imagine (Institute of molecular genetics and genetic engineering)
Not a member yet
3088 research outputs found
Sort by
Airborne Pathogens and Virulence Factors in Belgrade Metropolitan Area Metagenomic Insight
No full textIntroduction: Air pollution has been recognized as one of the greatest threats to human
health. The particulate matter-PM2.5, main air pollution indicator, is found to contain
numerous bacterial pathogens. Considering that Serbia is struggling with excessive
antibiotics use and misuse, a high percentage of multidrug-resistant (MDR) bacterial
isolates, and poor air quality, made Serbian capital Belgrade interesting research model
for the effects of air pollution on the abundance and diversity of airborne pathogens.
Objective: The presence of MDR pathogens, virulence factors as well as pathogen-host
interactions are studied in the airborne metagenome samples from seven locations of
Belgrade metropolitan area collected during the summer season of 2024.
Method: Air samples were collected from seven Belgrade locations using hydrophobic
polypropylene membrane filters (air volume of 30 m³). Total DNA was analysed by
shotgun metagenomic sequencing, followed by bioinformatic processing (Novogene,
UK). Taxonomic diversity/abundance, virulence factors and pathogen-host interactions
were analysed using MEGAN, VFDB, and PHI databases, respectively.
Findings: Important human and animal pathogens were found in analysed
metagenomes: Acinetobacter schindleri, Streptococcus pneumonie, Streptococcus
oralis, Staphylococcus aureus, Enterococcus faecalis, Klebsiella pneumoniae, Ewingella
americana, Burkholderia multivorans, Legionella pneumophila, Salmonella enterica and
Brachybacterium muris. Genes encoding for motility, adherence, immune modulation,
exotoxins and biofilm formation were also found. Additionally, PHI analysis revealed the
presence of different type of infections caused by pathogenic bacteria – nosocomial and
urinary tract infections, pneumonia, salmonellosis, soft rot.
Conclusion: This study indicated that Belgrade air could serve as a reservoir for
pathogenic bacteria, virulome and numerous types of infections.Book of abstract: FEMS MICRO 2025: Congress & Exhibition, taking place in Milan, Italy, from 14-17 July 2025
Establishment and characterization of induced pluripotent stem cells from patients with 22q11.2 Duplication Syndrome
No full textBackground: 22q11.2 Duplication Syndrome (22q11.2DupS), caused by microduplication
of the q11.2 region on chromosome 22, is associated with a high risk of developing
neurodevelopmental disorders (NDDs), including autism spectrum disorders, and
attention-deficit/hyperactivity disorder. On the other hand, in individuals affected by
schizophrenia, 22q11.2DupS occurs less compared to the general population, suggesting
that this microduplication could have protective effect against schizophrenia. Thus,
22q11.2DupS represents an adequate model system for studying the molecular
mechanisms underlying NDDs.
Material and Methods: iPSCs were generated by reprogramming peripheral blood
mononuclear cells using CytoTune™-iPS 2.0 Sendai Reprogramming Kit. Established iPSC
lines were characterized by examining their morphology, genomic integrity, pluripotency
and the ability to differentiate into cells of three germ layers.
Results: iPSCs were generated from three patients with an inherited form of
22q11.2DupS, their mothers that carry the microduplication, and three healthy individuals.
Genotyping showed that some of the iPSC lines have additional copy number variations.
All generated iPSC lines express pluripotency markers and are capable of differentiating
into the cells of three germ layers (ectoderm, mesoderm, and endoderm).
Conclusion: iPSCs were successfully generated from patients with 22q11.2DupS, their
carrier mothers, and healthy individuals, providing a valuable tool for gaining insights into
the molecular mechanisms underlying NDDs.Book of absctracts: European Society of Human Genetics (ESHG) 2025
Conference, May 24–27, 2025. Milan, Ital
Transcriptomic profiling of iPSC-derived astrocytes from patients with 22q11.2 deletion syndrome
No full textBackground: Neurodevelopmental disorders represent considerable public health
challenges.One of the syndromes associated with heightened risk of NDDs is 22q11.2
Deletion Syndrome (22q11.2DS), caused by microdeletion in chromosomal region
22q11.2. However, molecular mechanisms underlying NDDs are largely unidentified. Here
we analyzed transcriptomic profiling of 22q11.2DS astrocytes.
