imagine (Institute of molecular genetics and genetic engineering)
Not a member yet
3088 research outputs found
Sort by
Benzo[4,5]thieno[2,3-d]pyrimidine phthalimide derivative, one of the rare noncompetitive inhibitors of dipeptidyl peptidase-4
No full textA small library of benzo[4,5]thieno[2,3-d]pyrimidine phthalimide and amine derivatives was evaluated for inhibitory activity against dipeptidyl peptidase-4 (DPP-4). The phthalimide derivatives exhibited better activity than the amine precursors, with 2-(2-(3-chlorobenzyl)-5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidin-4-yl)isoindoline-1,3-dione (compound 14) as the most effective inhibitor (IC50 = 34.17 +/- 5.11 mu M). The five most potent selected inhibitors did not show cytotoxicity to a greater extent on Caco-2 cells, even at a concentration of 250 mu M. Compound 14 is considered as a novel representative of the rare noncompetitive DPP-4 inhibitors. Molecular docking and dynamics simulation indicated the importance of the Tyr547, Lys554, and Trp629 residues of DPP-4 in the formation of the enzyme-inhibitor complex. These observations could be potentially utilized for the rational design and optimization of novel (structurally similar, with phthalimide moiety, or different) noncompetitive DPP-4 inhibitors, which are anyway rare, but favorable in terms of the saturation of substrate competition
Supplementary information for the article: Filipović, N., Veselinović, L., Ražić, S., Jeremić, S., Filipič, M., Žegura, B., Tomić, S., Čolić, M., Stevanović, M., 2019. Poly (ε-caprolactone) microspheres for prolonged release of selenium nanoparticles. Materials Science and Engineering C 96, 776–789. https://doi.org/10.1016/j.msec.2018.11.073
Get PDF1. Experimental details for ICP-OES measurements; 1.1. Instrumental and operating conditions; 1.2.Solutions and Reagents; 1.3. Microwave assisted acid digestion; 1.4. Calibration curve 2. Experimental details for biocompatibility investigations of PCL/SeNPs; 2.1. Cell culture; 2.2.Determining citotoxicity of samples - MTT assay; 2.3. Determination of intracellular reactive oxygen species formation – DCFH-DA assay; 2.4. DNA damage (comet assay) Figure 1. SEM image of blank PCL microspheres Figure 2. XRD pattern of commercial PGA used in experiments Figure 3. Interaction with PCL/SeNPs in vivo by infiltrating cells. PCL/SeNPs (4mg/animal) were injected into sterile polyvinyl sponges implanted subcutaneously. The infiltrating cells were collected from the sponges after 3h and stained to anti-CD45/IgG Alexa 488 (Green) and Syto59 nuclear stain. PCL/SeNPs were detected as brightly scattering particles sized about 1-4 μm after 546nm laser excitation either intracellularly within granulocytes (A) or extracellularly (B). Note that some cells expressed strongly CD45 on the membrane and the cytoplasm, whereas others displayed a weak membrane expression and a strong expression in the granular ER at the nucleus level. Table 1. Melting temperatures Tm and corresponding enthalpies (heat) of fusion ΔHf of PCL/SeNPs samples taken after different time from different degradation mediums Table 2. Melting temperatures and corresponding enthalpies of PCL/SeNPs samples taken after different degradation periods from P. aeruginosa CFE mediumRelated to the published version: [https://imagine.imgge.bg.ac.rs/handle/123456789/1613]Supplementary material for: [https://doi.org/10.1016/j.msec.2018.11.073]Related to the accepted manuscript: [https://imagine.imgge.bg.ac.rs/handle/123456789/1614
Phylogeography, phenology and morphology of balkan snowdrops (Galanthus spp.)
