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Five-Year Retrospective Analysis of Superficial Fungal Infections: Insights from Hospital Experience
No full textThis study aimed to assess the incidence and distribution of dermatomycetes in patients at the Medical Military Academy (MMA) with suspected superficial skin infections over a five-year period (October 2017 to October 2022) and to analyze variations in fungal infections based on factors such as gender, body part, and time, particularly influenced by the COVID-19 pandemic. A total of 3993 samples were analyzed. Collected data were statistically analyzed with two tests. A total of 1048 samples were positive for fungal infections. Over the study period, Trichophyton mentagrophytes and Trichophyton rubrum were the predominant taxa, while Microsporum canis and Candida albicans were frequently observed. Statistical analysis indicated significant annual variations for T. mentagrophytes, T. rubrum, and M. canis, with monthly differences for T. mentagrophytes in June and August and M. canis in October and December. Gender-based analysis showed that T. rubrum and T. mentagrophytes were more common in males, while M. canis, C. albicans, Candida spp., and Geotrichum candidum were more prevalent in females. Analysis by body part revealed that Trichophyton rubrum and Microsporum canis showed significant differences between surface types. These findings can help improve diagnostic, therapeutic, and preventative strategies
Mixture design as an innovative tool to optimize antibacterial activity of three selected essential oilsin dairy preservation
No full textFoodborne diseases remain a global health challenge, with dairy products frequently implicated
in outbreaks caused by pathogenic bacteria such as Listeria monocytogenes and Escherichia coli
O157:H7 [1, 2]. In response, growing concerns over the safety and consumer acceptance of
synthetic preservatives have increased interest in natural antimicrobial alternatives for food
preservation.
This study introduces an innovative approach using mixture design methodology to optimize the
antibacterial activity of three essential oils (EOs) derived from Petroselinum crispum,
Helichrysum plicatum, and Origanum vulgare. The essential oils were chemically characterized
using GC-MS: H. plicatum EO was rich in α-pinene, neryl acetate, and sylvestrene, with italidione
also detected; P. crispum EO contained 1,3,8-p-menthatriene, α-pinene, and β-pinene, along with
phenylpropanoids such as myristicin; O. vulgare EO was dominated by carvacrol and thymol, while
o-cymene and γ-terpinene were present in lower amounts. Using a simplex mixture design,
12 combinations of the essential oils were formulated, and tested for their antimicrobial
properties, revealing proportion-dependent inhibitory effects. Simultaneous optimization was
conducted to determine the best compromise for enhancing antimicrobial responses against
tested L. monocytogenes and E. coli strains, resulting in a tertiary combination consisting of 17.03%
of P. crispum EO, 9.78% of H. plicatum EO, and 73.19% of O. vulgare EO. The method was used to
overcome potential sensory limitations in dairy products as well as to enhance antimicrobial
efficacy at reduced concentrations.
Optimized formulation offers a natural approach to dairy preservation that aligns with clean-label
consumer preferences and regulatory limits on synthetic additives. By combining mixture design
with traditional plant compounds and modern modeling, this work promotes safer, more
sustainable innovation in food technology.Book of Abstracts: International FoodTec Conference: Shaping the Future of Sustainable Food Ecosystems; 2025 Oct 27-29; Bragança, Portugal. Bragança, Portugal: Instituto Politécnico de Bragança Campus de Santa Apolónia; 2025. p. 316
Comparative analysis of honeybees (Apis mellifera) and bumblebees (Bombus terrestris) as environmental indicators of potentially toxic elements contamination
