imagine (Institute of molecular genetics and genetic engineering)
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Phenolic composition, antioxidant, and enzyme inhibitory activities of Saxifraga paniculata Mill.
No full textAlpine saxifrage (Saxifraga paniculataMill.), a rosette-forming subshrub, isdistributed inNorthAmerica, Europe, andAsia. This studyaimed to investigate the chemical compositionand invitro biological activities of themethanolic extract obtained fromthe aerialpartsoftheplant.Thephenolicprofileandcontentofthefour compounds were determined by LC-DAD-ESI-MS. Total phenolic content (TPC), totalflavonoidcontent (TFC), antioxidantpotential aswellasenzymeinhibitorycapacitywereevaluatedspectrophotometrically. The antimicrobial activitywas tested by the brothmicrodilutionmethod,whilethecytotoxiceffectwasexaminedby theMTTassay.Theanalysisrevealedthepresenceof23compounds, including quercetin, six quercetin glycosides, seven kaempferol glycosides, two caffeoylquinic acids, three galloylhexosides, two digalloylhexosides, galloylquinic acid, and (epi)gallocatechingallate. Themost abundant constituentwas quercitrin (1.62%), followed by chlorogenic acid (0.28%), kaempferol deoxyhexoside (0.12%), and rutin (0.07%). TPC and TFCwere 26.71% and 2.12%, respectively. The demonstrated antioxidant activitywas significant (DPPH test, IC50=9.30 μg/mL; FRAP test, FRAP value=2.89mmol Fe2+/g; ABTS test, IC50=34.22 μg/mL). The extract potently inhibitedα-glucosidase (IC50=1.86μg/mL) and, toa lesser extent, 15-lipooxygenase (IC50=49.60μg/mL) andαamylase (IC50=9.95mg/mL).Theminimuminhibitoryconcentration(MIC) againstAcinetobacter baumanniiwas1000μg/mL;MIC washigheragainsttheothersixtestedstrainsofmicroorganisms.No effect on cancer cell lines (HeLa, HT-29, HCT-116, LS174) and healthy cells (MRC-5)was observed. Thefindings expanded the scarceliteraturedataonS.paniculataandhighlighteditspotential, warrantingfurtherinvestigation.International Congress on Natural Products Research 2024 (ICNPR2024
Meat Industry as Our Best Chance for Controlling Transmission of a Tiny but Deadly Tapeworm
No full textEchinoccocosis is a zoonotic disease which can have severe, even lethal consequences for the host, yet multiple effective means of controlling transmission and thus preventing infection, are available. Echinococcus granulosus and E. multilocularis are the two tapeworm species which are clinically most relevant in Europe and both are endemic in the Balkans. The life cycle of the tapeworm includes intermediate and dead-end hosts, ungulates and humans, as well as definitive hosts, the Canidae and Felidae. Disease caused by E. multilocularis, alveolar echinococcosis (AE) is more severe and also difficult to diagnose, especially in animals. Echinococcosis in livestock leads to morbidity and mortality, thereby facilitating economic losses at various levels. Reporting is mandatory, yet under-reporting is a common occurrence as is the failure to identify relevant geographical transmission foci in a timely manner, thus contributing to continuous tapeworm transmission and spread. As echinococcosis is a foodborne disease in animals and humans, the food industry, and particularly the meat industry, are key stakeholders in raising awareness, lobbying government authorities for control measures and improved diagnostics
Plastic bioremediation potential of groundwater microbiomes
No full textGlobal increase in the production of highly recalcitrant plastic materials and their accumulation in the environment are creating the urgent need to develop new sustainable strategies to mitigate this type of pollution. Microbial biotechnology, including blue biotechnology, is a promising option for the sustainable disposal of plastic waste. Despite both soil and water representing potential sources of novel biocatalysts, studies unraveling the biotechnological potential of soil microbial communities considerably outnumber studies of aquatic microbiota. Although traditionally focused on marine resources, the scope of blue biotechnology has expanded to include all aquatic ecosystems and the focus is shifting towards underexplored aquatic environments. Environmental niches that are not readily accessible for sampling, representing reservoirs of enzymatic activities with potential biotechnological applications, such as subsurface environments, especially its core component groundwater, are still relatively understudied. Despite a proposed overlap between microbial enzymatic capacity to degrade one of the substrates that has been available on Earth for a very long time—lignocellulosic biomass and a relatively newly available substrate, introduced only about a century ago by humans—plastics, studies investigating potential for the transformation of fossil and bio-based plastics by lignocellulose-degrading microorganisms and their consortia and bioprospecting for microorganisms capable of degrading both, are still relatively scarce. Here, we discuss and summarize recent advances in decoding plastolytic and lignocellulolytic potential of relatively rarely examined groundwater bacteria
