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Bioactive metabolites isolated from Hericium erinaceus (Bull.) Pers.
No full textHericium erinaceus is considered one of the most promising medicinal fungi for the treatment
of neurodegenerative diseases, containing a wide variety of bioactive compounds, including
hericenones, erinacines, and many other related molecules, which exhibit significant
neurotrophic and neuroprotective properties. Previous studies demonstrated that erinacine
enriched mycelium of H. erinaceus can delay neuronal cell death in animal models of
neurodegenerative conditions such as Alzheimer’s disease and ischemic stroke. [1] Our research
focus on the isolation of a diverse range of bioactive compounds from employing conventional
chromatographic techniques, followed by their structural characterization predominantly
through nuclear magnetic resonance (NMR) spectroscopy. Hericene A was isolated from the
ethyl acetate extract of the H. erinaceus fruiting bodies by performing gravitational
chromatography, followed by additional purification on semi-dry flash chromatography.
Furthermore, erinacines A and C were isolated from the ethyl acetate extract of the mycelium.
The extract was initially processed through semi-dry flash chromatography, followed by
subsequent purification utilizing preparative high-performance liquid chromatography (HPLC).
The structures of isolated compounds were determined using 1D (¹H NMR and 13C) and 2D
(COSY, JRES, HMBC, HSQC and NOESY) NMR experiments. These purification methodologies
demonstrated high efficiency and cost-effectiveness, resulting in the isolation of pure natural
compounds appropriate for subsequent pharmacological investigations. Future investigations
will prioritize the optimization of isolation protocols for various metabolites derived from both
the mycelium and fruiting bodies of H. erinaceus, alongside comprehensive characterization and
systematic evaluation of their biological activities, with the aim of elucidating potential
therapeutic applications in the treatment of neurodegenerative disorders, particularly
Alzheimer's disease.Bregović N, Namjesnik D, Novak P, Parlov Vuković J, editors. Book of Abstracts: Adriatic NMR Conference; 2025 Sep 18-20; Vodice, Croatia. Zagreb: Croatian Chemical Society; 2025. p. 52
Chemical profile and enzyme-inhibitory activity from different parts of blue poppy (Papaver somniferum)
No full textDue to the significant side effects associated with synthetic drugs, there has been a growing interest in
natural agents in recent years. In an effort to prevent numerous diseases, plant-based natural
therapeutics are receiving increasing scientific attention. Poppy (Papaver somniferum) is one of the
oldest plants used in traditional and alternative medicine, primarily due to its rich chemical composition.
In this study, different parts of the poppy were examined, namely: stems, leaves, root, capsule, pressedcake (PC) and defatted pressed-cake (DPC). The main focus of this research was to determine the
chemical profile and potential enzyme-inhibitory activity of extracts obtained from different parts of
poppy. Chemical composition of obtained extracts was determined by liquid chromatography/mass
spectroscopy (LC-MS). Enzyme inhibitory effect was investigated against α-amylase, α-glucosidase,
tyrosinase and acetylcholinesterase.
To obtain extracts from different parts of poppy, conventional extraction (CE) was used, where different
percentages of ethanol (0, 20, 40, 60, 80, and 96% w/v) were used. The extraction lasted for 24 hours,
at room temperature, and the shaking speed was 180 rpm. One portion of PC was defatted using
Soxhlet extraction with methylene-chloride as a solvent, and then extracted by CE.
