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Nonlinear laser imaging for unravelling biomonitor potential: from insect down to fungi
No full textNonlinear laser scanning microscopy (NLSM) is an advanced powerful technique for deep tissue high resolution imaging. It is based on interaction of ultrashort (femtosecond) laser pulses with specimen material. The image is made upon detection of nonlinear optical effects, two photon excitation fluorescence (TPEF), second or third harmonic generation (S/THG) referred as three imaging modalities. Each of the modalities provide different information on specimen: TPEF on fluorescent molecules (both, exogenous and endogenous), SHG on ordered structures such as starch, collagen and myosin, while THG, as the most versatile, but yet the least selective, on distribution of indices of refraction changes. While NLSM is indispensable technique for in vivo imaging on large laboratory animals (mice and rats), there is plenty of literature on the various applications, from animal cells and tissues to plants [1] In this work we present the application of NLSM for imaging and laser processing on the insects and hyphae as the specimens that are potentially interesting for biomonitoring field. Insects are mostly built of chitin, polysaccharide which exhibit strong blue fluorescence when excited in near UV region by conventional single photon excitation. The same spectroscopic properties are valid for chitosan. This feature might be used for imaging, for instance by confocal microscopy under 405 nm excitation, but due to strong absorption of laser light by chitin structures, the imaging is limited only to outer morphology and shape of the specimen without deep insight and structural details. This is overcome by TPEF imaging since the excitation is achieved by 700-800 nm femtosecond laser pulses, penetrating deeply into specimen without interaction except in the tiny focal volume. Utilizing this advantage high resolution images form in depth structures and 3D models are reported [2]. Since chitin, along with chitosan, is the main constituent of the hyphal cell wall imaging possibilities by TPEF are utilized for hyphae, along with THG modality which provides more details with less selectivity [3]. Since THG is produced at steep changes of index of refraction, it is used mostly to visualise lipids (e.g. lipid droplets) that has significantly different index of refraction from the surrounding cytoplasm. These two techniques in combination have been employed for studying lipid metabolism in live hyphae of Phycomyces blakesleeanus and proved to be reliable, providing more details than conventional biochemical methods. In addition to TPEF signal form chitinous cell wall, other endogenous structures like NADH and FAD can produce detectable signal that might be used for imaging, as well. NLSM as a microscopic technique is not limited to imaging only, but for sample processing as well, in terms of laser cell surgery of hyphal cell wall. This method in combination with TPEF imaging has been utilized to enable patch clamping on the bare hyphal membrane and electrophysiological measurements on adult hyphae in controlled and reproducible manner [4]. This will open the possibility for a golden standard method such as patch clamp to be applied to whole kingdom of live organisms, which was not possible so far. Since both, insects and fungi are sensitive to pollution and might be used as biomonitors, utilization of NLSM and its modalities might be a turnover in their characterization by advanced techniques. The results shown in the literature and this presentation might be considered as the first step for utilization of NLSM’s modalities in the biomonitoring and in the environment protection in general.Zarić N, Aničić Urošević M, editors. Book of Abstracts: The 3rd Conference of the International Association for Biomonitoring of Environmental Pollution: IABEP2025; 2025 Oct 13-15; Belgrade, Serbia. Belgrade: Institute of Physics; 2025. p. 56
Artificial light at night and influence on eye structure in Triturus newts (Urodela, Salamandridae)
No full textLight pollution is one of the modern anthropogenic impacts on the environment that is
growing rapidly. The disruption of the natural day-night regime imposed by artificial
light at night (ALAN) places significant pressure on biological systems. So far it has
been observed that ALAN causes disturbances in the metabolism, reproduction, and
behavior of animals, but it is less known how it affects the visual system, especially in
amphibians. Newts are mostly nocturnal animals with complex lifecycles and they
may be sensitive to changes in the environment such as ALAN. This research aimed to
examine how different intensities of long-term exposure to Light Emitting Diodes
(LEDs) light during the night can affect retina of larvae, and metamorphosed juveniles
of Triturus sp.