Material and Methods: Total RNA was isolated from iPSCs-derived astrocytes of two
cases with familial 22q11.2DS with 1.5Mb microdeletion and one healthy control. Paired-
end RNA-seq was carried out on Illumina NovaSeq 6000 sequencer. Bioinformatic
processing of raw data was conducted via NVIDIA platform, while differential gene
expression analysis was performed in RStudio using DESeq2 R package. The obtained list
of DEGs was used for pathway enrichment analysis by employing EnrichR and
WikiPathways.
Results: 125 DEGs with lower expression and 287 DEGs with higher expression in
22q11.2DS astrocytes compared to the control were obtained. For genes with lower
expression in 22q11.2DS astrocytes, 22q11.2 Copy Number Variation Syndrome and Axon
Guidance were the top enriched pathways, while for genes with higher expression in
22q11.2DS astrocytes we did not identify biological pathways that are enriched in DEG
lists more than would be expected by chance. We found 178 DEGs with lower expression
and 205 DEGs with higher expression in astrocytes of symptomatic child with 22q11.2DS
compared to oligosymptomatic mother with 22q11.2DS. Employing EnrichR and
WikiPathways we did not identify biological pathways that are enriched in DEG lists more
than would be expected by chance.
Conclusion: We provide preliminary evidence for an altered transcriptomic landscape of
22q11.2DS astrocytes.Book of absctracts: European Society of Human Genetics (ESHG) 2025 Conference, May 24–27, 2025. Milan, Ital
Expression of miR-185 during neural differentiation of induced pluripotent stem cells from patients with 22q11.2 Deletion Syndrome
No full textBackground: The 22q11.2 Deletion Syndrome (22q11.2DS) is the most common
microdeletion syndrome in humans and it is associated with high rates of
neurodevelopmental disorders (NDDs). Literature data revealed downregulation of miR-
185 expression, located in the 22q11.2 region, in the whole blood of patients with
22q11.2DS. Here we analyze miR-185 expression during neural differentiation of induced
pluripotent stem cells (iPSCs), derived from patients with 22q11.2DS, and search for miR-
185 potential target genes associated with NDDs.
Material and Methods: The expression of miR-185 was assessed by Taqman qRT-PCR
using specified Taqman assay for miR-185. miRNet platform was used to elucidate miR-
185 function in NDDs. miR-185 target genes were predicted by using miRDB, mirWalk and
TargetScan. The selection of miR-185 target genes related to NDDs was conducted using
online available tools GeneTrack, miRNet and Metascape.
Results: Decreased expression of miR-185 was detected in iPSCs derived from patients
with 22q11.2DS, compared to healthy control. In addition, the expression of miR-185 was
significantly downregulated in neurons from patients with 22q11.2DS compared to
expression detected in neurons of healthy control. Using online available tools, we
selected three potential target genes of this miRNAs related to NDDs -
CSMD2, DLG2 and HIC2.
Conclusion: Obtained results indicate that miR-185 and it target
genes CSMD2, DLG2 and HIC2 could serve as novel therapeutic targets. Future studies are
needed to evaluate their relationship and involvement in neurodevelopmental outcome
seen in patients with 22q11.2DS.Book of absctracts: European Society of Human Genetics (ESHG) 2025 Conference, May 24–27, 2025. Milan, Ital
Respirometric analysis for monitoring microbial activity during terephthalic acid transformation
No full textPlastic pollution is a major challenge, with polyethylene
terephthalate (PET) among the most prevalent. Terephthalic acid (TPA), a key
monomer from PET depolymerization, can be toxic and persistent, posing
environmental risks. However, some microbial species have demonstrated the
ability to utilize TPA, opening avenues for bioremediation and recycling. This
work investigates microbial consortium and pure microbial strains isolated from
polluted soil and sediment to transform TPA. Microbial activity was monitored
using a respirometer, and TPA transformation using HPLC. Elucidating
pathways for TPA breakdown can enhance bioremediation processes and
promote sustainable plastic waste management.3rd INTERNATIONAL SYMPOSIUM ON BIOTECHNOLOGY XXX Savetovanje o biotehnologiji sa međunarodnim učešćem, 13–14 March 202
Bio-Upcycling of PET plastic waste into bacterial nanocellulose reinforced by poly(vinyl alcohol) as a platinum nanoparticles support efficient in methanol oxidation
No full textEffective polymer waste management is essential for mitigating the environmental impact of plastic waste
pollution. Bio-upcycling is an innovative, end-of-life strategy that leverages microbial bio-catalysis to convert pretreated
