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Establishment of a patient-derived induced pluripotent stem cells with 22q11.2 microdeletion: a model system for studying neurodevelopmental disorders
No full textBackground: Neurodevelopmental disorders (NDDs), including autism spectrum
disorders, intellectual disability and schizophrenia, represent a public health challenge in
modern societies. However, molecular mechanisms underlying NDDs are still unknown.
22q11.2 Deletion Syndrome (22q11.2DS), caused by microdeletion 22q11.2, is one of the
syndromes with a high risk for NDDs; it is one of the strongest known risk factors for
development of psychiatric illness and one of the highest known genetic risks for
schizophrenia.
Material and Methods: Peripheral blood mononuclear cells from patients with 22q11.2DS
and control subjects were reprogrammed using CytoTune™-iPS 2.0 Sendai
Reprogramming Kit. Characterization of generated iPSC lines were done by analyzing
their morphology, genomic integrity, pluripotency and the ability to differentiate into three
germ layers.
Results: Peripheral blood mononuclear cells from seven patients with 22q11.2
microdeletion and three healthy controls were reprogrammed. Genotyping revealed that
some of the iPSC lines contain additional CNVs. All generated iPS cell lines express
markers of pluripotency and have ability to differentiate into three germ layers.
Conclusion: iPSC lines derived from patients with 22q11.2DS and healthy controls were
successfully established. They represent a powerful model system for studying molecular
mechanisms underlying NDDs.Book of absctracts: European Society of Human Genetics (ESHG) 2025 Conference, May 24–27, 2025. Milan, Ital
National Lactic Acid Database development: current status and future direction
No full textIntroduction: Lactic acid bacteria (LAB) are gram-positive bacteria that
primarily utilise carbohydrates as a carbon source and exhibit high tolerance
to low pH. They are commonly found in raw milk and dairy products.
Although LAB encompasses over 60 genera, the most prevalent in food
fermentation include Lactobacillus, Lactococcus, Streptococcus,
Enterococcus, Pediococcus, Leuconostoc, and Weissella. The key metabolic
characteristics of LAB consist of acid and aroma production, protein
hydrolysis, exopolysaccharide synthesis, and antimicrobial compound
production. The indigenous microbiota of raw milk directly influences the
sensory properties of its products. LAB are classified as Generally
Recognized as Safe (GRAS) and are widely employed in fermented foods.
The WHO recommends including fermented dairy, meat, and vegetable
products in the daily diet because of LAB's beneficial impact on human
health.
Experimental: Our decades of research demonstrate that each LAB strain
possesses unique characteristics, offering specific health benefits for both
humans and animals. Given the limited biodiversity of commercial starter
cultures used in the fermented product industries, which have reached their
technological and probiotic potential, our distinct collection of natural LAB
isolates from Serbia (and the surrounding region) provides essential
resources for addressing future challenges in human and animal health,
agriculture, and the food industry. The IMGGE’s LAB collection comprises
approximately 5,000 different isolates collected from artisanal dairy, meat,
and other fermented products across Serbia, Montenegro, Bosnia and
Herzegovina, Croatia, Bulgaria, Iran, Azerbaijan, and the Russian
Federation. Our LAB strains are utilised in scientific research (doctoral and
master's theses) and industry (spinout companies, licensing agreements),
fostering both scientific excellence and innovation.Key findings: Our strategic project aims to establish the LABbank database,
which will catalog the technological and probiotic characteristics of each
LAB strain in our collection. The project's first phase involved developing
the LABbank database for the first 1,000 strains, implementing
administration features, and adding filter and search options based on strain
characteristics and taxonomy. Among these 1000 strains, the most dominant
genera are Lactobacillus sp. (52.85%), Enterococcus sp. (26.53%),
Lactococcus sp. (8.5%), Streptococcus sp. (6.22%) and Leuconostoc sp.
(5.49%). For now, eighteen strains are deposited in the BCCM culture
collection, and three are under international patent protection (PCT). Our
database will support researchers in their studies and provide easier access
for companies in the fermented food and pharmaceutical industries to
identify our LAB strains suitable for industrial applications.Book of Abstracts: International conference, Biobased future: green bioprocessing for innovative bioactive product
Effect of doxorubicin and quercetin combined treatment on osteosarcoma model systems
No full textIntroduction
Osteosarcoma (OS) is a highly aggressive bone tumor
primarily affecting pediatric patients. Standard treatments
include surgical resection, chemotherapy, and radiation
for tumors that cannot be surgically removed. Although
the 5–year survival rate is 65.5%, patients with
metastases and recurrence have a significantly lower
survival rate of ~30%. Despite this concerning statistic,
the treatment for OS has remained largely unchanged
over the past three decades. This stagnation in treatment
innovation highlights the urgent need for further research
and development in therapies tailored specifically for OS.