According to LC-MS analysis, 62 compounds were identified in different parts of the poppy. Some of the
compounds which are present in all parts of the plant were: glutaric acid, malic acid, p-coumaric acid,
narcein. As a result, for α-amylase and α-glucosidase, the extracts from PC and leaves showed the
greatest inhibitory power on these enzymes. Tyrosinase was most effectively inhibited by the extract
from the DPC, while the capsule exhibited the best activity in the case of acetylcholinesterase. It can be
concluded that poppy represents a plant rich in bioactive compounds, and as such can be used as an
alternative to synthetic drugs.Popović-Đorđević J, Kostić A, editors. Book of Abstracts: 3rd European Symposium on Phytochemicals in Medicine and Food (3-EuSPMF); 2025 Jul 1-4; Belgrade, Serbia. Belgrade: University of Belgrade, Faculty of Agriculture; 2025. p. 92
Pulmonaria mollis: Rediscovering a forgotten edible plant through phytochemical profiling and bioactivity assessment
No full textPulmonaria species were historically valued as traditional remedies for respiratory ailments and as wild food, yet their culinary and medicinal applications have largely been forgotten. Among them, Pulmonaria mollis remains underinvestigated, with scarce phytochemical and pharmacological data. This study aimed to fill this gap by investigating the phenolic composition and biological activities of methanol extracts from the aerial parts (PMA) and roots (PMR) of P. mollis. UHPLC–MS4 Orbitrap analysis identified 59 compounds, mostly phenolic acids. Quantitative profiling by UHPLC/DAD/(−)HESI-MS/MS showed PMA was rich in rosmarinic acid (4508.27 μg/g), p-hydroxybenzoic acid (605.70 μg/g), and caffeic acid (325.42 μg/g), whereas PMR contained higher levels of salvianolic acids A (2311.09 μg/g) and B (4697.98 μg/g). Along with significant antioxidant activity confirmed by standard spectrophotometric assays, where PMR showed slightly stronger radical scavenging capacity, both extracts effectively protected DNA from hydroxyl- and peroxyl-induced oxidative damage. In enzyme inhibition assays, both extracts demonstrated α-amylase inhibitory effects, while PMR showed particularly potent α-glucosidase inhibition (IC50 = 14.00 μg/mL), outperforming acarbose. Moderate antibacterial and antifungal activities were also observed, with PMR generally more effective, whereas cytotoxicity testing indicated selective activity against A431 (IC50 values of 178 ± 11 μg/mL and 64 ± 3 μg/mL for PMA and PMR, respectively) cells without affecting normal cells. Overall, the findings highlight P. mollis as a promising candidate for development into functional foods and nutraceuticals aimed at chronic disease prevention, providing valuable phenolic compounds and biological properties that support its potential use as an alternative to the commonly used Pulmonaria officinalis
Bioherbicidal Evaluation of Methanol Extract of Sorghum halepense L. Rhizome and Its Bioactive Components Against Selected Weed Species
No full textSorghum halepense (L.) Pers. (common name Johnson grass) is a perennial invasive weed that causes great harm worldwide, and its allelopathy has been demonstrated in a series of experiments. The present study offers new insights into its organ-specific phytochemical profiles using state-of-the-art metabolomic technology and explores the effects of a methanol extract of S. halepense rhizomes (ShER) and its major bioactive compounds (p-hydroxybenzoic acid and chlorogenic acid) on three noxious weed species. The phytotoxic effects of ShER are reflected through the inhibition of seed germination and reduced seedling growth, which are accompanied by changes in the antioxidant system of seedlings. Phytotoxicity is species specific and concentration dependent, and it is more pronounced against Chenopodiastrum murale (L.) S. Fuentes, Uotila & Borsch and Datura stramonium L. than highly tolerant Amaranthus retroflexus L. Catalase (CAT) is most likely the major mediator in the removal of reactive oxygen species, which are generated during germination and early seedling growth of Ch. murale exposed to ShER. The results of the present study imply the high potential of ShER in the management of amaranthaceous and solanaceous weeds, such as Ch. murale and D. stramonium, respectively. The present study offers an environmentally friendly solution for the biological control of weeds belonging to the families Amaranthaceae and Solanaceae. Also, the results of this research highlight the possibility of effective management of S. halepense by using it as a feedstock for bioherbicide production
Synergistic antimelanoma effect of shikonin-mediated glycolysis inhibition and chloroquine-induced lysosomal destabilization in A375 melanoma cells
No full textBackground: Malignant melanoma is the most aggressive form of skin cancer, for which a fully effective therapy is
still lacking. Melanoma cells upregulate glycolysis to sustain rapid growth and proliferation, while increased lysosomal
volume and elevated activity of lysosomal enzymes cathepsins contribute to their invasiveness and chemoresistance.
