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Assessment of chemical pollution of selected locations on the Nišava and the Štira rivers based on SPEAR parameters
No full textHemijski zagađivači imaju značajan uticaj na pad biodiverziteta slatkovodnih ekosistema na globalnom nivou. U ovom radu korišćen je SPEAR parametara za procenu uticaja pesticida i organskih zagađivača na vodene ekosisteme. Ovi parametri su zasnovani na osobinama vodenih makrobeskičmenjaka koje su osetljive na pesticide i organske zagađivače. Analizirana je zajednica makrobeskičmenjaka na 3 lokaliteta na reci Štiri i 4 na reci Nišavi. Na osnovu ispitivane zajednice i dobijenih vrednosti SPEAR parametara može se zaključiti da su lokaliteti u blizini i nizvodno od gradova pod većim pritiskom i uticajem pesticida i organskog zagađenja.Chemical pollutants have a significant impact on the decline of biodiversity in freshwater ecosystems on a global scale. In this work, the SPEAR parameters was used to assess the impact of pesticides and organic pollutants on aquatic ecosystems. These parameters are based on the characteristics of aquatic macroinvertebrates that are sensitive to pesticides and organic pollutants. The macroinvertebrate community was analysed at 3 sites on the Štira River and 4 sites on the Nišava River. Based on the analysed community and the determined values of SPEAR parameters values, it can be concluded that sites near and downstream of cities are exposed to greater pressure and influence of pesticides and organic pollution.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. 71-78
Caloric Restriction and Frailty: The Role of Onset, Duration, and Sex
No full textFrailty reflects an increased vulnerability to adverse health outcomes and
progressively worsens with age. The clinical frailty index (FI) is one of the methods
for assessing frailty, based on accumulation of physiological deficits in both
humans and animals. Given that caloric restriction (CR) is the most widely studied
non-pharmacological intervention for delaying age related changes, this study
aimed to examine how the timing of CR onset and sex influence its effects on
body weight, survival and frailty levels. Male and female Wistar rats were used in
this study. A 40% reduction in ad libitum (AL) food intake was implemented either
at six months of age and maintained until 18 or 24 months (long-term CR, LTCR) or
at 15 or 21 months of age and maintained until 18 or 24 months (short term CR,
STCR). FI was calculated using 21 physiological parameters reflecting the
condition of various organ systems, along with two cognitive parameters. FI
increased with aging in both sexes, though females reached peak frailty earlier. In
18- month-old rats, both LTCR and STCR improved survival and reduced FI
compared to age-matched AL controls. In 24-month-old animals, LTCR increased
survival in both sexes but reduced FI only in females. STCR had opposing effects
on survival at 24 months, increasing survival in males but decreasing it in females.
These findings highlight the importance of a personalized approach when
implementing CR as a strategy for promoting healthy aging, emphasizing the
impact of sex and intervention timing on its outcomes.Dragić M, editor. Book of abstracts: 9th Congress of Serbian Neuroscience Society; 2025 Oct 6-8; Belgrade, Serbia. Belgrade: Serbian Neuroscience Society; 2025
Gut microbiota composition combined with reduced intestinal fatty acid uptake prevents hepatic steatosis in obesity-resistant mice fed a high-fat diet
No full textThe metabolic response to excessive caloric intake varies and some individuals remain resistant to
obesity with different tendencies to develop hepatic steatosis. Gut microbiota may play a crucial role
in these differences by influencing energy utilization.The aim of this study was to investigate gut
microbiota composition that may underlie variations in the occurrence of hepatic steatosis between
obese and obesity-resistant mice. After 14 weeks on a high-fat diet (60 kcal% fat), the male C57BL/6J
mice with the highest body mass were categorized as obese, while the animals without weight gain
were selected as obesity-resistant. Despite similar caloric intake, obesity-resistant mice had lower
liver mass and serum triglyceride levels compared to obese animals.The gut microbiota profile of the
obesity-resistant mice was characterized by an increased abundance of RF39 and Lactobacillus, which
was negatively correlated with triglyceride levels, and a lower abundance of Helicobacter and Rothia.
