imagine (Institute of molecular genetics and genetic engineering)
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Fresh hyperalkaline groundwaters contain polyurethane-degrading Flavobacterium
Get PDFThe examined groundwaters, orginating from the fractured type of aquifer formed by Pre-
Middle Jurassic ultramafic rocks, i. e. harzburgites, dunites, lherzolites and serpentinites, belong to the
fresh hyperalkaline cold groundwaters of OH, Cl – Ca, Na + K genetic type. Studied occurrence was
characterized by the presence of aerobic cultivable bacteria with ability to grow in a pH range of 7-12.
Bacterial isolates were screened using plastic,cellulosic and hemicellulosic polymeric substrates as
predominantcarbon source to assess their biotechnological potential. All 16 screened isolates (100%)
demonstrated growth and/or activity on at least one tested substrate. One isolate belonging to genus
Flavobacterium, produced prominent zone of clearing on Impranil® DLN-SD, a model substrate for
polyurethanes (PU). Plastic-degrading potential has been previously documented for members of
genus Flavobacterium. However, this is the first report of their PU-degrading activity. Accumulation of
PU in the environment is of special concern, due to its toxicity, high production rates and lack of
efficient recycling strategies. In the future, biotechnological potential of alkalophilic bacteria from
groundwater environments should be explored more intensly, especially from the aspect of their
application in the management of plastic waste and bioremediation treatments of plastic-polluted
groundwater resources.XIII International Conference Industrial Engineering and
Environmental Protection (IIZS 2023), Zrenjanin, 5-6th October 2023
Asocijacija varijanti u genu za desaturazu masnih kiselina 1 (FADS1) sa težinom bolesti COVID-19
No full textOmega-3 masne kiseline su adekvatan kandidat u borbi protiv COVID-19 zbog svojih
imunomodulatorskih svojstava, odnosno zbog sposobnosti rezolucije inflamacije, česte
posledice COVID-19. Pokazano je da su EPA i DHA inverzno asocirani sa težinom COVID-19
ili da imaju protektivno dejstvo na SARS-CoV-2 (6,33). Imajući u vidu promenu ishrane u korist
omega-6 masnih kiselina i sve veći nedostatak omega-3 masnih kiselina u zapadnim
populacijama, od velike je važnosti skrenuti pažnju na važnost adekvatnog unosa ovih
nutrijenata u borbi protiv COVID-19 (32). Takođe je potrebno da se uzmu u obzir i genetičke
varijacije, budući da je više studija otkrilo njihov uticaj na nivo cirkulišućih omega-3 masnih
kiselina (17,18,21).
Uzimajući u obzir posledice pandemije COVID-19 i manjka cirkulišućih omega-3 masnih
kiselina u organizmu osobe obolelih od COVID-19, kao i važne imunološke uloge omega-3
masnih kiselina i ulogu genetičkih faktora u zastupljenosti omega-3 masnih kiselina, ciljevi
ovog rada su:
1. Da se ispita da li varijante rs174547 i rs174561 utiču na verovatnoću za hospitalizaciju
kod pacijenata obolelih od COVID-19
2. Da se ispita da li varijante rs174547 i rs174561 utiču na verovatnoću za razvijanje teške
kliničke slike kod pacijenata obolelih od COVID-1
Exploring Microorganisms from Plastic-Polluted Sites: Unveiling Plastic Degradation and PHA Production Potential
Get PDFThe exposure of microorganisms to conventional plastics is a relatively recent occurrence, affording limited time for evolutionary adaptation. As part of the EU-funded project BioICEP, this study delves into the plastic degradation potential of microorganisms isolated from sites with prolonged plastic pollution, such as plastic-polluted forests, biopolymer-contaminated soil, oil-contaminated soil, municipal landfill, but also a distinctive soil sample with plastic pieces buried three decades ago. Additionally, samples from Arthropoda species were investigated. In total, 150 strains were isolated and screened for the ability to use plastic-related substrates (Impranil dispersions, polyethylene terephthalate, terephthalic acid, and bis(2-hydroxyethyl) terephthalate). Twenty isolates selected based on their ability to grow on various substrates