imagine (Institute of molecular genetics and genetic engineering)
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Assessment of hemostatic disturbances in women with established rheumatoid arthritis
Get PDFObjectives This study was aimed to assess hemostatic disturbances in female patients with established rheumatoid arthritis (RA) in relation to menopausal status and disease activity. Method Ninety women were included in the study, 42 patients and 48 age-matched healthy controls. There were no differences between the investigated groups regarding the presence of traditional cardiovascular risk factors. Two global hemostatic assays were employed, namely endogenous thrombin potential (ETP) and overall hemostasis potential (OHP). The parameters of the ETP assay (ETP, C-max, t-lag, t-max) and OHP assay (overall coagulation potential (OCP) and overall fibrinolytic potential (OFP)) were assessed. Moreover, the parameters of the fibrin clot (lag time, Max Abs, and slope) were measured by clot turbidity and scanning electron microscopy (SEM). Both patients and controls were divided into four subgroups according to menopause status. Results The premenopausal controls differed significantly from all other subgroups in terms of diminished levels of ETP (p = 0.02), C-max (p = 0.01), OCP (p = 0.02), OHP (p = 0.001), and Max Abs (p = 0.008), while OFP (p = 0.0001) was increased. This tendency was not seen in the premenopausal RA patients compared with the postmenopausal RA patients. SEM images showed denser clots composed of thinner fibers in samples from RA patients. The disease activity measured by DAS28 correlated with OCP and OHP (r = 0.54; p = 0.001 and r = 0.44; p = 0.003, respectively) indicating persistent hypercoagulable condition in the whole group of RA patients. Conclusions Our results point towards coagulation activation in premenopausal women with established RA. The patients were well characterized, which enabled assessment in a real-life setting
Technical overview of nuclear microsatellites for Fagus sp., and their utility in F. sylvatica from the central Balkans (Serbia)
No full textSelectively neutral nuclear microsatellites (SSRs) are available for Fagus sp., and have been used in F. sylvatica, an ecologically and economically most important European Fagus species. We provide a comprehensive technical overview of available Fagus sp. SSRs (185 loci), carry out meta-analysis of loci used in F. sylvatica (62 loci employed in 62 surveys), and validate a set of 16 loci in 45 individuals of this species from the central Balkans (Serbia). Erroneous usage of marker's names/authors is rather frequent, and loci successfully used in a number of studies are characterized by other authors by high prevalence of null alleles and even multilocus amplification products. Frequent occurrence of null alleles at FS4-46, used in 26 surveys to date, most likely indicates a failure to record multiple alleles at this locus. Twelve loci are reliable/informative in F. sylvatica from the Balkans (5-18 alleles/locus, H-E ranging from 0.523 to 0.850), while four loci are characterized by high prevalence of null alleles (sfc0161 and sfc1063) and multilocus amplification products (FS4-46 and Fagsyl_007038). Our findings are important for future population genetics and studies on adaptation of F. sylvatica to its environment, because the latter rely on both selectively neutral and adaptive markers under selection
Biodegradation of poly(epsilon-caprolactone) (PCL) and medium chain length polyhydroxyalkanoate (mcl-PHA) using whole cells and cell free protein preparations of Pseudomonas and Streptomyces strains grown on waste cooking oil
No full textPetrochemical plastics are generally recalcitrant to microbial degradation and accumulate in the environment. Biodegradable polymers obtained synthetically like poly(epsilon-caprolactone) (PCL) or poly-hydroxyalkanoates (PHA), obtained biotechnologically, have shown great potential as a replacement for petroleum-based plastics. Nevertheless, their biodegradation and environmental faith have been less examined. In this study, thin films of PCL (200 mu m) and medium chain length PHA (mcl-PHA, 70 M fraction of 3-hydroxyoctanoate and 30 M fraction of 3-hydroxydecanoate, 600 mu m) were exposed to total protein preparations (extracellular proteins combined with a crude cell extract) of soil isolates Pseudomonas chiororaphis B-561 and Streptomyces sp. BV315 that had been grown on waste cooking oil as a sole carbon source. Biodegradation potential of two polyesters was evaluated in buffer with total protein preparations and in a laboratory compost model system augmented with selected bacteria. Overall, PCL showed better biodegradation properties in comparison to mcl-PHA. Both materials showed surface erosion after 4-weeks of exposure to total protein preparations of both strains, with a moderate weight loss of 1.3% when P. chlororaphis13-561 was utilized. In laboratory compost model system PCL and mcl-PHA showed significant weight loss ranging from 13 to 17% when Streptomyces sp. BV315 culture was used. Similar weight loss of PCL and mcl-PHA was achieved for 4 and 8 weeks, respectively indicating slower degradation of mcl-PHA. Growth on waste cooking oil as a sole carbon source increased the potential of both tested strains to degrade PCL and mcl-PHA, making them good candidates for augmentation of compost cultures in waste management of both waste cooking oils and biodegradable polymers.This is the peer-reviewed version of the article: [https://imagine.imgge.bg.ac.rs/handle/123456789/1760