No full textThe genus Galanthus L. (Amaryllidaceae) comprises 22 species, which are perennial bulbous
petaloid monocots, commonly known as snowdrops. It is distributed throughout Europe,
Asia Minor and the Near East. Galathus spp. are cherished garden plants and the world’s
most traded wild-sourced ornamental bulb genus, threatened by illegal collecting and
habitat destruction and thus listed in Appendix II of the CITES. However, species delimitation
is problematic and the infrageneric classification uncertain, mainly due to the overall
morphological similarity within the genus, and lack of easily discernible distinguishing
characters. In order to shed light on taxonomy and evolutionary history of Galathus species
from the Central Balkans, we analyzed variability of three plastid regions (rps16-trnK, trnLtrnF
and trnE-trnT) in 119 individuals from 40 populations representing six species (G. nivalis,
G. elwesii and G. reginae-olgae occurring naturally in this region and three allochthonous
species, phenology in 21 populations representing two species (G. nivalis and G. elwesii), and
variability of 13 quantitative characters in 510 individuals from 17 populations belonging to
one species (G. nivalis). Although evolutionary relationships among species were not fully
resolved, they were concordant with those based on morphological data (sensu Davis),
with the exception of G. gracilis which was not grouped with G. elwesii and G. woronowii.
Interspecific hybridization between G. nivalis and G. reginae-olge was detected. We observed
a phylogeographic structure in the Balkans, with eight lineages distributed throughout this
region. The most recent common ancestor of G. nivalis lineages was dated to the 1.683 Mya
(1.089 Mya - 2.376 Mya), and diversification of more or less all G. nivalis lineages took place
during the past 0.500 Mya. Phenological differences were observed among six G. nivalis
lineages, of which one was characterized also by specific morphological traits.Abstracts of the 6th CONGRESS OF THE SERBIAN GENETIC SOCIETY Vrnjačka Banja; October 13 to 17, 2019
Sequencing of Streptomyces sp. Np10 genome – a key to the treasure chest
No full textIn post genomic era the whole genome sequencing (WGS) is only a start point that
offers an alternative, but complementary approach to standard biodiscovery. In that
light, sequencing genome of Streptomyces sp. NP10, a strain from our laboratory collection,
not only supported the previous research on its secondary metabolites but
spurred further discovery of important enzymes. NP10 stands out by its ability to
produce exceptionally high amounts of fatty acids (FA) that have characteristics of
secondary metabolites, some of them detected in living organisms for the first time.
Owing to WGS it was possible to pinpoint synthesis of these FA to an unusual cluster
C12-2 whose inactivation almost completely abolished their synthesis. Further,
genome mining confirmed the presence of the genetic elements that corresponded to
metabolic and physiological features of NP10: DNase and hemolytic activity, cellulase,
endoglucanase, and catalase activity, hydrolysis of urea, degradation of arabinose
and xylose, high salt and low temperature tolerance. By carefully narrowing
our search and utilizing different bioinformatics tools and databases, we were able to
isolate a highly active cutinase/lipase. This enzyme, important in biocatalysis, can be
repurposed for plastic modification and degradation. Finally, specific search for biosynthetic
gene clusters (BGC) revealed over 30 of them spanning from polyketides
and non-ribosomal peptides synthesis to terpene and ectoine production. Most of these
small molecules cannot be detected in the cultivation medium during standard cultivation,
but once identified, BCGs could be activated by biotechnological engineering
or heterologously expressed. However, our inability to detect cyclic dipeptide BGC
(although we were able to detect cyclic dipeptides in the cultivation broth) shows that
in spite of tremendous advances in bioinformatic predictions that make genome mining
a reliable approach to enzyme discovery, there are a few shortcomings that still
need to be conquered.
