imagine (Institute of molecular genetics and genetic engineering)
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Influence of different concentrations of Zn-carbonate phase on physical-chemical properties of antimicrobial agar composite films
No full textAgar-based composites with different Zn-carbonate mineral phase content were prepared by in situ mineralization and the solvent casting method. The mineral phase within the composite films was identified as hydrozincite, Zn-5(CO3)(2)(OH)(6). The presence of the mineral phase improved, both mechanical and water vapor permeability properties of the obtained composite films, in a concentration-dependent manner. The release of zinc ions from composite films is in accepted levels (up to 2.5%), and sufficient to provide complete inhibition growth of S. Aureus. The results of this study suggest that agar/Zn-carbonate composites could be potentially used as affordable, eco-friendly and functional materials with tunable properties for food packaging, agriculture or biomedical application. In situ procedure offers possibilities for tailoring the physical-chemical properties of composite films, by varying the Zn-mineral phase load
Relationship between interleukin-23 and gastrointestinal involvement in patients with systemic sclerosis
Get PDFExpression of miRNA-210 in human bone marrow-derived mesenchymal stromal cells under oxygen deprivation
Get PDFA major limitation in the development of efficient clinical protocols for mesenchymal stromal cell (MStroC)-based tissue regeneration therapy is the low retention and survival of MStroC in injured tissue after therapeutic administration. Low oxygen concentration preconditioning (LOP) during ex vivo cultivation of MStroC, as a method for mimicking oxygenation in their physiological microenvironment, has been shown to be beneficial in clinical trials using MStroC. Introducing hypoxia-mimicking molecules into MStroC during cultivation could be an advantageous LOP strategy. MicroRNA (miRNA) drugs are good candidates for this approach. Analysis of the expression of miRNA-210 in human bone marrow-derived MStroC in conditions of acute and extended hypoxia (24 to 72 h) was performed using RT-qPCR methodology. HIF-1 alpha and HIF-2 alpha gene knockdown cell lines were generated using lentiviral transduction of short hairpin RNA (shRNA) in order to examine whether miRNA-210 expression is regulated by transcription factor HIF-1 and/or HIF-2. We detected a significant increase in miRNA-210 expression in hypoxic conditions at time points of 24, 48 and 72 h (p lt 0.05). Knocking down of HIF-1 alpha and HIF-2 alpha genes indicated involvement of both transcription factors in the elevation of miRNA-210 expression. These results point to miRNA-210 as a good candidate for a hypoxia-mimicking molecule in LOP strategy
Antimicrobial and anti-biofilm activity and biological decontamination efficiency of ED-1 emulsion
Get PDFAntimicrobial and antibiofilm activity, as well as biological decontamination potential of emulsion ED-1, highly efficient in radiological decontamination of metal surfaces contaminated with uranium isotopes, was assessed. The antimicrobial potency of ED-1 was evaluated against 10 different microorganisms including four Gram-negative bacteria (Acinetobacter baumannii ATCC 19606, Escherichia coli NCTC 9001, Klebsiella pneumoniae ATCC 13803 and Pseudomonas aeruginosa NCTC 10662), four Gram-positive bacteria (Enterococcus faecalis ATCC 29212, Enterococcus faecium ATCC 6057, Listeria monocytogenes NCTC 11994, Staphylococcus aureus NCTC 6571) and two fungi (Candida albicans ATCC 10231 and Candida parapsilosis ATCC 22019). Although without strong bactericidal and fungicidal properties in standard agar diffusion assays, ED-1 effectively inhibited the growth of P. aeruginosa cells in liquid culture and more importantly, showed high potential to disperse P. aeruginosa biofilms. ED-1 was also capable to efficiently remove Bacillus subtilis ATCC 6633 spores in quantitative and a semi-quantitative biological decontamination tests on metal surfaces. Antimicrobial and antibiofilm activity and biological decontamination efficiency of ED-1 was comparable to and better than that of calcium hypochlorite solution or commercial decontaminant BX-24. This study highlighted the possibility to use ED-1, with up to 5-fold reduced amounts of calcium hypochlorite in comparison to currently used methodology, for both biological and radiological decontamination, resulting in both environmental and financial benefits
Sox genes: for better or for worse…
No full textThe SOX genes encode a group of transcription factors that act as key regulators of diverse
cellular processes. Members of the SOX gene family show dynamic and diverse expression
patterns during development playing important roles ranging from blastocyst formation to
differentiation into adult tissues and organs. SOX transcription factors are involved in multiple
events from the maintaining of stem cells pluripotency and cell fate decision to driving
terminal differentiation of cells into specialized cell types. In addition, during adulthood SOX
transcription factors control various physiological processes. Mutations in SOX genes have
been associated with severe clinical disorders, while misregulation of their expressions cause
a broad range of pathological condition. Accumulating evidence suggests that SOX proteins
act as oncogenes and recent evidence points toward pro-proliferative, prosurvival and/or
antidifferentiation roles of the SOX proteins.