Subregion-specific Protective Effects of Fluoxetine and Clozapine on Parvalbumin Expression in Medial Prefrontal Cortex of Chronically Isolated Rats
No full textDysregulation of GABAergic system is becoming increasingly associated with depression, psychiatric disorder that imposes severe clinical, social and economic burden. Special attention is paid to the fast-spiking parvalbumin-positive (PV+) interneurons, GABAergic neurons which are highly susceptible to redox dysregulation and oxidative stress and implicated in a variety of psychiatric diseases. Here we analyzed the number of PV+ and cleaved caspase-3-positive (CC3+) cells in the rat medial prefrontal cortical (mPFC) subregions following chronic social isolation (CSIS), an animal model of depression and schizophrenia. Also, we examined potential protective effects of antidepressant fluoxetine (FLX) and atypical antipsychotic clozapine (CLZ) on the number of these cells in mPFC subregions, when applied parallel with CSIS in doses that correspond to therapeutically effective ones in patients. Immunofluorescence analysis revealed decreased number of PV+ cells in cingulate cortex area 1, prelimbic area (PrL), infralimbic area (IL) and dorsal peduncular cortex of the mPFC in isolated rats, which coincided with depressive- and anxiety-like behaviors. In addition, CSIS-induced increase in the number of CC3+ cells was detected in aforementioned subregions of mPFC. Treatments with either FLX or CLZ prevented behavioral changes, decrease in PV+ and increase in CC3+ cell numbers in PrL and IL subregions in isolated rats. These results indicate the importance of intact GABAergic signaling in these areas for resistance against CSIS-induced behavioral changes, as well as subregion-specific protective effects of FLX and CLZ in mPFC of CSIS rats
Production of bacterial nanocellulose (BNC) and its application as a solid support in transition metal catalysed cross-coupling reactions
Get PDFBacterial nanocellulose (BNC) emerged as an attractive advanced biomaterial that provides desirable properties such as high strength, lightweight, tailorable surface chemistry, hydrophilicity, and biodegradability. BNC was successfully obtained from a wide range of carbon sources including sugars derived from grass biomass using Komagataeibacter medellinensis ID13488 strain with yields up to 6 g L-1 in static fermentation. Produced BNC was utilized in straightforward catalyst preparation as a solid support for two different transition metals, palladium and copper with metal loading of 20 and 3 wt%, respectively. Sustainable catalysts were applied in the synthesis of valuable fine chemicals, such as biphenyl-4-amine and 4'-fluorobiphenyl-4-amine, used in drug discovery, perfumes and dye industries with excellent product yields of up to 99%. Pd/BNC catalyst was reused 4 times and applied in two consecutive reactions, Suzuki-Miyaura cross-coupling reaction followed by hydrogenation of nitro to amino group while Cu/BNC catalyst was examined in Chan-Lam coupling reaction. Overall, the environmentally benign process of obtaining nanocellulose from biomass, followed by its utilisation as a solid support in metal-catalysed reactions and its recovery has been described. These findings reveal that BNC is a good support material, and it can be used as a support for different catalytic systems.This is the peer-reviewed version of the article: Jeremic, S., Djokic, L., Ajdačić, V., Božinović, N., Pavlovic, V., Manojlović, D. D., Babu, R., Senthamaraikannan, R., Rojas, O., Opsenica, I., & Nikodinovic-Runic, J. (2019). Production of bacterial nanocellulose (BNC) and its application as a solid support in transition metal catalysed cross-coupling reactions. International Journal of Biological Macromolecules, 129, 351–360. [https://doi.org/10.1016/j.ijbiomac.2019.01.154][https://www.sciencedirect.com/science/article/abs/pii/S0141813018349213?via%3Dihub
Autistic traits and cognitive dysfunctions in children with patogenic copy number variants: a pilot study from Serbia
No full textIdentification and Characterization of New Laccase Biocatalysts from Pseudomonas Species Suitable for Degradation of Synthetic Textile Dyes