GENOME MINING,Abstracts of the 6th CONGRESS OF THE SERBIAN GENETIC SOCIETY Vrnjačka Banja; October 13 to 17, 2019
The role of intracellular traficking in reconstitution of oxidative damaged population of Ustilago maydis
No full textUstilago maydis is a phytopathogenic fungus exhibiting extreme resistance to UV
and ionizing radiation. Even regarding this distinctive feature that marks U. maydis
out among eukaryotes, the molecular knowledge that would have a direct bearing
on our understanding of its ecological success is still fragmentary. Our results show
that after heavy exposure of a population of U. maydis cells to oxidative damage a
great increase in viability is observed if the treated cells are incubated for prolonged
period in distilled water (starvation). This restitution of viability is achieved through
cell multiplication of survivors by reabsorption of the intracellular compounds
released from the killed cells. This finding suggests that U. maydis must possess
cellular mechanisms involved in recycling (reabsorption, processing and reuse)
of the leaked material for cell growth and multiplication. By screening mutants
defective in capability to use these compounds we identified four genes (kel1, slm1,
snf8 and did4) that contribute to the process. Interestingly, these mutants are also
sensitive to different genotoxic agents implying that the gene products are involved
in genome protection. Two of the genes, kel1 and slm1, are involved in cytoskeleton
reorganization and vesicle movement. The protein products of the other two genes,
snf8 and did4, are parts of ESCRT-II and ESCRT- III complexes, which are known to
play roles in vesicular trafficking and in destruction of damaged proteins. Obtained
results will have a strong impact on our understanding of the cellular mechanisms
that enable U. maydis populations to reabsorb and process the leaked, potentially
harmful cellular waste.Abstracts of the 6th CONGRESS OF THE SERBIAN GENETIC SOCIETY Vrnjačka Banja; October 13 to 17, 2019
DNA fingerprinting of Serbian grapevine varieties by means of nuclear microsatellites
No full textGrapevine, Vitis vinifera L., cultivated in some regions of Serbia since the Roman Empire, is
nowadays grown on c. 20.000 ha. Along with modern vineyards established using varieties
with certified identity, old and sometimes abandoned vineyards, which represent a possible
reservoir of local and potentially unknown varieties, are also common in Serbia. The Serbian
Ministry of Agriculture, Forestry and Water Management has recognized the importance of
characterizing local varieties at morphological and molecular levels and safeguarding those
that are threatened by extinction. A procedure for grape DNA fingerprinting by means of
nuclear microsatellites, established in the frame of several European research projects, has
been implemented in Serbia at the Institute of Molecular Genetic and Genetic Engineering
(IMGGE) in the frame of the of the EU IPA II Twinning Project “Strengthening capacities of
phytosanitary sector in the field of plant varieties registration, including improvement of
variety testing authorities”. We genotyped 12 Serbian grape varieties grown in the germplasm
repository of Sremski Karlovci whose genetic profiles have been compared against 3.888
genotypes available at the Vitis International Variety Catalogue (VIVC, http://www.vivc.de/).
We demonstrate that this methodology represents a powerful tool not only for distinguishing
Serbian grapevine varieties but also for resolving synonymy/homonymy, which is rather
common in grapevines, and in paternity and kinship analysis. We highlight the importance of
joint future efforts of molecular geneticists, breeders, vinegrowers and policy makers for the
valorization of grapevine varieties available in Serbia, which will help breeders in exploiting
local genetic resources and foster the production of quality grapes and wines in Serbia.Abstracts of the 6th CONGRESS OF THE SERBIAN GENETIC SOCIETY Vrnjačka Banja; October 13 to 17, 2019
A contribution to the estimation of berry fruits quality
No full textThis study aimed to estimate the quality (in terms of standard chemical parameters, total antioxidative activity /AO/ and thermal properties) of the fresh strawberry (Fragaria xananassa L. Duch.), raspberry (Rubus idaeus L.) and blackberry (Rubus fruticosusL.) samples grown in the Western Serbia. The examined blackberry variety was found to exhibit the most potent AO (DPPH /2,2-diphenyl-1-picrylhydrazyl/ assay, EC50 4.05 +/- 0.22 mu g and ARP 0.25 +/- 0.03 mu g(-1); Hydroxo-Perhydroxyl Mercury(II) Complex /HPMC/ assay, 0.072 +/- 0.007%/mu L), due to the enriched contents of both total phenolics and anthocyanins. It was actually followed by the raspberry and strawberry samples. Unlike the aforementioned trend for AO, thermal behaviour of all samples was similar. This is first time ever that a DC polarographic HPMC assay has been applied for the characterisation of the fresh berry fruits. Further studies aiming to screen total AO of the soft fruit during growth, storage and processing might be well facilitated by the application of this low cost, rapid, easy-to-handle and reliable electrochemical assay
