imagine (Institute of molecular genetics and genetic engineering)
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Nitrate inhibits primary root growth by reducing accumulation of reactive oxygen species in the root tip in Medicago truncatula
No full textIn Medicago truncatula, nitrate, acting as a signal perceived by NITRATE TRANSPORTER1/PEPTIDE TRANSPORTER FAMILY 6.8 (MtNPF6.8), inhibits primary root growth through a reduction of root cell elongation. Since reactive oxygen species (ROS) produced and converted in root tip (O-2(center dot-) - gt H2O2 - gt (OH)-O-center dot) have been reported to control cell elongation, the impact of nitrate on the distribution of these ROS in the primary root of M. truncatula was analyzed. We found that nitrate reduced the content of O-2(center dot-), H2O2 and (OH)-O-center dot in the root tip of three wild type genotypes sensitive to nitrate (R108, DZA, A17), inhibition of root growth and O-2(center dot-) accumulation being highly correlated. Nitrate also modified the capacity of R108 root tip to produce or remove ROS. The ROS content decrease observed in R108 in response to nitrate is linked to changes in peroxidase activity (EC1.11.1.7) with an increase in peroxidative activity that scavenge H2O2 and a decrease in hydroxylic activity that converts H2O2 into (OH)-O-center dot. These changes impair the accumulation of H2O2 and then the accumulation of (OH)-O-center dot, the species responsible for cell wall loosening and cell elongation. Accordingly, nitrate inhibitory effect was abolished by externally added H2O2 or mimicked by KI, an H2O2 scavenger. In contrast, nitrate has no effect on ROS production or removal capacities in npf6.8-2, a knockdown line insensitive to nitrate, affected in the nitrate transporter MtNPF6.8 (in R108 background) by RNAi. Altogether, our data show that ROS are mediators acting downstream of MtNPF6.8 in the nitrate signaling pathway.Peer-reviewed manuscript: [https://imagine.imgge.bg.ac.rs/handle/123456789/1628
Methanolic Extract of the Herb Ononis spinosa L. Is an Antifungal Agent with no Cytotoxicity to Primary Human Cells
Get PDFOnonis spinosa L. is a plant traditionally used as folk remedy. There are numerous studies regarding chemical constituents and health beneficial properties of Ononidis Radix. The following study was designed to investigate chemical composition and antifungal potential of the methanolic extract obtained from the O. spinosa L. herb. Chemical analyses regarding phenolic compounds of O. spinosa were performed by liquid chromatography with mass spectrometry (LC-DAD-ESI/MSn). Antifungal activity, antibiofilm properties and antifungal mode of action of the extract were evaluated, as well as cytotoxicity. Chemical analyses revealed the presence of flavonoids, isoflavonoids and phenolic acids in O. spinosa, with kaempherol-O-hexoside-pentoside being the most abundant compound (5.1 mg/g extract). Methanolic extract was active against all of the tested microfungi with Penicillium aurantiogriseum being the most sensitive to the extract inhibitory effect at 0.02 mg/mL; and effectively inhibited biofilms formed by Candida strains. Minimum fungicidal concentrations of extract rose in the presence of ergosterol and leakage of cellular components was detected. The extract showed no cytotoxicity to human gingival fibroblast (HGF-1) cells. This study significantly contributes to overall knowledge about medicinal potential of O. spinosa herbal extract and enlightens previously unrevealed properties. O. spinosa aerial parts seem to be an interesting candidate for the development of antifungal preparations, non-toxic to human cells
Optimization of extraction yield and chemical characterization of optimal extract from Juglans nigra L. leaves
No full textThe extraction yield of Juglans nigra L. leaves was assessed at different ethanol concentrations (0-96% (v/v)) and solvent-to-solid ratios (5-20 kg kg(-1)). The response surface methodology (RSM) and artificial neural network with genetic algorithms (ANN-GA) were developed to optimize the extraction variables. The RSM and ANN-GA models determined 50% (v/v) ethanol concentration and 20 kg kg(-1) solvent-to-solid ratio as optimal conditions, ensuring an extraction yield of 27.69 and 27.19 g 100 g(-1) of dry leaves. The phenolic compounds in optimal extract were quantified: 3-O-caffeoylquinic acid (2.27 mg g(-1)of dry leaves), quercetin-3-O-galactoside (10.99 mg g(-1) of dry leaves) and quercetin 3 0 rhamnoside (15.07 mg g(-1)of dry leaves) using high-performance liquid chromatography (HPLC). The minerals in optimal extract were quantified: macro-elements (the relative order by content was: K gt Mg gt Ca) using inductively coupled plasma optical emission spectrometry (ICP-OES) and micro-elements (the relative order by content was: Zn gt Rb gt Mn gt I gt Sr gt Ni gt Cu gt Co gt V gt Ag gt Se) using inductively coupled plasma mass spectrometry (ICP-MS). The extraction coefficients for minerals were determined and were highest for K (64.3%) and I (53.5%). Optimization of extraction process resulted in high extraction yield from J. nigra leaves and optimal extract containing different phytochemical compounds
