imagine (Institute of molecular genetics and genetic engineering)
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Enzymes for detection of per- and polyfluoroalkyl (PFAS) chemicals (PFASens)
Get PDFPer- and polyfluoroalkyl substances (PFAS) comprise a wide range of synthetic compounds characterized
by a carbon-fluorine bond, which is known to be one of the strongest and shortest chemical bonds. Due
to their unique properties, these compounds are widely used in consumer and industrial products. The
widespread use of PFAS chemicals, combined with their persistence, bioaccumulation, and mobility, has
resulted in severe pollution of air, soil, and water sources worldwide. The adverse health effects
associated with exposure to PFAS underscore the urgent need for immediate action and the development
of novel strategies to address these “forever chemicals”. In addition to effective remediation of PFAS,
there is a crucial need for improved tools to detect them. Currently, there is no PFAS sensor on the market
capable of detecting low concentrations in real time. The aim of this project is to investigate the potential
of unique bacterial enzymes for PFAS detection. The enzymes that can react with PFAS would have the
potential to be used as a biorecognition element in a specific product – an innovative PFAS biosensor.
Biosensors are affordable, portable, and rapid-response devices designed to detect specific analytes in a
sample. Such a device would enable on-site quantification and eliminate the need for complex equipment.
The envisioned biosensor, equipped with novel enzymes, could be used to detect PFAS chemicals in
environmental samples (water and soil) as well as in human samples (blood, breast milk). The
development of a cost-effective, user-friendly, and innovative solution such as a biosensor for PFAS
detection would be of great importance for environmental management and health.Principal Investigator: Dr Mina Mandić, IMGGEDuration period: 2024-202
Chemical Composition, Antimicrobial Efficacy, and Sensory Characteristics of Kombucha Beverages from Lentinus edodes and Coriolus versiclolor Medicinal Mushrooms
No full textIn contemporary times, food products are increasingly expected to demonstrate
positive health effects on the human body, leading to heightened interest in their role in
disease prevention. Beverages consist of different ingredients dissolved in water and are
considered as good way to provide the human body with nutrients and bioactive compounds
[1]. Incorporating mushrooms into beverages offers a significant opportunity for both
mushroom and beverage producers to meet the growing demand for innovative, enjoyable
products that positively impact quality of life. Kombucha is a fermented tea beverage created
by the symbiotic activity of different types of yeasts and acetic acid bacteria. It is traditionally
produced with black tea; however, the possibility of producing this beverage has also been
tested using different fermentation media [2]. Despite the proven pharmacological
properties of various mushrooms and their biologically active compounds such as
polysaccharides, polypeptides, proteins, phenols, flavonoids, sterols and other, their
utilization in food systems remains limited. Therefore, two medicinal mushrooms were
chosen for the development of novel kombucha beverage production. The first, Lentinus
edodes a medicinal and edible mushroom, better known as shiitake mushroom, is the leading
cultivated edible mushroom in the world, because of its appealing aroma and medicinal
properties. The second, C. versicolor mushroom is not edible, but was chosen because its
application in traditional medicine was based on the preparation of tea from the fruiting
body, as well as hot water extracts [3].
Cultivation of mushrooms fruiting bodies was conducted on the substrates composed
from agricultural wastes. Produced fruiting bodies were further crushed and extracted with
hot water under pressure. After the addition of sucrose, fermentation with kombucha
culture was conducted. Kombucha beverages compositions were analyzed using ATR-FTIR
and spectroscopic analysis. Their antimicrobial potentials were tested by determining
minimal inhibitory and minimal bactericidal activity. Sensorial properties of novel bioactive
beverages were evaluation by a method that uses a hedonic scale with nine levels.Extraction of medicinal mushrooms in the preparation of the medium for
fermentation of kombucha culture proved as suitable from the nutritional aspect for
achieving adequate conditions and successfully obtaining fermented kombucha products.
ATR-FTIR analysis of the qualitative chemical composition kombucha beverages revealed that
the dominant polysaccharide fraction was present in the samples, as well as proteins, lipids,
phenolic compounds and primary alcohols. The analysis of the quantitative chemical
composition showed that there was a statistically significant difference in the content of total
polyphenols among the samples of kombucha beverages made from different mushrooms. It
was found that the antibacterial activity of kombucha mushroom beverages does not come
only from acetic acid, but also from components extracted from the mushrooms or formed
during the fermentation process. Kombucha made from L. edodes mushroom received the
best rating based on sensorial properties. The aromatization of the samples with the aroma
of coconut contributed to achieving better results with the kombucha sample from C.
versicolor mushrooms.