were identified as Streptomyces, Bacillus, Enterococcus, and Pseudomonas spp. Morphological features were recorded, and the 16S rRNA sequence was employed to construct a phylogenetic tree. Subsequent assessments unveiled that 5 out of the 20 strains displayed the capability to produce polyhydroxyalkanoates, utilizing pre-treated post-consumer PET samples. With Priestia sp. DG69 and Neobacillus sp. DG40 emerging as the most successful producers (4.14% and 3.34% of PHA, respectively), these strains are poised for further utilization in upcycling purposes, laying the foundation for the development of sustainable strategies for plastic waste management
Conversion of mixed plastic waste containing PET into biopolymer bacterial nanocellulose
Get PDFThe rapid increase in global plastics production is
also causing an accelerated environmental
pollution. Recently, biotechnological solutions and
enzymatic recycling of poly(ethylene terephthalate)
(PET) waste stream have been put forward and
commercialized1. Increasing recycling and
upcycling rates is the most effective model
approach to plastic circularity. However, mixed
plastic waste is still quite a challenge for both
recycling and upcycling technologies. This study is
focused on the eco-conversion of plastic waste
containing poly(ethylene terephthalate), PET, into
biopolymer, bacterial nanocellulose. Polymer mix
contained selection of commercial biodegradable
plastics (poly(lactic acid), PLA, poly(ε-caprolactone),
PCL, poly(hyoxyl butyrate), PHB) and PET. This
mixture was hydrolysed under aqueous conditions
and hydrolysate was used as carbon source for
Komagataeibacter medellinensis ID13488 and
bacterial nanocellulose (BNC) production. HPLC
analysis confirmed the presence of monomers and
dimers of polymer mix components indicating
existence of potential substrates for BNC
production. BNC production by K. medellinensis
was investigated and optimized in ter of the
amount of carbon source and growth conditions.
Under the most efficient rate in ter of yield, BNC
production was scaled up and the obtained
biopolymer was characterized. The structure of
produced BNC was confirmed by FTIR analysis,
thermal properties by DSC/TG analysis, and the
morphology of material by optical microscopy and
SEM analysis. This research demonstrates how to
put the mixed plastic waste stream into a circular
loop through the biotechnological conversion into
valuable biopolymer.10th International Conference of MIKROBIOKOSMOS, Larissa from 30 Novewmber to 2 December 2023
Varijante i transkripcija gena koji kodiraju komponente leptinskog signalnog puta, inflamacije i antioksidativne zaštite u patogenezi multiple skleroze
Get PDFMultipla skleroza (MS) je hronična autoimunska bolest centralnog nervnog sistema (CNS) čiji osnovni
patogenetski mehanizmi obuhvataju neuroinflamaciju, demijelinizaciju i neurodegeneraciju. Glavno
patofiziološko obeležje MS je hiperaktivacija određenih komponenti imunskog sistema, koja dovodi do
nastajanja i akumulacije strukturno-funkcionalnih oštećenja CNS kao posledice hronične inflamacije i
oksidativnog stresa. Adipocitokin i hormon leptin potencijalni je molekularni marker MS, budući da
učestvuje u patogenezi ove bolesti delovanjem na regulaciju imunskog i odgovora na oksidativni stres.
Varijante i promene u transkripciji gena koji kodiraju komponente leptinskog signalnog puta, i sa njima
povezane komponente inflamacije i antioksidativne zaštite, asocirane su sa nastankom i/ili kliničkim tokom
MS kao i odgovorom na terapiju. Danas u Srbiji ima preko 9000 obolelih od MS, a samo 1 od 10 pacijenata
dobija trenutno dostupnu konvencionalnu imunomodulatornu terapiju. Mogućnost modulisanja nivoa
cirkulišućeg leptina sugeriše njegov potencijalni značaj u budućem razvoju personalizovanog pristupa u
lečenju MS baziranog na poboljšavanju efekata ili supstituciji postojeće konvencionalne terapije.Multiple sclerosis (MS) is a chronic autoimmune disease of the central nervous system (CNS) with underlying
pathogenetic mechanisms that include neuroinflammation, demyelination and neurodegeneration.
