imagine (Institute of molecular genetics and genetic engineering)
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Pollen–Pistil Interactions in Autochthonous Balkan Sweet Cherry Cultivars—The Impact of Genotype and Flowering Temperature
No full textThe efficacy of sweet cherry production is highly dependent on the regularity of flowering events and genetic-determined relations between female sporophyte and male gametophyte, which became even more important with higher flowering temperatures caused by climate change. Special attention is paid to the genetic diversity that provides essential sources of potential temperature-tolerance genes. Our study aimed at the genetic and reproductive characterization of Balkan cherry cultivars of autochthonous origin (‘Canetova’, ‘G-2’, ‘Dolga Šiška’ and ‘Ohridska Crna’), and six potential pollenizers. To identify S-haplotypes, the polymerase chain reaction (PCR) method was used to detect the S-ribonuclease (S-RNase) and S-haplotype-specific F-box protein (SFB) alleles, combined with fragment analysis and S-RNase sequencing. Pollination experiments were performed at three Balkan localities over two flowering seasons, and the fluorescence microscopy method was used to assess the cultivars’ male/female reproductive behaviour. A novel S-RNase allele S40 was identified in ‘Ohridska Crna’ for the first time. ‘Ohridska Crna’ also demonstrated the best adaptability to higher temperatures regarding primary ovule longevity. This feature makes it desirable from the aspect of breeding new cultivars that can withstand the impacts of climate change. The findings on male-female relations and their temperature dependence open up the possibility for yield prediction and smart horticultural decisions that can be made to guide cherry production
Significance of GSTM1 and GSTT1 Gene Deletions in Glioma Patients in Polish Population: Pilot Study
No full textBackground:Detoxification enzymes of the glutathione S-transferase (GST) family are cytosolic phase II detoxification enzymes and play an important role in the normal functioning of the human antioxidant system. When the normal function of GST is disturbed or absent, there can be disturbances in cell metabolism, proliferation, and apoptosis. Deletions in the GSTM1 and GSTT1 genes have been observed in several different diseases as well as in the development of cancer. There is a need to analyze the relationship between glioma and GSTM1 and GSTT1 gene deletion to better understand the relationship between brain tumors and GST polymorphisms, which is crucial for adopting a multidisciplinary approach to prognosis and treatment of brain tumors.Methods:In a cross-sectional clinical-laboratory study, gene deletions were examined in 34 patients with brain tumors originating from glial cells—gliomas and 88 healthy individuals. All participants were of Polish nationality and were not related.Results:An increase in GSTM1 and GSTT1 gene deletions was observed in glioma patients compared with the control group. The greatest increase showing a marked rise of 10 times (11.8% vs 1.14%, P < .05) is in the null genotype of both genes (GSTM1−/GSTT1) [odds ratio [OR] = 0.86; 95% confidence interval [CI] = 0.09-0.802] but less in the genotype with deletion of 1 GST gene (GSTM1−/GSTT1+ and GSTM1+/GSTT1−). In addition, the findings indicated a decrease in the non-deletion genotype of both genes (GSTM1+/GSTT1+) in healthy individuals. This study showed a higher frequency of GST gene deletion in glioma patients in the studied population.Conclusions:Based on the obtained findings, it can be said that the examination of the selected detoxification enzymes can be a useful marker in the diagnosis of glioblastoma
A Novel ST1 Lactonase Reduces Virulence and Inhibits Biofilm Forma=on and Extends the Lifespan of Caenorhabdi9s elegans Infected with Pseudomonas aeruginosa MMA83
No full textBackground: Bacterial biofilms represent a major challenge in clinical seNngs due to their resistance to
an3bio3cs and immune responses. Pseudomonas aeruginosa is a significant opportunis3c pathogen
responsible for chronic infec3ons, par3cularly in immunocompromised pa3ents. Quorum quenching
(QQ) enzymes, such as lactonases, show promise in disrup3ng biofilm forma3on and reducing bacterial
virulence. This study examines the effects of ST1-YtnP lactonase from thermophilic Bacillus licheniformis
on biofilm forma3on and virulence of P. aeruginosa MMA83.
Methods: The an3biofilm effect of recombinant ST1-YtnP lactonase on mul3-drug resistant P.
aeruginosa MMA83 was analyzed using fluorescence microscopy. CaenorhabdiLs elegans infec3on
model was used to evaluate the enzyme’s effect on virulence and host survival. The liquid killing assay
was employed using the C. elegans AU37 mutant strain to quan3ta3vely evaluate host survival in
response to ST1-YtnP lactonase treatment.