Get PDFLaccases are multicopper-oxidases with variety of biotechnological applications. While predominantly used, fungal laccases have limitations such as narrow pH and temperature range and their production via heterologous protein expression is more complex due to posttranslational modifications. In comparison, bacterial enzymes, including laccases, usually possess higher thermal and pH stability, and are more suitable for expression and genetic manipulations in bacterial expression hosts. Therefore, the aim of this study was to identify, recombinantly express, and characterize novel laccases from Pseudomonas spp. A combination of approaches including DNA sequence analysis, N-terminal protein sequencing, and genome sequencing data analysis for laccase amplification, cloning, and overexpression have been used. Four active recombinant laccases were obtained, one each from P. putida KT2440 and P. putida CA-3, and two from P. putida F6. The new laccases exhibited broad temperature and pH range and high thermal stability, as well as the potential to degrade selection of synthetic textile dyes. The best performing laccase was CopA from P. putida F6 which degraded five out of seven tested dyes, including Amido Black 10B, Brom Cresol Purple, Evans Blue, Reactive Black 5, and Remazol Brilliant Blue. This work highlighted species of Pseudomonas genus as still being good sources of biocatalytically relevant enzymes
SOX3 can promote the malignant behavior of glioblastoma cells
No full textPurposeGlioblastoma is the most common and lethal adult brain tumor. Despite current therapeutic strategies, including surgery, radiation and chemotherapy, the median survival of glioblastoma patients is 15months. The development of this tumor depends on a sub-population of glioblastoma stem cells governing tumor propagation and therapy resistance. SOX3 plays a role in both normal neural development and carcinogenesis. However, little is known about its role in glioblastoma. Thus, the aim of this work was to elucidate the role of SOX3 in glioblastoma.MethodsSOX3 expression was assessed using real-time quantitative PCR (RT-qPCR), Western blotting and immunohistochemistry. MTT, immunocytochemistry and Transwell assays were used to evaluate the effects of exogenous SOX3 overexpression on the viability, proliferation, migration and invasion of glioblastoma cells, respectively. The expression of Hedgehog signaling pathway components and autophagy markers was assessed using RT-qPCR and Western blot analyses, respectively.ResultsHigher levels of SOX3 expression were detected in a subset of primary glioblastoma samples compared to those in non-tumoral brain tissues. Exogenous overexpression of this gene was found to increase the proliferation, viability, migration and invasion of glioblastoma cells. We also found that SOX3 up-regulation was accompanied by an enhanced activity of the Hedgehog signaling pathway and by suppression of autophagy in glioblastoma cells. Additionally, we found that SOX3 expression was elevated in patient-derived glioblastoma stem cells, as well as in oncospheres derived from glioblastoma cell lines, compared to their differentiated counterparts, implying that SOX3 expression is associated with the undifferentiated state of glioblastoma cells.ConclusionFrom our data we conclude that SOX3 can promote the malignant behavior of glioblastoma cells
Synthesis and Characterization of 3-(1-((3,4-Dihydroxyphenethyl)amino)ethylidene)-chroman-2,4-dione as a Potential Antitumor Agent
Get PDFThe newly synthesized coumarin derivative with dopamine, 3-(1-((3,4-dihydroxyphenethyDamino)ethylidene)-chroman-2,4-dione, was completely structurally characterized by X-ray crystallography. It was shown that several types of hydrogen bonds are present, which additionally stabilize the structure. The compound was tested in vitro against different cell lines, healthy human keratinocyte HaCaT, cervical squamous cell carcinoma SiHa, breast carcinoma MCF7, and hepatocellular carcinoma HepG2. Compared to control, the new derivate showed a stronger effect on both healthy and carcinoma cell lines, with the most prominent effect on the breast carcinoma MCF7 cell line. The molecular docking study, obtained for ten different conformations of the new compound, showed its inhibitory nature against CDKS protein. Lower inhibition constant, relative to one of 4-OH-coumarine, proved stronger and more numerous interactions with CDKS protein. These interactions were carefully examined for both parent molecule and derivative and explained from a structural point of view
Radiation effects on early phase of NT2/D1 neural differentiation in vitro
Get PDFPurpose: 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.This is the peer reviewed version of the paper: Stanisavljević Ninković, D., Popović, J., Petrović, I., Davidović, S., Atkinson, M. J., Anastasov, N., & Stevanović, M. (2019). Radiation effects on early phase of NT2/D1 neural differentiation in vitro. International Journal of Radiation Biology, 95(12), 1627–1639. [https://doi.org/10.1080/09553002.2019.1665207]Published version: [https://imagine.imgge.bg.ac.rs/handle/123456789/1206