Diversity and Biotechnological Potential of Limestone and Serpentinite Microorganisms
No full textThe diversity and biotechnological potential of microorganisms cultivated from 10 micritic limestone, limestone and serpentinite (protolith harzburgite) rock samples collected in the Zlatibor Jurassic Ophiolitic Massif of western Serbia was assessed in this study. The most abundant cultivated rock bacteria belong to genera Stenotrophomonas, Arthrobacter, Bacillus, Paenarthobacter and Microbacterium, while the Archaeorhizomyces, Aspergillus, Mortierella, Termitomyces and Pleurotus were the most represented rock fungi after cultivation. All 120 rock isolates, screened on 7 lignocellulosic and plastic substrates, demonstrated the ability to grow and/or completely degrade at least 3 of the tested substrates. Ninety-five isolates (79.2%) grew on and/or degraded all tested substrates; 104 isolates (86.7%) grew on all tested plastic substrates, while 105 isolates (87.5%) grew on carboxymethyl cellulose (CMC), xylan and lignin. Twenty isolates (16.7%) demonstrated complete degradation of CMC. The majority of 32 isolates (87.5%) that were additionally screened on arabinoxylan grew on this substrate. Live dyeing of textile materials with a pigment-producing limestone isolate, identified as Arthrobacter, showed mixed results. While the process of coloration was successful, the color intensity after sterilization and washing was low. Potential applications of collected rock isolates in the biomass degradation, bioremediation of plastic-polluted geo-environments and textile dyeing should be further explored. Investigated rocks are of the micritic limestone, limestone and serpentinite (protolith harzburgite) type.Calcite is the main mineral phase in limestones, while lizardite is in serpentinites.The most abundant cultivable rock bacteria belong to genera Stenotrophomonas, Arthrobacter, Bacillus, Paenarthobacter and Microbacterium.Archaeorhizomyces, Aspergillus, Mortierella, Termitomyces and Pleurotus were the most represented rock fungi after cultivation.16.7% of screened rock microorganisms degraded carboxymethyl cellulose, while one limestone Arthrobacter isolate produced pigment. Investigated rocks are of the micritic limestone, limestone and serpentinite (protolith harzburgite) type. Calcite is the main mineral phase in limestones, while lizardite is in serpentinites. The most abundant cultivable rock bacteria belong to genera Stenotrophomonas, Arthrobacter, Bacillus, Paenarthobacter and Microbacterium. Archaeorhizomyces, Aspergillus, Mortierella, Termitomyces and Pleurotus were the most represented rock fungi after cultivation. 16.7% of screened rock microorganisms degraded carboxymethyl cellulose, while one limestone Arthrobacter isolate produced pigment
A simple alginate-based 3D in vitro model for preclinical anticancer drug evaluation
No full textThe development of novel anticancer drugs is a slow and costly process, with only 5 out of 5,000 compounds ultimately reaching the market. A major limitation lies in the low predictability of traditional preclinical methods, such as two-dimensional (2D) cell cultures and animal experiments, which often fail to replicate the cellular responses to drugs observed in patients. Therefore, three-dimensional (3D) in vitro models have emerged as valuable tools for drug screening. The aim of this study was to establish a 3D in vitro model based on alginate microfibers suitable for immobilizing various cancer cell types for anticancer drug testing. Murine osteosarcoma cells K7M2-wt (cell density: 4×106 cells cm-3) were immobilized in 2 wt.% Ca-alginate microfibers (diameter: 500−600 µm) with or without 2 wt.% hydroxyapatite (HAP) particles, while human non-small cell lung carcinoma cells NCI-H460 (cell density: 4×106 cells cm-3) were immobilized in 2 wt.% Ba-alginate microfibers (diameter ~ 550 µm) by manual extrusion. During long-term cultivation, cells in microfibers stayed viable and metabolically active, forming aggregates over time. Anticancer drug testing revealed that the half-maximal inhibitory concentration (IC50) values for 3D cultures of both K7M2-wt and NCI-H460 cells were up to 50-fold higher compared to those for 2D cultures. Additionally, the gene expression analysis showed a significant upregulation of drug resistance–related genes in 3D cultures. Overall, the obtained results align with the observed higher resistance to anticancer drugs in patients compared to traditional preclinical models, highlighting the potential of the proposed model to improve in vitro drug screening reliability.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
Isolation and solubility of flavonoids rutin and hesperidin from plant waste material
No full textThe aim of this research was to isolate flavonoids from plant waste material in the best possible
quality. The isolated flavonoids could be used for the formulation of powders or tablets, which can
be used for the treatment of varicose veins and hemorrhoids. This would not only contribute to
the pharmaceutical industry, but also to sustainable waste management in Serbia.
