imagine (Institute of molecular genetics and genetic engineering)
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Short-term exposure to perfluorooctanoic acid induces oxidative stress and necrotic cell death in human HepG2 hepatocytes
No full textPerfluorooctanoic acid (PFOA) is a synthetic chemical belonging to the class of per - and
polyfluoroalkyl substances known for their environmental persistence and bioaccumulative
potential. Due to its resistance to degradation, PFOA persists in human tissues and has been
associated with immunotoxicity, hepatotoxicity, endocrine disruption, and increased carcinogenic
risk, yet its acute cellular effects remain unclear. In this study, we characterized early
hepatocellular responses to 48 h PFOA exposure using human HepG2 cells. HepG2 cells were
exposed to vehicle (0.05% DMSO) or 1, 10, and 100 μM PFOA – concentrations comparable to
serum levels in highly exposed workers. Assessed endpoints included metabolic activity
(alamarBlue™ assay), cell death pathways (annexin V/propidium iodide flow cytometry), cell
cycle distribution (propidium iodide staining), intracellular reactive oxygen species (ROS;
dichlorofluorescein fluorescence), lipid peroxidation (thiobarbituric acid reactive substances
assay), and neutral lipid accumulation (Oil Red O staining). PFOA at 1 μM and 10 μM had no
measurable impact on any endpoint. In contrast, 100 μM PFOA elevated ROS by ~40%, decreased
viable cells from 92% to 84%, and more than doubled the number of necrotic cells, while metabolic
activity, cell cycle profile, lipid peroxidation, and lipid accumulation remained unchanged. These
data indicate that short-term, high-level PFOA exposure primarily induces oxidative stress and
shifts cell fate toward necrosis rather than apoptosis, without immediate disruption of lipid
metabolism or proliferation. These findings provide an integrated snapshot of acute PFOA toxicity
in human hepatocytes and establish baseline parameters for ongoing mechanistic investigations of
longer-term exposure.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
High Level Fluoroquinolone Resistance and Multidrug Resistance in Salmonella Spp. Isolated from Poultry, Turkey Floks and Slaughterhouses in Algeria
No full textIn this work, Salmonella spp. was detected in poultry and turkey farms, slaughterhouses
and hatcheries in the Sétif Province in Algeria. Eighty single isolates per farm were
analyzed by establishing the resistotype and detected resistance genes underlining the
mechanism of resistance. In one case, serotypes S. Virchow and S. Ivory were found
in the same sample and both isolates were resistant to nalidixic acid. S. Enteritidis
was detected in four broiler breeder fl ocks, three hatcheries, 12 fl ocks of layers, 12
broiler fl ocks while fi ve slaughterhouses yielded 10 isolates. The wide distribution of S.
Enteritidis in the primary production and food chain in Algeria requires special measures
in the management practice on poultry farms. All isolates except fi ve were resistant
to nalidixic acid and pefl oxacin which means that these salmonellae phenotypically
express reduced sensitivity to ciprofl oxacin. Five isolates were multidrug resistant.
Two Salmonella Galinarum biotype gallinarum isolates from fl ocks of laying hens were
resistant to quinolones, aminoglycosides and sulfonamides. One of these isolates was
also resistant to trimethoprim alone and in combination with sulafmethoxazole. One
S. Enteritidis isolate was resistant to ampicillin, nalidixic acid, pefl oxacin and colistin.
Especially worrying is the high level of resistance to ciprofl oxacin in nine isolates (six,
Salmonella Galinarum biotype gallinarum, two, S. Kentucky and one Salmonella enterica
subsp. enterica isolate) due to mutations in the enzymes DNA gyrase and topoisomerase
IV. Resistance genes were identifi ed in 21 isolates. All resistance genes detected are
commonly conferring resistance to ampicillin, streptomycin, gentamicin, tetracycline,
sulfonamides and trimethoprim antibiotic
ENGINEERING MICROBIAL PLATFORMS FOR SIMULTANEOUS BIODEGRADATION AND UPCYCLING OF PCL
No full textNowadays, the accumulation of plastic waste in the
environment has become a significant global
concern, with long-lasting ecological and health
impacts. This has driven the urgent need for novel
strategies, particularly those targeting the
degradation of polyesters, which are widely used
but poorly degraded in natural environments [1].