The results of long-term research of the structure, expression regulation and the function of
selected SOX genes will be presented. It will include data obtained by studying the roles of
SOX genes in in vitro neural differentiation of pluripotent embryonal carcinoma cells, as well
as interaction of SOX transcription factors with signaling pathways active during neurogenesis
and oncogenesis. Special focus will be made on ongoing research focused on the roles of SOX
genes in promotion of malignant phenotype of cancer cells and maintaining of cancer stem
cells. Obtained results provide novel insights into the functions of SOX genes in cancer that
will help in designing novels strategies for cancer treatment.Abstracts of the 6th CONGRESS OF THE SERBIAN GENETIC SOCIETY Vrnjačka Banja; October 13 to 17, 2019
Interactions of genetic and environmetal factors in chronic obstructive pulmonary disease
No full textChronic obstructive pulmonary disease (COPD) is featured by abnormal inflammatory
response of the lungs to toxic particles or gases, such as cigarette smoke. High heterogeneity
and complex etiology of the disease have made the genetic components and the mechanisms
of COPD pathogenesis difficult to unravel. Although recent advances in genetic technologies
have enabled revelation of various genetic factors as a cause of the disease, a huge portion
of estimated COPD heritability is still unknown. However, the ”missing heritability” of COPD
is thought to be hidden in unidentified interactions of genetic and environmental factors.
The aim of our research was to identify interactions between genes involved in xenobiotic
metabolism, antioxidative defense, matrix remodeling, inflammation and vascular function,
tagged by functional variants, and cigarette smoking in COPD using a case-control model.
Functional effects of gene-gene and gene-environment interactions were assessed by
measuring the levels of marker of systemic oxidative stress urinary 8-oxo-7,8-dihydro-2’-
deoxyguanosine (8-oxodG), the relation with biochemical pathways or relevant cell culture
model. We identified interactions of GSTM1-GSTP1 (p=0.029) and CYP1A1-GSTM1-EPHX
(p=0.025) in COPD and cumulative effect of risk variants on the levels of 8-oxodG (p=0.026)
in patients. Further, interactions of eNOS-ACE (p=0.041) and eNOS-smoking (p=0.013)
are revealed in COPD and related to protective markers e.g. caveolin. Furthermore, geneenvironment
interaction of MMP9-smoking is detected in COPD occurrence (p=0.005) and
severity (p=0.001), showing higher expression of MMP9 risk variant in response to cigarette
smoke extract (p<0.05) using macrophage cell line. Our research is the first to reveal several
interactions of genetic and environmental components as a possible cause of lung damage
contributing to “missing heritability” of COPD and ultimately improving our understanding
of the mechanisms of COPD pathogenesis.Abstracts of the 6th CONGRESS OF THE SERBIAN GENETIC SOCIETY Vrnjačka Banja; October 13 to 17, 2019
Characterization of a novel cutinase from Streptomyces sp. BV286
No full textCutinases are members of the α/β- hydrolase superfamily. They possess an enormous
potential for a variety of industrial applications based on their ability to hydrolyze a
wide range of substrates. The majority of information regarding the characteristics
of cutinases refers to fungal cutinases with significantly less information on bacterial
cutinases. The advantages of prokaryotic enzymes in industrial application and
the large number of predicted bacterial cutinases opened up possibilities for the
discovery of novel cutinases with desirable properties. Therefore, a novel cutinase
(designated CUT286) from Streptomyces sp. BV286 was identified, expressed and
biochemically characterized.