These changes paralleled the improved gut morphology and decreased protein levels of free fatty acid
transporters in the small intestine and liver of the obesity-resistant mice. In summary, gut microbiota
composition of obesity-resistant animals in combination with the reduced intestinal capacity for fatty
acid uptake can influence the lipid influx into the liver and ultimately prevent ectopic fat accumulation
Invasive alien species (IAS) in the Danube River Basin and Western Black Sea Coast
No full textThis chapter presents a brief overview of aquatic invasive alien species (IAS) in the Danube River Basin (DRB) and Western Black Sea Coast (WBSC), including species lists, pathways of introduction and spread, impact, and management strategies. The compiled lists include 48 freshwater species (32 invertebrates and 16 fish) alien to the DRB, 36 species of Ponto-Caspian origin (30 invertebrates and 6 fish) translocated to the Upper and Middle DRB, and 219 marine taxa (12 fungi, 95 algae, 103 invertebrates and 9 vertebrates) alien to the Black Sea. Ten of all listed species are IAS of Union concern, 21 are of DRB concern, and 28 are alien species of priority for management in countries of South-Eastern Europe. Some recent introductions, establishment, and spread of alien species pose a potential impact on native biota. Joint efforts at different levels (European Commission [EC], EU Strategy for the Danube Region [EUSDR], International Commission for the Protection of the Danube River [ICPDR], Black Sea Commission [BSC], International Association for Danube Research [IAD], and other regional and national organizations) are needed to overcome the numerous challenges in the management of the aquatic IAS and the different pathways and vectors of their introductions. The Danube Region Invasive Alien Species Network (DIAS) promotes an improved coordination among scientists, authorities, and stakeholders in the field of IAS.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. 165-93
Structure and hormonal output of the adrenal gland after experimental estrogenization of male rats
No full textOrchidectomy and estrogenization of the male represent a procedure that is applicable in sex reassignment or in prostate cancer therapy. This approach has an influence on the hypothalamic-pituitary-adrenal axis and thus affects cardiovascular function and metabolism. We utilized orchidectomized rats to evaluate the effects of estradiol on the structure and hormonal output of the adrenal gland. Adult Wistar rats were divided into sham-operated (SO; n=7), orchidectomized (Orx; n=7), and estradiol-treated orchidectomized (Orx+E; n=7) groups. Estradiol-dipropionate (0.625 mg/kg b.m.) was administered subcutaneously for three weeks, while the SO and Orx groups received vehicle alone. Set objectives were achieved using histochemistry/immunohistochemistry, stereology, and immunoassays. In Orx+E rats, the hormonal milieu was characterized by decreased testosterone and increased ACTH, compared with the Orx group. Also, orchidectomy and estradiol treatment provoked a significant increase in adrenal cortex volume and volume of ZF per se, with increased cell and nuclei volumes in all three adrenocortical zones (ZG, ZF, and ZR), in comparison with Orx rats. Concentrations of aldosterone in blood, as well as corticosterone in blood and adrenal tissue were increased, while circulating DHEA was decreased (with increased immunoexpression of adrenocortical CYP 17 enzyme), all in Orx+E compared with Orx animals. The wide zonal distribution of VEGF and the pronounced blood supply within the ZF of Orx+E animals acted to support the synthesis and secretion of corticosteroids. These results seem cautionary in the context of young male estrogenization, given the negative impact of high mineralocorticoids and glucocorticoids on cardiovascular function and metabolism
Targeting Glycolysis with 2-Deoxy-D-Glucose and Lysosomal Integrity with L-Leucyl-L-Leucine Methyl Ester as Antimelanoma Strategy