The extraction of rutin from the flower buds of Sophora japonica was carried out with hexane and
methanol. The dry extract was dissolved in warm water and filtered. The extraction of hesperidin
from orange peel was carried out in a similar manner using hexane and methanol. The dry extract
was mixed with 6% acetic acid, dried in a desiccator and then recrystallized with dimethyl
sulfoxide and washed with isopropanol. Chemical characterization was performed by LC-MS and
NMR spectroscopy. Solid dispersions were formed using the solvent method and by adding PVP
VA 64 and poloxamer P188.
A yield of 6.15% rutin and 0.265% hesperidin was obtained. The relative purity of rutin was
95.51% and of hesperidin 80.28%. In an in vitro dissolution test, over 90% of the rutin was released
from the formulation in less than 15 minutes, and the release from the hesperidin dispersion was
about 10%.
The extraction of rutin was easier to perform and the yield and purity were better than hesperidin.
Rutin showed a good dissolution rate. For hesperidin, the possibility of using other polymeric
excipients needs to be investigated.The aim of this research was to isolate flavonoids from plant waste material in the best possible
quality. The isolated flavonoids could be used for the formulation of powders or tablets, which can
be used for the treatment of varicose veins and hemorrhoids. This would not only contribute to
the pharmaceutical industry, but also to sustainable waste management in Serbia.
The extraction of rutin from the flower buds of Sophora japonica was carried out with hexane and
methanol. The dry extract was dissolved in warm water and filtered. The extraction of hesperidin
from orange peel was carried out in a similar manner using hexane and methanol. The dry extract
was mixed with 6% acetic acid, dried in a desiccator and then recrystallized with dimethyl
sulfoxide and washed with isopropanol. Chemical characterization was performed by LC-MS and
NMR spectroscopy. Solid dispersions were formed using the solvent method and by adding PVP
VA 64 and poloxamer P188.
A yield of 6.15% rutin and 0.265% hesperidin was obtained. The relative purity of rutin was
95.51% and of hesperidin 80.28%. In an in vitro dissolution test, over 90% of the rutin was released
from the formulation in less than 15 minutes, and the release from the hesperidin dispersion was
about 10%.
The extraction of rutin was easier to perform and the yield and purity were better than hesperidin.