However, lysosomal membrane permeabilization (LMP), followed by cathepsin release into the cytoplasm, can trigger
cell death. Shikonin (SH) is a natural inhibitor of the glycolytic enzyme pyruvate kinase M2 (PKM2). Chloroquine (CQ),
an antimalarial drug, can induce LMP and promote the release of cathepsins B and D into cytoplasm. The aim of this
study was to evaluate the antimelanoma effects of combined glycolysis inhibition by SH and lysosomal destabilization
by CQ in A375 melanoma cells.
Material and Methods: The expression levels of PKM2, cathepsin B (CTSB), and cathepsin D (CTSD) genes were
analyzed in patient-derived normal skin, benign nevi, and malignant melanoma samples from the publicly available
GEO dataset GSE3189 using the Mann-Whitney U test in GraphPad Prism. Cell viability was assessed by mitochondrial
dehydrogenase reduction assay and crystal violet staining. The type of cytotoxic interaction between the two treatments
was determined using the interaction coefficient (α), calculated as α = SFsh × SFCQ/SFsh+cq, where SF represents
the survival fraction of the indicated treatment. Apoptosis and necrosis were evaluated by flow cytometry following Annexin V-FITC and propidium iodide staining.
Results: Expression of PKM2, CTSB, and CTSD was significantly higher in malignant melanoma than in normal skin or
benign nevi. Both SH and CQ reduced A375 cell viability in a dose-dependent manner, with their combination exhibiting
enhanced cytotoxicity. The calculated α value was greater than 1, indicating a synergistic interaction between SH and
CQ. Annexin V/PI flow cytometry revealed that both agents induced apoptosis, which was significantly amplified by the
combined treatment.
Conclusions: Dual targeting of melanoma-upregulated PKM2 with SH and cathepsins B/C with CQ resulted in synergistic
proapoptotic effects in A375 melanoma cells, suggesting that this combined approach may represent an effective
therapeutic strategy for 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. 157-8. (Oncology Insights; No. 3)
Developing under artificial light is not so bright: Oxidative stress as a physiological response to light pollution across amphibian life stages
No full textLight pollution has emerged as a new environmental challenge. Natural light has long been a selecting force in shaping physiological, morphological and behavioral traits across species. The introduction of artificial light at night (ALAN) disrupts the natural light-dark cycle, affecting a broad range of biological processes. In amphibians, ALAN has been suggested as a potential driver behind demographic declines in recent years. In this study, we investigated how exposure to cold LED light affects the development, body length, and oxidative status of the European tree frog (Hyla arborea). Individuals were exposed to ALAN from egg hatching (GS25) to two developmental endpoints: the larvae (GS38) and the juveniles. The experiment included three night-time light conditions: a natural dark control (<0.3 lx), LED light intensity at 20 lx, and LED light intensity at 90 lx. Larvae reared under artificial light reached metamorphosis in a shorter period and exhibited larger body sizes. Nocturnal light also induced changes in oxidative status across life stages. Tadpoles under both 20 lx and 90 lx levels experienced greater oxidative damage of lipids. Likewise, juveniles had higher oxidative damage of proteins and elevated activities of the first-line antioxidant defense enzymes (SOD, CAT and GSH-Px) in response to both light intensities. Our results show that exposure to ALAN shapes development and body size of amphibians, which appears to be accompanied by disturbances in redox balance. Overall, we highlight that ALAN can be a factor affecting amphibian populations, and thus it should be taken into consideration in future conservation strategies.This is the peer reviewed version of the following article: Petrović T, Gavrić-Čampar JP, Burraco P, Gavrilović B, Despotović S, Radovanović T, Radmilović A, Mirč M, Tomašević Kolarov N, Prokić M. Developing under artificial light is not so bright: Oxidative stress as a physiological response to light pollution across amphibian life stages. in Comparative biochemistry and physilogy. Part A, Molecular & integrative physiology. 2025;302:111819. [https://doi.org/10.1016/j.cbpa.2025.111819