We used LED lights with intensities of 5 and 30 lux, which correspond to the actual
intensities of ALAN in area inhabited by amphibians, and two light color temperatures:
warm (2700K), and cold (6000K) light. After hatching and up to the 62 stage, larvae
were divided into five experimental groups. the control group (artificial daylight + dark
at night) and four treatment groups with LED lights. 5lux 2700K, 5lux 6000K, 30lux
2700K, and 30lux 6000K during night. The animals were kept under these condition for
three months. We also used metamorphosed luveniles which were divided
immediately after metamorphosis into two groups - control and treatment (30lux
6000K), kept under these conditions for two months. Eye tissue was studied using
histological methods of H&E staining as well as immunofluorescence for visual
pigment rhodopsin detection. Our preliminary results indicate a higher intensity of the fluorescent signal for rhodopsin in individuals that were treated in comparison with the
control group, especially in juveniles. Further data analysis should provide a clearer
insight into how ALAN affects different layers in the retina especially quantity and
distribution of rhodopsin in rod cells.Abstract booklet: Oral & Poster presentations: 23rd European Congress of Herpetology: SEH2025; 2025 Sep 8-12; Bonn, Germany. Societas Europaea Herpetologica; 2025. p. 290-1
The effect of French marigold (Tagetes patula L.) essential oil volatile compounds on the production of reactive oxygen species and defense responses to biotic stress in potato (Solanum tuberosum L.)
No full textBiljke su u neprestanoj interakciji sa organizmima iz svog okruženja, a isparljiva organska jedinjenja koja ispuštaju u atmosferu smatraju se „jezikom“ kojim biljke ostvaruju tu interakciju. Međutim, o uticaju koji ova isparljiva jedinjenja imaju na biljke iz susedstva se veoma malo zna. Kadifica (Tagetes patula L.) je aromatična biljka poznata po svom repelentnom dejstvu na brojne štetočine. Tradicionalno se uzgaja uz različite povrtarske kulture, uključujući i krompir (Solanum tuberosum L.). U cilju ispitivanja efekata isparljivih jedinjenja etarskog ulja (EU) kadifice na metabolizam i fiziologiju krompira, biljke krompira su izlagane njihovom dejstvu u laboratorijskim uslovima tokom različitih vremenskih perioda i praćen je odgovor krompira, odnosno njegov redoks, antioksidativni, fitohormonski i odbrambeni status, kao i uticaj na primarni metabolizam šećera. Analiza transkriptoma mikroerej metodom, pokazala je da je 2796 transkripata diferencijalno eksprimirano nakon izlaganja EU, potvrdivši značajan uticaj ovog EU na različite procese krompira. Pokazano je da u krompiru izloženom EU dolazi do akumuliranja reaktivnih kiseoničnih vrsta, ali i njihovog intenzivnog uklanjanja putem aktivacije antioksidativnih mehanizama. Povećana ekspresija gena koji kodiraju enzime uključene u regulaciju glutationskog metabolizma ukazala je na značajnu ulogu glutationa u inicijalnom odgovoru krompira na isparljiva jedinjenja, ali i na njegovu potencijalnu ulogu u intercelularnoj sekvestraciji usvojenih isparljivih molekula EU. Izmenjeni fitohormonski profil, kao i izmenjeni metabolizam šećera ukazuju na aktiviranje odbrambenih odgovora krompira, međutim povećan nivo šećera pogodovao je razviću krompirove zlatice (Leptinotarsa decemlineata Say.), koja se njima hranila. S druge strane, krompir izložen EU je pokazao povišenu otpornost na fitopatogenu gljivu Alternaria alternata (Fr.) Keissl. sa pet puta manjim procentom površine listova pod infekcijom.Plants are in constant communication with their environment. The volatile organic compounds they release into the atmosphere are considered the "language" plants use to communicate. However, the impact of these volatile compounds on recipient plants has not been extensively studied. French marigold (Tagetes patula L.) is an aromatic plant known for its pest-repellent properties. Traditionally, it is cultivated alongside various vegetable crops, including potatoes (Solanum tuberosum L.). To investigate the effects of French marigold essential oil volatile compounds on potato metabolism