imagine (Institute of molecular genetics and genetic engineering)
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Preanalyzed data for the article: Lukić, J., Vass, N., Ardó, L., Stanivuk, J., Lengyel-Kónya, É., Golić, N., Jakabné Sándor, Z., & Ljubobratović, U. (2024). Evaluation of weaning diets for sustainable indoor largemouth bass (Micropterus salmoides) larviculture. Czech Journal of Animal Science, 69(12), 471–483. https://doi.org/10.17221/129/2024-CJAS
No full text***Dataset contents*** Pond reared_13jun2023..csv (8.859Kb)
Morphometry..csv (34.89Kb)
Proteins_Tris..csv (11.50Kb)
Trypsin..csv (23.83Kb)
Alkaline phosphatase..csv (23.12Kb)
Lipase..csv (21.33Kb)
Chymotrypsin..csv (22.66Kb)
Phospholipase A2..csv (18.57Kb)
Proteins_PBS..csv (12.51Kb)
Amylase..csv (16.83Kb)
T4..csv (15.62Kb)
CCK..csv (18.33Kb)
T3..csv (14.23Kb)
Experimental design..csv (15.76Kb)
First experiment..csv (14.61Kb)
Summary gene expression..csv (14.85Kb)
qPCR_7..csv (26.97Kb)
qPCR_6..csv (27.00Kb)
qPCR_5..csv (26.70Kb)
qPCR_4..csv (28.13Kb)
qPCR_3..csv (26.99Kb)
qPCR_2..csv (25.80Kb)
qPCR_Repetition..csv (22.85Kb)
Standard curves..csv (13.21Kb)
PL assay..csv (14.23Kb)
Digestibility assay..csv (17.61Kb)
Sheet2..csv (12.34Kb)
LMB_intestine_trypsine_activity..csv (9.805Kb)
Total protein_with CMC..csv (11.56Kb)
Digestibility assay with CMC..csv (14.07Kb)
Leaching proteins..csv (12.99Kb)
Leaching amino acids..csv (12.78Kb)
Preanalyzed data_10.172211292024-CJAS (1).xlsx (335.6Kb)Preanalyzed data for:[http://dx.doi.org/10.17221/129/2024-CJAS]Related to published version: [https://imagine.imgge.bg.ac.rs/handle/123456789/2576]Related to supplementary information:[https://imagine.imgge.bg.ac.rs/handle/123456789/2758
Razvoj biokatalizatora za razgradnju poli(uretana)
No full textPlastični polimeri predstavljaju nezamenjivu sirovinu u današnjem društvu. Velika količina
proizvedenih plastičnih produkata na kraju upotrebe završe u životnoj sredini zagađujući je
(Bishop et al., 2020). Iz ovog razloga naučna zajednica ulaže sve veće napore u pronalasku
adekvatnog rešenja. Plastični polimeri su materijali koji se ne mogu lako razgraditi prirodnim
putem. Abiotički procesi razgradnje plastike traju stotinama godina, a zbog spore razradnje
plastični polimeri se akumuliraju u životnoj sredini,otpuštajući mikroplastiku i druge hemikalije,
što dodatno povecava potrebu pronalaska rešenja opisanog problema (MacLeod et al., 2021).
Jedan od najčešće korišćenih tipova plastičnih polimera su poliuretani (PU). Poliuretani
predstavljaju raznovrstan tip plastike, a po hemijskoj strukturi oni su estri karbamatne kiseline.
Sama sinteza poliuretana je kompleksna, a variranjem koncentracije reaktanata dobijaju se
proizvodi koji mogu biti u formi fleksibilnih i rigidnih sunđera, elastomera i adheziva (Ionescu,
2005). Raznolikost u strukturi poliuretana omogućuje njihovu široku primenu u različitim
industrijama poput industrije nameštaja, automobilske industrije, tekstilne industrije i građevine
(Gama et al., 2018)
Tracking antimicrobial resistance in diverse ecological niches - one health perspective (TRACE)
Get PDFThe rise of antimicrobial resistance (AMR) has become a global public health crisis. The AMR
evolvability is mainly influenced by the flow of bacteria, antimicrobial resistance genes (ARGs),
and active antimicrobials through hospital settings, farm animals, and water sources. Thus, the
One Health approach (OHA) is best suited to address the existing and emerging threats of AMR,
particularly in multidrug-resistant (MDR) Gram-negative bacteria (GNB) as global priority AMR
pathogens widely distributed in three interconnected sectors.
