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 paniculata Mill.), a rosette-forming subshrub, is distributed in North America,
Europe, and Asia. This study aimed to investigate the chemical composition and in vitro biological activities
of the methanolic extract obtained from the aerial parts of the plant. The phenolic profile and content of the
four compounds were determined by LC-DAD-ESI-MS. Total phenolic content (TPC), total flavonoid content
(TFC), antioxidant potential as well as enzyme inhibitory capacity were evaluated spectrophotometrically.
The antimicrobial activity was tested by the broth- microdilution method, while the cytotoxic effect was
examined by the MTT assay. The analysis revealed the presence of 23 compounds, including quercetin,
six quercetin glycosides, seven kaempferol glycosides, two caffeoylquinic acids, three galloylhexosides,
two digalloylhexosides, galloylquinic acid, and (epi)gallocatechin gallate. The most abundant constituent
was quercitrin (1.62%), followed by chlorogenic acid (0.28%), kaempferol deoxyhexoside (0.12%),
and rutin (0.07%). TPC and TFC were 26.71% and 2.12%, respectively. The demonstrated antioxidant
activity was significant (DPPH test, IC50=9.30 μg/mL; FRAP test, FRAP value=2.89 mmol Fe2+/g; ABTS
test, IC50=34.22 μg/mL). The extract potently inhibited α-glucosidase (IC50=1.86 μg/mL) and, to a lesser
extent, 15-lipooxygenase (IC50=49.60 μg/mL) and α-amylase (IC50=9.95 mg/mL). The minimum inhibitory
concentration (MIC) against Acinetobacter baumannii was 1000 μg/mL; MIC was higher against the other
six tested strains of microorganisms. No effect on cancer cell lines (HeLa, HT-29, HCT-116, LS174) and
healthy cells (MRC-5) was observed. The findings expanded the scarce literature data on S. paniculata and
highlighted its potential, warranting further investigation.Book of absctract: International congress on natural products research Kraków 2024, July 13-1
Modulating the Release Kinetics of Natural Product Actinomycin from Bacterial Nanocellulose Films and Their Antimicrobial Activity
No full textThe present study aimed to create a more sustainable and controlled delivery system based on natural biopolymer bacterial nanocellulose (BNC) and bacterial natural product actinomycin (Act), with the applicative potential in the biomedical field. In order to provide improved interaction between BNC and the active compound, and thus to modulate the release kinetics, the TEMPO oxidation of BNC support was carried out. A mix of actinomycins from bacterial fermentation (ActX) were used as natural antimicrobial agents with an established bioactivity profile and clinical use. BNC and TEMPO-oxidized BNC films with incorporated active compounds were obtained and analyzed by FTIR, SEM, XPS, and XRD. The ActX release profiles were determined in phosphate-buffer solution, PBS, at 37 °C over time. FTIR analysis confirmed the improved incorporation and efficiency of ActX adsorption on oxidized BNC due to the availability of more active sites provided by oxidation. SEM analysis indicated the incorporation of ActX into the less-dense morphology of the TEMPO-oxidized BNC in comparison to pure BNC. The release kinetics of ActX were significantly affected by the BNC structure, and the activated BNC sample indicated the sustained release of active compounds over time, corresponding to the Fickian diffusion mechanism. Antimicrobial tests using Staphylococcus aureus NCTC 6571 confirmed the potency of this BNC-based system for biomedical applications, taking advantage of the capacity of modified BNC to control and modulate the release of bioactive compounds
LABBANK – THE FIRST BIOBANK OFAUTOCHTHONOUS PROBIOTIC LACTIC ACID BACTERIA
No full textLactic acid bacteria (LAB) are a group of bacteria that are generally recognized as safe (GRAS) and
are widely used in fermented foods. They are widespread in nature mainly in raw milk and dairy
products. The indigenous microbiota of raw milk directly affects the sensory properties of the raw
milk products. The WHO recommends to the use of fermented dairy, meat and vegetables products
in the daily diet, as LAB have a great impact on human health.