This research demonstrated the great potential of medicinal mushrooms for
developing innovative, health-promoting beverages in the food industry.The 12th International Medicinal Mushrooms Conference (IMMC12), September 24−27, 2024, Bari, Ital
Association of the methionine sulfoxide reductase A rs10903323 gene polymorphism with functional activity and oxidative modification of alpha-1-antitrypsin in COPD patients
Get PDFObjectiveChronic obstructive pulmonary disease (COPD) is multi–factorial disorder which results from environmental influences and genetic factors. We aimed to investigate whether methionine sulfoxide reductase A (MSRA) rs10903323 gene polymorphism is associated with COPD development and severity in Serbian adult population.MethodsThe study included 155 patients with COPD and 134 healthy volunteers. Genotyping was determined performing home-made polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The difference between the inhibitory activities of normal and oxidized Alpha-1-Antitrypsin (A1AT) against elastase and trypsin was used for determination of Oxidized Alpha-1-Antitrypsin (OxyA1AT) (expressed as % and g/L). Functional activity of A1AT was presented as a specific inhibitor activity to elastase (SIA-Elastase, kU/g).ResultsFrequencies of the genotypes AA, AG and GG were 80.0%, 20.0%, 0% in COPD patients and 80.5%, 18.5% and 1.5% in the control group, and there was no significant difference in genotype or allele distributions between groups. Serum level of A1AT (g/L) and OxyA1AT was significantly higher in COPD patients than in the control group, but functional activity of A1AT (SIA-Elastase) was significantly lower in COPD patients than in the control group. In COPD group, increased level of OxyA1AT was present in G allele carriers who were smokers relative to G allele carriers who were not smokers. In the smoker group of patients with severe and very severe COPD (GOLD3+4), significant increase in OxyA1AT level was present in G allele carriers compared to AA homozygotes.ConclusionThese findings suggest that MSRA rs10903323 gene polymorphism is probably not a risk for COPD by itself but could represent a COPD modifier, since minor, G allele, is associated with an increased level of oxidized A1AT, indicating impaired ability of MSRA to repair oxidized A1AT in COPD-smokers, and in severe form of COPD
Green chemistry approaches for late-stage diversification of bacterial natural products
Get PDFBakterijski prirodni proizvodi (BPP) su od ključnog značaja u savremenoj medicini
zahvaljujući svojoj efikasnosti i širokoj kliničkoj primeni, a pored toga imaju veliki
potencijal i kao kandidati za razvoj novih lekova i za hemijske inovacije. Perspektiva
korišćenja biokatalize za diverzifikaciju BPP-a predstavlja privlačnu alternativu totalnoj
hemijskoj sintezi. Uprkos sve većem prihvatanju biokatalize kao ekološki prihvatljivog
pristupa, značajan deo sekvenciranih genoma, povezanih sa metaboličkim putevima za
biosintezu BNP-a i njihovim modifikacijskim enzimima, a kojima funkcija nije ispitana,
ostaje uglavnom neiskorišćen. Ova studija ističe ispitivanje zelenih biotransformacija 1-
hidroksifenazina i aktinomicina D, što može pomoći u pronalaženju novih strukturnih
optimizacija bioaktivnih molekula i unapređenju razvoja lekova.Bacterial natural products (BNPs) are crucial in modern medicine due to their effectiveness
and diverse applications, as evidenced by their extensive clinical use, and also as potential
drug leads and for chemical innovation. The prospect of employing biocatalysis for latestage
diversification of BNPs presents an appealing alternative to total chemical synthesis.
Despite the growing acceptance of biocatalysis as an environmentally friendly approach, a
significant portion of orphan sequenced genomic data associated with metabolic pathways
for BNP biosynthesis and their modifying enzymes remains largely untapped. This study
highlights an examination of the green biotransformations to of 1-hydroxyphenazine and
actinomycin D, which may aid in identifying novel structural optimizations of bioactive
molecules and ultimately advancing drug development60. SAVETOVANJE SRPSKOG HEMIJSKOG DRUŠTVA Niš, 8. i 9. jun 2024.
Book of abstracts: 60th MEETING OF THE SERBIAN CHEMICAL SOCIETY
Niš, Serbia, 8-9 June 202
Way2Drug Platform: From Biological Activity Prediction to Systems Pharmacology
No full textGlobal chemical space is extremely vast and finding a molecule with the required
pharmacotherapeutic properties is a formidable challenge. Starting from analysis of big
chemical-biological data obtained in silico, in vitro, in vivo and in clinics it is necessary to
finish with one active pharmaceutical ingredient possessing the needed safety and
efficacy. Combining information extracted from the curated datasets of active/inactive
compounds available through World Wide Web and AI/ML tools, investigators are surfing
from global to local scales in pharmaceutical R&D, enabling faster and more efficient
development of new therapeutic remedies.