No full textUnderstanding the movement of potentially toxic elements (PTEs) through ecological compartments remains a major challenge in environmental pollution research. We applied in situ sampling of soil, Salvia nemorosa (roots, leaves, and flowers), and two pollinator species—Apis mellifera and Bombus terrestris to trace the exposure pathway of PTEs from soil to pollinators, simultaneously exploring the potential of bumblebees alongside honeybees as bioindicator species. This approach provides a more comprehensive and realistic assessment of element transfer in terrestrial ecosystems.Sampling was conducted in both polluted mining sites (Mining and Metallurgical Complex Bor) and unpolluted natural areas. Concentrations of Cd, Cr, Cu, Fe, Li, Ni, Pb, and Zn were determined using ICP-OAES and metallothionein gene expression in honeybees was assessed as a biomarker of PTE exposure.Results revealed distinct accumulation patterns and exposure pathways between pollinators. Li, Fe, Zn, and Cu were primarily accumulated via plants, whereas Cd, Pb, Ni, and Cr were mainly introduced through atmospheric deposition. S. nemorosa efficiently accumulated PTEs, facilitating their transfer to pollinators via pollen and floral tissues. A. mellifera demonstrated greater efficacy and stability as bioindicators, while B. terrestris was more suited as a sensitive indicator of local pollution. Strong positive correlations in element concentrations between both species support their combined use in biomonitoring.This study highlights the value of a multi-organism, integrative approach to environmental monitoring. By tracking the full pathway of pollutant transfer from soil to pollinators, the study underscores the critical role pollinators play in assessing the ecological impacts of mining-related pollution.This is the peer reviewed version of the following article: Tanasković M, Matić M, Pavlović D, Dimitrijević M, Patenković A, Erić K, Erić P, Pavlović P, Davidović S. Comparative analysis of honeybees (Apis mellifera) and bumblebees (Bombus terrestris) as environmental indicators of potentially toxic elements contamination. Environ Pollut. 2025 Oct 13;386:127258. [http://dx.doi.org/10.1016/j.envpol.2025.127258
Mechanistic insights into synergistic antimelanoma activity of glycolysis inhibition and lysosomal destabilization
No full textBackground: Melanoma cells exhibit high glycolytic activity and increased lysosomal fragility. Cathepsins, overexpressed
in melanoma, support tumor progression but trigger cell death upon lysosomal membrane permeabilization (LMP).
This study investigated the mechanisms by which the glycolytic inhibitor 2-deoxy-D-glucose (2DG) synergized with LMP
inducer L-leucyl-L-leucine methyl ester (LLOMe), to induce death of human A375 melanoma cells.
Materials and Methods: Cell viability was measured by the crystal violet test. Lysosomal integrity was evaluated using
acidophilic dyes LysoTracker and acridine orange via fluorescent microscopy and flow cytometry. Fluorometric assays
were used to measure mitochondrial membrane potential (JC-1), mitochondrial superoxide production (MitoSOX™
Red), glycolytic activity (pH-Xtra™ Glycolysis Assay), oxidative phosphorylation (MitoXpress® Xtra Oxygen Consumption
Assay), and intracellular ATP levels (ATP assay kit). Cells were pretreated with cysteine cathepsin inhibitors (E64d,
MG132), aspartic cathepsin inhibitor (Pepstatin A), lysosomal acidification inhibitor (bafilomycin A1), antioxidant NAC,
and energy booster L-carnitine to further dissect the contribution of specific pathways.
Results: LLOMe synergized with 2DG to induce cytotoxicity in A375 cells. LLOMe, with or without 2DG, induced LMP,
evidenced by reduced LysoTracker and acridine orange signals. Bafilomycin A1 rescued cell viability, likely by preventing
LLOMe accumulation and activation within lysosomes. E64d and MG132, but not Pepstatin A, partially restored
viability, implicating cysteine cathepsins in the observed toxicity. LLOMe, with or without 2DG, triggered mitochondrial
depolarization and mitochondrial superoxide production, which was prevented by bafilomycin A1 and MG132. NAC
rescued cells from 2DG+LLOMe-induced cytotoxicity. Metabolic assays showed that 2DG reduced glycolysis and LLOMe
suppressed oxidative phosphorylation, leading to synergistic ATP depletion, while L-carnitine protected cells from the
combined treatment.
Conclusions: LLOMe-induced LMP and cysteine cathepsin release lead to mitochondrial dysfunction and OXPHOS
inhibition, which, together with 2DG-mediated glycolysis blockade, result in energy collapse and cell death, highlighting
the therapeutic potential of dual glycolytic–lysosomal targeting.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. 158. (Oncology Insights; No. 3)
Engineered 3D in vitro model based on Ba-alginate microfibers for rapid anticancer drug screening
No full textBackground: Reliable preclinical models in cancer research are crucial for improving anticancer drug development.
