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Изолација протопласта код кичице (Centaurium erythraea Rafn.)
No full textИзолација протопласта је кључна техника у биљној биотехнологији која омогућава директно проучавање изолованих биљних ћелија, њихову генетичку трансформацију и регенерацију генетички униформних биљака. У овом раду, по први пут је успешно изведена изолација протопласта из кичице (Centaurium erythraea Rafn.), лековите биљке са значајним фармаколошким својствима. Кичицу такође карактерише велика развојна пластичност, због чега се позиционирала као значајна врста за проучавање развића и регенерације биљака. На кичицу и Arabidopsis, као контролну биљку и широко примењен модел-систем у биљној биологији, примењен је оптимизован протокол за изолацију протопласта из листова уз помоћ ензима Cellulase R10 и Macerozyme R10 за дигестију ћелијског зида, док су протопласти стабилизовани у МЕС пуферу са додатком манитола. Протопласти обе врсте су профилтрирани кроз филтере промера пора 70 µm, центрифугирани и ресуспендовани у пуферу, након чега су квантификовани уз помоћ хемоцитометра. Након утврђивања задовољавајуће бројности, протопласти су обојени флуоресцеин диацетатом (ФДА) ради процене вијабилности. Добијени резултати указују на ефикасну изолацију великог броја вијабилних протопласта. Успешна изолација протопласта из кичице отвара нове могућности за генетичко инжењерство ове врсте. Планирано је даље коришћење изолованих протопласта за трансформацију посредовану полиетилен гликолом (ПЕГ), са циљем убацивања векторске ДНК и рибонуклеопротеинских (РНП) комплекса у ћелије. Ова истраживања ће допринети развоју стратегија за боље разумевање механизама развојне пластичности кичице, кроз функционалну карактеризацију гена од интереса и регенерацију трансформисаних биљака без химерних својстава.Stanković M, Bojović B, Stefanović O, Kosanić M, Milutinović M, editors. Program i izvodi saopštenja: Druga konferencija srpskog biološkog društva "Stevan Jakovljević" Kragujevac; 2025 Sep 17-19; Kragujevac, Serbia. Kragujevac: Srpsko biološko društvo "Stavan Jakovljević"; 2025. p. 153
Book of abstracts / The International Conference on Light Pollution and Dark Sky Protection ENLIGHT; 2025 Nov 28-30; Novi Sad, Serbia
No full textTranscriptomic Landscape of Paclitaxel-Induced Multidrug Resistance in 3D Cultures of Colon Cancer Cell Line DLD1
No full textMultidrug resistance (MDR) significantly contributes to colon cancer recurrence, making it essential to understand its molecular basis for improved therapies. This study aimed to identify genes and pathways involved in resistance to standard chemotherapeutics by comparing transcriptome profiles of sensitive and paclitaxel-induced MDR colonospheres. Cell viability and growth were assessed following treatment with 5-fluorouracil, oxaliplatin, irinotecan, bevacizumab, and cetuximab. Drug concentrations in culture media posttreatment were measured using high-performance liquid chromatography (HPLC). RNA sequencing (RNA-seq) of untreated sensitive and resistant colonospheres identified differentially expressed genes linked to baseline resistance. Our results confirmed cross-resistance in the resistant model, showing highest oxaliplatin tolerance may involve mechanisms beyond efflux. Transcriptome analysis highlighted upregulation of PIGR and activation of the ribosomal signaling pathway as potential resistance mediators. Notably, AKR1B10, a gene linked to chemotherapeutic detoxification, was overexpressed, whereas genes related to adhesion and membrane transport were downregulated. The overexpression of ribosomal protein genes suggests ribosome biogenesis plays a key role in acquired resistance. These findings suggest that targeting ribosome biogenesis and specific deregulated genes such as PIGR and AKR1B10 may offer promising strategies to overcome MDR in colon cancer
Cold Plasma Treatment Alters the Morphology, Oxidative Stress Response and Specialized Metabolite Content in Yellow Iris (I. reichenbachii) Callus
No full textThe application of non-thermal (cold) plasmas is considered an environmentally friendly method that could affect plant metabolism and cellular development or can be used for the commercial production of natural products that cannot be chemically synthesized. In the present study, the non-embryogenic callus of iris (Iris reichenbachii Heuff.) was treated with a Radio Frequency (RF) plasma needle device using He as a working gas. We investigated short-term (up to seven days) and long-term (up to one year) changes on morphological, physiological and biochemical levels. An increased production of O2− and H2O2 was observed in the callus tissue after plasma treatment. The enzymes SOD and CAT represented the frontline in the antioxidant defense against reactive oxygen species (ROS) produced during the first hour of treatment, while POX was the leading antioxidant enzyme seven days after plasma treatment. Significant long-term morphological changes were observed in the calli due to the increased mitotic activity of the plant cells. In addition, three flavonoids (naringenin, apigenin and acacetin) and two isoflavonoids (irisolidone and irilone) were detected only in the plasma-treated tissue even one year after plasma treatment. The present study emphasizes the application of the plasma technique to promote meristematic activity and stimulate the production of specialized metabolites in iris calli
Surviving Allchar: arsenic and thallium tolerance and distribution in Viola metallophytes
No full textBackground and Aims
Viola tricolor subsp. macedonica and Viola arsenica are metallophytes from the abandoned Allchar mine site in the Republic of North Macedonia, an area extremely enriched in thallium and arsenic, with up to 5750 and 12 800 mg kg−1 in the soil, respectively. This study aimed to infer tolerance mechanisms evolved in the two Viola species by analysis of the tissue-level distribution of arsenic and thallium.
Methods
Seedlings of V. tricolor subsp. macedonica and V. arsenica were grown under different thallium and arsenic treatments in hydroponics. Synchrotron-based micro-X-ray fluorescence (µXRF) analysis was used to elucidate elemental distribution in hydrated plant organs and tissues.