Material and method
We identified the DEGs between bone (7 samples) and
OS (27 samples). To conduct in-depth study of the
obtained upregulated DEGs, we constructed a PPI
network and identified the most significant gene cluster.
We investigated the effects of the combined treatment
with doxorubicin and quercetin on SAOS-2 osteosarcoma
cells in 2D condition and immobilized in alginate microbeads
in 3D condition. We assessed the effects of treatments
on cell viability using MTT and the expression of
genes using qPCR.
Result and discussion
We have analysed DEGs between bone and human
osteosarcoma samples and identified 630 upregulated
genes. We extended the networks with information from DrugBank to identify potential therapeutics for osteosarcoma
focusing on the top 10% of interconnected genes
in cluster due to their important biological functions. The
identified cluster had enrichment in biological processes
connected to oxidative phosphorylation and we found
quercetin as a promising candidate for treating OS. We
analysed quercetin’s effect utilizing the Saos-2 in 2D and
3D on viability and gene expression, alone or in
combination with doxorubicin. Following treatment, we
assessed cell viability and the expression of genes. Our
results have shown that the combined treatment
statistically significantly decreased the viability of
SAOS-2 cells cultured in 2D and 3D conditions
compared to cells treated with doxorubicin. We analyzed
the expression of genes associated with poor prognosis in
patients such as pluripotency genes, an OS marker, and a
resistance-related gene. Collectively our results show
different responses to the combined treatment depending
on the model system used.
Conclusion
The combined treatment substantially reduced cell
viability in 2D and 3D models and decreased expression
of genes associated with poor prognosis compared to
doxorubicin alone in 3D models. We can hypothesize
that microenvironment-based mechanisms modulate cell
sensitivity to therapy and increase resistance to treatment
of osteosarcoma cells cultured in 3D condition. Understanding
the molecular mechanisms will significantly
contribute to the development and enhancement of
existing therapies, thereby facilitating advancements in
the treatment of osteosarcoma.EACR 2025 Congress Abstract
Structural Properties of Intrinsically Disordered Dehydrins Underlying Their Protective Role in Desiccation Tolerance
No full textDrought is a major threat to global food security, presenting a critical challenge for agriculture.
Resurrection plants exhibit extraordinary desiccation tolerance, surviving the loss of up to 98 % of their
water content for extended periods and fully recovering metabolic function upon rehydration.
Investigating the molecular mechanisms underlying this adaptation can offer valuable insights for
enhancing crop resilience to drought.
Dehydrins, a subclass of late embryogenesis abundant (LEA) proteins, accumulate in resurrection plant
leaves during desiccation. Although they have been implicated in the protection of proteins, membranes,
and nucleic acids, their precise physiological function remains unknown.
In this study, we recombinantly produced DHN152, a desiccation-induced dehydrin from Ramonda
serbica, an ancient resurrection species, and analysed its structural and functional properties. DHN152 is
a highly hydrophilic protein (GRAVY index: –1.29) with a high glycine content (22.6%) and abundant
charged residues (lysine, glutamate, and aspartate). In silico analyses of DHN152 revealed high disorder
propensity, which was confirmed by circular dichroism (CD) spectroscopy under physiological conditions.
Upon exposure to 2,2,2-trifluoroethanol (a desiccation-mimicking agent) and lipid-mimetic detergents,
DHN152 partially adopted an α-helical conformation, highlighting its structural plasticity. Given its lysine-
rich composition and predicted nuclear localisation, we examined its effect on DNA under desiccation
conditions. Additionally, we assessed its protective effects on enzyme activity and Escherichia coli growth
under osmotic and salt stress. Taken together, our findings provide novel insights into the structure-
function relationship of dehydrins in resurrection plants, laying the groundwork for bioengineering
strategies aimed at improving drought resistance in crops.Book of abstract: 15th EBSA Congress in Rome, Italy, from the 30th of June to the 4th of July 2025
LEA4 protein family group originated from Ramonda serbica:– Structural analysis
No full textAn extreme loss of cellular water or desiccation (5-10% of relative water content) leads to protein denaturation, aggregation, and degradation, and affects the fluidity of the membrane resulting in a loss of its integrity. Ramonda serbica is an ancient resurrection plant, that survives a long desiccation period and fully recovers metabolic functions upon watering. The key constituents of vegetative desiccation tolerance resurrection plants are Late Embryogenesis Abundant Proteins (LEAPs). This heterogeneous group of intrinsically disordered proteins forms mostly random conformation when fully hydrated, turning into compact α-helices during desiccation. In this study, we produced three LEA proteins belonging to the LEA4 group from R. serbica and purified with a purity higher than 95%. Using circular dichroism spectroscopy for structural characterisation, it has been shown that all three LEAPs were mostly disordered in physiological conditions and at different pH. The protein predominantly folds into an α-helical form when 2,2,2-trifluoroethanol and lipid mimetic detergent are present, suggesting the significance of structural plasticity. LEA protein protective role and their ability for structural disorder-to-order transitions might be crucial in the desiccation tolerance mechanism. We propose that the protective role of LEAPs in cells by this structural duality is most likely a result of interactions with membranes and proteins that are desiccation-sensitive. Our research provides new insights into the link between the structure and function of LEAPs in resurrection plants. It establishes the basis for future bioengineering techniques development to improve crops' resistance to drought.Book of abstract: 15th EBSA Congress in Rome, Italy, from the 30th of June to the 4th of July 2025
Optimization of Actinomycin D Structure for Antifungal Activity: Strategies to Improve Combat Against Fungal Infections
No full textActinomycin D (ActD), initially discovered as an antibiotic with anticancer potential, was
discontinued as a chemotherapeutic agent due to severe side effects, including bone
marrow and liver toxicity. However, ActD demonstrates significant antimicrobial activity,
with minimum inhibitory concentrations (MICs) of 0.15 μg/mL against Staphylococcus
and Bacillus species, as well as mild antifungal effects. These properties are especially
important in the context of rising invasive Candida infections, which affect over 200,000
patients annually and face growing concerns over drug resistance and limited treatment
options.