Biological approaches, particularly those utilizing
microorganisms, offer a sustainable alternative for
addressing plastic waste [2]. In this study, we
evaluated the ability of various microbial strains to
metabolize polycaprolactone monomers. These
included Ralstonia eutropha H16, newly isolated
pigmented Streptomyces isolates, Streptomyces
albus wild-type strain, as well as a strain evolved via
adaptive laboratory evolution, which was selected
to better utilize the PCL monomer as a sole carbon
source. Initial screening revealed variable growth
across the studied microorganisms, which are
known to produce valuable bioproducts, such as
bioplastics, biopigments, and antibiotics [3] [4].
Additionally, aiming to construct strains that
achieve polymer degradation and further
metabolize the monomers, plasmids harboring selected polyesterase genes, Se1JFR [5] or
DmPETase [6], were introduced into R. eutropha
and Streptomyces species. Transformed strains
were identified and further analyzed for their
polyester-degrading abilities by growth assessment
as well as the determination of esterase activity in
culture supernatants. Overall, this work contributes
to the broader field of microbial upcycling by
combining metabolic screening, genetic
engineering, and synthetic biology to construct
bacterial strains capable of degrading and isolating
synthetic polymers.Book of abstract: MikroBioKosmos Society & The Central and East Europe Symposium of Microbial Ecology (#mbkceesme2025), in Thessaloniki, Greece, between 22 and 24 September 2025
Petrography of high-pH fluids sedimentary assemblages and biotechnological potential of sediment Bacillus
No full textSerpentinization-driven high-pH fluids (pH > 11.0) represent exceedingly rare
environments on Earth. Sediment samples, originating from four distinct zones of emergence of
these low-mineralized springs within the Zlatibor Jurassic Ophiolitic Massif, were analyzed
visually and microscopically. Sediment analysis revealed a diverse range of petrographic
varieties within these relict environments. The unconsolidated clastic sediments are polymictic,
consisting of pebbles with a moderately uniform size and appearance. They exhibit a coarse-
grained psephitic texture, with colors ranging from brown to dark gray, with occasional hints of
green. The gravel-sized fraction of the sediments consists of well- to moderately-rounded clasts,
mainly ranging from 2 to 20 mm, while the poorly sorted sandy fraction ranges from fine to
coarse. Additionally, an investigation of the lignocellulolytic and plastolytic potential of
microorganisms from these sediments identified one Bacillus isolate of potential biotechnological
interest. This isolate was able to grow on polyurethane polymers as its sole carbon source and
degrade both carboxymethyl cellulose and more complex xylan substrates.3rd International Conference on Chemo and
BioInformatics, ICCBIKG 2025, September 25-26, 2025, Kragujevac, Serbi
Association Between Hypertension, Dipping Status, and ACE and AGTR1 Gene Polymorphisms in Adolescents with Type 1 Diabetes
No full textObjectives: This study aims to show the distribution of angiotensin-converting enzyme (ACE) rs1799752 (I>D) gene insertion/deletion (I/D) polymorphism and angiotensin II receptor type 1 (AGTR1) rs5186 (A>C) gene polymorphism in adolescents with hypertension (HT) and type 1 diabetes (T1D), as well as its association with hypertension and the diurnal variation of mean blood pressure (dipping phenomenon). Methods: A cross-sectional study was conducted involving 118 adolescents diagnosed with T1D who underwent clinical and laboratory investigations, genetic analyses, and 24 h ambulatory blood pressure monitoring. The genotype frequencies were compared between adolescents with HT and those with normal blood pressure. Additionally, the genotype frequencies were compared between dippers and non-dippers. Results: Patients with HT were more likely to be female and exhibited significantly poorer glycemic control and higher triglycerides, along with increased body mass index and daily insulin dosage. The prevalence of ACE rs1799752 genotypes in the hypertensive group was 20% II, 66.7% ID, and 13.3% DD, which did not significantly differ from the normal blood pressure group with 29.1% II, 53.4% ID, and 17.5% DD (p = 0.625). The prevalence of AGTR1 rs5186 genotypes in the hypertensive group was 53.3% AC, 40% AA, and 6.7% CC, which also did not significantly differ from the normal blood pressure group with 39.8% AC, 52.4% AA, and 7.8% CC (p = 0.608). A total of 46% of the patients exhibited non-dipping phenomena. The prevalence of non-dippers among the ACE genotypes was 13% DD, 33.3% II, and 53.7% ID (p = 0.369), while for the AGTR1 genotypes, it was 50% AA, 42.6% AC, and 7.4% CC (p = 0.976). Conclusions: Our results indicate that in our adolescents with T1D, clinical and metabolic factors such as higher body mass index, triglycerides, suboptimal glycemic control, and female gender are more indicative of the development of hypertension than ACE and AGTR1 gene polymorphisms. A potential reason for this finding could be the young age of the patients or the relatively small size of the study group. Future research involving larger sample sizes is needed to further investigate the genetic predisposition for the development of hypertension
FROM HIVE TO HEALTH: PROBIOTIC POTENTIAL OF APILACTOBACILLUS KUNKEEI AND ITS EFFECT ON HUMAN VITAMIN D RECEPTOR EXPRESSION
No full textObjective: The Vitamin D receptor (VDR) plays a key role in
maintaining calcium and phosphate homeostasis, modulating the
immune response, and mediating interactions between the gut
microbiota and the host. Dysfunction of the VDR has been linked
to alterations in gut microbiota composition and the development
of autoimmune diseases. While traditional probiotics are
typically isolated from fermented dairy products, there is growing
interest in alternative sources, including bee products and the
gut microbiota of honeybees. Among bee-associated bacteria,
the fructophilic lactic acid bacterium Apilactobacillus kunkeei
has emerged as a promising candidate for probiotic applications.