The main pathophysiological feature of MS is the hyperactivation of certain immune system components,
which leads to formation and accumulation of structural and functional damage in the CNS as a result
of chronic inflammation and oxidative stress. The adipocytokine and hormone leptin is a potential molecular
marker of MS, since it participates in the pathogenesis of this disease by acting on the regulation of
immune system and response to oxidative stress. Variants and transcriptional changes of genes coding
for components of the leptin signaling pathway, and associated components of inflammation and antioxidant
protection, are associated with the onset and/or clinical course of MS as well as response to therapy.
Today, there are over 9,000 MS patients in Serbia, and only 1 out of 10 patients receives currently available
conventional immunomodulatory therapy. The possibility of modulating the level of circulating leptin
suggests its potential importance in the future development of a personalized approach in treatment of
MS based on improving the effects or substituting the existing conventional therapy
Unique pipeline for the assessment of novel genetic variants leads to confirmation of PCD diagnosis
Get PDFBackground/Objectives: Primary ciliary dyskinesia (PCD) is a
disease caused by impaired ciliary motility and mainly affects the
lungs and reproductive organs. Inheritance is autosomal recessive
and X-linked with more than 40 disease-causing genes, wherefore
PCD patients have diverse clinical manifestations, thus making
diagnosis difficult. The utility of next-generation sequencing (NGS)
technology for diagnostic purposes allows a better understanding
of the PCD genetic background. However, the identification of
specific disease-causing variants is challenging. The objective of
this study was to create a unique guideline that will enable the
standardization of the assessment of novel variants within PCD
associated genes.
Methods: The study included designing a pipeline for the classification
of the rare genetic variants detected using NGS. The pipeline
included in silico (translation, 3D-model, protein-protein interactions,
sequence conservation, posttranslational modifications) and functional
analysis (expressional analysis, Western Blot) of the variants.
Results: The designed pipeline consists of three steps: sequencing,
detection, and identification of genes/variants; classification of
variants according to their effect; and variant characterization using
in silico structural and functional analysis. The pipeline was validated
by the analysis of the variants detected in a disease-causing
gene (DNAI1) and the novel candidate gene (SPAG16).
Conclusion: The application of the pipeline resulted in the
identification of disease-causing variants, as well as pathogenicity
validation, through the analysis on transcriptional, translational,
and posttranslational levels.The application of created pipeline
leads to the confirmation of PCD diagnosis and enables a shift
from candidate to disease-causing gene.Abstracts from the 55th European Society of Human Genetics (ESHG) Conferenc
Molecular diagnosis of Fabry disease in patients with chronic renal failure of unknown etiology
Get PDFBackground/Objectives: Fabry disease (FD) is a rare X-linked
disorder caused by variants in the GLA gene leading to the deficiency
of lysosomal α-galactosidase-A and progressive accumulation
of globotriaosylceramide affecting the heart, nervous system,
and kidneys. FD has overlapping phenotypes and often remains
undiagnosed. Therefore, the precise molecular-genetic diagnosis
and the earliest possible treatment are essential to avoid significant
disease progression.
Methods: We analyzed 95 (34 female and 61 male) hemodialysis
patients with clinical suspicion of FD using Sanger sequencing
of all coding exons (7) and flanking intron regions of the GLA
gene, and measured the relative expression of the GLA gene in
available samples.
Results: The genetic analysis revealed 3 patients with a missense
variant (p.Asp313Tyr), and 10 patients with combinations of
non-coding variants, described as complex intronic haplotypes
(CIHs). CIH1 (c.-10C>T, c.370-81_370-77delCAGCC, c.640-16A>G,
c.1000-22C>T), the most frequent haplotype, was detected in 7
(7.4%) patients. Lyso-Gb3 biomarker levels were within the normal
range in each tested patient. However, RT-qPCR analysis revealed
decreased relative expression of GLA gene in PBMC of 2 female
patients with CIH1 and one female patient carrying only c.-10C>T
variant by 9,1%, 7,4%, 46,3%, respectively, pointing out that further
analyses are needed to confirm/exclude FD in these patients.