Results: Fluorescence microscopy showed a significant reduc3on in P. aeruginosa MMA83 biofilm
forma3on with ST1-YtnP lactonase, leading to a looser, less dense biofilm. Planktonic cell growth
remained unaffected. In C. elegans, treatment resulted in an almost 100% survival rate, while untreated
infected worms showed 0% survival within 24 hours. ST1-YtnP lactonase exhibited no toxicity, consistent
with previous studies on QQ enzyme safety in eukaryo3c models.
Conclusion: ST1-YtnP lactonase inhibits P. aeruginosa MMA83 biofilm forma3on and reduces virulence
in C. elegans, indica3ng its poten3al as a safe and effec3ve an3virulence agent for biomedical
applica3ons.Book of abstract: BIOFILMS 11, 13 – 15 May 2025, Mercure Cardiff Holland House
Hotel & Spa, Cardiff, U
Paraburkholderia phytofirmans PsJN triggers local and systemic transcriptional reprogramming in Arabidopsis thaliana and increases resistance against Botrytis cinerea
No full textFungal pathogens are one of the main causes of yield losses in many crops, severely affecting agricultural production worldwide. Among the various approaches to alleviate this problem, beneficial microorganisms emerge as an environmentally friendly and sustainable alternative. In addition to direct biocontrol action against pathogens, certain plant growth-promoting bacteria (PGPB) enhance the plant immune defense to control diseases through induced systemic resistance (ISR). Paraburkholderia phytofirmans PsJN has been shown as an efficient biocontrol agent against diseases. However, the specific mechanisms underlying these beneficial effects at both local and systemic level remain largely unknown. In this study, we investigated the transcriptional response of Arabidopsis thaliana at above- and below-ground levels upon interaction with P. phytofirmans PsJN, and after Botrytis cinerea infection. Our data clearly support the protective effect of P. phytofirmans PsJN through ISR against B. cinerea in plants grown in both soil and hydroponic conditions. The comparative transcriptome analysis of the mRNA and miRNA sequences revealed that PsJN modulates the expression of genes involved in abiotic stress responses, microbe-plant interactions and ISR, with ethylene signaling pathway genes standing out. In roots, PsJN predominantly downregulated the expression of genes related to microbe perception, signaling and immune response, indicating that PsJN locally provoked attenuation of defense responses to facilitate and support colonization and the maintenance of mutualistic relationship. In leaves, the increased expression of defense-related genes prior to infection in combination with the protective effect of PsJN observed in later stages of infection suggests that bacterial inoculation primes plants for enhanced systemic immune response after subsequent pathogen attack
Organic and natural deep eutectic solvents for the extraction of bacterial prodiginine biopigments
No full textEfficient bacterial cell wall disruption is a crucial step for obtaining valuable intracellular bacterial natural products like deeply colored prodiginines. Development of greener, environmentally friendly chemical methods is a preferred approach with respect to energy requirements and cost. In this study, we have re-examined extraction of bacterial prodiginines, aiming at making their biotechnological production more sustainable, particularly concerning downstream processing and recovery. For the first time, natural deep eutectic solvents (NADES) were employed to extract prodigiosin (PG) and streptorubin (SB) from lyophilized bacterial biomasses and their extraction efficiency was compared to green organic solvents. Bacterial biomasses of PG-producing Gram-negative Serratia marcescens ATCC 27117 and SB-producing Gram-positive Streptomyces sp. BPS43 were obtained by fermentation, followed by culture centrifugation and freeze-drying. Acidified ethyl acetate (1 % v/v HCl) was the most efficient for PG extraction, yielding 2.36 mg of PG per 100 mg of biomass, corresponding to predicted yields by UV–visible spectroscopy. NADES composed of glycerol:ascorbic acid in 1:1 molar ratio afforded a comparable amount of 2.14 mg PG/100 mg of biomass. For SB extraction, acidified ethyl acetate (1 % v/v HCl) was the most efficient, yielding 1.41 mg SB/100 mg of biomass, while the best NADES composed of betaine:lactic acid in 1:1 molar ratio extracted 17-fold less 0.0834 mg SB/100 mg biomass. While NADES were shown as a greener alternative for PG extraction, thick peptidoglycan layer of Gram-positive Streptomyces sp. proved difficult for treatment with NADES alone, indicating a need for combining NADES with other downstream processing techniques for effective SB extraction
IMMOBILIZATION OF LEAF BRANCH COMPOST CUTINASE (LCC) ONTO BACILLUS SUBTILIS SPORES FOR EFFICIENT PLASTIC DEGRADATION
No full textEnzymes provide an efficient, clean, and reliable means of performing specific chemical reactions and have
found use in the agricultural, food, chemical, and pharmaceutical industries. Despite the numerous advantages of
using enzymes, their application is hindered by a lack of stability in harsh conditions, cost of production, and
purification. A potential strategy for mitigating these shortcomings is the anchoring of enzymes onto bacterial
spores. Spores are structures produced by certain bacteria to help them survive extreme environmental
conditions. Fusing enzymes to the surface of spores via spore display systems can increase their thermal and pH
stability while substantially simplifying the enzyme purification process.