Methods: Ten strains of A. kunkeei isolated from the guts
of healthy honey bees (Apis mellifera) from the Vojvodina
region (Serbia) were tested for probiotic properties and safety
attributes for potential human use. Their effect on the expression
of VDR and VDR-related genes was examined using an in vitro
model of the human intestinal epithelial barrier (Caco-2 cell line).
Additionally, the genomes of two selected strains were analyzed.
Results: All examined strains exhibited strong antimicrobial
activity against human gut pathogens. Among them, only
strains A1, A4, A28, and A56 demonstrated mucin-binding
ability. Notably, A. kunkeei strains A1 and A55 were capable
of modulating the expression of VDR and tight junction-related
genes in the Caco-2 cell line. Both strains also showed sensitivity
to all tested antibiotics.
Conclusions: These findings highlight the potential of A. kunkeei
A1 and A55 strains to modulate VDR function and support their
further development as safe and effective probiotics for human
use
PULMONARY IN VITRO MODEL SYSTEM ENABLES EXPLORATION OF INNOVATIVE TREATMENT STRATEGIES FOR RARE RESPIRATORY DISEASES
No full textIntroduction: Hundreds of rare pulmonary
diseases have been identified, most of which
involve impaired mucociliary clearance, a
process essential for lung function. These
progressive, life-threatening disorders lack
effective treatments, with lung transplantation
often being the only option. This study focuses on
the molecular mechanisms of lung epithelial
formation and aims to develop an in vitro model
system for investigating novel therapeutic
strategies.
Methods: NHBE cells were cultured in an air–
liquid interface (ALI) system to induce
differentiation into multiciliated and goblet cells.
Validation included confocal microscopy (β-
tubulin, DNAI1, MUC5B, MUC5AC), qRT-PCR
of ciliogenesis markers (TP63, NOTCH1, CP110,
MCIDAS, GEMC1, CCNO, RFX3), and differentiated cell markers (FOXJ1, DNAI1,
TFF3, MUC5B, MUC5AC), along with Western
blot (acetyl-α-tubulin, DNAI1, TP63).
Results: Ciliated cells appeared around day 12
post-ALI. By day 28, a fully differentiated
pseudostratified airway epithelium was formed.
Confocal imaging confirmed motile cilia and
mucus, and protein analysis demonstrated the
presence of key ciliary proteins.