Group for Probiotics and Microbiota – Host Interaction within the Institute of Molecular Genetics
and Genetic Engineering, University of Belgrade has a large collection of LAB (about 5000 various
strains) isolated from artisanal autochthonous dairy products produced in a traditional way without
the addition starter cultures. The characterization of isolated LAB strains was done by classic
microbiological methods and identification by 16S rDNA sequencing. These LAB strains are
collected in previously 30 years and stored in refrigerator at -80℃.
Until now 23 distinctive LAB strains from LABbank collection have been deposited in the Belgian
Coordinated Collection of Microorganisms, University of Ghent, Belgium. The 13 strains have been
licensed through license agreements, as the subjects of innovations:
-HiraVet probiotic for prevention and treatment of intestinal infections in humans and animals.
-Diasolution probiotic for diabetes management.
-Lagendairy starter cultures for innovative dairy products for the production of soft white cheese,
cheese in brine, yogurt and sour cream.
-Sixteen LAB strains are currently the subject of innovations submitted as 2 PoC projects.
Our findings illustrate the importance of the research on natural isolates of LAB as a valuable
source of strains with novel properties, since they can provide a deeper and more complete insight
into the functioning and organization of the comprehensive metabolic system in these bacteria and
their impact on human and animal health.Book of Abstracts : The 3rd International UNIFood Conference, UNIFood2024 Conference, Belgrade, June 28-29, 202
Twinning Innovation Hub for Microbial Platforms in Plastic Upcycling (Twinn4MicroUP)
No full textPlastics have become indispensable in various industries today, but their widespread use has
resulted in a severe environmental crisis—plastic waste accumulation. Traditional methods of
plastic waste management are insufficient and recycling faces quality and degradation
challenges. Enter upcycling, a process gaining prominence in converting plastic waste into
valuable chemicals and materials in an eco-friendly manner. TwInn4MicroUp presents an
innovative approach to plastic waste upcycling, employing green bio/mechano/chemical
technologies to recover plastic monomers. Modern molecular techniques will be utilized to
create microbial cell factories capable of producing bioactive compounds from these
plasticderived feedstocks, revolutionizing industries related to bio-colorants, biotherapeutics,
bio-nutraceuticals, biosurfactants, and biomaterials.
TwInn4MicroUp is on a visionary mission to create the MicroUp HUB, an advanced Research and
Innovation Hub in Synthetic Microbial Biotechnology. In our pursuit of maximal impact, we aim
to establish strategic partnerships, strengthen NTUA's capabilities, enhance scientific excellence,
and promote entrepreneurship and innovation. Our overarching goal is to establish NTUA at the
core of the MicroUp HUB, leaving a lasting legacy by advancing scientific knowledge,
environmental protection, public awareness, economic development, and global collaboration.
Together, we are steering toward a sustainable and responsible future.Principal Investigator: Dr Evangelos Topakas, National Technical University of Athens, GreeceParticipants from IMGGE: Dr. Jasmina Nikodinović-Runić, Dr. Sandra Vojnović, Dr. Jelena
Milovanović, Dr. Ivana Aleksić, Dr Tatjana Ilić-Tomić, Vukašin Janković, Brana Pantelić, Lena
PantelićDuration period: 2024-202
Polycaprolactone-based electrospun films incorporating sage extract: From active food packaging application to accelerated biodegradation by Pseudomonas
No full textPolycaprolactone films incorporating sage extract (SE), developed by electrospinning and annealing, were tested as active, fast-degradable food contact materials. First, a release test with food simulants showed the films’ ability to release phenolics. This beneficial property was highlighted in the films’ application as pads for preventing raspberry quality deterioration. The highest SE containing film (20 %) demonstrated the highest preserving potential, suppressing raspberry spoilage, breakdown of soluble solids, maintaining color, and increasing phenolics content. Secondly, the degradation under natural environment conditions and conditions accelerating the degradation of the films was evaluated. Complete films’ bio-disintegration in compost occurred within three months, being catalyzed to four weeks by adding Pseudomonas aeruginosa, regardless of the SE incorporation. Visual appearance, light, scanning electron, and atomic force microscopy indicated an inhomogeneous surface degradation mechanism depended on the morphology of the films. Analysis of the structural properties and crystallinity showed that both amorphous and crystalline film regions were affected by biodegradation. The developed films as packaging materials can be beneficial from an economic and environmental point of view, as they can extend the shelf-life of fresh fruit, prevent the generation of food waste, reduce the accumulation of discarded packaging materials, protect the environment, and contribute to sustainable development
The unlimited potential of CRISPR/Cas9 technology in plant sciences and agriculture: an illustration through the example of mutagenesis of two small, highly homologous plant DSS1 genes
No full textPronalaženje novih ćelijskih komponenti koje leže u osnovi molekularnih mehanizama odgovora biljaka
na abiotički ili biotički stres može proisteći iz kombinacije genetičkih, biohemijskih i fizioloških pristupa.