No full textBackground/Objectives: Melanoma cells enhance glycolysis and expand lysosomes to support energy metabolism, proliferation, and metastasis. However, lysosomal membrane permeabilization (LMP) causes cathepsin leakage into cytosol triggering cytotoxicity. This study investigated the antimelanoma effect of 2-deoxy-D-glucose (2DG), an inhibitor of glycolytic enzyme hexokinase-2, in combination with cathepsin C-dependent LMP inducer L-leucyl-L-leucine methyl ester (LLOMe) and cathepsin C-independent LMP-inducers mefloquine and siramesine. Methods: The viability of A375 and B16 melanoma cells and primary fibroblasts was measured by crystal violet. Apoptosis, necrosis, and LMP were assessed by flow cytometry; caspase activation, mitochondrial depolarization, superoxide production, and energy metabolism were analyzed by fluorimetry, and expression of cathepsins and hexokinase-2 was evaluated by immunoblot. Appropriate inhibitors, antioxidant, and energy boosters were used to confirm cell death type and mechanism. Results: LLOMe triggered LMP, mitochondrial depolarization, and mitochondrial superoxide production, while suppressing oxidative phosphorylation. 2DG suppressed glycolysis and, together with LLOMe, synergized in ATP depletion, caspase activation, and mixed apoptosis and necrosis in A375 cells. Inhibitors of lysosomal acidification, cysteine cathepsins, and caspases, as well as antioxidant and energy boosters, reduced 2DG+LLOMe-induced toxicity. Cathepsins B, C, and D were lower, while hexokinase-2 was higher in A375 cells than fibroblasts. Accordingly, 2DG exhibited lower while LLOMe exhibited higher toxicity against fibroblasts than A375 and B16 cells. However, mefloquine and siramesine induced stronger LMP in A375 cells than in fibroblasts and showed melanoma-selective toxicity when combined with 2DG. Conclusions: 2DG-mediated glycolysis inhibition in combination with lysosomal destabilization induced by mefloquine and siramesine, but not with non-selectively toxic LLOMe, may be promising antimelanoma strategy
Evolutionary rate of adaptive chromosomal inversions in Drosophila subobscura (Diptera: Drosophilidae) in response to global warming
No full textIn the face of global warming, insects can employ various survival strategies. One possibility is that natural selection favors genetic combinations that are adaptive to this environmental situation. In some species, chromosomal inversions allow a particular combination of genes to be transmitted intact across generations, provided it is adaptive. Drosophila subobscura could therefore be considered a model species, given its rich chromosomal inversion polymorphism and its relationship with climate change. This polymorphism was studied in the Djerdap (Serbia) population in 2024, and its composition was found to have changed over time when the results were compared with those from 2001 and 2002. A significant decrease in 'cold' and an increase in 'warm' adapted inversions were observed comparing 2001 and 2024. Also, the CTI (chromosomal thermal index) value for the whole karyotype in 2001 (0.162) was significantly different to that in 2024 (0.332) (adjusted p = 0.0084). Of the inversions studied, nine presented an observed frequency outside the interval expected assuming genetic drift. However, a change in agreement with global warming was detected in only five of these inversions, while the frequency of two 'non-thermal' adapted inversions also increased. Finally, an evolutionary rate was calculated by dividing the difference in frequency between 2024 and 2001 by the number of years that had elapsed. The values obtained for the 'cold', 'warm' and 'non-thermal' adapted were -0.0030126, 0.0030148 and -2.609 x 10-6, respectively. These values are lower than those previously reported for the Apatin (Serbia) population
Agmatine suppresses excessive superoxide production in LPS-stimulated BV-2 microglia
No full textDuring neuroinflammation, activated microglia increase reactive oxygen species (ROS) production, especially superoxide, which is mainly generated extracellularly via NADPH oxidase (NOX) to eliminate pathogens. Concurrently, ROS produced by microglia also serve as signalling molecules that initiate and modulate its activation and polarisation. Excessive superoxide production and prolonged oxidative stress contribute to microglia shift into a chronic pro-inflammatory phenotype and promote neurodegeneration. Therefore, targeting superoxide-generating pathways represents a potential strategy for modulating microglial activation. In this study, we investigated whether agmatine pretreatment (Agm, 100 μM) affects superoxide-related responses in BV-2 microglial cells stimulated with lipopolysaccharide (Lps, 1 μg/ml). The expression of NOX2-related genes, Cybb and Nhe1, was analysed by qPCR. Superoxide levels and its sources were determined by nitroblue tetrazolium assay, MitoSOX staining and xanthine oxidase (XO) activity assay. The antioxidant response was assessed by analysing expression, protein level and activity of superoxide dismutase (SOD), the main superoxide-neutralising enzyme. Real-time cell analysis and F-actin staining were used to assess morphological