Molecular characterization of MDR GNB isolated from human, animal, and water samples (≥300)
will be done by multilocus sequence typing and whole-genome sequencing, while the long- and
short-reads-based metagenomics will be applied to assess bacterial diversity and resistomes of
animal and water samples. Post-sequencing analyses aimed at extracting phylogenetic and
functional information will be performed by advanced bioinformatic tools.
Hence, the “One health genomics“ to tackle AMR will be adopted for the first time in Serbia, a
South-Eastern European country with a substantial burden of AMR in clinical settings. Besides,
the Project will bring progress beyond the state-of-the-art in genomic analyses, enabling the
detection of emerging MDR GNB clones and their potential for further spread.
The scientific impact is reflected in obtaining a comprehensive and multidisciplinary insight into
the molecular diversity and spread of relevant MDR GNB. The long-term impact will be the
development of a research basis for rational public health practices, which will primarily be
directed to strengthening national capacities to address AMR towards OHA.
The expected results of the Project will be an in-depth insight into the clonal complexes and
genetic relatedness of the circulating MDR GNB, their transmission routes and point sources, and
the assessment of the selective pressure of antibiotics on ARGs and bacterial communities in
Serbian cities.Principal Investigator: Dr. Ina Gajić, Faculty of Medicine, University of BelgradeCoordinator for IMGGE: Dr. Radmila NovakovićDuration period: 2024-202
A novel thermostable YtnP lactonase from Stenotrophomonas maltophilia inhibits Pseudomonas aeruginosa virulence in vitro and in vivo
Get PDFInfections caused by multidrug-resistant pathogens are one of the biggest challenges facing the healthcare system today. Quorum quenching (QQ) enzymes have the potential to be used as innovative enzyme-based antivirulence therapeutics to combat infections caused by multidrug-resistant pathogens. The main objective of this research was to describe the novel YtnP lactonase derived from the clinical isolate Stenotrophomonas maltophilia and to investigate its antivirulence potential against multidrug-resistant Pseudomonas aeruginosa MMA83. YtnP lactonase, the QQ enzyme, belongs to the family of metallo-β-lactamases. The recombinant enzyme has several advantageous biotechnological properties, such as high thermostability, activity in a wide pH range, and no cytotoxic effect. High-performance liquid chromatography analysis revealed the activity of recombinant YtnP lactonase toward a wide range of N-acyl-homoserine lactones (AHLs), quorum sensing signaling molecules, with a higher preference for long-chain AHLs. Recombinant YtnP lactonase was shown to inhibit P. aeruginosa MMA83 biofilm formation, induce biofilm decomposition, and reduce extracellular virulence factors production. Moreover, the lifespan of MMA83-infected Caenorhabditis elegans was prolonged with YtnP lactonase treatment. YtnP lactonase showed synergistic inhibitory activity in combination with gentamicin and acted additively with meropenem against MMA83. The described properties make YtnP lactonase a promising therapeutic candidate for the development of next-generation antivirulence agents
Antimicrobial activity of Hyssopus officinalis L. subsp. aristatus (Godr.) Nyman (Lamiaceae) essential oils from Montenegro and Serbia
Get PDFIn this study, antimicrobial activity of essential oils extracted from the aerial flowering parts (herbs) of Hyssopus officinalis subsp. aristatus (Godr.) Nyman (Lamiaceae) collected from five different locations in Montenegro, or purchased in Serbia, were investigated. In addition, their antibacterial activity in combination with antibiotics was studied. The antimicrobial activity against selected standard bacterial and yeast strains was investigated using the broth microdilution method. Two standard antibiotics were used for comparison: the aminoglycoside antibiotic amikacin and the cephalosporin antibiotic ceftriaxone. The antimicrobial activity of the essential oil-amikacin combination was investigated using the checkerboard assay. The main components of the essential oils were 1,8-cineole, cis-pinocamphone, β-pinene and limonene in varying quantities. Most of the tested essential oils showed no significant antimicrobial activity. However, an essential oil rich in cis-pinocamphone showed moderate activity against both Staphylococcus aureus and Escherichia coli (MIC = 400 µg/mL). The overall effect of the essential oils and antibiotic combinations against E. coli or S. aureus ranged from additive (FICI = 0.625) to indifferent (FICI = 1.5), depending on the source of the essential oil