and physiology, potato plants were exposed to these volatiles under laboratory conditions for different time periods. The response of potato, in terms of redox balance, antioxidant activity, phytohormonal and defensive status, as well as the impact on primary sugar metabolism, was monitored. cDNA microarray revealed that 2796 transcripts were differentially expressed following exposure to the essential oil, confirming its significant impact on various physiological aspects of potato. It was shown that treated potatoes acumulate reactive oxygen species, which are subsequently neutralized by antioxidants. The increased expression of genes of glutathione metabolism highlighted the critical role of glutathione in the initial potato response to volatiles and its potential role in the sequestration of volatile molecules. Altered phytohormonal profiles and sugar metabolism indicated the activation of defense responses, however the elevated sugar levels proved beneficial to Colorado potato beetles (Leptinotarsa decemlineata Say.), which exhibited enhanced developmental performance when feeding on exposed plants them. On the other hand, treated potatoes exhibited enhanced resistance to the phytopathogenic fungus Alternaria alternata, with a fivefold reduction in the percentage of lesions in the total leaf area
Myxoid pattern in smooth muscle tumor of uncertain malignant potential (STUMP) is a significant prognostic parameter with difficult diagnostic dilemmas
No full textUterine smooth muscle tumors of uncertain malignant potential
(STUMPs) represent a heterogeneous group of tumors with varying histological and
biological characteristics. Tumors featuring a dominant myxoid stroma are uncommon
in the uterus. This case emphasizes the importance of recognizing the myxoid pattern in
smooth muscle tumors, especially within the STUMP category. Case: In this study, we
present a 39-year-old woman with a large transmural intrauterine nodule. The findings
indicated smooth muscle tumor tissue with a fascicular pattern, without significant
ischemic necrosis or cytological pleomorphism. The mitotic count was 1/10 High Power
Field (HPF). The tumor exhibited more than 50% myxoid histomorphology, indicative of
a rare and specific type of smooth muscle differentiation. Immunohistochemical analysis
showed positive reactions for smooth muscle differentiation, retained progesterone
receptor expression, and no aberrant tumor protein p53 (p53) or cyclin-dependent kinase
inhibitor 2A (p16) expressions. The marker of proliferation Ki-67 was low. Considering
all the characteristics, this myxoid smooth muscle tumor was diagnosed as a STUMP
myxoid neoplasm. The patient underwent a hysterectomy with bilateral salpingectomy
and bilateral ovarian conservation. The patient is currently under follow-up and has not
experienced any complications. Conclusions: This case illustrates an extremely rare
instance of STUMP, which continues to pose challenges in both diagnosis and treatment.
Myxoid patterns are exceptionally uncommon and often hard to detect, making them
surprising in smooth muscle tumors. Recognizing this distinct differentiation is crucial,
as accurate diagnostic criteria must be applied during the histopathological evaluation of
these tumors. The presence of a myxoid pattern suggests a worse prognosis, highlighting
the need for more vigilant follow-up in these patient
Pancreatin digestion of B-phycoerythrin from Porphyridium purpureum releases chromopeptides with inhibitory activity towards SARS-CoV-2 main protease
No full textUnique features of covalently attached tetrapyrrole chromophores give phycobiliproteins (PBPs) remarkable
optical and bioactive properties with significant potential for application in the food industry, medicine, and
biotechnology. This study investigated the potential of chromopeptides, released upon enzymatic digestion of
major PBPs from cyanobacteria Limnospira platensis (C-Phycocyanin) and red microalgae Porphyridium purpureum
(B-Phycoerythrin), to inhibit the main protease (Mpro) of SARS-CoV-2 virus. The digest obtained by pancreatin