Mutageneza, kojom se deaktivira određeni gen, predstavlja ključnu strategiju u utvrđivanju uloge datog
gena u određenim ćelijskim funkcijama. Kompleksnost biljnih genoma u poređenju sa životinjskim,
prisustvo velikog broja genskih familija, kao i pseudogena naučnicima je oduvek predstavljao problem
da uspešno generišu željene mutante. Revolucionarna CRISPR/Cas9 tehnologija donela je ogroman
napredak na polju biljne mutageneze i istraživanja funkcije biljnih gena. Ovaj rad daje kratki pregled
dosadašnjih znanja vezanih za primenu CRISPR/Cas9 tehnologije u biljnim funkcionalnim studijama, kao
i u unapređenju ekonomski značajnih useva. Takođe, na primeru mutageneze dva mala, visoko homologna
gena DSS1 koji kodiraju prirodno neuređene proteine, biće prikazane mogućnosti i rešenja koje donosi
ova savremena tehnologija kada su veličina gena i visoka identičnost sekvenci biljnih homologa
ograničavajući faktori za mutagenezu. Između ostalog, ovde su predstavljeni mnogobrojni izazovi u
izučavanju biljaka sa kojima smo se susreli prilikom CRISPR/Cas9 editovanja gena DSS1(I) i DSS1(V) vrste
Arabidopsis thaliana, kao što su: prevazilaženje efekata off-target mutacija, pronalaženje efikasnih metoda
za isporuku CRISPR/Cas9 kaseta u biljne ćelije, kao i načina za brzo pretraživanje velikih kolekcija biljaka u
potrazi za mutantnim biljkama sa željenim indelima.The search for new cellular components underlying the molecular mechanisms of plant response to abiotic
or biotic stress can be accomplished through a combination of genetic, biochemical, and physiological
approaches. Deactivating specific genes via mutagenesis is a key strategy for determining their roles in
specific cell functions. Due to the complexity of plant genomes compared to animal genomes the presence
of numerous gene families, and the presence of pseudogenes, generating desired mutants has always
been challenging for scientists. The revolutionary CRISPR/Cas9 technology has brought tremendous
progress in the field of plant mutagenesis and the study of plant gene functions. This study provides a
brief overview of the current state of knowledge regarding the application of CRISPR/Cas9 technology in
plant gene function research and the improvement of economically important crops. Also, the possibilities
and solutions offered by this modern technology will be shown when gene size and high sequence identity
of plant homologs are limiting factors in the example of mutagenesis of two small, highly homologous
DSS1 genes encoding intrinsically disordered proteins. Furthermore, we highlight challenges encountered
during CRISPR/Cas9 editing of the DSS1(I) and DSS1(V) genes in Arabidopsis thaliana, including overcoming
off-target effects, developing successful methods for transferring CRISPR/Cas9 cassettes into plant cells,
and efficiently screening large plant collections for gene-edited plants with desired indels
Microencapsulation of Origanum heracleoticum L. and Thymus vulgaris L. essential oils – Novel strategy to combat multi-resistant Acinetobacter baumannii
No full textWithin the global rise of antimicrobial resistance enhanced by the COVID-19 pandemic, where Acinetobacter baumannii has been distinguished as an emerging multi-resistant pathogen, essential oils, become the focus of novel therapeutic approaches. Hypothesizing that encapsulated Origanum heracleoticum L. and Thymus vulgaris L. essential oils could express multi-target approach against A. baumannii, this study aimed to develop microencapsulated systems with optimal technological qualities using 2-hydroxypropyl-β-cyclodextrin as a carrier, and to evaluate their pharmacological potential against A. baumannii, and their morphological and physicochemical characteristics, safety and stability profiles. The highest yield and encapsulation efficiency were obtained with 1:10 essential oil to carrier, and 1.5:10 carrier to water w/w ratios. The formation of inclusion complexes was confirmed by Fourier Transform Infrared Spectroscopy. Both microencapsulates achieved improved homogeneity, particle surface, and thermal stability compared with the pure carrier. Dominant bioactive compounds (carvacrol and p-cymene in O. heracleoticum essential oil, and thymol and p-cymene in T. vulgaris essential oil) remained the most abundant after encapsulation. While non-encapsulated essential oils revealed similar antimicrobial activity towards clinical A. baumannii isolates obtained from COVID-19 patients, encapsulated O. heracleoticum essential oil inhibited the bacterial growth at lower concentrations than T. vulgaris essential oil