imagine (Institute of molecular genetics and genetic engineering)
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Neuroprotective Role of Selected Antioxidant Agents in Preventing Cisplatin-Induced Damage of Human Neurons In Vitro
Get PDFChemotherapy-induced peripheral neuropathy (CIPN) is a side effect of platinum-based chemotherapy and decreases the quality of life of cancer patients. We compared neuroprotective properties of several agents using an in vitro model of terminally differentiated human cells NT2-N derived from cell line NT2/D1. Sodium azide and an active metabolite of amifostine (WR1065) increase cell viability in simultaneous treatment with cisplatin. In addition, WR1065 protects the non-dividing neurons by decreasing cisplatin caused oxidative stress and apoptosis. Accumulation of Pt in cisplatin-treated cells was heterogeneous, but the frequency and concentration of Pt in cells were lowered in the presence of WR1065 as shown by X-ray fluorescence microscopy (XFM). Transition metals accumulation accompanied Pt increase in cells; this effect was equally diminished in the presence of WR1065. To analyze possible chemical modulation of Pt-DNA bonds, we examined the platinum L-III near edge spectrum by X-ray absorption spectroscopy. The spectrum found in cisplatin-DNA samples is altered differently by the addition of either WR1065 or sodium azide. Importantly, a similar change in Pt edge spectra was noted in cells treated with cisplatin and WR1065. Therefore, amifostine should be reconsidered as a candidate for treatments that reduce or prevent CIPN
Expression pattern of long non-coding RNA growth arrest-specific 5 in the remission induction therapy in childhood acute lymphoblastic leukemia
Get PDFUvod: Ekspresija duge nekodirajuće RNK G AS5 je izme - njena u mnogim kancerima zbog njene uloge u apoptozi i inhibiciji rasta ćelije. G AS5 interaguje sa glukokortikoidnim receptorom, što je čini potencijalnim farmakotranskripcionim markerom značajnim za glukokortikoidnu terapiju. Naš cilj u ovoj studiji je bio da analiziramo ekspresiju GAS5 tokom indukcione terapije kod dečje akutne limfoblastne leukemije (ALL), u kojoj se koriste glukokortikoidni lekovi, i da te rezultate povežemo sa odgovorom na terapiju. Metode: Nivo ekspresije G AS5 u mononuklearnim ćelijama periferne krvi izolovanih od 29 dece obolelih od ALL, određen je metodologijom RT-qPCR, i to u momentu dijagnoze, 15. i 33. dana indukcione terapije. Rezultati: Naši rezultati su pokazali da postoje interindivi - dualne razlike u ekspresiji GAS5 kod pacijenata, i to u svim analiziranim tačkama. Kod svakog ALL pacijenta GAS5 ekspresija je 15. dana bila visa u odnosu na ekspresiju u momentu dijagnoze (p lt 0,0005). Nivo ekspresije GAS5 je 33. dana bio niži u poređenju sa 15. danom (p lt 0,0005), ali je i dalje bio značajno visi u odnosu na momenat dijagnoze kod većine pacijenata (p = 0,001). Pacijenti čiji je broj blasta u perifernoj krvi 8. dana bio ispod 100 po mikrolitru periferne krvi, imali su viši nivo ekspresije GAS5 (p = 0,016) i niži odnos ekspresija merenih 15. dana i u momentu dijagnoze (p = 0,009). Zaključak: Nasi rezultati ukazuju da bi nivo ekspresije GAS5 mogao da bude marker terapijskog odgovora u indukcionoj terapiji kod dece obolele od ALL.Background: Long non-coding RNA growth arrest-specific 5 (GAS5) is deregulated in many cancers because of its role in cell growth arrest and apoptosis. Additionally, GAS5 interacts with glucocorticoid receptor, making it a potential pharmacotranscription marker of glucocorticoid (GC) therapy. In this study, we aimed at analysing G AS5 expression in the remission induction therapy phase of childhood acute lymphoblastic leukemia (ALL), in which GCs are mandatorily used, and to correlate it with therapy response. Methods: G AS5 expression was measured in peripheral blood mononuclear cells taken from 29 childhood ALL patients at diagnosis, on day 15 and day 33 of remission induction therapy using RT-qPCR methodology. Results: Our results have shown interindividual