Impact of Mn and Zn deficiency on photosynthesis and pigments of Tilia tomentosa moench in urban environment
No full textSilver linden (Tilia tomentosa Moench) is a native, drought-tolerant, and pollution-resistant
species that is widely distributed in urban green spaces, and whose ability to respond to reduced
availability of essential micronutrients may indicate urban environmental stress. The aim of
this study was to evaluate its potential as a bioindicator of growing conditions in urban parks
of Belgrade (Obrenovac, Hala Pionir, Kosmaj) by analyzing the content of essential
micronutrients (Mn and Zn), photosynthetic efficiency (Fv/Fm), and total carotenoids (Tot
Carot). The leaves were analysed by ICP-OES (Mn and Zn), fluorometry (Fv/Fm), and
spectrophotometry (Tot Carot). The results showed that Mn was deficient at all sites, in the
range of 10.58-24.64 mg kg-1, while Zn was in the deficit range in Obrenovac and Kosmaj,
(10.22-20.71 mg kg-1), confirming micronutrient limitation in urban environment. The lowest
Fv/Fm value (0.698) was measured in Hala Pionir, indicating that Mn deficiency, which is
essential for PSII function and photosynthetic pigment synthesis, significantly affects
photosynthetic efficiency. The total carotenoid content (1.703-1.834 mg g-1) was below the
optimum, probably due to the combined deficiency of Mn and Zn, with Mn deficiency
impairing pigment synthesis and stability, and Zn deficiency weakening antioxidant protection
and accelerating oxidative pigment degradation. The results confirm that T. tomentosa clearly
responds to reduced micronutrient availability through changes in photosynthetic and pigment
parameters, emphasizing its potential as a sensitive bioindicator of urban environmental stress
and as a tool for environmental biomonitoring.Golub D, editor. Book of asbtracts: V Symposium of biologists and ecologists of Republic of Srpska SBERS2025; 2025 Nov 13-25; Banja Luka, Bosnia and Herzegovina. Banja Luka: Faculty of Natural Sciences and Mathematics, University of Banja Luka; 2025. p. 156
The Impact of Decreased GSK3β and S6K1 Expression in TNBC Patients
No full textBreast cancer is the most frequent and lethal type of cancer that affects women worldwide. Triple-negative breast cancer (TNBC) is the most aggressive type of breast cancer, having high rate of recurrence, metastasis, and mortality, with very limited options for treatment, and a tendency to develop resistance to conventional therapy. These circumstances mean that it is necessary to develop effective therapies for TNBC patients which would circumvent resistance mechanisms. The PAM and Wnt signaling pathways are among those responsible for therapy resistance in TNBC, as they also have major roles in different cellular processes such as metabolism, proliferation, metastasis, stemness, and survival. We analysed the expression of GSK3β and S6K1 as interacting components of the two pathways in order to examine the relation between them and determine whether they could be used as predictive markers in TNBC. The expression of mRNA was examined with real-time PCR and protein expression with immunohistochemistry. Our results showed that protein expression is in line with mRNA expression. We found a positive correlation between the mRNA expressions of GSK3β and S6K1, showing their coordinated transcription. We also showed that their simultaneous low expression is unfavorable for TNBC patients and could possibly be used as a predictive marker
A systematic review on the health effects of fermented wheat germ extract with emphasis on cancer
No full textFermented wheat germ extract (FWGE) is one of the few fermented food products
listed in the EFSA novel food Catalogue. It is derived from wheat germ, a by-product
of wheat processing, through fermentation with Sacchoromyces cerevisiae. The
most widely studied and patented form of FWGE is marketed as Avemar (also
referred to as MSC), standardized to contain methoxy-substituted benzoquinones.