Existing models, such as two-dimensional (2D) cell cultures and animal studies, often fall short in accurately replicating
human cancer biology, leading to significant differences in drug efficacy between preclinical studies and clinical trials.
In this work, a simplified 3D cancer cell culture model in the form of Ba-alginate microfibers with immobilized cancer
cells was developed, optimized and validated for anticancer drug screening.
Material and Methods: Alginate microfibers with immobilized human non-small cell lung carcinoma cells (NCI-460
cell line, ATCC® HTB-177™) were obtained by manual extrusion of cell-alginate suspension (4×106 cells cm-3, 2 wt. %
Na-alginate solution) through a 25-gauge needle into the gelling solution containing 45 mM Ba2+ (BaCl2∙2H2O). After
1 min of gelation, the obtained microfibers were washed with culture medium RPMI-1640 and cultured up to 7 days.
Cells in microfibers, as well as cells in 2D cultures, were treated with different concentrations of cisplatin (0.5−50 μM),
etoposide (0.5−500 μM), or vinorelbine (1−1000 μM) either 24 or 72 h after immobilization, while untreated cells
served as the control. Microfibers were evaluated for cell viability by live/dead staining and metabolic activity by MTT
assay, while the expression of drug resistance–related genes was assessed by using quantitative real-time PCR.
Results and Conclusions: The cells remained viable and metabolically active, forming cell aggregates during the 7-day
cultivation period. Analysis of cell sensitivity to anticancer drugs revealed increased drug resistance in 3D cultures
compared to 2D cultures across all treatment groups (cisplatin, etoposide and vinorelbine). Additionally, the gene
expression analysis showed a significant upregulation of drug resistance–related genes in 3D cultures, including a
3.2-fold increase in MDR1 and a 2.7-fold increase in ABCG2 expression following cisplatin treatment, and a 1.5-fold
increase in ABCC1 expression after vinorelbine exposure, whereas these changes were not observed in 2D cultures.
Overall, the obtained results demonstrate the potential of the proposed 3D cancer model for rapid, relevant and highthroughput
anticancer drug screening.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. 167-8. (Oncology Insights; No. 3)
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
The effect of salinity stress on total phenolic, flavonoid, and hydroxycinnamic acid content and the antioxidative capacity of centaury (Centaurium erythraea Rafn) shoots and roots grown in vitro
No full textCommon centaury (Centaurium erythraea Rafn) is an important plant species in traditional medicine because it has been used for the treatment of numerous diseases since ancient times. Moreover, centaury is rich source of antioxidants and antimicrobial compounds and, as such, can be considered as a potential food additive. Considering that, in nature, centaury can often be found on moderately saline soils, it was interesting to investigate the effects of salinity stress, caused by sodium chloride (NaCl), on total phenolic, flavonoid, and hydroxycinnamic acid content and antioxidative capacity, in shoots and roots grown in vitro. Centaury shoots were cultured on a solid ½MS medium supplemented with graded NaCl concentrations (0, 50, 100, 150, and 200 mM). Centaury roots were also grown in vitro but in liquid ½MS medium containing the same graded NaCl concentrations. After eight weeks, the content of total phenolic, flavonoid, and hydroxycinnamic acid content and antioxidative capacity were determined using colorimetric methods. The results obtained in this work showed that graded NaCl concentrations induced the accumulation of total phenols in both shoots and roots. Regarding total flavonoid content, the graded NaCl concentrations also induced the accumulation of total flavonoids in roots, while in shoots the highest content was detected in the presence of 50 mM NaCl. On the other hand, the increment of NaCl in nutrient media promoted the accumulation of total hydroxycinnamic acid content in both shoots and roots, with the exception of roots grown in the presence of 200 mM NaCl. Finally, the DPPH test has shown an increased antioxidative capacity in shoots and roots, which was dependent on applied NaCl concentration. It can be concluded that salinity stress affected shoots and roots’ total phenolic, flavonoid, and hydroxycinnamic acid content differently but also elevated antioxidative capacity.Plants 2025: From Seeds to Food Security; 2025 Mar 31 - April 2; Barcelona, Spain, 2025; Basel: MDPI; 2025. p. 21847