microencapsulate. All samples significantly reduced the formation of A. baumannii biofilm, for at least 53.90% towards the most infective isolate according to the Caenorhabditis elegans assay. Further, in silico molecular docking study revealed strong interaction pattern of carvacrol and thymol with the outer membrane protein A, which is the main factor for the A. baumannii biofilm formation. Cytotoxicity investigation on human lung A549 cells showed high survival rate in the presence of all tested concentrations, and the stability study revealed notable preservation of the bioactives’ content and pharmacological potential. Altogether, microencapsulated essential oils exhibited a multi-target approach towards A. baumannii, with satisfactory preserving capability during storage
mir-219 as a potential radiosensitier in glioblastoma cells
No full textGlioblastoma (GBM) is the most aggressive type of primary malignant brain tumor, and
is characterized by an extremely poor therapeutic response and overall survival.
Adjuvant radiotherapy remains the standard treatment after surgical resection.
Resistance to radiotherapy is frequently observed in patients with GBM, which limits
treatment efficacy. Finding drugs or molecules that can overcome this restrain is crucial
for finding novel more effective glioblastoma treatments. MicroRNAs (miRNAs) are a
class of small non-coding RNAs that are important regulatory molecules that can control
GBM radiosensitivity by affecting radiation-related signal transduction pathways, such
as proliferation, migration, senescence, and cell cycle regulation. Evolutionary
conserved miR-219 has specific expression in brain and it can act as tumour suppressor
in GBM. The main goal of this study is to analyze the effects of modulation of miR-219
expression on radiosensitivity of GBM cells.
First, in silico analyses show that the expression of miR-219 is downregulated in GBM
tumours and that many of the target genes of this miRNA have a critical role in the
modulation of key cellular pathways that mediate response to radiation. Additionally, our
results show increased expression of miR-219 upon irradiation of GBM cells. To further
investigate the function of mir-219 in GBM radiosensitivity, a stable miR-219-
overexpressed LN229 and U87 cell line was constructed using lentivirus transduction. In
addition, we analyze the effect of miR-219 overexpression on proliferation and
senescence of GBM cells. The results of this research will contribute to better
understanding how miR-219 affect radiosensitivity of GBM cells and may lead to
improvement of radiotherapy of GBM tumours making progress in treatment of this
aggressive and one of the deadliest forms of brain cancer.The 2 nd Biomedicine and HealthPhD Students Congress Faculty of Medicine University of Rijeka, Croatia 16.-18.5.2024
Functional Characterisation and Application in Biomaterials of a Fusarium oxysporum AA9 Lytic Polysaccharide Monooxygenase
Get PDFLytic polysaccharide monooxygenases (LPMOs) are recognized for their crucial role in the breakdown of plant biomass and are increasingly utilized in
biotechnological applications. This research focuses on a specific LPMO from family 9 of the auxiliary activities (AA9) from Fusarium oxysporum,
referred to as FoLPMO9A. FoLPMO9A was expressed in Pichia pastoris and assessed for its ability to interact with other enzymes from the auxiliary
activity families AA1, AA9, AA16, and with polyphenol oxidases (PPOs). The potential of FoLPMO9A for modifying cellulose was explored, aiming to
enhance the development of bio-based materials. This study explored two novel applications of FoLPMO9A. Initially, an enzymatic process was
developed for extracting microscale cellulose from pretreated wheat straw. Subsequently, FoLPMO9A was used to functionalize bacterial
nanocellulose, leveraging synergistic interactions with AA16 LPMOs. These applications highlight the broad catalytic potential across diverse
substrates
Genske terapije za retke bolesti
No full textEvery disease with a prevalence of less than 1 in
2000 people is defined as a rare disease (RD). So far,
over 6000 different RDs have been described, and
this number increases year by year. It is considered
that over 80% of RDs have a monogenic genetic
basis. Despite incredible progress in developing therapies
for RDs, still, for more than 95% of them, there
is no specific effective treatment. Next-generation
sequencing plays a significant role in making accurate
diagnoses and identifying new targets that will
serve as a foundation for developing innovative therapeutics.