This study explores strategies to optimize the structure of ActD for enhanced antifungal
activity against Candida species while minimizing its cytotoxicity. ActD’s peptide
component and chromophore were synthesized separately and biological screening,
including MIC determination and cytotoxicity tests, was performed. Additionally, organic
fragments with antimicrobial properties—curcumin, carvacrol, ferulic acid, and
cinnamic acid—were tested for their ability to improve ActD’s antifungal efficacy. We also
analyzed the peptide length and amino acid composition of ActD to identify structural
modifications that could enhance its activity.
Our findings show that the peptide component alone is biologically inactive, while the
chromophore exhibits antifungal activity against Candida parapsilosis at 250 μg/mL.
Notably, all fragments tested showed reduced cytotoxic effects on MRC-5 fibroblasts.
These preliminary results highlight the potential of the ActD chromophore as an
optimized antifungal agent, offering a promising strategy for combating Candida
infections and mixed Candida-Staphylococcus infections, addressing a critical unmet
medical need.Book of abstract: FEMS MICRO 2025: Congress & Exhibition, taking place in Milan, Italy, from 14-17 July 2025
Newly identified species from the dog dental plaque microbiome highlight little overlap with humans
No full textUnderstudied pet-associated microbiomes represent a rich source for the discovery of microbial taxa important for pet and human health. From a cohort of 23 dogs, we sampled and metagenomically sequenced 64 dental plaque microbiomes, generating 1945 metagenome-assembled genomes spanning 347 microbial species, including 277 undercharacterized species without cultivated representatives. Integration with human microbiome data revealed the dog plaque microbiome is more diverse than – and shows little overlap (5.9% species in common) with – the human plaque microbiome, even though some shared periodontal pathobionts arise as a potential concern
Synthesis of high-performance bio-polyurethanes using medium-chain length polyhydroxyalkanoates
No full textThe data and files contained in this dataset are related to research on the potential of using
microbial biopolymer polyhydroxyalkanoates (PHAs) and castor oil as the polyol for the synthesis of
polyurethanes (PUs), using hexamethylene diisocyanate (HMDI) as a crosslinking agent. PUs are
extensively used in various applications such as foams, coatings, elastomers, adhesives and biomedicine,
making them a significant part of the polymer market. Recent advancements in green PU synthesis aim
to replace fossil-based diisocyanates and polyols with bio-based alternatives to create environmentally
friendly polymers without sacrificing performance. In this study, the PHA was extracted from the
bacterial biomass of Pseudomonas putida CA-3 using a combination of commercially available enzymes
(Cellulase/Bromelain/Trypsin/Pancreatin/Savinase) and the PHA was characterized using Nuclear
Magnetic Resonance (NMR). Following an eco-friendly approach, bio-PU polymers were synthesized via
solvent casting by varying the ratios of castor oil (C) and mcl-PHA (P) (C:P 100:0, 80:20, 50:50, 20:80 and
0:100). The resulting products were characterized by Thermogravimetric Analysis (TGA) and X-Ray
Diffraction (XRD). Our findings demonstrate the potential of PHA-based synthesis of PUs as a promising
approach for green, sustainable polyurethane production, contributing to the circular bio-economy and
environmentally conscious plastics synthesis.readme.txt (7.073Kb)***Dataset contents***mcl-PHA 1H NMR.csv (1.118Mb) mcl-PHA 1H NMR.png(13.89Kb)
mcl-PHA 13C NMR.csv (1.081Mb)
mcl-PHA 13C NMR.png (42.80Kb)
TGA mcl-PUs.png (73.53Kb)
TGA PU1-mclPU.csv (86.00Kb)
TGA PU2-20-80 (C-P).csv (85.12Kb)
TGA PU3-50-50 (C-P).csv (85.52Kb)
TGA PU4-80-20 (C-P).csv (84.35Kb)
XRD mcl-PUs.png (97.37Kb)
XRD PU1-CO+HMDI.csv (15.08Kb)
XRD PU2-mcL-PHA.csv (15.28Kb)
XRD PU3-20-80 (C-P).csv (15.05Kb)
XRD PU4-50-50 (C-P).csv (15.09Kb)
XRD PU5-80-20 (C-P).csv (15.19Kb)File readme.txt (7.073Kb) is under licence public domain CC
Tuning Properties of Sustainable Castor Oil Based Polyurethanes With Bacterial Biomass as Fillers