Chemo- and biocatalytic esterification of marchantin A and cytotoxic activity of ester derivatives
Get PDFChemical and biocatalytic synthesis of seven previously undescribed marchantin A ester derivatives has been presented. Chemical synthesis afforded three peresterified bisbibenzyl products (TE1-TE3), while enzymatic method, using lipase, produced regioselective monoester derivatives (ME1-ME4). The antiproliferative activities of all prepared derivatives of marchantin A were tested on MRC-5 healthy human lung fibroblast, A549 human lung cancer, and MDA-MB-231 human breast cancer cell lines. All tested esters were less cytotoxic in comparison to marchantin A, but they also exhibited lower cytotoxicity against healthy cells. Monoesters displayed higher cytotoxic activities than the corresponding peresterified products, presumably due to the presence of free catechol group. Monohexanoyl ester ME3 displayed the same IC50 like marchantin A against MDA-MB-231 cells, but the selectivity was higher. In this way, regioselective enzymatic monoesterification enhanced selectivity of marchantin A. ME3 was also the most active among all derivatives against lung cancer cells A549 with the slightly lower activity and selectivity in comparison to marchantin A.Related to published version: [https://imagine.imgge.bg.ac.rs/handle/123456789/1367]This is the peer reviewed version of the paper: Novaković, M., Simić, S., Koracak, L., Zlatović, M., Ilić-Tomić, T., Asakawa, Y., Nikodinović-Runić, J., & Opsenica, I. (2020). Chemo- and biocatalytic esterification of marchantin A and cytotoxic activity of ester derivatives. Fitoterapia, 142.[ https://doi.org/10.1016/j.fitote.2020.104520
Bisaurones - enzymatic production and biological evaluation
Get PDFTheTrametes versicolorlaccase catalyzed oxidation of chalcone butein afforded four dimers of aurone sulfuretin (i.e.two regioisomeric pairs of diasteromers,1-4) as the major products. The formation of the dimers was explained by a two step process involving the initial cyclization of butein into aurone sulfuretin, followed by the combination of two molecules of sulfuretin. The coupling process occurred between the 2,10-double bond of one molecule of sulfuretin and the (3 ',4 ') catechol group of the other to yield a dimeric structure. This was confirmed by the experiment involving the laccase catalyzed oxidation of sulfuretin yielding the same dimeric bisaurones. Compounds1,3and4, were isolated using semipreparative HPLC and characterized by the detailed analysis of the NMR, MS, IR, and UV-vis data. The structure of compound2, isolated as a mixture containingca.25% of compound1,was proposed by the comparison of(1)H NMR data to compound1and by using LC-ESIMS analysis. The starting chalcone butein and the products of the biocatalytic transformation, aurone sulfuretin and sulfuretin dimers1,3and4,were evaluated for their cytotoxic and antioxidative propertiesin vitrousing a healthy human fibroblast (MRC5) cell line. The biotransformation products showed lower cytotoxicity but higher antioxidative properties. TheC. coggygriabark methanol extract rich in butein and sulfuretin was also biotransformed by laccase. The transformed extract exhibited significantly improved antioxidative activities.Related to published version: [https://imagine.imgge.bg.ac.rs/handle/123456789/1310]This is the peer reviewed version of the paper: Novakovi, M. M., Ilić-Tomić, T., Tešević, V., Simić, K., Ivanović, S., Simić, S., Opsenica, I., & Nikodinović-Runić, J. (2020). Bisaurones—Enzymatic production and biological evaluation. New Journal of Chemistry, 44(23), 9647–9655. [https://doi.org/10.1039/d0nj00758g
Effect of symbiosis with arbuscular mycorrhizal fungi on salt stress tolerance in GF677 (peach x almond) rootstock
No full textGenus Prunus is of great importance for cultivation, mainly because its main species provide fruits and seeds, valuable ornamental qualifies and timber. However, stone fruit trees and almonds, major cultivars of genus Prunus, are sensitive to salt stress. Such sensitivity causes losses in stone fruit and almond production (ca. 50% of regular yield at high salinity grounds). Having in mind that symbiosis with arbuscular mycorrhiza fungi (AMF) may improve plant tolerance to salt stress and that symbiosis effect should be tested in case-by-case approach, we tested salt stress response of AMF inoculated GF677 rootstock (Prunus dulcis x Prunus persica hybrid) in this study. Adding to that, we tested GF677 symbiosis with two AMF species R. intraradices and F. mosseae. Results showed that under salinity stress AMF inoculated GF677 plants displayed improved physiological parameters (chlorophyll, soluble sugars and proline content) and increased antioxidant enzymes activity in comparison to non-inoculated control plants. Comparison of two AMF species beneficial effects on tested parameters revealed that for total chlorophyll content