differences in G AS5 expression at all time points. For each ALL patient, G A S5 expression was higher on day 15 in comparison to its level at diagnosis (p lt 0.0005). On day 33, the level of G A S5 expression decreased in comparison with day 15 (p lt 0.0005), but it was still significantly higher than at diagnosis for the majority of patients (p=0.001). Patients whose number of blasts on day 8 was below 100 per mL of peripheral blood had a higher GAS5 expression at diagnosis (p=0.016), and lower ratio day 15/diagnosis (p=0.009). Conclusions: Our results suggest that the expression level of GAS5 could be a potential marker of therapy response in remission induction therapy of childhood ALL
Ankrd2 in Mechanotransduction and Oxidative Stress Response in Skeletal Muscle: New Cues for the Pathogenesis of Muscular Laminopathies
Get PDFAnkrd2 (ankyrin repeats containing domain 2) or Arpp (ankyrin repeat, PEST sequence, and proline-rich region) is a member of the muscle ankyrin repeat protein family. Ankrd2 is mostly expressed in skeletal muscle, where it plays an intriguing role in the transcriptional response to stress induced by mechanical stimulation as well as by cellular reactive oxygen species. Our studies in myoblasts from Emery-Dreifuss muscular dystrophy 2, a LMNA-linked disease affecting skeletal and cardiac muscles, demonstrated that Ankrd2 is a lamin A-binding protein and that mutated lamins found in Emery-Dreifuss muscular dystrophy change the dynamics of Ankrd2 nuclear import, thus affecting oxidative stress response. In this review, besides describing the latest advances related to Ankrd2 studies, including novel discoveries on Ankrd2 isoform-specific functions, we report the main findings on the relationship of Ankrd2 with A-type lamins and discuss known and potential mechanisms involving defective Ankrd2-lamin A interplay in the pathogenesis of muscular laminopathies
The influence of specific mutations in the AT gene (SERPINC1) on the type of pregnancy related complications
No full textBackground: Inherited antithrombin (AT) deficiency is a rare autosomal dominant disorder, caused by mutations in the SERPINC1 gene. The most common clinical presentation in AT deficient patients includes venous thrombosis and pulmonary embolism, while the association of AT deficiency and its effect on the development of pregnancy complications has been less studied. The aim of our research was to evaluate the effect of AT deficiency types, determined by genotyping, on pregnancy outcomes. Methods: A retrospective cohort study included 28 women with AT deficiency, and their 64 pregnancies were analyzed. Results: With regard to live birth rate, a significant difference was observed among women who were carriers of different SERPINC1 mutations, as the rate varied from 100% in cases of type I to the extremely low rate of 8% for women with type II HBS (AT Budapest 3) in the homozygous variant, P=0.0005. All pregnancies from the type I group, even untreated ones, resulted in live births. In women with AT Budapest 3 in homozygous variant the overall live birth rate increased to 28.5% in the treated pregnancies. In this group the highest incidence of fetal death was observed of 62%; repeated fetal losses in 30%; fetal growth restriction in 22% and placental abruption in 7% of all pregnancies. Conclusion: Our study results indicate a difference between type I and type II AT deficiency. The risk of pregnancy related VTE was equally present in both groups, except for AT Budapest 3 in the heterozygous variant, while adverse pregnancy outcomes were strictly related to type II, especially AT Budapest 3 in the homozygous variant
Characterization of the lactic acid bacteria producers of gama-aminobutyric acid and their role in the prevention and treatment of experimental autoimmune encephalomyelitis
