this line, yet this feature of the model has oftenbeen dismissed as an irrelevant artifact. However, hyperactivity in rodents mayreflect underlying neuropsychiatric traits, including those associated withpsychosis or attention-deficit/hyperactivity disorder (ADHD), which can bedistinguished using D-amphetamine challenge test. The aim of this study was toassess, using D-amphetamine challenge, whether the observed hyperactivity inthe JAX006823 strain reflects an ADHD-like hyperactivity. The sample consistedof 22 6-month-old male transgenic carriers, 23 of their non transgenic littermatesand 22 C57BL/6J mice (strain used in backcrossing for the colony maintenance).Spontaneous activity in an inescapable open arena was evaluated for 30 minutes,after which animals were injected with D-amphetamine (0.25 mg/kg, 2 mg/kg) orsaline and their behavior was monitored for an additional 120 minutes. Theincreased locomotor response to novelty was observed as an endophenotypictrait of the strain. Additionally, an inverse association between severity ofhyperactivity and the degree of calming effects of D-amphetamine (ratedependency) was observed in mice of the JAX006823 strain. This studycharacterized the observed behavior in the JAX006823 strain as ADHD relatedpsychomotor activity. The potential of using the D-amphetamine challenge indistinguishing between different neuropsychiatric features underlying animalbehavior is further discussed, as well as the translational potential of the mousemodel considering that adult ADHD may be part of the PD prodrome.(poster) Book of abstracts: 9th Congress of Serbian Neuroscience Society; 2025 Oct 6-8; Belgrade, Serbia. Belgrade: Serbian Neuroscience Society; 2025
Mimicking the Dutch protocol in rats: Model characterization and effects on the hippocampal gene expression and dendritic spine architecture
No full textClinical treatment for transgender adolescents follows the Dutch protocol (pubertal suppression and subsequent administration of gender-affirming hormones). The biological consequences of such interventions during puberty remain insufficiently understood. This study aimed to establish and characterize an experimental model inspired by the Dutch protocol, focusing on hormone-sensitive brain regions – hippocampus and hypothalamus.
On the day of preputial separation, Wistar rats received diphereline (2.9 mg/kg, i.m.). Seven days after diphereline treatment, estradiol treatment was initiated daily P.O. (estradiol in sesame oil with Nutella® cream; doses increased weekly: 10, 20, and 30 µg/kg/day). Appropriate control groups were used (intact control and diphereline-treated animals). After administering estradiol for three weeks, animals were sacrificed, and testicular mass, anogenital distance, and circulating testosterone levels were measured. Expression of genes in the hypothalamus and hippocampus was analyzed by RT-qPCR, and dendritic spine density in the hippocampus was assessed using Golgi staining.
Diphereline-treated groups showed significantly reduced testicular mass, anogenital distance, and testosterone levels. In the hypothalamus, Kiss1 and Gper were upregulated in diphereline-treated groups. Esr1 and Gnrh1 were selectively upregulated by estradiol. In the hippocampus, Ppp1r9b and Kalrn7 were downregulated by diphereline and estradiol respectively, while Syp, Nos1 and Shank1 were upregulated by diphereline. Stub-type dendritic spine density was significantly increased by estradiol.
These findings demonstrate that rodent models can present a valuable tool for understanding the effects of hormonal interventions on the brain during adolescence.Dragić M, editor. Book of abstracts: 9th Congress of Serbian Neuroscience Society; 2025 Oct 6-8; Belgrade, Serbia. Belgrade: Serbian Neuroscience Society; 2025
Harnessing artificial intelligence for real – time water quality insights
No full textImpacted by climate change and human activities, the increasing contamination of water bodies poses a growing threat to environmental and public health. Real-time water quality assessment, particularly at the source, is essential for effective monitoring and ensuring safe, contaminant-free drinking water. Artificial Intelligence (AI) and Machine Learning (ML) technologies have transformed this field by enabling efficient data processing, accurate detection of pollutants, and early warning systems.
Developing high-accuracy AI-based models can significantly enhance water monitoring systems, reduce operational costs, support eco-friendly treatment practices, and promote sustainable resource management.
The evaluation of biological indicators, such as Chlorophyll-a, offers valuable insights into water quality due to its strong correlation with chemical status. When paired with appropriate data collection frequencies, ML models can be used for real-time forecasting. These models can analyze Chlorophyll-a trends from remote sensing data and serve as early warning systems for harmful algal blooms and related environmental issues.
Incorporating diverse data sources - such as satellite imagery, water temperature, nutrient concentrations, and meteorological data - can significantly improve predictive accuracy. Developing a robust methodology for these models will empower water utilities and decision-makers in Serbia, allowing them to respond proactively to water quality threats.Višković M, Šunjević M, Zoraja B, editors. Proceedings: 3rd International EUROSA Conference; 2025 May 14-17; Vrnjačka Banja, Serbia. Novi Sad: Faculty of Technical Sciences, University of Novi Sad; 2025. p. 187-92
Environmental drivers of the diversity and distribution of black flies (Diptera: Simuliidae) in the Balkan Peninsula
No full text1. Black flies (simuliids) are an ecologically and medically important group of cosmopolitan insects whose distribution is influenced by climate and landscape changes.