Conclusion: The use of NHBE cells provides a
reproducible, accessible, and standardized
system, ideal for high-throughput drug screening
targeting mucociliary clearance and other
processes in the respiratory tract, while also
reducing variability associated with patientderived
samples.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
Investigating long non-coding RNA lnc-RNMT-2:5 indicates Stromal Origin and Putative Regulatory Function in Rectal Cancer
No full textRecent pan-cancer transcriptome analysis revealed differential activity of two alternative LDLRAD4 gene promoters in malignant vs. non-malignant rectal tissue. The promoter upregulated in rectal cancer (RC) drives expression of the long non-coding RNA lnc-RNMT-2:5, previously annotated as LDLRAD4-217 transcript (ENST00000592657). This study aimed to investigate the relevance of lncRNMT-2:5 for malignant transformation using in silico and in vitro approaches. Promoter binding site prediction using TFBIND, Motiflab, and CIIIDER identified several transcription factors, including GATA1/2/3, YY1, ARNT, and STAT3, which are known to regulate key cancer-related processes such as proliferation, invasion, metastasis, and therapeutic response in cancer. Additionally, miRNA target analysis predicted binding sites for miR-214-5p and members of the miR-30 family, both recognized as tumor suppressors in colorectal malignancies. These findings suggest that lncRNMT-2:5 may act as a competing endogenous RNA (ceRNA), promoting tumor progression by sequestering suppressive miRNAs. lncLocator predicts its predominant localization in the cytoplasm, further supporting its potential sponge-like function. Although previous studies reported differential expression of lnc-RNMT-2:5 in rectal tissue, its absence in gut epithelial cell lines (HCEC-1CT, HCT116, DLD1, and SW620) in our transcriptomic analysis suggests a stromal-specific expression pattern. This stromal origin is further supported by our qPCR validation, which confirmed its presence in the fibroblast cell line CCD-18Co and in healthy and inflamed colon mucosa. lnc-RNMT-2:5 likely represents a stromal-derived sponge lncRNA with a potential role in promoting colorectal cancer progression. Future studies should confirm its stromal-specific expression and further explore its molecular function and therapeutic potential
HSALR Mice Exhibit Co-Expression of Proteostasis Genes Prior to Development of Muscle Weakness
No full textMyotonic dystrophy type 1 (DM1) is a progressive multisystemic disease caused by a CTG repeat expansion in the DMPK gene. The toxic mutant mRNA sequesters MBNL proteins, disrupting global RNA metabolism. Although alternative splicing in DM1 skeletal muscle pathology has been extensively studied, early-stage transcriptomic changes remained uncharacterized. To gain deeper and contextual insight into DM1 transcriptome, we performed the first Weighted Gene Co-expression Network Analysis (WGCNA) on skeletal muscle RNA sequencing data from the widely used DM1 mouse model HSALR (~250 CTG repeats). We identified 532 core genes using data from 16-week-old mice, an age before the onset of muscle weakness. Additional differential expression analysis across multiple HSALR datasets revealed 42 common up-regulated coding and non-coding genes. Within identified core genes, the pathway gene-pair signature analysis enabled contextual selection of functionally related genes involved in maintaining proteostasis, including endoplasmic reticulum (ER) protein processing, the ubiquitin-proteasome system (UPS), macroautophagy and mitophagy, and muscle contraction. The enrichment of ER protein processing with prevailing core genes related to ER-associated degradation suggests adaptive chaperone and UPS activation, while core genes such as Ambra1, Mfn2, and Usp30 indicate adaptations in mitochondrial quality control. Coordinated early alterations in processes maintaining protein homeostasis, critical for muscle mass and function, possibly reflect a response to cellular stress due to repeat expansion and appears before muscle weakness development. Although the study relies exclusively on transcriptomic analyses, it offers a comprehensive, hypothesis-generating perspective that pinpoints candidate pathways, preceding muscle weakness, for future mechanistic validation
Genotype-sensitive dynamic model of the serotonin presynapse shows complex interplay between synapse metabolites
No full textSuicide attempts are prevalent among patients with bipolar disorder (BD), with an impaired
serotonin system playing a critical role in this pathology. Given the heritable nature of some
contributing factors, this study aims to develop a model of the serotonin presynapse that integrates
multiple genetic variants to assess their collective impact on serotonin metabolism.
We constructed a dynamic model that incorporates five genetic variants linked to serotonin
synthesis (TPH2), transport (SLC6A4), and degradation (MAOA). The model, comprised of eight
differential equations, quantifies changes in serotonin metabolite concentrations over time. We
validated the model using data from 140 healthy individuals and tested it with a sample of 101 BD
patients. We compared predicted metabolite levels between patients with and without a history of
suicide attempts.
The model demonstrated accurate predictions of metabolite concentrations in healthy
individuals, aligning well with experimental measurements. Among BD patients, those who had
attempted suicide showed significantly higher concentrations of free cellular serotonin (p=0.048)
and stored serotonin (p=0.047), along with a trend toward reduced levels of the degradation
product 5-hydroxy-3-indolacetic acid (p=0.054).
This model provides a novel approximation of the complex biological interactions within
the serotonin system and highlights the significant role of genetic variants in suicide risk among
BD patients. While the findings emphasize the model's potential for yielding quantitative insights
into individual predispositions, limitations such as sample size necessitate further investigation to
enhance stratification of suicide attempters. This approach holds promise for advancing
personalized suicide risk assessments beyond traditional genetic studies.Book of abstract: Belgrade Neuroscience Next Hub 2025 with international participation
February 27, 2025 in Belgrade, Serbi