Conclusion: Because the effects of CIHs are not yet fully
understood, our work highlights the importance of analyzing
intronic regions of the GLA gene as genetic modifiers and the
need to include expression analysis in the diagnostic algorithm.Abstracts from the 55th European Society of Human Genetics (ESHG) Conferenc
Interactions of ultrashort laser pulses with hemoglobin: Photophysical aspects and potential applications
Get PDFHemoglobin (Hb), a life-sustaining and highly abundant erythrocyte protein, is not readily fluorescent. A few studies have already reported Two-Photon Excited Fluorescence (TPEF) of Hb, however, the mechanisms through which Hb becomes fluorescent upon interaction with ultrashort laser pulses are not completely understood. Here, we characterized photophysically this interaction on Hb thin film and erythrocytes using fluorescence spectroscopy upon single-photon/two-photon absorption, and UV-VIS single-photon absorption spectroscopy. A gradual increase of the fluorescence intensity, ending up with saturation, is observed upon prolonged exposure of Hb thin layer and erythrocytes to ultrashort laser pulses at 730 nm. When compared to protoporphyrin IX (PpIX) and oxidized Hb by H2O2, TPEF spectra from a thin Hb film and erythrocytes showed good mutual agreement, broad peaking at 550 nm, supporting hemoglobin undergoes degradation and that same fluorescent specie(s) originating from the heme moiety are generated. The uniform square shaped patterns of the fluorescent photoproduct exhibited the same level of the fluorescence intensity even after 12 weeks from the formation, indicating high photoproduct stability. We finally demonstrated the full potential of the formed Hb photoproduct with TPEF scanning microscopy towards spatiotemporally controlled micropatterning in HTF and single human erythrocyte labelling and tracking in the whole blood
The influence of BCL2, BAX, and ABCB1 gene expression on prognosis of adult de novo acute myeloid leukemia with normal karyotype patients
Get PDFBackground: Deregulation of the apoptotic process underlies the pathogenesis of many cancers, including leukemia, but is also very important for the success of chemotherapy treatment. Therefore, the gene expression profile of main apoptotic factors, such as anti-apoptotic BCL2 (B-cell lymphoma protein 2) and pro-apoptotic BAX (BCL2-associated X), as well as genes involved in the multi-drug resistance (ABCB1), could have significant impact on the prognosis and could be used as targets for specific therapy.Patients and methodsWe analyzed the expression of BCL2, BAX, and ABCB1 in bone-marrow samples collected at diagnosis from 51 adult patients with acute myeloid leukemia with normal karyotype (AML-NK) using real-time polymerase chain reaction method, and examined their prognostic potential.ResultsIncreased expression of BCL2 (BCL2+) was associated with the presence of chemoresistance (p = 0.024), while patients with low BAX expression were more prone to relapse (p = 0.047). Analysis of the combined effect of BCL2 and BAX expression showed that 87% of patients with BAX/BCL2low status were resistant to therapy (p = 0.044). High expression of ABCB1 was associated with BCL2+ status (p < 0.001), and with absence FLT3-ITD mutations (p = 0.019).ConclusionsThe present analysis of BCL2, BAX, and ABCB1 gene expression profiles is the first study focusing solel
Exploring zebrafish swim bladder as a model for chronic lung diseases
Get PDFThis project aims to identify transgenic zebrafish lines within the Kawakami Laboratory collection,
exhibiting Gal4 expression patterns in the swim bladder, to establish a robust model for investigating
the impacts of e-cigarettes. The swim bladder, sharing an embryonic background with the mammalian
lung, serves as a pertinent model due to the expression of genes crucial for lung function in adult
zebrafish. Key signaling pathways involved in human lung development, including the Sonic Hedgehog
and Wnt/β-catenin pathways, play pivotal roles in zebrafish swim bladder development. The zebrafish
swim bladder features a monolayer of cells, supported by a basement membrane. Surface surfactant is
present, while beneath the membrane, connective tissues house collagen, elastin, immune cells, and a
layer of smooth muscle cells. The research seeks zebrafish lines expressing Gal4 throughout the entire
swim bladder and, ideally, within specific cell types or layers of the swim bladder. The overarching
objective of this project is to unravel the mechanistic pathways activated by e-cigarettes, potentially
contributing to the onset of lung diseases later in life. By utilizing transgenic zebrafish lines with targeted
Gal4 expression in the swim bladder, this study aims to provide insights into the specific cellular and
molecular responses induced by e-cigarette exposure, contributing valuable knowledge to the
understanding of lung disease development.Principal Investigator: Dr Aleksandra Divac Rankov, IMGGEDuration period: 2023-202