Rutin showed a good dissolution rate. For hesperidin, the possibility of using other polymeric
excipients needs to be investigated
PHARMACOGENOMICS OF CHILDHOOD ACUTE LYMPHOBLASTIC LEUKEMIA IN CHILDREN IN SERBIA
No full textPersonalized medicine uses modern treatment approach aiming to tailor therapy to the individual genetic
and molecular characteristics of each patient. In Serbia, pharmacogenomics research has demonstrated
the potential of this approach in children with acute lymphoblastic leukemia (ALL), the most common
malignancy in the pediatric population. Although the cure rate for ALL is high, treatment is often
accompanied by significant side effects and drug resistance, highlighting the need for further
improvement of therapeutic protocols. As part of a collaborative study between the Institute of Molecular
Genetics and Genetic Engineering and the University Children's Hospital in Belgrade, pharmacogenetic
markers associated with the response to thiopurine drugs, glucocorticoids, vincristine, and methotrexate
were analyzed in the Serbian pediatric population. Variants in candidate genes were identified using PCR
and Sanger sequencing methodologies. The resulting genetic data were retrospectively correlated with
clinical parameters and treatment outcomes. Additionally, a polygenic risk score (PRS)-based model was
developed to predict methotrexate-induced hepatotoxicity, offering a potential tool for individualized
treatment planning. The results of our study highlight the importance of tested molecular markers in
determining the efficacy and safety of therapy. Also, they indicate the need for additional research to
support the broader application of these findings in clinical practice in Serbia. Further recommendations
emphasize the implementation of advanced methods for detecting genetic variants, such as nextgeneration
sequencing (NGS), along with the integration of bioinformatics tools and machine learning
algorithms. Additionally, the use of patient-derived induced pluripotent stem cells model system for
research is proposed. These advancements will support the progress and implementation of personalized
medicine in Serbia.4TH CONGRESS OF GENETICISTS IN BOSNIA AND HERZEGOVINA WITH
INTERNATIONAL PARTICIPATION
2ND - 4TH OCTOBER 2025, BANJA LUKA, BOSNIA AND HERZEGOVIN
ALDOA-212 noncoding transcript – a potential biomarker for colorectal cancer
No full textBackground: Patients with locally advanced rectal cancer (LARC) often receive neoadjuvant chemoradiotherapy (nCRT)
based on 5-fluorouracil (5-FU). Cytochrome P450 (CYP) enzymes, involved in drug metabolism and carcinogenesis,
may influence response to therapy. Variability in CYP gene expression makes them promising biomarkers for predicting
treatment outcomes in rectal cancer.
Material and methods: Genomic DNA from 38 LARC patients treated with standard nCRT (fluorouracil + leucovorin
+ radiotherapy) was analyzed. Clinical response was assessed by tumor regression grade (TRG). Seven patients with
extreme responses – good (3 TRG1 and 2 TRG2) and poor (2 TRG5) were selected for whole-exome sequencing (WES).
Candidate predictive variants were identified based on their differential presence between responder and nonresponder
groups and biological relevance (localization in coding region or regulatory elements and enzyme-altering
effects). Selected variants were validated in 29 moderate-response patients (15 TRG3 and 14 TRG4) by targeted
sequencing.
Results: Two genetic variants were selected according to the criteria outlined above: rs149012039, which is located
within the CYP2D7 pseudogene and represents a frameshift variant, and rs3093200, which is located in the first exon
of the CYP4F2 gene and has a damaging effect on the protein. Validation in the remaining samples showed that neither
variant was present in patients with a moderate response to therapy.
Conclusions: The presence of variants rs149012039 and rs3093200 in individuals with a good clinical response and
their absence in individuals with a moderate or poor response supports their potential as predictive biological markers
for response to nCRT in rectal cancer
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
ESTABLISHMENT OF AN IN VITRO INSULIN RESISTANCE MODEL IN HepG2 CELLS THROUGH GLUCOSE AND INSULIN COTREATMENT
No full textInsulin resistance is a metabolic condition where
the body's cells do not respond effectively to
insulin, resulting in increased blood sugar levels.
It plays a key role in the development of type 2
diabetes, which causes over 6.7 million deaths
each year worldwide.
In vitro models of insulin resistance still
represent a powerful tool for studying various
aspects of this complex metabolic condition,
despite well-known limitations. The HepG2 cell
line preserves the insulin response pathways of
hepatocytes, making it an ideal model for
studying insulin resistance in liver cells.
Here we treated HepG2 cells with high
concentrations of glucose (3, 4.5 and 10g/L) and
various doses of insulin (10-5, 10-6, 10-7, 10-
8M) in order to establish the model of insulin
resistance. Cell viability was measured using
MTT test, and the qPCR was used for
determination of expression of IRS1, GLUT2,
AKT1 and G6PC1 genes in treated cells. Our results demonstrated a significant decrease
in cell viability at the highest insulin
concentration (10⁻⁵ M). Notably, neither insulin
alone nor high glucose concentrations
individually induced insulin resistance in HepG2
cells. However, the combined treatment with 4.5
g/L glucose and 10⁻⁶ M insulin lead to decreased
expression of IRS1, GLUT2, and AKT1 (0.4, 0.15,
and 0.01 times lower than control, respectively)
and increased expression of G6PC1 gene (2.0
times higher than control), which is expected in
insulin resistance model. Cell viability was not
affected with this treatment.
The established model of insulin resistance will
be further used for drug screening studiesBook 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