polymer waste into high-value biomaterials [1]. This approach assimilates the reduction of plastic waste
accumulation by enabling the development of innovative materials with advanced functionalities. In this study, a
strategy was demonstrated in which various types of PET plastic waste were processed through reactive extrusion and
enzymatic depolymerisation [2]. The resulting depolymerisation products were then transformed into biopolymer
bacterial nanocellulose (BNC), effectively closing the loop in a circular process. The obtained BNC was further utilized
as a suitable support for platinum (Pt) nanoparticles, forming an efficient catalyst with the great potential in methanol
oxidation reaction. To enhance catalytic performance while reducing the amount of loaded Pt, BNC was enriched with
poly(vinyl alcohol) (PVA), creating a novel BNC/PVA composite support. The use of BNC/PVA composite as a support
allowed the deposition of only 3 wt% of Pt, significantly lowering catalyst production costs while preserving high
catalytic activity. The structural, thermal, and morphological properties of the new Pt-BNC/PVA catalyst were
thoroughly characterized using FTIR, TGA, XRD, XPS, TEM, and SEM-EDX analysis. The developed catalyst
exhibited remarkable electrochemical performance in the methanol oxidation reaction, highlighting its potential for
application in direct methanol fuel cells (DMFCs). Moreover, the use of BNC/PVA composites as Pt nanoparticle
supports, offers a sustainable alternative to conventional, commonly used carbon-based materials. By integrating PET
plastic waste into a bio-upcycling loop, this study advances the development of cost-effective, environmentally friendly
catalysts for sustainable energy applications.E-book: 19th International Conference on Chemistry and the Environment ICCE 2025 Belgrade 8-12 June 202
Unveiling the antimicrobial, biofilm inhibition, and photoprotective potential of Bupleurum falcatum L. for dermatological applications
No full textBupleurum falcatum L. is known for its therapeutic properties, especially in treating fever, inflammation, and infectious diseases. However, its potential for dermatological applications remains mainly unexplored. Thus, the present study explores antimicrobial potential of B. falcatum against biofilm-associated infections, antibiotic-resistant strains, and UV-induced skin damage. This aligns with the growing interest in natural products as sources of bioactive compounds with skin-protecting features. Herein, we employed maceration (M) and ultra-sound-assisted extraction (USA) at 50 Hz and 100 Hz (USA 50 and USA 100) to obtain extracts from aerial parts of the plant. Chemical profiling was performed using UHPLC. Antimicrobial activity, biofilm inhibition, EPS and eDNA production were assessed using microdilution test, crystal violet, Congo red, and eDNA assays, respectively. Cytotoxicity and photoprotective effects were evaluated on human keratinocytes using the MTT assay. Chemical analysis identified 64 compounds, including benzoic and cinnamic acid derivatives, flavonoid glycosides, and saikosaponins. Extracts showed strong antimicrobial activity against Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus (MIC as low as 0.5 mg/mL). The M extract displayed moderate biofilm inhibition and reduced eDNA production. Cytotoxicity assays confirmed safety on keratinocytes, while M and USA 100 extracts demonstrated photoprotective effects. B. falcatum extracts showed promising potential in addressing biofilm-associated infections, antibiotic resistance, and UV-induced skin damage
DEVELOPMENT OF NOVEL FLUOROGENIC SUBSTRATES FOR RAPID AND SENSITIVE ASSESSMENT OF PHA DEPOLYMERASE ACTIVITY
No full textPolyhydroxyalkanoates (PHAs) are microbial polyesters synthesized naturally and commercialized as
biodegradable plastics, yet their biodegradation in the environment was shown to be slow (1). PHA
depolymerases (PHAses) are essential for their degradation. Currently known PHA depolymerases lack the
efficiency for larger-scale processing, highlighting the need for discovering and engineering more efficient
variants. The lack of a reproducible, high-throughput methodology hinders straightforward and reliable
monitoring and assessment of PHA depolymerases' activity (2).
In our previous research, we reported the synthesis of two chromogenic compounds derived from
polyhydroxyoctanoate (PHO) and their successful application in a continuous, quantitative spectrophotometric
assay. This assay enables the rapid evaluation of PHO depolymerase activities within just 10 minutes at
temperatures exceeding 45 °C, streamlining the comparison of PHAses and advancing the analysis of enzymatic
degradation processes (3).