In this work, we present the construction of a Bacillus subtilis spore display system for the immobilization of the
benchmark plastic-degrading enzyme LCCICCG using the spore coat protein CotG. We established the optimal
sporulation conditions for spore yield in terms of incubation temperature and time. The stability of spores and
the immobilized enzyme was tested at high temperatures, high salt concentrations and in organic solvents.
Additionally, we tested the esterase activity of spores produced at different temperatures and determined that
spores produced at 25°C for 24 h had the highest activity. The most active spores were used in poly(ethylene
terephthalate) (PET) and poly(caprolactone) (PCL) degradation experiments, proving the spores could be used as
efficient biocatalysts for plastic degradation. In addition to the previously mentioned benefits, the use of spores
as plastic-degrading biocatalysts opens up opportunities for coupling the degradation process with biosynthetic
processes for the up-cycling of released monomers. This innovative approach paves the way for sustainable
plastic waste management and biotechnological advancements.BOOK OF ABSTRACTS: 17th International Symposium on Biocatalysis and Biotransformations Basel, Switzerland, June 29-July 3, 202
Development of Bioactive Cotton, Wool, and Silk Fabrics Functionalized with Origanum vulgare L. for Healthcare and Medical Applications: An In Vivo Study
No full textBackground: This study presents an innovative approach to developing bioactive natural fabrics for healthcare and medical applications. Methods: An ethanol extract of Origanum vulgare L. (in further text: OE), exhibiting exceptional antioxidant (100%) and antibacterial activity (>99% against E.coli and S.aureus), was employed to biofunctionalize cotton, wool, and silk fabrics. Results: All biofunctionalized fabrics demonstrated strong antioxidant activity (>99%), while antibacterial efficacy varied by fabric: cotton > 54%, wool > 99%, and silk > 89%. OE-biofunctionalized wool possessed the highest release of OE’s bioactive compounds, followed by silk and cotton, indicating substrate-dependent release behavior. This tunable fabrics’ OE release profile, along with their unique bioactivity, supports targeted applications: OE-functionalized silk for luxury or prolonged therapeutic use (skin-care textiles, post-surgical dressings, anti-aging products), cotton for disposable or short-term use (protective wipes, minor wound coverings), and wool for wound dressings. The biocompatibility and cytotoxicity of OE-biofunctionalized wool were evaluated via in vitro assays using healthy human keratinocytes and in vivo testing in Wistar albino male rats. The obtained results revealed that OE-functionalized wool significantly accelerated wound closure (97.8% by day 14), enhanced collagen synthesis (6.92 µg/mg hydroxyproline), and improved tissue and systemic antioxidant defense while reducing oxidative stress markers in skin and blood samples of rats treated with OE-biofunctionalized wool. Conclusions: OE-biofunctionalized wool demonstrates strong potential as an advanced natural solution for managing chronic wounds. Further clinical validation is recommended to confirm its performance in real-world healthcare settings. This work introduces an entirely new application of OE in textile biofunctionalization, offering alternatives for healthcare and medical textiles
Degradation of Synthetic and Natural Textile Materials Using Streptomyces Strains: Model Compost and Genome Exploration for Potential Plastic-Degrading Enzymes
No full textGiven the environmental significance of the textile industry, especially the accumulation of nondegradable materials, there is extensive development of greener approaches to fabric waste management. Here, we investigated the biodegradation potential of three Streptomyces strains in model compost on polyamide (PA) and polyamide-elastane (PA-EA) as synthetic, and on cotton (CO) as natural textile materials. Weight change of the materials was followed, while Fourier-Transform Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy (SEM) were used to analyze surface changes of the materials upon biodegradation. The bioluminescence-based toxicity test employing Aliivibrio fischeri confirmed the ecological safety of the tested textiles. After 12 months, the increase of 10 and 16% weight loss, of PA-EA and PA, respectively, was observed in compost augmented with Streptomyces sp. BPS43. Additionally, a 14% increase in cotton degradation was recorded after 2 months in compost augmented with Streptomyces sp. NP10. Genome exploration of the strains was carried out for potential plastic-degrading enzymes. It highlighted BPS43 as the most versatile strain with specific amidases that show sequence identity to UMG-SP-1, UMG-SP-2, and UMG-SP-3 (polyurethane degrading enzymes identified from compost metagenome). Our results showcase the behavior of Streptomyces sp. BPS43 in the degradation of PA and PA-EA textiles in composting conditions, with enzymatic potential that could be further characterized and optimized for increased synthetic textile degradation
Abundance and diversity of airborne antibiotic resistome and mobilome in Belgrade metropolitan area during summer season: a metagenomic approach
No full textIntroduction: The importance of the environment for the transmission of antimicrobial
resistance is increasingly acknowledged. The airborne transmission route of antibiotic
resistance genes (ARGs) is particularly significant since ARGs-carrying bioaerosols could
travel long distances and remain in the atmosphere for a long period of time. In addition,
mobile genetic elements (MGEs) can promote the spread of ARGs in the environment.