This study aimed to analyse diversity, distribution and frequency of simuliid species
in the Balkan Peninsula, investigating how their ecological patterns relate to both
regional geographic factors and local habitat characteristics.
2. Total of n = 40,733 larvae and pupae (45 taxa) were collected from 256 sites from 2010 to 2021. Localities were categorised by catchment basin, hydro-ecoregion and predefined watercourse types. Distribution, land use and diversity measures were analysed. Altitude and 19 bioclimatic variableswere included as continuous environmental predictors.
3. Simulium ornatum, S. reptans, S. variegatum and S. balcanicum were the most frequent. Overall species richness ranged from 1 to 8 taxa, influenced by bioclimatic factors and land use, with peak richness observed under moderate anthropogenic disturbance. Species replacement was a significant component of beta diversity. Altitude significantly
influenced assemblage composition. The key bioclimatic drivers included precipitation
during driest/warmest periods and mean temperature of the wettest quarter.
4. Hierarchical clustering identified nine distinct simuliid assemblage groups, confirming
altitudinal niche differentiation consistent with Rapoport’s elevational rule. Higher altitudes
are dominated by the S. variegatum group and the genus Prosimulium, while subg.
Wilhelmia was characteristic of lowland sites. Diversity distances between simuliid
assemblageswere also correlatedwith geographic distances between sampling sites.
5. This study provides a valuable baseline for understanding the complex spatial and
environmental drivers of simuliid diversity in the Balkans, crucial for future ecological
and conservation research
Ekološki uticaji rudnika uglja i fotonaponskih elektrana na zajednicu makrobeskičmenjaka u akumulaciji Oslomej i reci Temnica (R. Severna Makedonija)
No full textCoal mining and photovoltaic power plant operation significantly impact the
macroinvertebrate communities in freshwater ecosystems due to changes in water quality
and habitat structure resulting from dramatic erosion processes. Therefore, monitoring the
macroinvertebrate community serves as an important indicator of ecological health. This
study presents the ecosystem conditions in Oslomej reservoir and river Temnica based on
the aquatic invertebrate community. The obtained results showed that Oslomej reservoir
and river Temnica suffer habitat degradation and put up with different pollutants,
introduced through the soil and sediment that entered the reservoir and the river during the
construction of the PV Oslomej 3, as well as from previous acid mine drainage from the
coal mine “Oslomej”.Iskopavanje uglja i rad fotonaponskih elektrana značajno utiču na zajednice
makrobeskičmenjaka u slatkovodnim ekosistemima zbog promena u kvalitetu vode i
strukturi staništa koje su rezultat dramatičnih procesa erozije. Stoga, praćenje zajednice
makrobeskičmenjaka služi kao važan indikator ekološkog zdravlja. Ova studija prikazuje
uslove ekosistema u Oslomejskom akumulaciji i reci Temnici na osnovu zajednice vodenih
beskičmenjaka. Dobijeni rezultati su pokazali da akumulacija Oslomej i reka Temnica trpe
degradaciju staništa i trpe različite zagađivače, unete kroz zemljište i sediment koji su
dospeli u akumulaciju i reku tokom izgradnje PV Oslomej 3, kao i prethodnim kiselim
rudničkim odvodom iz rudnika uglja „Oslomej“.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. 197-208
Aquatic biodiversity along the Danube River
No full textThe Danube River Basin (DRB) covers parts of four EU bioregions and nine different ecoregions. This chapter briefly gives an overview of major water-dependent groups of plants and animals (algae, phytoplankton, zooplankton, phytobenthos, macrozoobenthos, macrophytes, fish, amphibians, reptiles, birds, and mammals) found during surveys and research along the Danube and its floodplains. Due to the immense size and complexity of the Danube tributaries, this overview does not cover the complete biodiversity of the DRB. However, it provides insight into some novel methods for research and opens the question of the accuracy and reliability of species lists and their use in the protection and management of biodiversity. Finally, it summarizes critical drivers of pressures on the existing biodiversity, including the recent lack of water due to climate change.Bloesch J, Cyffka B, Hein T, Sandu C, Sommerwerk N, editors. The Danube River and The Western Black Sea Coast: Complex Transboundary Management. Elsevier; 2025. p. 83-105
Phytoremediation potential of metallophytes in Europe: progress, enhancement strategies, and biomass utilisation
No full textPhytoremediation is a plant-driven process, widely regarded as a cost-effective and environmentally friendly in situ approach for remediating contaminated soil and water by taking up contaminants including potentially toxic elements (PTEs). In the last two decades, substantial research has focused on elucidating the mechanisms of phytoremediation and enhancing its efficiency, primarily through the identification of optimal plant species and the use of various amendments. Nevertheless, real-scale application of phytoremediation remains rare, and several critical questions need to be addressed, including selection of most effective species, improved effectiveness of phytoremediation process, and managing the safe utilisation of contaminated biomass. This review specifically focuses on phytoremediation of potentially toxic metals and metalloids in major metallophyte groups (wild herbaceous species, trees, and agricultural crops) recognizing the most efficient species for the anthropogenically influenced soils in Europe. It summarises the current state of knowledge regarding the use of respective plant species, highlighting the phytoremediation efficiency, critically examining existing and novel phytoremediation enhancement strategies and biomass utilisation pathways for each particular group. Future perspectives and research needed to refine the efficiency and economic viability of the phytoremediation process in Europe lay in better recognition of underlying physiological mechanism for metal stress tolerance, particularly among the most effective species and genera, application of synergistic enhancing techniques for delineated group of metallophytes and development of sustainable and cost-effective biomass utilization routes.This is the peer reviewed version of the following article: Ranđelović D, Jakovljević K, Zeremski T, Pošćić F, Baltrėnaitė-Gedienė E, Noulas C, Mašková P, Jurković J, Baragaño Coto D, Milićević T, Cornu JY, Habyarimana E, Sukiaysan A, Bert V, Renella G, Carletti P, Saletović M, Maletić S. Phytoremediation potential of metallophytes in Europe: Progress, enhancement strategies, and biomass utilisation. J Environ Manage. 2025 Jul 9;391:126516. [https://doi.org/10.1016/j.jenvman.2025.126516
Multifaceted role of AMPK in autophagy: more than a simple trigger?
