imagine (Institute of molecular genetics and genetic engineering)
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Quantification of volatile fatty acids to safeguard reclaimed water quality
No full textEscalating water scarcity has driven the global expansion of water reuse, yet trace-level volatile
organic compounds, including volatile fatty acids (VFAs), challenge public acceptance and
ecosystem health. Their sour odors at low concentrations signal treatment barrier breakthrough or
contamination, threaten aquatic ecosystems, promote eutrophication, and contribute to greenhouse
gas emissions. Conversely, VFA-rich side streams from biological treatment can be valorized
within circular water–wastewater biorefineries: recovering VFAs enables polyhydroxyalkanoate
production, enhances biological phosphorus removal, and feeds anaerobic co-digestion for biogas.
By coupling validated analytics with by-product recovery and upcycling, operators align water
reuse with circular-economy principles, thus protecting human and environmental health while
generating marketable polymers and energy.
This study developed and validated a suppressed-ion chromatography protocol using an anionexchange
column under isocratic conditions for routine quantification of nine VFAs (lactic,
formic, acetic, propionic, isobutyric, butyric, isovaleric, valeric, and caproic acid) in reclaimed
water. Calibration standards spanning 1–250 mg L⁻¹ were analyzed in triplicate to determine linear
range, detection limits, precision, accuracy and recovery. The method demonstrated excellent
linearity (R² > 0.995), low detection limits in the single-mg L⁻¹ range, and intra- and interday
relative standard deviations below 5%, with recoveries of 70–120% for most analytes in both
municipal wastewater and permeate samples. The protocol is cost -effective, time-efficient, and
requires no extraction or organic solvents. These performance metrics confirm its robustness for
real-time quality control in water reuse scenarios.BeCELS 2025: Belgrade Conference for Early-Career Life Scientists, taking place on Friday, September 5, 2025, at the Institute of Molecular Genetics and Genetic Engineering (IMGGE) in Belgrad
Effects of herbicide S-metolachlor on survival and hatching of shrimp Artemia salina
No full textMetolachlor is a commonly used herbicide for weed control in agriculture. This chemical
frequently enters aquatic ecosystems and the negative impacts on non-target species encompass
both physiological and morphological deformities. Here, we investigated the effects of Smetolachlor
(Mont 960 EC; Agromarket d.o.o., Kragujevac) on mortality and hatching rate in the
brine shrimp, Artemia salina L, 1758. The cysts were hatched within 72 hours in artificial seawater
at 25°C with continuous lightning and aeration. We transferred 10-15 nauplii to each well
containing 900 μl of freshly prepared artificial seawater and 100 μl of test supstance. We used the
following concentrations: 1 mg/l, 2 mg/l, 5 mg/l and 10 mg/l of Mont 960 EC. Positive control
was K₂Cr₂O₇ and artificial seawater was negative control (100 μl). The experiments were
performed with 4 replicates/each concentration in triplicate, and the average percentage mortality
was recorded after 24 and 48 hours of exposure. For hatching experiment, we added 10-20
hydrolised cysts in the predetermined concentrations and recorded percentage of hatching after 24
and 48 hours of exposure. The toxicity of Mont 960 EC increased with time in our research, but
not enough for calculation of LD50. We didn`t observe dose-dependent response. At 48hrs all
treatments showed increased hatching rate compared to negative control. In positive control, we
observed inhibition of hatching rate. At tested concentrations, the toxicity was low, and the
hatching rate was mildly stimulated. Considering these results, further research requires testing the
effects of higher concentrations on possible morphological abnormalities and oxidative stress
response.BeCELS 2025: Belgrade Conference for Early-Career Life Scientists, taking place on Friday, September 5, 2025, at the Institute of Molecular Genetics and Genetic Engineering (IMGGE) in Belgrad
Expression patterns and potential clinical relevance of ASAH1 in rectal cancer
No full textAcid ceramidase (ASAH1) is a central enzyme in sphingolipid
metabolism with a key role in maintaining the balance between
cell survival and apoptosis. Its elevated activity may contribute
to reduced sensitivity to therapy in rectal cancer. This study
aimed to evaluate the gene and protein expression of ASAH1 in
rectal cancer patients and investigate their associations with the
apoptotic status of tumor tissue and response to therapy.
