imagine (Institute of molecular genetics and genetic engineering)
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Determination of Rare Genomic Variants Leading to Hematological Malignancies Using Next-Generation Sequencing
Get PDFRare hematological malignancies are a heterogeneous group
of the disease. Malignant transformation of a hematopoietic stem
cell is a result of gradual accumulation of mutations in a number of genes involved in basic cellular processes. One of the main
goals in the study of hematological malignancies is the definition
of genomic markers crucial for the development of the disease, as
well as for recognition of multiple entities characterized by distinct
prognosis and outcome of the disease. Application of new highthroughput technologies has enabled better insight into genomic
landscape of hematological malignancies. The most important
achievement of genome-based medicine is more precise classification of patients with hematological malignancies, based on newly
discovered molecular markers, and molecular–targeted therapy,
tailored to genomic profile of a disease.
In our studies we applied amplicon based next generation sequencing (NGS) approach, using TruSeq Amplicon Cancer Panel
(Illumina, Inc) for analysis of 48 cancer-related genes in primary
diffuse large B-cell lymphoma of central nervous system (DLBCL
CNS), acute myeloid leukemia (AML) and acute lymphoblastic
leukemia (ALL).
Our findings strongly suggested that the TP53 and ATM genes
were involved in the molecular pathophysiology of primary DLBCL CNS. Mutations in the PTEN and SMO genes affect survival of
the patients. Additionally, we found that AML and ALL contain
small number of genetic alterations, contrary to lymphomas.
While protein-changing variants were found in tyrosine kinase
genes, genes encoding tyrosine kinase associated proteins (JAK3,
ABL1, GNAQ, and EGFR) and in the methylation and histone
modifying genes (IDH1, IDH2, and SMARCB1) in patients with
AML, the mutations detected in ALL patients are related to key
signaling pathways, primarily on Ras/RTK cascade.
Application of next-generation sequencing technology resulted
in the information about genetic profile of each patient. Individual
genetic profiling leads to highly specific personalized therapy of
hematological malignancies
Polymorphism RAD51 172G > T in Serbian patients with colorectal cancer
No full textPurpose: The RAD51 gene plays an important role in homologous strand exchange in DNA repair. Two common single nucleotide polymorphisms in this gene, 135G gt C and 172G gt T, were associated with altered gene transcription. While 135G gt C was already linked to breast and colorectal cancers in certain populations, 172G gt T is far less investigated, although sporadic studies showed it could be a prognostic factor for some cancer lesions. The purpose of this study was to investigate RAD51 172G gt T polymorphism in Serbian population, its association with colorectal carcinoma, as well as correlation with disease characteristics and response to neoadjuvant chemoradiotherapy. Methods: The 172G gt T polymorphism was evaluated by PCR-RFLP method in blood samples of 209 colorectal cancer patients and 43 healthy subjects who served as controls. The distribution of genotypes was also analyzed in respect to several tumor characteristics in cases where histopathological data were available. Results: A significant association between the RAD51 172G gt T polymoprhism and desmoplastic reaction of colorectal cancer was demonstrated. The 172G allele was found to be significantly more frequent in patients with more intensive desmoplastic response of the tumor tissue. Conclusions: The results of our study suggest that the 172T allele of RAD51 may be a favorable prognostic factor in Serbian patients with colorectal cancer, although larger prospective studies are required to confirm this finding
New Insight into Biofilm Formation Ability, the Presence of Virulence Genes and Probiotic Potential of Enterococcus sp Dairy Isolates