Genotype phenotype correlation in a pediatric population with antithrombin deficiency
No full textInherited antithrombin (AT) deficiency is a rare autosomal dominant disorder, caused by mutations in the AT gene (SERPINC1). Considering that the genotype phenotype relationship in AT deficiency patients remains unclear, especially in pediatric patients, the aim of our study was to evaluate genotype phenotype correlation in a Serbian pediatric population. A retrospective cohort study included 19 children younger than 18 years, from 15 Serbian families, with newly diagnosed AT deficiency. In 21% of the recruited families, mutations affecting exon 4, 5, and 6 of the SERPINC1 gene that causes type I AT deficiency were detected. In the remaining families, the mutation in exon 2 causing type II HBS (AT Budapest 3) was found. Thrombosis events were observed in 1 (33%) of those with type I, 11 (85%) of those with AT Budapest 3 in the homozygous respectively, and 1(33%) in the heterozygous form. Recurrent thrombosis was observed only in AT Budapest 3 in the homozygous form, in 27% during initial treatment of the first thrombotic event. Abdominal venous thrombosis and arterial ischemic stroke, observed in almost half of the children from the group with AT Budapest 3 in the homozygous form, were unprovoked in all cases. Conclusion: Type II HBS (AT Budapest 3) in the homozygous form is a strong risk factor for arterial and venous thrombosis in pediatric patients.What is Known:center dot Inherited AT deficiency is a rare autosomal dominant disorder, caused by mutations in the SERPINC1gene.center dot The genotype phenotype correlation in AT deficiency patients remains unclear, especially in pediatric patients.What is New:center dot The genetic results for our paediatric population predominantly showed the presence of a single specific mutation in exon 2, that causes type II HBS deficiency (AT Budapest 3).center dot In this group thrombosis mostly occurred as unprovoked, in almost half of them as abdominal thrombosis or stroke with high incidence of recurrent thrombosis, in 27% during initial treatment
Prothrombin 3'end Gene Variants in Patients With Sporadic Colon Adenocarcinoma
Get PDFBackground/ Aim: Thrombin plays significant roles in various types of cancer. However, the expression levels of prothrombin, the thrombin precursor, in cancer remain unclear. Variants of the 3'end of the prothrombin gene lead to increased prothrombin expression. This study aimed to analyze prothrombin 3'end gene variants in colon tumor and adjacent normal tissue samples. Materials and Methods: The study group consisted of 93 patients suffering from colon adenocarcinoma. The 3'end of the prothrombin gene was analyzed by DNA sequencing. Results: Three variants, all previously associated with increased prothrombin expression were detected. Frequency of the FII 19911G allele was 46.77% and 47.85% in tumor and normal tissue, respectively. For the FII 20210A allele, the detected frequencies were 2.15% and 1.61%, respectively. The frequency of the FII c.1824T allele was 0.54% in both tissues. Four patients showed different genotypes in tumor and normal tissue. Conclusion: Prothrombin 3' end gene variants may play a role in colorectal cancer
N-Benzyl Derivatives of Long-Chained 4-Amino-7-chloro-quionolines as Inhibitors of Pyocyanin Production in Pseudomonas aeruginosa
No full textPseudomonas aeruginosa is a leading cause of nosocomial infections that are becoming increasingly difficult to treat due to the occurrence of antibiotic resistant strains. Since P. aeruginosa virulence is controlled through quorum sensing, small molecule treatments inhibiting quorum sensing signaling pathways provide a promising therapeutic option. Consequently, we synthesized a series of N-octaneamino-4-aminoquinoline derivatives to optimize this chemotype's antivirulence activity against P. aeruginosa via inhibition of pyocyanin production. The most potent derivative, which possesses a benzofuran substituent, provided effective inhibition of pyocyanin production (IC50 = 12 mu M), biofilm formation (BFIC50 = 50 mu M), and motility. Experimentally, the compound's activity is achieved through competitive inhibition of PqsR, and structure-activity data were rationalized using molecular docking studies