Way2Drug (https://www.way2drug.com/dr/) is a quickly expanding web portal focused
at integrating of demanded in silico tools for drug discovery. Way2Drug currently hosts
services for predicting the biological activities (PHARMA), toxicity (TOX), metabolism
(META) and physicochemical characteristics (ADME) of drug-like molecules. All tools are
freely available for non-commercial academic research.
In addition to the predictive web-services, several informational resources are available
at the Way2Drug portal, including WWAD (World-Wide Approved Drugs, https://www.
way2drug.com/wwad/), phytocomponents of the Russian officinal medicinal plants
Phyto4Health (https://www.way2drug.com/p4h/), host gut microbiota metabolism
xenobiotics database HGMMX (https://www.way2drug.com/hgmmx/).
Way2Drug is evolving as the basis for development of computational platform for
efficient analysis and interpretation of the extensive biomedical and clinical data,
comparative analysis of information extracted from this data to differentiate the normal
and pathological states, obtaining new knowledge to identify potential pharmacological
targets and biomarkers, designing potential pharmacological substances with the
required properties, determining the optimal approach to therapy taking into account the
individuality of patients.Book of abstracts: 5th Belgrade Bioinformatics Conference, Serbia, Belgrade,17-20 june 2024
Exploring protein translation dynamics in autism spectrum disorder-associated mutations
No full textAutism spectrum disorder (ASD) is a complex and highly prevalent neurodevelopmental disorder characterized by significant heterogeneity in genotypes and phenotypes. Recent research has identified dysregulation of protein synthesis as a potential converging mechanism in ASD. To explore altered translation in ASD, we used induced pluripotent stem cell (iPSC)-derived iNeurons and cortical brain organoids. CRISPR/Cas9 technology was employed to introduce ASD-causing mutations in the FMR1, PTEN, and TSC2 genes in iPSCs from healthy donors. These modified iPSCs were then differentiated into either iNeurons or cortical brain organoids. Using label-free data-independent acquisition (DIA), we identified about 8,000 proteins. Principal component analysis (PCA) revealed that mutations in TSC2 and PTEN caused a wide range of consistent changes in both iNeurons and organoids. These changes included up-regulated translation and mitochondrial processes and down-regulated transcriptional processes. Notably, the down-regulated proteins were strongly enriched in genes known to be associated with ASD, indicating that the proteomic data can help explain the broad genetic diversity of ASD. In summary, our integrated proteomic analyses of iPSC-derived iNeurons and organoids shed new light on the pathophysiological processes underlying ASD.HUPO World Congress 2024 from October 20 - 24 in Dresden, German
Laboratory evolved microbial consortium towards arabinoxylan can degrade polyurethane
No full textBiodegradation of hydrocarbon based lignocellulose and plastics is often more efficient by
natural and synthetic microbial consortia than by individual microorganisms, due to their
extended enzymatic repertoire. Despite structural similarities of some plastics and plant-derived
polymers and a proposed overlapping microbial degradation activity, the potential for the
plastics degradation by lignocellulose-degrading microbial consortia still remains
underexplored.
Synthetic consortia (SC) combining 35 environmental Streptomyces isolates with confirmed
lignocelullolytic activities (SC0) was subjected to 3 rounds of enrichment on insoluble wheat
arabinoxylan (AXYL) as sole carbon source. Reduction in biomass and morphological diversity
was observed through rounds of enrichment for SC1 to SC3. In SC2 and SC3, only yellow (Y)
and white (W) colonies could be observed. These were identified as Ochrobacterium, a genera
with lignolytic and cellulolytic activity and Stenotrophomonas, a genera known for their high
hydrolytic activity and likely represent a background contamination enriched on AXYL.
SC1 demonstrated multiple clearance zones on Impranil SD, a model compound for degradation
of polyurethanes. Eight selected isolates from SD clearance zones were identified as
Stenotrophomonas (3), Pseudomonas (3), Streptomyces and Lysinibacillus. Isolates grew on all
8 tested polyester-based plastic and lignocellulosic substrates. Four isolates produced clearance
zones on carboxymethyl cellulose, while SD degradation was confirmed for 5 isolates. Isolates
with simultaneous plastolytic and lignolytic capabilities may be of special biotechnological
interest and should be further explored.VII Congress of the Serbian Genetic Society Zlatibor; October 2 to 5, 2024
CHARACTERIZATION OF INDUCED PLURIPOTENT STEM CELLS FROM PATIENTS WITH 22Q11.2 DUPLICATION SYNDROME
No full textThe 22q11.2 Duplication Syndrome (22q11.2DupS), caused by a microduplication in the q11.2
region of chromosome 22, is associated with a high risk of developing neurodevelopmental
disorders (NDDs). Autism spectrum disorder (ASD) occurs in 14-25% of individuals with
22q11.2DupS, making it one of the genetic syndromes with the highest prevalence of ASD.