Given its predominant use in research, this systematic narrative review focused
primarily on FWGE use within the medical application. The objective of this review
was to systematically evaluate the functionality and potential health benefits of
FWGE, following the Study Protocol-S8 developed under COST Action CA20128 –
PIMENTO and registered on the Open Science Framework (https://osf.io/fq53j/).
A systematic literature search of human studies was conducted through PubMed,
Scopus, and Cochrane Library. Studies involving adult participants who received
FWGE interventions were included, with primary clinical endpoints selected according
to the main indications for FWGE. Additional outcomes were reported when
available. Out of the 51 records identified by the literature search, six studies met
the inclusion criteria. Data from these studies were extracted and synthesized
in summary tables. Supplementary information on the functionality of FWGE
was retrieved through a non-systematic search of animal and in vitro studies.
Furthermore, this review highlights the potential bioactive constituents of FWGE,
particularly benzoquinones, peptides, and phenolic compounds, as mediators of
its anticancer and anti-inflammatory properties. Among its proposed mechanisms,
FWGE may suppress cancer cell proliferation by disrupting the glucose-related
metabolic pathways. While the findings suggest that FWGE may possess therapeutic
potential, especially in oncology, the strength of evidence remains limited. Of the
six included human studies, only four employed proper control groups and only
two demonstrated high methodological quality. As such, the current evidence
base is insufficient to draw definitive conclusions, and well-designed clinical trials
are needed to strengthen this evidence. Moreover, future research should also explore the development of novel FWGE formulations with enhanced bioactive profiles, optimized by modulating fermentation conditions, including such as microbial strain, pH, temperature, and duration
A systematic review of health promoting effects of consumption of whey-based fermented products on adults
No full textIntroduction: Fermented whey-based products show significant potential as
functional foods, owing to their rich nutritional profile and the generation of
bioactive compounds during fermentation. This systematic narrative review
evaluates the health effects of fermented-whey consumption based on evidence
from human studies in adults.
Methods: A systematic literature search was conducted using electronic
databases including, PubMed, Scopus, and Cochrane Library for studies
published between 1.1.1970 and 31.12.2024. All human clinical studies
conducted with adults over 18 years old were included in this study. Inclusion
criteria were randomized controlled trials and clinical studies involving adults
consuming fermented whey products. Data extraction and quality assessment
were performed using CADIMA software and standardized protocols. Studies
identified by the search strategy and extracted data were screened independently
by 2 reviewers using the CADIMA software. Risk of bias was assessed using the
Risk of Bias 2 tool.
Results: After screening 1852 titles and abstracts and assessing 20 articles for
eligibility, a total of 12 human intervention studies met the inclusion criteria and
were included in the systematic narrative review. Consumption of fermented
whey products was associated with improvements in muscle mass, glycemic
control, lipid profiles (notably triglycerides and LDL cholesterol), immune
function (e.g., increased natural killer cell activity), and reductions in oxidative stress and inflammation. Some studies also reported benefits for gastrointestinal and urinary tract health. The health effects were attributed to increased bioavailability of branched-chain amino acids, bioactive peptides, and microbial metabolites such as exopolysaccharides and short-chain fatty acids. Most interventions were well tolerated, with no serious adverse effects reported.
Conclusion: Fermented whey products demonstrate promising health benefits across multiple physiological systems. While current evidence supports their use as functional food ingredients, further large-scale, long-term clinical trials are needed to confirm efficacy and elucidate mechanisms of action. Fermented whey appears to be a safe and versatile option for enhancing adult nutrition and health