Streptomyces sp. BV286 was isolated from the rhizosphere of Papaver rhoeas and
displayed cutinase activity in enzyme assays. Cutinase gene that was identified from
the genome of this strain showed similarity with the cutinase from Malbranchea
cinnamomea. pRSET B vector was used for cloning and expression of cutinase in E.
coli ROSETTA (DE3) cells. CUT286 was partially purified using affinity chromatography
and its esterase activity was determined in the standard cutinase assay with paranitrophenilbutyrate
(pNPB) as a substrate. Activity of CUT286 was highest in neutral
to slightly acidic environments (pH 5.0 - 7.0) with a sudden drop of activity at pH
levels higher than 8.0. CUT286 exhibited optimal activity at pH 5.0 and 60 °C, and high
stability at 80 °C retaining 85% of its original activity after two hours. Additionally,
CUT286 was stable in 15% v/v of the following organic solvents: dimethyl sulfoxide
(DMSO), dimethylformamide (DMF), methanol and acetonitrile. All these properties,
mainly thermostability, make cutinase from Streptomyces sp. BV286 suitable for
potential industrial applications.Abstracts of the 6th CONGRESS OF THE SERBIAN GENETIC SOCIETY Vrnjačka Banja; October 13 to 17, 2019
Chokeberry polyphenols preservation using spray drying: effect of encapsulation using maltodextrin and skimmed milk on their recovery following in vitro digestion
No full textAim: Microencapsulation of chokeberry extracts was performed in order to improve functionality, stability, and bioavailability of extracted polyphenols. Methods: Chokeberry fruits and juice by-product (waste) extracts were spray-dried by using two carriers, maltodextrin and skimmed milk. Morphological and physicochemical characteristics of the obtained powders were analysed. In vitro simulated digestion model was used as an indicator of polyphenolics bioavailability. Results: The moisture content varied between 3.39 and 4.61%, zeta potential had negative values (35-39 mV), maltodetrin powders were smaller (4.27-5.12 mu m) compared to skimmed ones (8.50-11.01 mu m). All microparticles exhibited high encapsulation efficiency of total polyphenols and anthocyanins (73-97% and 63-96%, respectively). For both extract types, maltodextrin powders released higher phenolics content compared to skimmed milk. During in vitro digestion, maltodextrin exhibited a higher protective effect on both active compounds. Conclusion: Taking into account the obtained results, chokeberry polyphenols stability might be improved using spray drying technique, and maltodextrin showed better properties
Different coordination abilities of 1,7-and 4,7-phenanthroline in the reactions with copper(II) salts: Structural characterization and biological evaluation of the reaction products
No full textThe reactions between equimolar amounts of CuX2 (X = NO3- and CF3SO3-) and two aromatic nitrogen-containing heterocycles differing in the position of nitrogen atoms, 1,7- and 4,7-phenanthroline (1,7-and 4,7-phen), were performed in ethanol/methanol at room temperature. When CuX 2 salts were mixed with 4,7-phen, two copper(II) complexes, [Cu(NO3)(2)(4,7-Hphen)(2)](NO3)(2) (1) and [Cu(CF3SO3)(4,7-phen)(2)(H2O)(2)]CF3SO3 (2), were formed. On the other hand, in the reaction of CuX2 salts with 1,7-phen, only 1,7-HphenNO(3) (3a/b) and 1,7-HphenCF(3)SO(3) (4) were obtained as the final products. The obtained products 1-4 were characterized by spectroscopic and X-ray diffraction techniques. In the copper(II) complexes 1 and 2, the coordination geometry around the Cu(II) ion is distorted octahedral and square pyramidal, respectively. The antimicrobial potential of the copper(II) complexes 1 and 2 and corresponding compounds used for their synthesis were assessed against four different bacterial species and Candida albicans, displaying moderate growth inhibiting activity. The cytotoxic properties of the investigated complexes were also evaluated against the normal human lung fibroblast cell line (MRC-5) indicating moderate, yet more pronounced cytotoxicity than antimicrobial properties
Characterization of mid-intestinal microbiota of farmed Chinook salmon using 16S rRNA gene metabarcoding
Get PDFWith the growing importance of aquaculture worldwide, characterization of the microbiota of high-value aquaculture species and identification of their shifts induced by changes in fish physiology or nutrition is of special interest. Here we report the first 16S rRNA gene metabarcoding survey of the mid-intestinal bacteria of Chinook salmon (Oncorhynchus tshawytscha), an economically important aquacultured species. The microbiota of 30 farmed Chinook salmon from a single cohort was surveyed using metabarcode profiling of the V3-V4 hypervariable region of the bacterial 16S rRNA gene. Seawater, feed and mid-intestinal samples and controls were sequenced in quadruplicate to assess both biological and technical variation in the microbial profiles. Over 1000 operational taxonomic units were identified within the cohort, providing a first glimpse into the mid-intestinal microbiota of farmed Chinook salmon. The taxonomic distribution of the salmon microbiota was reasonably stable, with around two thirds of individuals dominated by members of the family Vibrionaceae. We anticipate that the workflow presented in this paper could be applied in other aquacultured fish species to capture variation or dysbiosis occurring as a result of changes in feed, health or environmental conditions