Primary tumor tissue, adjacent healthy mucosa, and serum
samples were collected from 27 patients with locally advanced
rectal cancer before undergoing neoadjuvant chemoradiotherapy
(nCRT). The relative mRNA expression of ASAH1, pro-
apoptotic BAX, and anti-apoptotic BCL2 in tissue samples was
assessed by quantitative real-time polymerase chain reaction,
while serum ASAH1 levels were measured using enzyme-linked
immunosorbent assay. ASAH1 gene expression significantly
differed between tumor and non-tumor tissues (p = 0.022). In
67% of patients, the expression of ASAH1 was 1.1-35-fold
decreased in tumor tissues compared to non-tumor tissues. While
ASAH1 expression showed no correlation with apoptotic
markers, serum ASAH1 levels were negatively correlated with the
BAX/BCL2 ratio in tumor tissue (rs = -0.537, p = 0.026). We
found no association of analyzed parameters with response to
nCRT. Receiver operating characteristic (ROC) analysis
demonstrated limited predictive value, with ASAH1 gene
expression in tumor tissue showing an AUC of 0.608 and serum
ASAH1 an AUC of 0.558. Our results suggest potential
association of serum ASAH1 levels with an anti-apoptotic status
of tumor tissues. However, ASAH1 expression in tissue and
serum showed no significant correlation with therapy response.
Larger and more comprehensive studies are needed to further
evaluate the role of ASAH1 in predicting treatment outcomes
Eco-Friendly Formulation Based on Polyhydroxyalkanoates and Natural Polymeric Surfactants for Fabrics’ Dyeing with the Bacterial Pigment Prodigiosin
No full textBacterial prodigiosins are naturally occurring red alkaloids characterized by a linear
tripyrrole chemical structure, which confers their distinctive biological activity [1].
Prodigiosin is primarily synthesized during the late stages of bacterial growth, and its
production is highly dependent on environmental conditions. It has attracted
significant attention for its therapeutic properties and demonstrates potential for a
wide range of applications, including use as a natural colorant [1,2].
Polyhydroxyalkanoates (PHAs) are natural, non-toxic, and biodegradable
thermoplastic linear biopolymers. Their barrier properties make them a promising
coating agent for textiles, offering a potential approach to optimize material
performance [3]. In this study, a newly isolated bacterial strain, Serratia sp. VJ20, was
grown in Medium E* [4] supplemented with glycerol at a concentration of 40 g/L,
yielding pink-pigmented biomass. The pigment was identified to belong to tri-pyrrole
prodigiosin family. Multi-fiber strips and different types of fabrics (cotton, hemp and
linen) were dyed using a formulation comprising PHA powder (extracted from bacterial
cells by enzymatic treatment [5]) and the freeze-dried pigmented biomass, which was
mixed with an aqueous solution of a surfactant/suspending agent and emulsified at
10000 rpm, for 10 min. The formulation’s composition and the conditions for its
preparation were optimized. Natural polymeric surfactants were tested to replace
synthetic ones, thus rendering the formulation eco-friendlier. The formulation was cast
on test fabrics’ strips that were subjected to a fixation involving drying at temperatures
of at least 100 ºC, followed by hot-pressing at 130-180 ºC and 1-9 ton. The conditions
were adjusted to obtain homogeneous coating of the samples and a good pigment
fixation, assessed by applying several washing cycles and evaluating the color
retention. A good prodigiosin fixation on the fabrics was achieved, which renders this
procedure highly promising for textile uses.12th ESBP will be held from October 1st to 3rd, 2025, in Lisbon, Portuga
Identification and characterization of microRNAs involved in the progression from normal oral mucosa to oral cancer
No full textThe development of oral cancer is a complex, multistep process involving the transformation of normal
oral mucosa into premalignant lesions, such as oral leukoplakia with dysplasia, and eventually into oral cancer. A key
feature of this transformation is the deregulation of small non-coding RNA molecules, microRNAs (miRNAs). The
identification of miRNAs that are consistently deregulated at different stages of oral carcinogenesis is crucial to gaining
deeper insights into the molecular changes involved. Our aim was to identify and characterize commonly deregulated
miRNAs in normal oral mucosa, oral leukoplakia, and oral cancer using publicly available databases.