To further improve sensitivity and enable measurements at lower substrate concentrations, we designed and
synthesized for the first time three novel fluorogenic substrates: 4-methylumbelliferone esters (4-mU) of
3-hydroxy butyric acid (4-mUHB), 3-hydroxyoctanoic (4-mUHO) acid and octanoic acid (4-mUO). These
substrates were fully characterized using NMR, IR, and LC/MS, confirming their structure and suitability for
enzymatic assays. Novel fluorogenic substrates were then used in the enzymatic activity assay with the
benchmark PHO depolymerase from Pseudomonas fluorescens GK13. This activity was compared to
commercially available Candida antarctica lipase B (CALB). A clear distinction in activity was observed
between PHOase and CALB lipase when tested with these substrates. These assays were rapid and highly
sensitive, requiring low substrate concentrations facilitating the discovery and optimization of more efficient
PHA depolymerases for industrial and environmental applications.BOOK OF ABSTRACTS: 17th International Symposium on Biocatalysis and Biotransformations Basel, Switzerland, June 29-July 3, 202
Eriodictyol and Diosmetin Protective Potential in Skin Infection: Antimicrobial Action, Gene and Molecular Targets, and Keratinocyte Protection Against Bacteria-Induced Damage
No full textEriodictyol and diosmetin are bioactive flavonoids. This study explored their antimicrobial activities and antibiofilm potential along with the effect on pyocyanin and protease production and virulence-linked gene expression, followed by in silico molecular target predictions. Moreover, keratinocytes were used for the evaluation of cytotoxicity and protective antioxidant and anti-inflammatory effects in the infected cells. Both compounds have shown significant antibacterial capacity towards skin pathogens (minimal inhibitory concentrations 0.025–0.2 mg/mL). Their ability to prevent biofilm formation of Pseudomonas aeruginosa was drastic, as well as the impact on other virulence factors, proteases, and pyocyanin production. RT-qPCR determined downregulation of almost all genes examined (lasI, lasR, lasB, rhlI, rhlR, rhlC, pqsH, pqsR, pvdS, pvdF, phzM, and algK), while molecular docking predicted strong binding affinities to the LasI, LasR, PqsR, and QscR quorum-sensing proteins. Moreover, both compounds were not toxic to HaCaT and were able to reduce damage induced by P. aeruginosa in this cell line. Precisely, eriodictyol reduced levels of secreted IL-6 (from 335.32 to 261.76 pg), while both compounds reduced the formation of superoxide. Both eriodictyol and diosmetin displayed remarkable antimicrobial potential while employing a wide array of antimicrobial mechanisms, making them attractive candidates for further assessment and eventual incorporation into novel therapeutic strategies
Harnessing synthetic microbial biotechnology for efficient plastic waste upcycling
No full textPlastics have become indispensable in various industries today, but their extensive use
has resulted in a severe environmental crisis—plastic waste accumulation. Traditional
methods of plastic waste management are insufficient, facing persistent pitfalls, such as
overall material downgrading and release of hazardous microplastics and chemicals into
the environment. Plastic upcycling, on the contrary, is gaining prominence in converting
plastic waste into valuable chemicals and materials in an eco-friendly manner. The
current project presents an innovative approach to plastic waste upcycling: employing
green bio/mechano/chemical technologies for the hydrolytic degradation of plastic
polymers into oligomers and their constituent building blocks, is followed by the creation
of microbial cell factories capable of producing industrially relevant compounds from
these plastic-derived feedstocks. Successful microbial upcycling hinges on efficient
carbon assimilation from plastic feedstocks and the presence of functional intracellular
biosynthetic pathways to generate valuable bioproducts. To achieve this, a wide variety
of microorganisms with natural capabilities for these processes are identified through
bioprospecting, and synthetic biology is employed to engineer or enhance the other
necessary functions. Alternatively, some well-established microbial hosts including
Bacillus spp., Pseudomonas spp. or Yarrowia spp. serve as suitable chassis. By
integrating these advanced methodologies, we aim to create sustainable and efficient
microbial systems for plastic waste valorization. Our ultimate goal is to establish a
research and innovation hub in Synthetic Microbial Biotechnology that would advance
scientific knowledge, raise public awareness, promote environmental protection, and
boost economic development, all the while minimizing humanity’s reliance on finite
fossil fuel resources.Book of abstract: FEMS MICRO 2025: Congress & Exhibition, taking place in Milan, Italy, from 14-17 July 2025