Get PDFEnterococci have controversial status due to their emerging role in nosocomial infections and transmission of antibiotic resistance genes, while some enterococci strains are used as probiotics for humans and animals and starter cultures in dairy industry. In order to improve our understanding of factors involved in the safe use of enterococci as potential probiotics, the antibiotic susceptibility, virulence and probiotic traits of 75 dairy enterococci isolates belonging to Enterococcus durans (50), En. faeciurn (15), En. faecalis (6), En. italious (3), and En, hirae (1) were evaluated. The results revealed that ciprofloxacin resistance and biofilm formation are correlated with isolates originated from Golija mountain (Serbia), while gelatinase activity was more common in isolates from Prigorje region (Croatia), pointing to uncontrolled use of antibiotics and anthropogenic impact on dairy products" microbiota in these regions. The virulence genes were sporadically present in 13 selected dairy enterococci isolates. Interestingly, biofilm formation was correlated with higher ability of strains to reduce the adhesion of E. coil and Salmonella Enteritidis to HT29-MTX cells. To our knowledge this is the first study reporting the presence of the esp gene (previously correlated with pathogenesis) in dairy enterococci isolates, mostly associated with the genes involved in adhesion property. Hence, the results of this study revealed that the virulence genes are sporadically present in dairy isolates and more correlated to adhesion properties and biofilm formation, implicating their role in gut colonization rather than to the virulence traits
Influence of decreased fibrinolytic activity and plasminogen activator inhibitor-1 4G/5G polymorphism on the risk of venous thrombosis
No full textObjective of our study is to determine whether decreased fibrinolytic activity or plasminogen activator inhibitor (PAI)-1 4G/5G polymorphism influence the risk of venous thrombosis. Our case-control study included 100 patients with venous thrombosis, and 100 random controls. When patients were compared with random controls, unconditional logistic regression was used to calculate odds ratios (ORs) with 95% confidence intervals (CIs). Decreased fibrinolytic activity yielded a 2.7-fold increase in risk for venous thrombosis than physiological fibrinolytic activity (OR 2.70; 95% CI 1.22-5.98), when comparing patients with random controls. Adjustment for several putative confounders did not change the estimate (OR 3.02; 95% CI 1.26-7.22). Analysis of venous thrombotic risk influenced by PAI-1 genotype, showed no influence of PAI-1 4G/5G gene variant in comparison with 5G/5G genotype (OR 0.57 95% CI; 0.27-1.20). Decreased fibrinolytic activity increased, whereas PAI-1 4G/5G polymorphism did not influence venous thrombosis risk in this study
Association of interleukin-17 and interleukin-23 MRNA and protein expression profiles with clinical presentation in patients with systemic sclerosis
No full textBioavailability of Nutritional Resources From Cells Killed by Oxidation Supports Expansion of Survivors in Ustilago maydis Populations
Get PDFAfter heavy exposure of Ustilago maydis cells to clastogens, a great increase in viability was observed if the treated cells were kept under starvation conditions This restitution of viability is based on cell multiplication at the expense of the intracellular compounds freed from the damaged cells. Analysis of the effect of the leaked material on the growth of undamaged cells revealed opposing biological activity, indicating that U. maydis must possess cellular mechanisms involved not only in reabsorption of the released compounds from external environment but also in contending with their treatment-induced toxicity. From a screen for mutants defective in the restitution of viability, we identified four genes (adr1, did4, kel1, and tbp1) that contribute to the process The mutants in did4, kel1, and tbp1 exhibited sensitivity to different genotoxic agents implying that the gene products are in some overlapping fashion involved in the protection of genome integrity The genetic determinants identified by our analysis have already been known to play roles in growth regulation, protein turnover, cytoskeleton structure, and transcription. We discuss ecological and evolutionary implications of these results
Collaboration in the actions of Brh2 with resolving functions during DNA repair and replication stress in Ustilago maydis
No full textCells maintain a small arsenal of resolving functions to process and eliminate complex DNA intermediates that result as a consequence of homologous recombination and distressed replication. Ordinarily the homologous recombination system serves as a high-fidelity mechanism to restore the integrity of a damaged genome, but in the absence of the appropriate resolving function it can turn DNA intermediates resulting from replication stress into pathological forms that are toxic to cells. Here we have investigated how the nucleases Mus81 and Gen1 and the helicase Blm contribute to survival after DNA damage or replication stress in Ustilago maydis cells with crippled yet homologous recombination-proficient forms of Brh2, the BRCA2 ortholog and primary Rad51 mediator. We found collaboration among the factors. Notable were three findings. First, the ability of Gen1 to rescue hydroxyurea sensitivity of dysfunctional Blm requires the absence of Mus81. Second, the response of mutants defective in Blm and Gen1 to hydroxyurea challenge is markedly similar suggesting cooperation of these factors in the same pathway. Third, the repair proficiency of Brh2 mutant variants deleted of its N-terminal DNA binding region requires not only Rad52 but also Gen1. and Mus81. We suggest these factors comprise a sub pathway for channeling repair when Brh2 is compromised in its interplay with DNA.Peer-reviewed manuscript: [https://imagine.imgge.bg.ac.rs/handle/123456789/1626