Material and Methods: The National Center for Biotechnology Information Gene Expression Omnibus (NCBI GEO) was
utilized to identify databases containing miRNA expression data from normal oral mucosa, oral leukoplakia, and oral
cancer. Data from the GSE246050 dataset were analyzed using GEO2R software. Raw fastq files were retrieved from
Sequence Read Archive (SRP468035). HISAT2 and StringTie were used for identification and quantification of miRNA
transcripts. A list of miRNAs commonly deregulated was further characterised by miRNet v2.0 software.
Results: In total 112 genes were identified as commonly differentially deregulated across normal oral mucosa,
leukoplakia with dysplasia and oral cancer. Among them were 6 microRNA genes: MIR429, MIR6726, MIR135B,
MIR204, MIR139 and MIR211. Analysis of expression profiles of mature transcripts of these miRNA genes, revealed
significant changes in expression of hsa-miR-429, hsa-miR-135b-5p, hsa-miR-135b-3p, hsa-miR-204-5p, hsa-miR-139-
5p, hsa-miR-139-3p and hsa-miR-211-5p. We observed progressive upregulation of hsa-miR-429 and hsa-miR-135b-
5p/3p, while downregulation of hsa-miR-204-5p, hsa-miR-139-5p/3p and hsa-miR-211-5p in progression from normal
oral mucosa to oral cancer. To investigate the function of all genes regulated by miRNAs, a hypergeometric test and the
KEGG database were used. Pathways in cancer were significantly enriched with the largest number of genes.
Conclusions: Progression from normal oral mucosa to oral cancer might be characterized by up-regulation of hsamiR-
429, hsa-miR-135b-5p/3p and down-regulation of hsa-miR-204-5p, hsa-miR-139-5p/3p and hsa-miR-211-5p.
These results are good starting point for further validation of miRNA candidates in oral carcinogenesis in a larger group
of clinical samples
Integrated profiling of essential metals, phenolic compounds, anti-inflammatory and hydroxyl radical scavenging activities for five medicinal plants
No full textMedicinal plants are valuable sources of bioactive compounds. In this study, we analyzed the levels of essential metals (Fe, Cu, Zn, Mn) in the leaves of Punica granatum, Teucrium montanum, Chaerophyllum coloratum, Petteria ramentacea, and Menyanthes trifoliata using atomic absorption spectrophotometry. Ethanolic leaf extracts were assessed for anti-inflammatory activity via bovine serum albumin BSA and egg albumin (EgA) denaturation assays and for antioxidant potential by hydroxyl radical scavenging using EPR (Electron Paramagnetic Resonance) spectroscopy. Metal concentrations varied across species: Fe (33–93 mg/kg), Cu (4.29–11.61 mg/kg), Zn (12.39–105 mg/kg), and Mn (1.38–1005 mg/kg), with T. montanum showing the highest Fe and M. trifoliata with highest Cu, Zn and Mn levels. Total phenolic content (TPC) ranged from 32 to 117 mg/gDW, with the highest in P. granatum, which also demonstrated the strongest hydroxyl radical scavenging. The highest BSA inhibition (~ 80%) was observed in Ch. coloratum, P. granatum, and T. montanum at 0.01 µg/mL TPC. Egg albumin inhibition ranged from 28–50%, with Ch. coloratum being the most active. Significant correlations were found between Cu, Mn, and anti-inflammatory activity, and between Zn, TPC, and both antioxidant and anti-inflammatory effects. These findings suggest a synergistic role of phenolics and trace elements and support the potential use of these plants as dietary supplements against inflammation and oxidative stress
GENE EXPRESSION PROFILING OF IPSC-DERIVED NEURAL PROGENITORS FROM PATIENTS WITH 22Q11.2 DUPLICATION SYNDROME
No full textThe molecular mechanisms underlying neurodevelopmental disorders (NDDs) are understudied.