Key Results
Plants dosed with increasing concentrations of arsenic and thallium had higher accumulation of these elements, especially in the roots. In V. arsenica, thallium mainly accumulated in the shoots, with the mature leaves being the main site of deposition. In the leaves of V. tricolor subsp. macedonica, the highest thallium concentrations occured around the stomata.
Conclusions
Foliar accumulation of thallium is the main tolerance strategy in V. arsenica, whereas the limited translocation into the shoot and potentially excreting excess thallium through the stomata in V. tricolor subsp. macedonica appears to be an important mechanism for survival in the extremely toxic habitat at the Allchar site
Horseradish (Armoracia rusticana L.) leaf juice encapsulated within polysaccharides-blend-based carriers: Characterization and application as potential antioxidants in mayonnaise production
No full textThis study aimed to encapsulate cold-pressed horseradish leaf juice within maltodextrin/alginate (MD/AL), maltodextrin/guar gum (MD/GG), and maltodextrin/gum Arabic (MD/GA) by spray-drying, to characterize the encapsulates, and to test their potential as mayonnaise oxidation-preventing ingredients. The encapsulates exhibited desirable physicochemical, morphological, structural, and thermal properties, highlighting MD/GA-containing encapsulates, especially regarding high encapsulation yield (78.50 %). Also, encapsulates contained a significant amount of phenolics, which were stable during freezer storage. The encapsulates successfully delayed the mayonnaise oxidation: 31.91–38.94 % more than the synthetic antioxidant ethylenediaminetetraacetic acid, especially highlighting MD/AL-containing encapsulates. Also, the encapsulates improved product quality with a higher pH and lower acidity after storage compared to the controls. Overall acceptability of encapsulates-containing mayonnaises and commercial mayonnaise did not differ significantly. This study contributes to sustainable development by providing new insights into the valorization of horseradish leaves, as a promising alternative to synthetic additives to prolong the oxidative stability and shelf-life of high-oil-containing foods
Ergothioneine improves healthspan of aged animals by enhancing cGPDH activity through CSE-dependent persulfidation
No full textErgothioneine (ET), a dietary thione/thiol, is receiving growing attention for its possible benefits in healthy
aging and metabolic resilience. Our study investigates ET’s effects on healthspan in aged animals, revealing lifespan extension and enhanced mobility in Caenorhabditis elegans, accompanied by improved stress resistance and reduced age-associated biomarkers. In aged rats, ET administration enhances exercise endurance, muscle mass, and vascularization, concomitant with higher NAD+ levels in muscle. Mechanistically, ET acts as an alternative substrate for cystathionine gamma-lyase (CSE), stimulating H2S production, which increases protein persulfidation of more than 300 protein targets.Among these, protein-persulfidation-driven activation of cytosolic glycerol-3-phosphate dehydrogenase (cGPDH) primarily contributes to the ETinduced NAD+ increase. ET’s effects are abolished in models lacking CSE or cGPDH, highlighting the essential role of H2S signaling and protein persulfidation. These findings elucidate ET’s multifaceted actions and provide insights into its therapeutic potential for combating age-related muscle decline and metabolic perturbations
Evaluation of the Impact of Selected Metallic Contaminants on Wild Rutilus rutilus Through Integrated Antioxidant Biomarker Responses
No full textThe application of biomarkers is often used to better understand the assessment of the toxicity in aquatic organisms within their natural environment. To this end, we examined the gills and liver of roach (Rutilus rutilus) to measure changes in the activity of the antioxidant enzymes superoxide dismutase, catalase (CAT), glutathione peroxidase, glutathione reductase and the phase II biotransformation enzyme glutathione-S-transferase (GST), as well as changes in the concentration of lipid peroxides (LPOs). Roach were collected at two localities, Veliko Ratno ostrvo and Višnjica (VIS), on the Danube River, where the concentrations of dissolved metals (Cu, Cd, Zn, Fe, Mn, Hg, Ni, As, Pb) in both fish tissues were quantified. This paper describes the integrated reaction of antioxidant enzymes and LPO levels of roach from two sampling localities in the Danube River. Principal component analysis revealed obvious differences of the antioxidant biomarkers investigated between tissues and localities, while integrated biomarker response showed that the liver of the VIS locality had the stronger antioxidant biomarker response. Despite the concordant hypothesized antioxidant induction at the VIS locality in both tissues studied, the effects as a sign of contaminant exposure appear to be mediated in part by suppression of the antioxidant system, with CAT and GST as potential candidate tissue toxicity biomarkers of contaminants
European distribution and intramuscular pathogenicity of divergent lyssaviruses West Caucasian bat virus and Lleida bat lyssavirus
No full textAmong lyssaviruses, West Caucasian bat virus (WCBV) and Lleida bat lyssavirus (LLEBV) raise concern as their divergence from rabies virus leads to the inefficacy of available prophylactic agents. Both viruses were described in the bat Miniopterus schreibersii. We investigated the European distribution of WCBV and LLEBV by screening sera fromMiniopterus schreibersii across eight countries, finding widespread serological evidence and positivity up to 70%.We evaluated the intramuscular lethality of wild type isolates in Syrian hamsters.WCBV induced 100% lethality and a clinical disease compatible with furious rabies. All animals infected with LLEBV remained healthy for 40 days, despite one individual testing positive in the brain. We confirmed LLEBV’s intramuscular a-pathogenicity using mice. Infected hamsters developed antibodies by day seven, regardless the virus and the clinical outcome. This study highlights the widespread circulation ofWCBV and LLEBV in Europe and suggests differences in neuro-invasiveness and/or pathogenesis that are crucial for risk assessment