No full textγ-aminobuterna kiselina (GABA) je biogeni amin prisutan kod biljaka, životinja imikroorganizama i ostvaruje brojne fiziološke funkcije: inhibitor je neurotransmisije usinapsama, ima umirujući efekat i ublažava hipertenziju i važan je imunosupresivni molekul. γ-aminobuterna kiselina nastaje ireverzibilnom dekarboksilacijom L-glutamata aktivnošćuglutamat dekarboksilaze (GAD).Ovo istraživanje imalo je za cilj ispitivanje potencijala sojeva mlečnokiselinskih bakterija(MKB) izolovanih iz mlečnih proizvoda Zapadnog Balkana kao i iz proizvoda sa Kavkaza i izAzerbejdžana za produkciju γ-aminobuterne kiseline, selekciju sojeva sa navedenih područja kojiprodukuju najviše ovog metabolita i njihovu detaljnu probiotičku karakterizaciju. Nakonselekcije soja, koji produkuje najveću koncentraciju γ-aminobuterne kiseline ispitivan je efekatživog soja i supernatanata kultura ovog soja na modelu inflamacije in vitro i na poboljšanjekliničke slike na modelu eksperimentalnog autoimunskog encefalomijelitisa, životinjskogmodela multiple skleroze. Korišćenjem standardnih prajmera (CoreR/CoreF) prisustvo gadBgena, gena koji kodira glutamat dekarboksilazu, detektovano je kod 124 soja i to: 83 (27,4 %)laktobacila, 15 (4,9 %) laktokoka, 11 (3,6 %) leukonostoka, 8 (2,64 %) streptokoka, 6 (1,98 %)enterokoka i 1 (0,33 %) pediokoka od testirana 303 soja. TLC metodom je produkcija ovogmetabolita detektovana je kod 4,6 % sojeva, odnosno 14 sojeva, od 124 testirana soja, a kojipripadaju vrstama Lactobacillus brevis i Lb. plantarum, dok je HPLC metodom izmerenakoncentracija produkovane γ-aminobuterne kiseline. Simulirane uslove gastrointestinalnog traktaod 7 testiranih sojeva uspešno su preživela 4 soja vrste Lb. brevis BGLMM10, BGLMM11,BGZLS10-17 i BGZLS30-2. Ispitivanje bezbednosnog statusa sojeva pokazalo je da je sojBGZLS10-17 je bio osetljiv na sve testirane antibiotike, BGLMM11 je bio rezistentan nakanamicin, BGLMM10 je bio rezistentan na kanamicin i tetraciklin, dok je soj BGZLS30-2 biorezistentan na ampicilin, kanamicin i tetraciklin, dok ni jedan od 4 ispitivana soja nije ispoljio nihemolitičku ni želatinoznu aktivnost...microorganisms. GABA functions as an inhibitory neurotransmitter, has a calming effectalleviates hypertension, and is an effective immunosuppressive molecule. GABA is synthesizedby irreversible decarboxylation of L-glutamate by glutamate decarboxylase (GAD) activity.The research goal of this study was to examine the potential of lactic acid bacteria (LAB),isolated from dairy products from the Western Balkans and from the products from the Caucasusand Azerbaijan, for the production of GABA, selection of the best GABA-producing strains, andtheir detailed probiotic characterization. After selecting the best GABA-producing strain nextobjective was to examine the effects of live strain, as well as, its culture supernatants on an invitro inflammation models and on the clinical picture on experimental autoimmuneencephalomyelitis (EAE), an animal multiple sclerosis (MS) model. Using standard(CoreR/CoreF) primers, the presence of the gadB gene, which encodes glutamate decarboxylase,was detected at 83 (27,4 %) Lactobacillus spp., 15 (4,9 %) Lactococcus spp., 11 (3,6 %)Leuconostoc spp., 8 (2,64 %) Streptococcus spp., 6 (1,98 %) Enterococcus spp., and 1 (0,33 %)Pediococcus spp out of 303 tested strains. By TLC method, the production of GABA wasdetected in 4.6% of the strains, ie 14 strains, out of 124 tested strains, belonging to the speciesLactobacillus brevis and Lb. plantarum, while the concentration of produced GABA wasmeasured by HPLC. The simulated gastrointestinal conditions successfully survived 4 out of 7tested Lb. brevis strains: BGLMM10, BGLMM11, BGZLS10-17 and BGZLS30-2. Examinationof the four Lb. brevis strains safety status showed that strain BGZLS10-17 was sensitive to alltested antibiotics, BGLMM11 was resistant to kanamycin, BGLMM10 was resistant tokanamycin and tetracycline, strain BGZLS30-2 was resistant to ampicillin, kanamycin andtetracycline, while none of the 4 tested strains showed hemolytic or gelatinous activity. TheBGLMM10 strain exhibited the most potent antimicrobial activity, on 11 out of 18 pathogens,while BGZLS10-17 strain did not exhibit antimicrobial activity..