Also, basic research, such as the functional
characterization of genetic variants and exploring the
molecular mechanisms of disease onset, are necessary
prerequisites for developing various molecular
therapeutics, including gene therapy. Gene therapy
involves introducing genetic material into cells to compensate for an inborn genetic defect and to
enable the continuous synthesis of a functional protein
and cure the disease. The genetic material introduced
can be a functional copy of a gene (DNA
molecule), an oligonucleotide (a short non-coding
RNA molecule) that binds by complementarity to
mRNA or pre-mRNA and leads to the desired modulation
of protein expression (e.g., modulation of splicing,
blocking of translation, or activation of mRNA
degradation), or short RNAs that precisely correct a
gene within the cell itself (CRISPR/Cas9). Some of
the diseases for which gene therapy has been registered
so far include adenosine deaminase deficiency,
spinal muscular atrophy, retinal dystrophy, and many
clinical trials are ongoing. The importance of basic
research is undeniable, as is its rapid translation into
medical practice.Svaka bolest ~ija je u~estalost manja od 1 u 2000
ljudi defini{e se kao retka bolest (RB). Do sada je
opisano preko 6000 razli~itih RB i taj broj se iz
godine u godinu pove}ava. Smatra se da preko 80%
RB ima monogensku geneti~ku osnovu. Uprkos
neve rovatnom napretku u razvoju terapija za RB, i
dalje za >95% njih ne postoji specifi~an efikasan tretman.
Ovo su razlozi za{to su znanja iz molekularne
genetike od neprocenjivog zna~aja za istra`ivanje
molekularne osnove RB. Sekvenciranje nove gene -
racije ima veliku ulogu u postavljanju ta~ne dijagnoze
i identifikaciji novih meta koje }e poslu`iti kao osnov
za razvoj inovativnih terapeutika. Tako|e, ba zi~na
istra`ivanja, poput funkcionalne karakterizacije
geneti~kih varijanti i istra`ivanje molekularnih mehanizama
nastanka bolesti su neophodni pred uslovi za
razvoj razli~itih molekularnih terapeutika, pa tako i genske terapije. Genska terapija podra zumeva
uno{enje geneti~kog materijala u }elije kako bi se
nadomestio uro|eni geneti~ki nedostatak i kako bi se
omogu}ila kontinuirana sinteza funkcionalnog proteina
i izle~ila bolest. Geneti~ki materijal koji se unosi
mo`e biti funkcionalna kopija gena (DNK molekul),
oligonukleotid (kratak nekodiraju}i RNK molekul)
koji se po principu komplementarnosti vezuju za
iRNK ili pre-iRNK i dovodi do `eljene mo dulacije
ekspresije proteina (npr. modulacija iskrajanja, blokiranje
translacije ili aktivacija degradacije iRNK) ili
kratke RNK koje precizno koriguju gen unutar same
}elije (CRISPR/Cas9). Neke od bolesti za koje je do
sada registrovana genska terapija su deficijencija
adenozin deaminaze, spinalna mi{i}na atrofija, retinalna
distrofija, a mnoga klini~ka ispitivanja su u
toku. Nesumnjiva je va`nost bazi~nih istra`ivanja, ali
i njihova brza translacija u medicinsku praksu.Book of abstract: XIII International Serbian Congress of Medical Biochemistry and Laboratory Medicine. 16-18 September 2024, Belgrade, Serbia