changes reflecting the microglial phenotype. Agm did not affect Lps-driven upregulation of Cybb and Nhe1. However, Agm decreased Lps-triggered superoxide-producing activity, confirmed by reduced mitochondrial superoxide accumulation and XO activity. Furthermore, Agm had no effect on Lps-induced SOD2 transcription and activity, but increased SOD2 protein level. Agm prevented Lps-induced cell enlargement, an effect also evidenced by delayed and reduced increase in cell index. These results indicate that Agm reduces Lps-induced superoxide production by targeting cytoplasmic and mitochondrial sources, suggesting its potential to attenuate oxidative stress and modulate microglial activation.Dragić M, editor. Book of abstracts: 9th Congress of Serbian Neuroscience Society; 2025 Oct 6-8; Belgrade, Serbia. Belgrade: Serbian Neuroscience Society; 2025
Recent Advances in On-Tissue Chemical Derivatization Strategies for Enhancing MALDI-MSI
No full textMatrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) has rapidly advanced in biomedical research, enabling label-free, untargeted spatial detection of metabolites, lipids, proteins, and glycans in tissue sections. However, challenges such as low ionization efficiency and chemical instability limit the detection of certain molecules. To address these issues, on-tissue chemical derivatization (OTCD) has been widely applied as an effective strategy to enhance imaging capabilities. This review systematically summarizes the development of derivatization reagents targeting different reactive functional groups and their applications in MALDI-MSI, including strategies for the derivatization of amines, carbonyls, carboxyls, double bonds, hydroxyls, thiols, and platinum-based drugs. Particular attention is given to how these derivatization reagents enhance the detection range and biological relevance by increasing molecular weight, improving ionization efficiency, and reducing background noise interference. Additionally, we explore the application of OTCD in various biological samples and discuss challenges related to experimental workflows, derivatization efficiency, and tissue integrity. This review provides important theoretical support for the advancement of MSI technology and highlights its broad potential applications in biomedical research
Sclareol-based natural nanoparticles with adamantane moieties exert anticancer effects against non-small cell lung carcinoma cells
No full textSynthesis of novel derivatives of sclareol bearing an adamantane moiety was achieved aiming to improve cytotoxicity of sclareol. Novel compounds exhibited up to 100 times more pronounced effects compared to the sclareol against non-small cell lung carcinoma cell lines NCI-H460 and its multidrug resistant variant NCI-H460/R. Nine derivatives (11b, 11c, 11i, 12a, 12b, 12c, 12e, 12f, and 12 g) caused cell growth inhibition below 50 % at 1 μM concentration in NCI-H460 cells, and four derivatives (11c, 11i, 12b and 12e) showed similar activity against NCI-H460/R cells. Notably, compound 12b exhibited selectivity towards cancer cells, as well as collateral sensitivity being more potent against MDR cells. Sclareol, as well as novel derivatives 12a, 12b, 12c, 12e, 12f, and 12 g increased accumulation of rhodamine 123 which suggested their potential to modulate P-glycoprotein (P-gp) activity. Compound 12b emerged as inhibitor of P-gp and 12e and 12f as P-gp substrates. Furthermore, 12b, 12e and 12f induced reversal of doxorubicin resistance. Cell death analysis indicated necrosis as a primary type of cell death in NCI-H460 with significant increase in late apoptosis in MDR cells. We discovered for the first time that sclareol can spontaneously form negatively charged nanoparticles by being dissolved in ultrapure water. Adamantyl derivatives 12a-c and 12e-g and parental diamine derivatives 8c-f formed positively charged nanoparticles, implying that they can bind to the negatively charged cellular membrane and penetrate cancer cells. The unique nanoparticle characteristics combined with the significant cytotoxicity of the novel adamantane-sclareol derivatives underscore their potential as promising candidates for advanced anticancer nanotherapeutic applications.This is the peer reviewed version of the following article: Stojković P, Lupšić E, Nataša TJ, Stepanović A, Nedialkov P, Podolski- Renić A, Pešić M, Opsenica IM. Sclareol-based natural nanoparticles with adamantane moieties exert anticancer effects against nonsmall cell lung carcinoma cells. in Bioorganic Chemistry. 2025;165:108967. [https://doi.org/10.1016/j.bioorg.2025.108967