Proteomic examination of polyester-polyurethane degradation by Streptomyces sp. PU10: Diverting polyurethane intermediates to secondary metabolite production
Get PDFGlobal plastic waste accumulation has become omnipresent in public discourse and the focus of scientific research. Ranking as the sixth most produced polymer globally, polyurethanes (PU) significantly contribute to plastic waste and environmental pollution due to the toxicity of their building blocks, such as diisocyanates. In this study, the effects of PU on soil microbial communities over 18 months were monitored revealing that it had marginal effects on microbial diversity. However, Streptomyces sp. PU10, isolated from this PU-contaminated soil, proved exceptional in the degradation of a soluble polyester-PU (Impranil) across a range of temperatures with over 96% degradation of 10 g/L in 48 h. Proteins involved in PU degradation and metabolic changes occurring in this strain with Impranil as the sole carbon source were further investigated employing quantitative proteomics. The proposed degradation mechanism implicated the action of three enzymes: a polyester-degrading esterase, a urethane bond-degrading amidase and an oxidoreductase. Furthermore, proteome data revealed that PU degradation intermediates were incorporated into Streptomyces sp. PU10 metabolism via the fatty acid degradation pathway and subsequently channelled to polyketide biosynthesis. Most notably, the production of the tri-pyrrole undecylprodigiosin was confirmed paving the way for establishing PU upcycling strategies to bioactive metabolites using Streptomyces strains
YTNP LACTONASE IMPROVES THE ABILITY OF CAENORHABDITIS ELEGANS TO SURVIVE PSEUDOMONAS AERUGINOSA MMA83 INFECTION
Get PDFPseudomonas aeruginosa is a Gram-negative
pathogen responsible for frequent hospital-acquired
infections of the bloodstream, the respiratory
tract, and the urinary tract. Quorum
quenching enzymes are recognized as an alternative
antivirulence approach targeting pathogenic
bacteria. The efficacy of YtnP lactonase in
reducing the virulence of P. aeruginosa MMA83
in vivo using Caenorhabditis elegans as a model
system was investigated. The recombinant YtnP
lactonase exhibits no cytotoxicity, demonstrated
by its lack of harmful effects on both the
immortalized human HaCaT cell line and two
strains of C. elegans (AU37 and N2 wild-type). In
a toxin-mediated killing liquid assay, the survival
rates of C. elegans AU37 mutant and N2 wildtype
strains infected with the clinical isolate P.
aeruginosa MMA83 significantly increased when
pre-treated with YtnP lactonase, compared to
untreated controls. Considering that virulence
factors expression is regulated by quorum sensing
(QS) signaling it is hypothesized that YtnP
lactonase prolongs the life span of C. elegans
by downregulating the QS and expression of
virulence factors of MMA83. The protective effects
of YtnP lactonase against MMA83-induced
pathogenicity in C. elegans, coupled with its absence
of cytotoxicity, position YtnP lactonase as
a promising prophylactic agent with antivirulence
properties.Book of abstract: From biotechnology to human and planetary health XIII congress of microbiologists of Serbia with international participation Mikromed regio 5, ums series 24: 4th – 6th april 2024, Mona Plaza hotel, Belgrade, Serbi
Silver(I) complexes containing antifungal azoles: significant improvement of the anti-Candida potential of the azole drug after its coordination to the silver(I) ion
Get PDFInspired by the emergence of resistance to currently available antifungal therapy and by the great potential of metal complexes for the treatment of various diseases, we synthesized three new silver(I) complexes containing clinically used antifungal azoles as ligands, [Ag(ecz)2]SbF6 (1, ecz is econazole), {[Ag(vcz)2]SbF6}n (2, vcz is voriconazole), and [Ag(ctz)2]SbF6 (3, ctz is clotrimazole), and investigated their antimicrobial properties. The synthesized complexes were characterized by mass spectrometry, IR, UV-vis and 1H NMR spectroscopy, cyclic voltammetry, and single-crystal X-ray diffraction analysis. In the mononuclear complexes 1 and 3 with ecz and ctz, respectively, the silver(I) ion has the expected linear geometry, in which the azoles are monodentately coordinated to this metal center through the N3 imidazole nitrogen atom. In contrast, the vcz-containing complex 2 has a polymeric structure in the solid state in which the silver(I) ions are coordinated by four nitrogen atoms in a distorted tetrahedral geometry. DFT calculations were done to predict the most favorable structures of the studied complexes in DMSO solution. All the studied silver(I) complexes have shown excellent