Objective: To reveal the potential of the atmosphere in the dissemination of ARGs and
MGEs, the antibiotic resistome and mobilome of airborne metagenomes collected
during summer season 2024 at seven locations in the Belgrade metropolitan area were
analysed.
Method: Outdoor air was sampled at seven locations using hydrophobic polypropylene
membrane filters (air volume of 30 m3). Airborne DNA samples were subjected to shotgun
metagenome sequencing (Illumina Novaseq X plus) and bioinformatic analysis
(Novogene, UK). ARGs and MGEs were annotated using CARD, Integrall, Isfinder and
Plasmid databases, respectively.
Findings: The tested locations were selected according to the air pollution during heating
season (highly polluted: Borca, Despota Stefana, Lestane, Zemun Ugrinovacka, Veliki
Crljeni; low polluted: Barajevo, Zeleno brdo). Metagenomic analysis revealed that the air
sampled at Zemun Ugrinovacka showed the highest abundance of ARGs, while the
lowest level was observed in sample Barajevo. The most abundant ARG was blaTEM, while
transposon TnSwi1 was dominant in four samples. MGEs content profiles varied among
the samples, although Veliki Crljeni and Lestane showed similar patterns of MGEs
relative abundance.
Conclusion: This study revealed that Belgrade air could be a potential dissemination
route for antimicrobial resistance.Book of abstract: FEMS MICRO 2025: Congress & Exhibition, taking place in Milan, Italy, from 14-17 July 2025
Biodentine Stimulates Calcium-Dependent Osteogenic Differentiation of Mesenchymal Stromal Cells from Periapical Lesions
No full textBiodentine, a tricalcium silicate cement, has emerged as a retrograde root-end filling material to promote periapical lesion (PL) healing after apicoectomy. However, its underlying mechanisms remain unclear. This study tested the hypothesis that Biodentine stimulates the osteogenic differentiation of mesenchymal stromal cells (MSCs) derived from PLs. The Biodentine extract (B-Ex) was prepared by incubating polymerized Biodentine in RPMI medium (0.2 g/mL) for three days at 37 °C. B-Ex, containing both released microparticles and soluble components, was incubated with PL-MSCs cultured in either a basal MSC medium or suboptimal osteogenic medium. Osteogenic differentiation was assessed by Alizarin Red staining and the expression of 20 osteoblastogenesis-related genes. Non-cytotoxic concentrations of B-Ex stimulated the proliferation of PL-MSCs and induced their osteogenic differentiation in a dose-dependent manner, with a significantly enhanced effect in suboptimal osteogenic medium. B-Ex upregulated most early and late osteoblastic genes. However, the differentiation process was prolonged, as indicated by the delayed expression of wingless-type MMTV integration site family member 2 (WNT2), bone gamma-carboxyglutamate protein (BGLAP), bone morphogenic protein-2 (BMP-2), growth hormone receptor (GHR), and FOS-like 2, AP-1 transcription factor subunit (FOSL2), compared with their expression under optimal osteogenic conditions. The stimulatory effect of B-Ex was primarily calcium dependent, as it was reduced by 85% when B-Ex was treated with the calcium-chelating agent EGTA. In conclusion, Biodentine promotes the osteogenic differentiation of PL-MSCs in a calcium-dependent manner, supporting its stimulatory role in periapical healing