Intra-articular injection of autologous adipose-derived mesenchymal stem cells in the treatment of knee osteoarthritis
Get PDFBackgroundOsteoarthritis (OA) is a chronic degenerative joint disease and is considered to be the fourth leading cause of disability and the second cause of inability to work in men. Recently, adipose-derived mesenchymal stem cells (AD-MSCs) came into focus for regenerative medicine as a promising tool for the treatment of OA. The administration of stem cells into impaired joints results in pain relief and improves quality of life, accompanied by restoration of hyaline articular cartilage. MethodsIn the present study, nine patients (including two patients with bilateral symptoms) diagnosed with osteoarthritis (International Knee Documentation grade B in 5 and grade D in six knees) were treated using a single injection of AD-MSCs at a concentration of 0.5-1.0 x 10(7) cells and were followed up for 18months. During follow-up, all the cases were evaluated clinically by Knee Society score (KSS), Hospital for Special Surgery knee score (HSS-KS), Tegner-Lysholm (T-L) score and visual analogue scale (VAS) of pain, as well as by plain radiography and by magnetic resonance imaging visualization with 2D Magnetic Resonance Observation of Cartilage Repair Tissue (MOCART) score assessment. ResultsSignificant improvement of all four clinical scores was observed within the first 6months (KSS for 41.4 points, HSS-KS for 33.9 points, T-L score for 44.8 points, VAS of pain from 54.5 to 9.3) and improvement persisted throughout the rest of the follow-up. MOCART score showed significant cartilage restoration (from 43 7.2 to 63 +/- 17.1), whereas radiography showed neither improvement, nor further joint degeneration. ConclusionsThe results obtained in the present study provide good basis for prospective randomized controlled clinical trials with respect to the use of AD-MSCs in the treatment of osteoarthritis.This is the peer reviewed version of the paper : Spasovski, D., Spasovski, V., Baščarević, Z., Stojiljković, M., Vreća, M., Anđelković, M., & Pavlović, S. (2018). Intra-articular injection of autologous adipose-derived mesenchymal stem cells in the treatment of knee osteoarthritis. The Journal of Gene Medicine, 20(1), e3002. [https://doi.org/10.1002/jgm.3002
Perioperativna anafilaksa kod dece u Srbiji
Get PDFINTRODUCTION: Several studies have identified neuromuscular blocking agents as the most common cause of
anaphylaxis during general anesthesia. The reported data vary considerably between countries.
AIM: In this study we have investigated anaphylactic reactions to neuromuscular blocking agents, general
anaesthetics and latex in children from Serbia.
METHODS: This case series study has encompassed hypersensitivity testing in 78 children over a period of twelve
consecutive months. The diagnostic protocol consisted of a case history, immunoglobulin E (IgE) testing and skin
tests for hypersensitivity to neuromuscular blocking agents, general anesthetics and latex.
RESULTS: Of 78 children, 63% were hypersensitive to neuromuscular blocking agents, 28% to latex, 13% to
hypnotics, 10% to opioids and 15% to coloids. Sixty cases (77%) of anaphylaxis were IgE-mediated. Among
children who underwent more than one intervention, the presence of cross-sensibilisation for neuromuscular
blocking agents commercially available in Serbia was identified in 77% of cases.
CONCLUSION: Neuromuscular blocking agents were the most frequent causal agents of anaphylaxis in Serbian
children, as verified by skin tests and specific IgE tests. High frequencies of allergies to neuromuscular blocking
agents and latex obtained in this study underscore the importance of carrying out an allergy study to detect an
allergic cause in perioperative reactionsUVOD: Nekoliko studija je identifikovalo neuromuskularne blokatore kao najčešći uzrok anafilakse tokom opšte
anestezije, ali prijavljeni podaci znatno variraju između zemalja.