No full textA novel series of bio-based polyurethanes (bio-PUs) were synthesized from castor oil using hexamethylenediisocyanate (HMDI) as crosslinking agent by solvent casting method without any catalyst and further reinforced with bacterial biomass as bio-fillers. For the first time, biomasses from the biopolymer polyhydroxyalkanoate (PHA) production process, containing medium chain length biopolymer, mcl-PHA (F1) and residual bacterial biomass after the biopolymer extraction (F2), as well as bacterial biomass from the biopigment prodigiosin production process (F3) were applied as bio-fillers, resulting in PU-F1 to PU-F3 materials, respectively. The resulting functional bio-polyurethanes were characterized by various techniques including ATR-FTIR spectroscopy, SEM, X-ray diffraction, mechanical tests, transparency, water contact angle, but also cytotoxicity tests and shape memory ability were evaluated to open their applicative potential. The FTIR spectroscopy analysis confirmed the formation of polyurethane linkage. Bacterial biomass particles size and distribution reflected on the PUs properties suggesting that the type and the dispersion of the filler play an important role in the modulation of new PU materials. The water contact angle measurements revealed that PU-F1, containing mcl-PHA biopolymer exhibits higher hydrophobicity than other bio-PUs, that further reflected on better biofilm attachment in comparison to other bio-PUs. The addition of bacterial biomass containing biopigment resulted in purple dyed material of stable color over time and with the proved absence of toxicity (PU-F3). All synthesized bio-PUs appeared as non-toxic materials for human healthy fibroblast cell line MRC5. Shape memory ability was observed for the bio-PUs. The addition of variety of bacterial biomass into polyurethane matrix is a significant step towards the green conversion of resources and circular bioeconomy for plastics
Innovating Thyroid Health with Genomics and Predictive Algorithms through Collaborative Excellence (InnoThyroGen)
No full textGlobal research indicates that millions of people suffer from thyroid disorders, such as hypothyroidism, hyperthyroidism, autoimmune diseases, and tumors, with nearly 60% of them unaware of their condition. Autoimmune thyroid diseases, notably Hashimoto's thyroiditis and Graves' disease, are the most prevalent, affecting about 5% of the population. Thyroid cancers are also on the rise, with significant increases in detection over recent decades. Despite various treatment and diagnostic options, many face misdiagnosis, ineffective treatments, and diminished quality of life. Levothyroxine, a synthetic thyroid hormone, is widely prescribed in Europe and the USA, yet 20-50% of patients experience improper dosing, necessitating frequent adjustments. Antithyroid medications can cause severe side effects, underscoring the need for careful management of thyroid diseases to avoid increased healthcare costs and lost productivity. Current treatment approaches, often not personalized, overlook genetic predispositions and environmental factors. Advances in genomics, such as next-generation sequencing (NGS) and the polygenic risk score (PRS), offer new opportunities for personalized medicine. These tools, combined with electronic health records (EHR) and lifestyle data, can improve disease prediction and prevention strategies. Pharmacogenomics further enables tailored drug treatment based on individual genetic profiles, promising more effective and personalized care. The design of thyroid disease gene panels and the development of innovative and digital solutions based on artificial intelligence stand as pivotal advancements. These could markedly accelerate the integration of more personalized and cost-effective approaches into clinical practice, heralding a new era of precision medicine for thyroid disorders.Principal Investigator: Dr. Nikolina Pleic, University of Split School of Medicine, Split, CroatiaParticipants from IMGGE: Dr Branka Zukic, dr Natasa Tosic, dr Nikola Kotur, dr Biljana Stankovic,
dr Vladimir Gasic, dr Teodora Karan Djurasevic, dr Ivana Grubisa, dr Bojan Ristivojevic, dr Irena
Marjanovic, Marina Jelovac, Djordje Pavlovic, Isidora Curic, Katarina Krstajic, Mihajlo StasukDuration period: 2025 - 202