digestion of B-Phycoerythrin exhibits the highest inhibitory activity with estimated IC50 values in the low
micromolar range. Mass spectrometry was used to determine the sequence of RP-HPLC-purified chromopeptides,
and small peptides (2–7 amino acid residues) were identified. The inhibitory activity of purified chromopeptides
confirmed that chromopeptides are the major components in B-Phycoerythrin digest and are responsible for Mpro
inhibition. The binding of chromopeptides to Mpro was evaluated using molecular docking, while molecular
dynamics simulations were employed to propose a plausible mechanism of Mpro inhibition
Pre-sowing seed exposure to static magnetic field can improve cabbage yield
No full textThe static magnetic field (SMF) has been shown to influence living organisms, for example, plants, fungi, insects, mice, and rats. The effects caused by SMF depend on the intensity of the applied field and exposure duration; however, discovered dependence on these parameters is non-linear. Numerous studies that used SMF for interaction with plants reported increased germination, seedling growth, length and biomass of plants, as well as drought and salinity tolerance of different plants. However, majority of the published studies rely on experiments performed in the laboratory conditions. In order to investigate if and how the conditions of pre-sowing exposure of cabbage seeds (Brassica oleracea var. capitata f. alba) to SMF affect the yield of crops we took the experiment into the agricultural field and followed plant development from sowing to harvesting. Pre-sowing seeds exposures for one hour to highly homogeneous field of 50, 100, and 350 mT were considered and the plants were analyzed two and 3.5 months after sowing. In the transplant stage, seed exposure to 100 mT was found to produce 21.5% more leaves and plant weight increase of 30.7% compared to the control group. In the mature stage, the seed treatment with 350 and 100 mT caused statistically significant increase of the plant weight and head weight. In the group exposed to 350 mT the increase of plant and head weight was 22.7 and 24.8%, respectively, whereas for the 100 mT exposure the corresponding increases were 16.9 and 18.3%, respectively. Consequently, the pre-sowing 1 h exposure of cabbage seeds to 350 mT was found to be the most effective of the three studied SMF treatments, since it can produce 24.8% larger cabbage yield compared to the non-treated control
Nove antimikrobne strategije u liječenju infekcija uzrokovanih multirezistentnim sojevima gram pozitivnih bakterija
No full textInfections caused by gram-positive, multidrug-resistant bacteria,
especially streptococci, coagulase-positive staphylococci, and vancomycin-resistant
enterococci, have been a major medical and economic problem for decades, leading to
increased morbidity, mortality, and healthcare costs. Multidrug-resistant bacteria are
often found in hospitals and long-term care facilities, infecting mainly people with a
weakened immune system, such as patients on chronic hemodialysis and with post
operative wound infections, as well as solid organ transplant recipients. The World
Health Organization categorizes these bacteria into critical, high, and medium priority
groups depending on the urgency of developing new antibiotics. This review aims to
summarize the latest findings on the research and development of new and necessary
strategies to prevent the spread of antimicrobial resistance in gram-positive bacteria.
A literature search on this topic over the last two decades was conducted in the
PubMed, Scopus, and Google Scholar databases. As the number of newly developed
and approved antibiotics decreases, great efforts are being made to find new
antimicrobial strategies to treat infections caused by multidrug-resistant strains. These
include (1) modulation of commercial antibiotics to enhance their effect, (2)
development and application of specific monoclonal antibodies to neutralize bacteria,
(3) blocking the production of virulence factors and inhibition of quorum sensing
especially in biofilm, (4) modulation of the innate immune response by probiotics and