Green solution for lead pollution: phytoremediation with Festuca rubra and brushite-aluminosilicate geopolymer material
No full textABSTRACTAluminosilicate materials are known for their high removal efficiency of lead (Pb) ions from aqueoussolutions, whereas in Pb-contaminated soils they can immobilise lead ions and improve soil quality,contributing to more efficient remediation. Brushite-aluminosilicate geopolymer materials synthesisedfrom an abandoned, raw kaolinite clay with the addition of 2, 4, and 6 wt% of brushite, were tested fortheir efficiency in removing Pb ions with Festuca rubra, which is known to tolerate its high concentrationin soil. The highest efficiency for Pb immobilisation in the soil was observed with the addition ofbrushite-metakaolin-based geopolymer GPB2% and GPB6%, whereas physiological and biochemicalparameters indicate a reduction in metal stress with increasing brushite content, as the concentrationsof proline, total phenolic content and antioxidant activity in the shoots of the plants to which GPB6%was added were even lower than in some samples grown on uncontaminated soil. Moreover, in thissample the highest concentrations of photosynthetic pigments were found. This study suggests that thebrushite-metakaolin geopolymer material improves the potential of F. rubra in remediation, whereas itsstability and harmlessness to the environment, further recommend its use in the field.This is an Accepted Manuscript version of the following article, accepted for publication in Plant Biosystems on April p, 2025, available at: [https://doi.org/10.1080/11263504.2025.2485975
The role of the gut microbiota in high-fat diet-induced hepatic steatosis in juvenile and adult mice
No full textBackground and aims: A high-fat diet (HFD) disrupts hepatic lipid
metabolism and contributes to steatosis, with the gut microbiota
playing a key role through bacterial metabolites. As the composition
of the gut microbiota changes with age, the timing of HFD
introduction could influence hepatic lipid metabolism. Therefore,
the aim of this study was to investigate the role of gut microbiota
composition in hepatic lipid metabolism disturbances in juvenile and
adult mice on HFD.
Method: Male C57BL/6J mice were divided into three groups: a
control group on a standard diet (10 kcal% fat, D12450J, Research
Diets) and two groups on a HFD (60 kcal% fat, D12492, Research
Diets,) starting either in the juvenile (3rd week) or adult phase (9th
week). All animals were sacrificed at 17 weeks of age. Liver sections
were analyzed histologically and levels of markers of hepatic lipid
metabolism were determined by Western blot or qPCR. The
composition of the gut microbiota was determined by 16S rRNA
sequencing. Metabolites were quantified by HPLC, while their effects
on lipid metabolism were assessed by measuring the levels of Gprotein-coupled receptor 43 (GPR43) and farnesoid X receptor (FXR)
in the liver.
Results: HFD-induced hepatic steatosis occurred in both juvenile and
adult mice, although the mechanisms underlying this pathology
differed between the two groups. Adult mice showed increased de
novo lipogenesis and decreased β-oxidation, whereas juvenile mice
showed increased fatty acid uptake and decreased triglyceride
secretion. The composition of the gut microbiota and their
metabolites differed between groups, resulting in lower total SCFA,
acetate, and predicted bile acid production in adults. These results
were accompanied by decreased expression of GPR43 and FXR, which
could contribute to altered hepatic lipid metabolism in adult mice.
Conclusion: The results show that changes in gut microbiota
composition induced by HFD are more detrimental in adult mice,
since it is associated with increased de novo lipogenesis and
decreased β-oxidation in the liver, as well as reduced production of
beneficial microbial metabolites. These results pinpoint that the
effect of high-calorie diets on the gut microbiota is age-specific, with
more prominent metabolite-driven effects on lipid metabolism in
adult miceAbstract book of EASL Congress 2025; 2025 May 7-10; Amsterdam, the Netherlands. Amsterdam: Elsevier; 2025. p. S280. (Journal of Hepatology; Vol. 82. Suppl. 1)