22q11.2 Duplication Syndrome (22q11.2DupS) is associated with an increased risk of NDDs,
including autism spectrum disorders, developmental delay, and intellectual disability, while
potentially having protective effect against schizophrenia. Here we performed transcriptomic
profiling of 22q11.2DupS neural progenitors (NPCs). Total RNA was isolated from
iPSCs-derived NPCs of a family trio consisting of a child with 22q11.2DupS, a carrier unaffected
mother, and a healthy control father. Paired-end RNA-sequencing was carried out by
commercial services. FASTQ files were processed on an NVIDIA platform and differential gene
expression analysis was carried out in RStudio using the DESeq2 R package. The resulting lists
of differentially expressed genes (DEGs) was utilized for pathway and gene ontology enrichment
analysis using clusterProfiler in RStudio. 60 DEGs with lower expression and 58 DEGs with
higher expression were obtained in 22q11.2DupS NPCs compared to both carrier and control
NPCs. For genes with lower expression in 22q11.2DupS NPCs, “miRNA targets in ECM and
membrane receptors” was the top enriched pathway. On the other hand, for genes with higher
expression in 22q11.2DupS NPCs, no biological pathways were identified as being significantly
enriched in the DEG list beyond what would be expected by chance. However, for genes with
higher expression in 22q11.2DupS NPCs, we obtained statistically significant enrichment of
molecular functions such as Histone demethylase activity. 22q11.2DupS NPCs exhibit altered
signaling pathways and molecular functions, providing preliminary insights into the impact of this
microduplication on neural differentiation.Abstract book: FENS Regional Meeting 2025, Oslo, Norway, 16-19 June 202
UNRAVELLING THE CAUSE OF RECURRENT VENOUS THROMBOSIS IN A DABIGATRANTREATED PATIENT
No full textThis study aimed to explore the cause of recurrent
venous thrombosis in female patient treated with
dabigatran.
The patient’s first thrombotic event occurred at
the age of 59, presenting as deep vein thrombosis
with massive pulmonary embolism. Following
heparin and one month of dabigatran, recurrent
thrombosis occurred. The patient was switched to
enoxaparin and acenocoumarol, used without
complications for eight years. Haemostatic
balance was assessed using the overall
haemostatic potential (OHP) assay. Wholeexome sequencing was conducted. The
coagulation-related genes were selected based on
literature and STRING-DB database. Gene
variants were identified using ClinVar, ACMG,
UniProt, as well as MetaSVM, MetaLR,
MetaRNN, REVEL, ClinPred, and AlphaMissense scores. AlphaFold were used to
visualize impact of identified variants on protein
structures. Dabigatran-related ABCB1 and CES1
variants were also explored.
Elevated OHP and the overall coagulation
potential levels, accompanied by reduced the
overall fibrinolysis potential levels, were
observed. Highly damaging heterozygous
variants in THBS1 (p.Gln1089His) and MYH9
(p.Arg1877Trp) genes were identified. The
variant in THBS1 was predicted to affect the
protein structure. In addition, multiple mutations
in ABCB1 gene were observed.
Findings suggest a notable influence of
underexplored genetic factors in case of RVT and
further functional analyses.Book of abstract: 15th Balkan congress of human genetics and 3rd Alpe Adria meeting of human genetics
9 - 11 October 2025, Rikli Balance Hotel ,Bled, Sloveni
PHARMACOGENOMIC LANDSCAPE OF THE SERBIAN POPULATION
No full textPurpose: Pharmacogenomics offers the
opportunity to predict drug response by analyzing
an individual’s genetic profile. Highthroughput
sequencing technologies facilitated identification
and interpretation of variants in many
pharmacogenes simultaneously. The integration
of pharmacogenomics into clinical practice is still
scarce. The aim of our study was to assess the
most comprehensive pharmacogenomics
landscape of the Serbian population so far.