fecal transplantation, (5) use of bacteriophages due to their pronounced ability to
inhibit multidrug-resistant bacteria that cause wound infections, (6) combined use of
bacteriophages and antibiotics, and (7) repurposing of non-antibiotic drugs. Despite
the potential of the aforementioned antimicrobial strategies, the focus will continue to
be on finding new strategies to more effectively address the challenge of antimicrobial
resistance and improve public health.Infekcije uzrokovane gram-pozitivnim, multirezistentnim bakterijama,
posebno streptokokama, koagulaza pozitivnim stafilokokama i vankomicin rezistentnim
enterokokama su decenijama veliki medicinski i ekonomski problem, što je dovelo do
povećanog morbiditeta, mortaliteta i troškova zdravstvene zaštite. Multirezistentne
bakterije su često prisutne u bolnicama i ustanovama za dugotrajnu njegu, inficirajući
uglavnom osobe sa oslabljenim imunološkim sistemom, kao što su pacijenti na
hroničnoj hemodijalizi i sa postoperativnim infekcijama rana, kao i primaoci
transplantata solidnih organa. Svjetska zdravstvena organizacija kategoriše ove
bakterije u grupu kritičnog, visokog i srednjeg prioriteta u zavisnosti od hitnosti
razvoja novih antibiotika. Ovaj pregled ima za cilj da sumira najnovije nalaze o
istraživanju i razvoju novih i neophodnih strategija za sprječavanje širenja
antimikrobne rezistencije kod gram-pozitivnih bakterija. Pretraživanje literature o ovoj
temi u posljednje dvije decenije obavljeno je u bazama podataka PubMed, Scopus i
Google Scholar. Kako se broj novorazvijenih i odobrenih antibiotika smanjuje, ulažu
se veliki napori u pronalaženju novih antimikrobnih strategija za liječenje infekcija
uzrokovanih multirezistentnim sojevima. Ovo uključuje (1) modifikaciju komercijalnih
antibiotika da bi se pojačao njihov efekat, (2) razvoj i primjenu specifičnih
monoklonskih antitela za neutralizaciju bakterija, (3) blokiranje proizvodnje faktora
virulencije i inhibiciju quorum sensing, posebno u biofilmu, (4) modulaciju urođenog
imunološkog odgovora probioticima i fekalnom transplantacijom, (5) upotrebu
bakteriofaga zbog izražene sposobnosti inhibicije multirezistentnih bakterija koje
izazivaju infekcije rana, (6) kombinovanu primjenu bakteriofaga i antibiotika i (7)
prenamjenu lijekova koji ne pripadaju antibioticima. Uprkos potencijalu navedenih
antimikrobnih strategija, fokus će i dalje biti na pronalaženju novih strategija za
efikasnije rješavanje izazova antimikrobne rezistencije i poboljšanje javnog zdravlja.Grujić R, Stijepić М, editors. Scientific Proceedings: Scientific Conference: SANUS 2025. Book 4; 2025 Sep 18-20; Prijedor, Bosnia and Herzegovina. Prijedor : College of Health Sciences; 2025. p. 305-13. (Scientific Conference; Vol. 3. No. 3)
Developing a method for direct detection of erinacine A in mycelium of Hericium erinaceus (Bull.) Persoon
No full textHericum erinaceus represents one of the most promising functional foods, with potential
benefits in the treatment of neurodegenerative diseases, such as Alzheimer's disease. In
addition to neuroprotective activity, H. erinaceus exhibits several other pharmacological
effects, such as anti-hypercholesterolemic, anti-tumor, and antioxidant activities, which
have been well documented [1]. Neuroprotective potential of the mycelium is best
demonstrated on the two main compounds, erinacine A, and C, stimulators of nerve
growth factor (NGF) synthesis [2]. Numerous Hericium-based supplements are currently
marketed for the treatment of dementia; however, their therapeutic efficacy largely
depends on the concentration of bioactive constituents, such as erinacines, underscoring
the importance of rigorous standardization. In the present study, two complementary
analytical methods were integrated as a quality control approach for the detection of
erinacine A in H. erinaceus mycelium extracts. Specifically, nuclear magnetic resonance
(NMR) spectroscopy was employed alongside semi-preparative high-performance liquid
chromatography (HPLC). The presence of erinacine A was initially confirmed through
comprehensive 2D NMR spectroscopy and detailed analysis of the resultant mixture.