No full textAMP-activated protein kinase (AMPK) is a key sensor and regulator of intracellular energy balance. During energy stress, AMPK helps restore cellular ATP levels by preventing anabolic and promoting catabolic processes, such as autophagy. AMPK activates autophagy both posttranslationally and transcriptionally, by suppressing the mechanistic target of rapamycin complex 1 activity and stimulating the activation of unc-51 like autophagy activating kinase (ULK), autophagosome-lysosome fusion, and expression of autophagy-related genes. Recent research, however, suggests an unexpected role of AMPK in energy stress, where AMPK inhibits ULK and suppresses ATP-consuming autophagic response, possibly to save energy and maintain the autophagic machinery for subsequent activation once the stress subsides. The present review elucidates this dual nature of AMPK in autophagy regulation while highlighting its molecular mechanisms and importance for therapeutic approaches involving AMPK modulation
Targeting glycolysis with 2-Deoxy-D-Glucose and lysosomal integrity with L-Leucyl-L-Leucine methyl ester as antimelanoma strategy
No full textBackground: Malignant melanomas are characterized by enhanced glycolysis, which supplies energy and biosynthetic precursors for rapid proliferation. Melanomas also exhibit increased lysosomal fragility and elevated levels of lysosomal
enzymes cathepsins. Cathepsins promote metastasis and immune evasion, but when released into the cytosol upon
lysosomal membrane permeabilization, they trigger cell death. This study aimed to evaluate the antimelanoma
potential of 2-deoxy-D-glucose (2DG), an inhibitor of the glycolytic enzyme hexokinase, in combination with L-leucyl-L
leucine methyl ester (LLOMe), a lysosome-destabilizing agent.
Materials and Methods: Gene expression data from melanoma patients and normal skin (GEO dataset GSE3189)
were analyzed using the Mann-Whitney U test. Human A375 melanoma cells were treated with 2DG, LLOMe, or their
combination. Cell viability was assessed by MTT assay. Flow cytometry was used to assess apoptosis by detecting
phosphatidylserine externalization (Annexin V/PI staining) and DNA fragmentation (sub-G1 DNA content analysis of PI
stained cells). Caspase activation was measured fluorometrically (ApoStat staining). The type of 2DG+LLOMe-induced
cell death was determined by assessing viability of cells pretreated with inhibitors of caspases, necroptosis, ferroptosis,
and autophagy.
Results: GEO analysis revealed elevated expression of glycolytic enzymes (hexokinase, phosphoglucose isomerase,
platelet-type phosphofructokinase, aldolase, lactate dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase,
triosephosphate isomerase, pyruvate kinase) and lysosomal proteases (cathepsins B, D, Z) in melanoma relative
to normal skin. Both 2DG and LLOMe reduced the viability of A375 cells, with a markedly enhanced effect when
combined, indicating a synergistic interaction as demonstrated by α-index >1. This cytotoxicity involved caspase
activation, phosphatidylserine exposure, and DNA fragmentation. Caspase inhibitors, but not inhibitors of necroptosis,
ferroptosis, or autophagy, reduced 2DG+LLOMe-induced cell death.
Conclusions: By demonstrating elevated expression of glycolytic enzymes and cathepsins during melanoma progression,
along with the synergistic pro-apoptotic effect of the glycolytic inhibitor 2DG and lysosomal destabilizing agent LLOMe
in melanoma cells, our study supports further investigation of dual targeting of glycolysis and lysosomal stability as a
therapeutic strategy in melanoma.Čavić M, editor. Proceedings book: The 1st Regional SDIR-HDIR-MOKAD Congress; 2025 Oct 8-10; Belgrade, Serbia. Belgrade: Serbian Association for Cancer Research; 2025. p. 156-7. (Oncology Insights; No. 3)