Conversely, in individuals affected by schizophrenia, 22q11.2DupS occurs less frequently than
in the general population, suggesting that this microduplication could have a protective effect
against schizophrenia.
Peripheral blood mononuclear cells from 22q11.2DupS patients and healthy controls were
reprogrammed into induced pluripotent stem cells (iPSCs) using CytoTuneTM-iPSC2.0 Sendai
Reprogramming Kit. Genotyping of iPSCs was performed to identify potential additional
pathogenic CNVs. Pluripotency markers' expression was analyzed by RT-PCR. The ability of
iPSCs to differentiate into the cells of three germ layers was tested using STEMdiff Trilineage
Differentiation Kit.
Successful establishment of iPSCs from three 22q11.2DupS patients, their mothers that carry
the microdeletion, and three healthy individuals was confirmed by expression of pluripotency
markers. Generated iPSCs have the ability to differentiate into cells of all three germ layers.
Additional CNVs in some of the iPSC lines were revealed by genotyping.
iPSCs from 22q11.2DupS patients, their carrier mothers, and healthy controls were generated
representing a valuable model for studying the molecular mechanisms underlying NDDs.VII Congress of the Serbian Genetic Society Zlatibor; October 2 to 5, 2024
Low soil nitrogen and phosphorus enhances primary and secondary metabolites in grape berries and wine quality of Cabernet Sauvignon
No full textThe concept of terroir encompasses how the environment of a vineyard influences the distinct
characteristics and complexity of wine. In this study, we investigated how micro soil heterogeneity
(micro-soil terroir) affects primary (sugars and organic acids) and secondary (phenolics)
metabolites in the berries of grapevine (Vitis vinifera L.) cultivar Cabernet Sauvignon, grafted on
P1103 rootstock. Using aerial multispectral imaging, specifically the Normalized Difference Red
Edge Index (NDRE), we identified two distinct areas, one comprising 27% and the other comprising
73% of the total 2.82 ha vineyard. Further soil analyses revealed significant differences in soil
characteristics between these areas, with lower concentrations of total nitrogen (N) and available
phosphorus (P) in the smaller one. The vine plants in the area with lower soil N and P exhibited
lower leaf N, but higher P concentrations. Although total sugar content in grape juice did not differ
significantly between the areas, those from sections with lower soil N and P exhibited higher concentrations
of yeast-assessable nitrogen (YAN), anthocyanins, and volatile organic acids in grape
juice, and higher lactic acid content in wine. Furthermore, wine sensory evaluation using a 5-point
interval scale indicated higher overall quality of wine produced from the lower N and P sections.
Our findings suggest that the enhanced wine quality is linked to increased biosynthesis of organic
acids and anthocyanins, which may be attributed to grape vine adaptations to low soil fertility and
in particular to low P conditions.Book of abstract: 5th International Conference
on Plant Biology
(24th SPPS Meeting
Tumor engineering: Biomimetic 3D models for reliable osteosarcoma research and drug discovery
No full textIntroduction
Osteosarcoma (OS) is a highly aggressive malignant bone tumor that primarily affects children and teenagers
with a tendency to spread rapidly to the lungs and other bones. The current treatment includes chemotherapy
before and after surgery to remove the tumor with a 5-year survival rate of approximately 60% for localized
tumors [1], while the survival rate decreases to 20-30% in cases of metastases [2]. Many research groups are
attempting to enhance and streamline current therapies for OS, but the progress has been slow due to
inadequate research methodologies that rely on two-dimensional (2D) cell cultures and animal models. One of
the approaches to overcome this problem is to develop artificial in vivo-like three-dimensional (3D) tumor
models by applying tissue engineering principles. Therefore, the aim of this work was to develop a simple,
robust, relevant and reliable 3D cell cultures able to recapitulate key features of osteosarcoma, based on
alginate hydrogels with mineral particles (hydroxyapatite particles - HAP) as cell carriers, mimicking
extracellular matrix (ECM) of bone in vitro and a biomimetic perfusion bioreactor (“3D Perfuse”, Innovation
Center of the Faculty of Technology and Metallurgy, Belgrade, Serbia) that provides efficient mass transfer
and adequate levels of hydrodynamic shear stresses. Specifically, two advanced 3D cell culture models were
created: one simple based on alginate microfibers containing HAP particles with immobilized cells for rapid
and reliable drug screening, and another utilizing macroporous composite alginate scaffolds embedded with
HAP particles as bone cell carriers and biomimetic perfusion bioreactor that closely mimics the highly
vascularized environment of bone tissue, providing a relevant platform for osteosarcoma research.Book of proceedings: BIOMATERIALS AND NOVEL TECHNOLOGIES FOR HEALTHCARE 4th Biennal International Conference BioMaH, October 15th-18th, 2024 Rome, Ital