Methods: Genomic data of 881 individuals from
Serbia obtained by clinical and whole exome
sequencing was used. Raw sequencing files were
processed using an in-house pipeline. The
PharmCAT and Stargazer tools were used for
annotation of pharmacogenetics star alleles and
determination of phenotypes. Star allele and
phenotype frequencies were calculated and compared to worldwide and European
populations.
Summary of results: The greatest differences in
pharmacogenomic profiles were noted between
the Serbian and worldwide populations. In the
Serbian population, the most relevant
pharmacogenes were CYP2B6, NAT2, SLCO1B1,
UGT1A1 and VKORC1. Significant differences in
frequencies of pharmacogenetic phenotypes that
influence response to several drug categories
including statins and antidepressants indicate that
pharmacogenomics testing would be beneficial in
the Serbian population. Implementation of
pharmacogenetic testing could be achieved
through analysis of clinical and whole exome
sequencing data.Book of abstract: 15th Balkan congress of human genetics and 3rd Alpe Adria meeting of human genetics, 9 - 11 October 2025, Rikli Balance Hotel ,Bled, Sloveni
Outer membrane vesicles (OMVs) secreted by Paraburkholderia phytofirmans PsJN: characterization of associated RNA content and interaction with plant cells
No full textPlant growth-promoting bacteria (PGPB) support plant development, increase productivity, and
improve resilience to stress. These benefits arise from complex interactions between species,
involving the exchange of various signaling and effector molecules. Extracellular vesicles (EVs) are
an important means of signaling between cells, organisms, and even across biological kingdoms.
Among EVs, outer membrane vesicles (OMVs)—produced by Gram-negative bacteria—can elicit
both immune-activating and immune-suppressing responses in plants. However, direct interaction
between OMVs and plant cell membranes has been shown for only a single phytopathogenic
bacterial strain. Even less is known about the role of OMVs in symbiotic relationships between
plants and beneficial bacteria, with existing data mostly limited to rhizobia-legume systems.
To explore OMV-mediated interactions between non-rhizobial plant-beneficial bacteria and plant
cells, we isolated and characterized OMVs from Paraburkholderia phytofirmans PsJN—a wellknown
plant growth-promoting and stress-mitigating strain effective across various plant species.
Using OMVs labeled with the fluorescent lipid-binding dye Vybrant™ DiD and visualization via
confocal laser scanning microscopy, we demonstrated that PsJN-OMVs directly interact with the
root hairs and epidermal cells of Arabidopsis thaliana and tomato. Using the membrane fusion
probe octadecyl-rhodamine B chloride (R18), we confirmed that PsJN-OMVs can fuse with plant
cell membranes. We examined the RNA content of PsJN-OMVs. The OMV-associated RNA,
which was mostly sized up to 200 nt, was sequenced using the DNBSEQ Sequencing Platform.
Prior to library synthesis, samples were separated into two subsamples: one up to 40 nt and the
other from 40 to 200 nt. These were then sequenced using the SE50 and PE100 approaches,
respectively. The same procedure was used for small RNA isolated from PsJN cells. Comparison
of RNA sequences from PsJN OMVs and cells showed substantial differences in their size
distribution, biotypes, and genome regions of origin. The results indicate that the packaging of
RNA into PsJN-OMVs is not random, but involves the selective enrichment of certain RNA species
in OMVs, while others are predominantly retained within the cells. Previous studies suggested
that EVs may both remove unnecessary cellular RNAs and transfer RNAs to other cells, as
signaling functions. The molecular mechanisms for selective loading of eukaryotic EVs are only
partially described, while those responsible for bacterial vesicles are poorly understood. The
data on RNA associated with OMVs secreted by plant beneficial bacteria are even more scarce.
Our results make a significant step toward understanding the role of OMVs in the initial
phases of root colonization and symbiosis establishment. Advancing knowledge in this field
may lead to the development of innovative, environmentally friendly strategies to address
agricultural challenges and promote sustainable crop production.Book of abstract: Arturo Falaschi Conference “Cell-cell communication in bacteria: new insights and future trends”, 28-30 October 2025 | Trieste, ITAL