antifungal and good to moderate antibacterial activities with minimal inhibitory concentration (MIC) values in the ranges of 0.01–27.1 and 2.61–47.9 μM on the selected panel of fungi and bacteria, respectively. Importantly, the complexes 1–3 have exhibited a significantly improved antifungal activity compared to the free azoles, with the most pronounced effect observed in the case of complex 2 compared to the parent vcz against Candida glabrata with an increase of activity by five orders of magnitude. Moreover, the silver(I)-azole complexes 2 and 3 significantly inhibited the formation of C. albicans hyphae and biofilms at the subinhibitory concentration of 50% MIC. To investigate the impact of the complex 3 more thoroughly on Candida pathogenesis, its effect on the adherence of C. albicans to A549 cells (human adenocarcinoma alveolar basal epithelial cells), as an initial step of the invasion of host cells, was studied
Phosphorylated neurofilament heavy chain in cerebrospinal fluid and plasma as a Nusinersen treatment response marker in childhood-onset SMA individuals from Serbia
Get PDFIntroductionBiomarkers capable of reflecting disease onset and short- and long-term therapeutic effects in individuals with spinal muscular atrophy (SMA) are still an unmet need and phosphorylated neurofilament heavy chain (pNF-H) holds significant promise.MethodsWe conducted a longitudinal prospective study to evaluate pNF-H levels in the cerebrospinal fluid (CSF) and plasma of 29 individuals with childhood-onset SMA treated with Nuinersen (SMA type 1: n = 6, 2: n = 17, 3: n = 6). pNF-H levels before and during treatment were compared with the levels of controls (n = 22), patients with Duchenne muscular dystrophy (n = 17), myotonic dystrophy type 1 (n = 11), untreated SMA individuals with chronic type 3 disease (n = 8), and children with presymptomatic SMA (n = 3).ResultsSMA type 1 showed the highest mean CSF pNF-H levels before treatment initiation. All Nusinersen-treated individuals (types 1, 2, and 3) showed significantly elevated mean baseline CSF pNF-H compared to controls, which inversely correlated with age at disease onset, age at first dose, disease duration and the initial CHOP INTEND result (SMA type 1 and 2). During 22 months of treatment, CSF pNF-H levels declined during loading doses, stabilizing at reduced levels from the initial maintenance dose in all individuals. Baseline plasma pNF-H levels in type 1 and 2 SMA were significantly increased compared to other cohorts and decreased notably in type 1 after 2 months of treatment and type 2 after 14 months. Conversely, SMA type 3, characterized by lower baseline pNF-H levels, did not show significant fluctuations in plasma pNF-H levels after 14 months of treatment.ConclusionOur findings suggest that CSF pNF-H levels in untreated SMA individuals are significantly higher than in controls and that monitoring of CSF pNF-H levels may serve as an indicator of rapid short-term treatment response in childhood-onset SMA individuals, irrespective of the subtype of the disease, while also suggesting its potential for assessing long-term suppression of neurodegeneration. Plasma pNF-H may serve as an appropriate outcome measure for disease progression and/or response to treatment in types 1 and 2 but not in type 3. Presymptomatic infants with SMA may show elevated pNF-H levels, confirming early neuronal degeneration
A model compounds for the discovery of novel PHA depolymerases – a computational study
Get PDFPolyhydroxyalkanoates (PHA) represent a sustainable alternative to conventional plastics due
to their microbial biological origin, biodegradability, biocompatibility, and structural diversity.
The primary challenge in the industrial enzymatic recycling of PHA lies in the relatively slow
biodegradation rates, spanning several months to years, depending on environmental
conditions and the characteristics of both PHA and the PHA-degrading enzymes.
This study involves experimental and computational testing of monomeric and dimeric units
of polyhydroxy octanoate (PHO) as suitable substrates for continuous spectrophotometric
monitoring of enzymatic reactions. Additionally, we explore the specificity of these novel model
substrates for polyhydroxyalkanoate depolymerases (PHOase) in comparison to other esterases
(lipase and cutinase).
The experimental findings are complemented by computational evaluations. We have
modeled the most active enzyme, poly(3-hydroxyoctanoate) depolymerase from Pseudomonas
fluorescens, followed by cutinase LCC, which exhibits lower activity on tested substrates, and
lipase CalB that doesn't show detectable activity. Our computational insights provide a rationale
for these experimental observations