Subsequently, HPLC analysis was performed using both the conventional 210 nm
wavelength and an additional wavelength specific to erinacine A. The applied
methodology represents a reliable and promising approach for the determination of
erinacines, offering enhanced speed, improved efficiency, and increased reliability.Book of abstracts: 11th Conference of Young Chemists of Serbia; 2025 Oct 25; Kragujevac; Serbia. Belgrade: Serbian Chemical Society; 2025. p. 107
Impact of urbanisation on local amphibian diversity: an example from Belgrade city and suburbia
No full textUrbanisation fragments habitats, increases isolation, hinders amphibian migration and causes population decline. Urban pollution significantly impacts amphibian physiology, behaviour, ecology and health. Tolerant species dominate, as sensitive ones often cope poorly with environmental change. We surveyed 13 Belgrade urban sites (2022–2023) - an area of 1720 ha - for amphibians and found 611 individuals from 7 species.
Site attributes, including the 500 m surrounding area, were analysed in QGIS. Attributes were corelated using Kendall Tau, excluding but one if ρ ≥ 0.9. Observed individual numbers were correlated with site attributes per species using Kendall Tau, applying 20% FDR and LOOCV for stability. Due to the small sample size, only strong, stable correlations (ρ ≥ 0.5; LOOCV ≥ 0.7) are reported. All lengths are in km, all areas in ha.
For Rana dalmatina presence, a strong correlation was found with the number of lakes at a site (ρ = 1.00; LOOCV = 92.31%). Green frogs presence correlated with: Urban environment area around site (ρ = -0.70); roads length around site (ρ = -0.67); distance to nearest natural environment (ρ = -0.63); distance to nearest river (ρ = -0.58); footpaths quantity at site (ρ = -0.53); rivers length around site (ρ = 0.51); running water length around site (ρ = 0.51); running waters number around site (ρ = 0.53); streams length at site (ρ = 0.54); rivers number at site (ρ = 0.54); running water length at site (ρ = 0.55); green areas size around site (ρ = 0.58); rivers number around site (ρ = 0.59); standing water area at site (ρ = 0.62); streams number at site (ρ = 0.64); Running water number at site (ρ = 0.73) – all LOOCV = 100%. No significant results for Bombina variegata, Bufo bufo, Lissotriton vulgaris and Salamandra salamandra.Abstract booklet: Oral & Poster presentations: 23rd European Congress of Herpetology: SEH2025; 2025 Sep 8-12; Bonn, Germany. Societas Europaea Herpetologica; 2025. p. 283
A bite in a summer night: nocturnal feeding of dice snakes on the shore of Prespa lake
No full textDuring long-term ecological monitoring of dice snakes (Natrix tessellata) on Golem Grad Island—a strictly protected site in Lake Prespa, North Macedonia—we consistently observed nocturnal activity, particularly during summer months. Over 25 night surveys (from 2009 to 2024) resulted in the capture of 368 individuals. Observed night activities, such as swimming and foraging, in July and August (~between 21:00 and 23:00), suggest a partial shift to nocturnal activity, likely to reduce heat stress, dehydration, and predation risk. During one session in July 2024, coinciding with the spawning of the endemic Prespa bleak (Alburnus belvica), 28 snakes were captured in just 30 minutes along a five-meter shoreline stretch. Of these, 23 were females— including five gravid—and five were males. A total of 18 fish, averaging 10 cm in length and 9 g in mass, were regurgitated. Notably, four gravid females had recently fed, challenging the assumption that gravid snakes cease feeding. The presence of exhausted fish in shoreline spawning pools likely provides valuable foraging opportunities. These observations underscore the ecological link between dice snakes and Lake Prespa’s endemic fish, both of which face increasing threats from human activity. Poaching is a major concern, as illegal nets often trap and drown snakes, including gravid females, causing severe population decline. Our data show a steady drop from a population once estimated at over 10,000 individuals. These findings highlight the urgent need for stronger enforcement of protective measures on and around Golem Grad Island. Nocturnal activity has been documented in Natrix snakes, but most reports are based on citizen science rather than systematic research, leaving this aspect of their behavior understudied.Abstract booklet: Oral & Poster presentations: 23rd European Congress of Herpetology: SEH2025; 2025 Sep 8-12; Bonn, Germany. Societas Europaea Herpetologica; 2025. p. 24-5