inoculation with F. mosseae has prevailed, while for total soluble sugars and proline content R. intraradices has prevailed. Finally, GF677 in symbiosis with F. mosseae was selected for molecular studies of salinity response. Since many of plants' genes involved in simultaneous response to salt stress and AMF colonization remained unidentified so far, we performed bioinformatics analysis of freely online available data to find differentially expressed genes common to these two responses. Upon GO classification and networking analysis of genes identified as common to both responses, we selected two most prominent ones (UDPGT73C6 and CYP707A3) and tested their expression profile in leaves and roots of F. mosseae inoculated GF677 rootstocks under salt stress. Even though specific roles of UDPGT73C6 and CYP707A3 are un-characterized in Prunus tree species, results suggested their involvement in response to salt stress and AMF inoculation of GF677 plants, which is in concordance with a scarce knowledge on their roles in other plant species. Based on this study finding we may conclude that symbiosis of GF677 rootstocks with AMF increases plants tolerance to salinity stress, which should be considered in other Prunus tree species as well
Characterization, Antibiofilm, and Depolymerizing Activity of Two Phages Active on Carbapenem-ResistantAcinetobacter baumannii
Get PDFAcinetobacter baumanniiis a leading cause of healthcare-associated infections worldwide. Its various intrinsic and acquired mechanisms of antibiotic resistance make the therapeutic challenge even more serious. One of the promising alternative treatments that is increasingly highlighted is phage therapy, the therapeutic use of bacteriophages to treat bacterial infections. Two phages active against nosocomial carbapenem-resistantA. baumanniistrain 6077/12, vB_AbaM_ISTD, and vB_AbaM_NOVI, were isolated from Belgrade wastewaters, purified, and concentrated using CsCl gradient ultracentrifugation. The phages were screened against 103 clinical isolates ofA. baumanniifrom a laboratory collection and characterized based on plaque and virion morphology, host range, adsorption rate, and one-step growth curve. Given that phage ISTD showed a broader host range, better adsorption rate, shorter latent period, and larger burst size, its ability to lyse planktonic and biofilm-embedded cells was tested in detail. Phage ISTD yielded a 3.5- and 2-log reduction in planktonic and biofilm-associated viable bacterial cell count, respectively, but the effect was time-dependent. Both phages produced growing turbid halos around plaques indicating the synthesis of depolymerases, enzymes capable of degrading bacterial exopolysaccharides. Halos tested positive for presence of phages in the proximity of the plaque, but not further from the plaque, which indicates that the observed halo enlargement is a consequence of enzyme diffusion through the agar, independently of the phages. This notion was also supported by the growing halos induced by phage preparations applied on pregrown bacterial lawns, indicating that depolymerizing effect was achieved also on non-dividing sensitive cells. Overall, good rates of growth, fast adsorption rate, broad host range, and high depolymerizing activity, as well as antibacterial effectiveness against planktonic and biofilm-associated bacteria, make these phages good candidates for potential application in combatingA. baumanniiinfections
Identification of novel potent and non-toxic anticancer, anti-angiogenic and antimetastatic rhenium complexes against colorectal carcinoma
Get PDFCombination therapy targeting both tumor growth and vascularization is considered to be a cornerstone for colorectal carcinomas (CRC) treatment. However, the major obstacles of most clinical anticancer drugs are their weak selective activity towards cancer cells and inherent inner organs toxicity, accompanied with fast drug resistance development. In our effort to discover novel selective and non-toxic agents effective against CRC, we designed, synthesized and characterized a series of rhenium(I) tricarbonyl-based complexes with increased lipophilicity. Two of these novel compounds were discovered to possess remarkable anticancer, anti-angiogenic and antimetastatic activity in vivo (zebrafish-human HCT-116 xenograft model), being effective at very low doses (1-3 mu M). At doses as high as 250 mu M the complexes did not provoke toxicity issues encountered in clinical anticancer drugs (cardio-, hepato-, and myelotoxicity). In vivo assays showed that the two compounds exceed the anti-tumor and anti-angiogenic activity of clinical drugs cisplatin and sunitinib malate, and display a large therapeutic window.This is the peer reviewed version of the paper: Delasoie, J., Pavic, A., Voutier, N., Vojnovic, S., Crochet, A., Nikodinovic-Runic, J., & Zobi, F. (2020). Identification of novel potent and non-toxic anticancer, anti-angiogenic and antimetastatic rhenium complexes against colorectal carcinoma. European Journal of Medicinal Chemistry, 204, 112583.