Pd(II) complexes with N-heteroaromatic hydrazone ligands: Anticancer activity, in silico and experimental target identification
No full textAnticancer activity of Pd complexes 1-5 with bidentate N-heteroaromatic hydrazone ligands was investigated on human acute monocytic leukemia (THP-1; cells in a suspension) and human mammary adenocarcinoma (MCF-7; two-dimensional layer and three-dimensional spheroid tumor model) cell lines. For the Pd(II) complexes with condensation products of ethyl hydrazainoacetate and quinoline-8-carboxaldehyde (complex 1) and 2-for-mylpyridine (complex 3), for which apoptosis was determined as a mechanism of anticancer activity, further investigation revealed that they arrest the cell cycle in G0/G1 phase, induce generation of reactive oxygen species and inhibit Topoisomerase I in vitro. In silico studies corroborate experimental findings that these complexes show topoisomerase inhibition activity in the micromolar range and indicate binding to a DNA's minor groove as another potential target. Based on the results obtained by circular dichroism and fluorescence spectroscopy measurements, the most active complexes are suitable to be delivered to a blood stream via human serum albumin
4-Aminoquinoline-based compounds as antileishmanial agents that inhibit the energy metabolism of Leishmania
No full textAmong neglected tropical diseases, leishmaniasis is one of the most relevant with an estimated 30,000 deaths annually. Existing therapies have serious drawbacks in safety, drug resistance, field-adapted application and cost; therefore, new safer and shorter treatments are needed for this disease. Here we report on the synthesis of novel 4-amino-7-chloroquinoline-based compounds with leishmanicidal activity, together with deeper insight into the mechanism of action of our previously published hit compound 1. New derivatives showed comparable activity to 1 against both promastigote and intracellular amastigote forms of Leishmania infantum, with IC50 lt 1 mu M. Furthermore, we have determined that compound 1 induced a decrease of intracellular ATP levels, as well as a mitochondrial depolarization, together with an alteration of plasma membrane permeability and a significant ROS production. The inhibition of the energy metabolism of Leishmania plays an important role in the leishmanicidal mechanism of this compound. In all, these results support the consideration of compound 1 for the future development of new leishmanicidal drugs
Novel sodium alkyl-1,3-disulfates, anionic biosurfactants produced from microbial polyesters
No full textA sodium alkyl disulfate mixture (SADM) synthesised from microbially produced 3-hydroxy fatty acids methyl esters (HFAMEs), showed 13-fold surface tension decrease when compared with the reference surfactant sodium dodecyl sulfate (SDS). Polyhydroxyalkanoates, accumulated by bacteria intracellularly when supplied with a mixture of fatty acids derived from hydrolysed rapeseed oil, were isolated, depolymerised and methylated to produce HFAMEs in very high yield (90%). A sequential chemical reduction and sulfation of the HFAMEs produced the sodium alkyl disulfates in high yields ( gt 65%). SADM performs also 1.3-times better than dodecyl (1,3) disulfate, in surface tension tests. SADM shows also the formation of a specific critical micelle concentration (CMC) at a concentration 21-fold lower than SDS. The wettability of the SADM mixture is similar to SDS but the foaming volume of SADM is 1.5-fold higher. The foam is also more stable with its volume decreasing 3 times slower over time compared to SDS at their respective CMC values. Established sulfation technologies in chemical manufacturing could use the 3-hydroxy fatty acids methyl esters moiety (3-HFAME) given its origin from rapeseed oil and the extra OH residue on 3-position in the molecule, which affords the opportunity to produce disulfate surfactants with a proven superior performance to monosulphated surfactants. Thus, not only addressing environmental issues by avoiding threats of deforestation and monocultivation associated with palm oil use but also achieve a higher performance with lower use of surfactants