CILJ: U ovoj studiji ispitivane su anafilaktičke reakcije na neuromuskularne blokatore, opšte anestetike i lateks
kod dece iz Srbije.
METODE: Ova studija serije slučajeva obuhvatila je testiranje preosetljivosti kod 78 dece tokom perioda od
dvanaest uzastopnih meseci. Dijagnostički protokol se sastojao od istorije slučajeva, testiranja imunoglobulina E
(IgE) i kožnih testova za preosetljivost na neuromuskularne blokatore, opšte anestetike i lateks.
REZULTATI: Od 78 ispitivane dece, 63% je preosetljivo na neuromuskularne blokatore, 28% na lateks, 13% na
hipnotike, 10% na opioide i 15% na koloide. U 60 slučajeva (77%) anafilaksija je posredovana IgE-om. Među
decom koja su prošla više od jedne intervencije, prisustvo unakrsne senzibilizacije za neuromuskularne bokatore
komercijalno dostupne u Srbiji identifikovano je u 77% slučajeva.
ZAKLJUČAK: Neuromuskularni blokatori najčešći su uzročni agensi anafilaksije kod srpske dece, što je potvrđeno
testovima kože i specifičnim IgE testovima. Visoke učestalosti alergija na neuromuskularne blokatore i lateks
dobijene u ovoj studiji naglašavaju važnost sprovođenja studija alergije kako bi se otkrili alergijski uzroci u
perioperativnim reakcijama
Mononuclear silver(I) complexes with 1,7-phenanthroline as potent inhibitors of Candida growth
Get PDFMononuclear silver(I) complexes with 1,7-phenanthroline (1,7-phen), [Ag(NO3-O,O') (1,7-phen-N7)(2)] (1) and [Ag(1,7-phen-N7)(2)]X, X = ClO4- (2), CF3SO3- (3), BF4- (4) and SbF6- (5) were synthesized and structurally characterized by NMR (H-1 and C-13), IR and UV-Vis spectroscopy and ESI mass spectrometry. The crystal structures of 1, 3 and 4 were determined by single-crystal X-ray diffraction analysis. In all these complexes, 1,7-phen coordinates to the Ag(I) ion in a monodentate fashion via the less sterically hindered N7 nitrogen atom. The investigation of the solution stability of 1-5 in DMSO revealed that they are sufficiently stable in this solvent at room temperature. Complexes 1-5 showed selectivity towards Candida spp. in comparison to bacteria, effectively inhibiting the growth of four different Candida species with minimal inhibitory concentrations (MIC) between 1.2 and 11.3 mu M. Based on the lowest MIC values and the lowest cytotoxicity against healthy human fibroblasts with selectivity index of more than 30, the antifungal potential was examined in detail for the complex 1. It had the ability to attenuate C. albicans virulence and to reduce epithelial cell damage in the cell infection model. Induction of reactive oxygen species (ROS) response has been detected in C. albicans, with fungal DNA being one of the possible target biomolecules. The toxicity profile of 1 in the zebrafish model (Danio rerio) revealed improved safety and activity in comparison to that of clinically utilized silver(I) sulfadiazine. (C) 2018 Elsevier Masson SAS. All rights reserved.This is peer-reviewed version of the following article: Savić, N. D.; Vojnovic, S.; Glišić, B. Đ.; Crochet, A.; Pavic, A.; Janjić, G. V.; Pekmezović, M.; Opsenica, I. M.; Fromm, K. M.; Nikodinovic-Runic, J.; et al. Mononuclear Silver(I) Complexes with 1,7-Phenanthroline as Potent Inhibitors of Candida Growth. Eur. J. Med. Chem. 2018, 156, 760–773. [https://doi.org/10.1016/j.ejmech.2018.07.049]Supplementary material: [https://imagine.imgge.bg.ac.rs/handle/123456789/2229