[https://doi.org/10.1016/j.ejmech.2020.112583
Effect of composition and method of preparation of 2-hydroxyethyl methacrylate/gelatin hydrogels on biological in vitro (cell line) and in vivo (zebrafish) properties
No full textWe have studied the effect of compositions and methods of preparation on the physico-chemical and biocompatible behavior of the hydrogel matrices. Hydrogel matrices are synthesized by free radical polymerization of 2-hydroxyethyl methacrylate net and with gelatin. Highly porous hydrogel structures were obtained by porogenation, and by cryogenic treatment followed by freeze-drying. All samples were characterized for structural, morphological, absorption, degradation and in vitro (healthy human fibroblast cell line) and in vivo (zebrafishDanio rerio) biocompatible properties. The obtained results show that cryo samples, especially with gelatin show better, favorable absorption, morphological and biocompatible properties in comparison with PHEMA samples, which makes these materials highly attractive for biomedical uses
Pharmacogenomics landscape of COVID-19 therapy response in Serbian population and comparison with worldwide populations
Get PDFUvod: Kako ne postoje odobreni terapeutici za lečenje pacijenata sa COVID-19, mogućnost upotrebe postojećih lekova je postala važna. U nedostatku vremena za testiranje farmakogenomskih markera kod pojedinaca, populaciona farmakogenomika bi mogla biti od koristi u predviđanju povećanog rizika za pojavu neželjenih reakcija i neuspeha lečenja kod pacijenata sa COVID-19. Cilj naše studije bio je identifikovanje farmakogena i farmakogenomskih markera povezanih sa lekovima koji se preporučuju za lečenje COVID-19, hlorokin/hidroksihlorokin, azitromicin, lopinavir i ritonavir, u populaciji Srbije i drugim svetskim populacijama. Metode: Podaci o genotipu 143 osobe srpskog porekla dobijeni su iz baze podataka prethodno formirane analizama genoma korišćenjem TruSight One Gene Panel (Illumina). Podaci o genotipu pojedinaca iz različitih svetskih populacija dobijeni su iz Projekta 1000 genoma. Fišerov egzaktni test korišćen je za poređenje učestalosti alela. Rezultati: Identifikovali smo 11 potencijalnih farmakogenomskih markera u 7 farmakogena značajnih za lečenje COVID-19. Na osnovu visoke alterativne učestalosti alela u populaciji Srbije i funkcionalnog efekta varijanti, ABCB1 rs1045642 i rs2032582 mogu biti značajne za smanjeni klirens lekova azitromicina, lopinavira i ritonavira, a varijanta UGT1A7 rs17868323 za hiperbilirubinemiju kod bolesnika sa COVID-19 koji se leče ritonavirom. SLCO1B1 rs4149056 je potencijalni marker odgovora na lopinavir, posebno u populaciji Italije. Naši rezultati potvrdili su da se farmakogenomski profil afričke populacije razlikuje od ostatka sveta. Zaključak: Uzimajući u obzir farmakogenomski profil specifičan za populaciju, preventivno testiranje farmakogena značajnih za lekove koji se koriste u lečenju COVID-19 moglo bi doprineti boljem razumevanju interindividualnih razlika u odgovorima na terapiju i poboljšanju ishoda lečenja pacijenata sa COVID-19.Background: Since there are no certified therapeutics to treat COVID-19 patients, drug repurposing became important. With lack of time to test individual pharmacogenomics markers, population pharmacogenomics could be helpful in predicting a higher risk of developing adverse reactions and treatment failure in COVID-19 patients. Aim of our study was to identify pharmacogenes and pharmacogenomics markers associated with drugs recommended for COVID-19 treatment, chloroquine/hydroxychloroquine, azithromycin, lopinavir and ritonavir, in population of Serbia and other world populations. Methods: Genotype information of 143 individuals of Serbian origin was extracted from database previously obtained using TruSight One Gene Panel (Illumina). Genotype data of individuals from different world populations were extracted from the 1000 Genome Project. Fisher's exact test was used for comparison of allele frequencies. Results: We have identified 11 potential pharmacogenomics markers in 7 pharmacogenes relevant for COVID-19 treatment. Based on high alternative allele frequencies in population and the functional effect of the variants, ABCB1 rs1045642 and rs2032582 could be relevant for reduced clearance of azithromycin, lopinavir and ritonavir drugs and UGT1A7 rs17868323 for hyperbilirubinemia in ritonavir treated COVID-19 patients in Serbian population. SLCO1B1 rs4149056 is a potential marker of lopinavir response, especially in Italian population. Our results confirmed that pharmacogenomics profile of African population is different from the rest of the world. Conclusions: Considering population specific pharmacogenomics landscape, preemptive testing for pharmacogenes relevant for drugs used in COVID-19 treatment could contribute to better understanding of the inconsistency in therapy response and could be applied to improve the outcome of the COVID-19 patients