Radiation effects on early phase of NT2/D1 neural differentiation in vitro
No full textPurpose: Widespread medical use of radiation in diagnosis, imaging and treatment of different central nervous system malignancies lead to various consequences. Aim of this study was to further elucidate mechanism of cell response to radiation and possible consequence on neural differentiation. Materials and methods: NT2/D1 cells that resemble neural progenitors were used as a model system. Undifferentiated NT2/D1 cells and NT2/D1 cells in the early phase of neural differentiation were irradiated with low (0.2 Gy) and moderate (2 Gy) doses of gamma radiation. The effect was analyzed on apoptosis, cell cycle, senescence, spheroid formation and the expression of genes and miRNAs involved in the regulation of pluripotency or neural differentiation. Results: Two grays of irradiation induced apoptosis, senescence and cell cycle arrest of NT2/D1 cells, accompanied with altered expression of several genes (SOX2, OCT4, SOX3, PAX6) and miRNAs (miR-219, miR-21, miR124-a). Presented results show that 2 Gy of radiation significantly affected early phase of neural differentiation in vitro. Conclusions: These results suggest that 2 Gy of radiation significantly affected early phase of neural differentiation and affect the population of neural progenitors. These findings might help in better understanding of side effects of radiotherapy in treatments of central nervous system malignancies.Peer-reviewed manuscript: [https://imagine.imgge.bg.ac.rs/handle/123456789/1629
Neutrophil extracellular traps-associated markers are elevated in patients with systemic lupus erythematosus
No full textNeutrophil extracellular traps (NETs) are the main source of autoantigens in systemic lupus erythematosus (SLE). The aim of this study was to evaluate the clinical importance of NETs-associated markers in SLE. We compared NETs-associated markers in SLE patients (n = 111) with healthy controls (n = 50). Moreover, in 35 patients with drug-naive SLE (n = 35), we investigated correlation between NETs-associated markers [DNase I concentration, myeloperoxidase (MPO) activity, anti-MPO antibodies, cell-free DNA (cfDNA), NETolytic activity] with serological parameters [anti-dsDNA antibodies, C3, C4 and B-cell activating factor (BAFF) levels] and disease activity measured by modified SLE Disease Activity Index (M-SLEDAI-2K). In comparison with healthy controls, SLE patients had higher cfDNA, MPO activity, anti-MPO antibodies (p lt 0.001), BAFF and DNase I concentration (p lt 0.01). Contrary, NETolytic activity was lower in SLE patients (p lt 0.05), despite higher concentration of DNase I. MPO activity and cfDNA levels showed correlation with DNase I concentration (p lt 0.001, p lt 0.01, respectively). BAFF levels correlated with cfDNA, DNase I concentration and MPO activity (p lt 0.05). Anti-dsDNA antibodies showed correlation with MPO activity (p lt 0.01), cfDNA and BAFF levels (p lt 0.001). Anti-dsDNA and C3 levels were independent predictors of M-SLEDAI-2K in multivariate analysis (p lt 0.01). We demonstrated that sera of SLE patients have decreased NETolytic activity, leading to increased levels of various NETs-associated markers, which correlate with anti-dsDNA antibodies in drug-naive SLE. We showed that BAFF participates in a complex relationship between NETosis and anti-dsDNA antibodies production. These findings have important implications for a better understanding of SLE pathogenesis and development of therapy that inhibits NETs persistence and disease progression