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Native bovine hemoglobin reduces differentiation capacity of mesenchymal stromal cells in vitro
No full textWe have tested in vitro effects of hemoglobin from bovine slaughterhouse blood (BHb) on stromal cells of mesodermal origin, with an aim to explore its use as a component of cell culture media. Human peripheral blood mesenchymal stromal cells (PB-MSCs) and three mouse cell lines (ATDC5, MC3T3-E1 and 3T3-L1) were employed to study BHb effects on their growth and migration. The cells multilineage differentiation capacity in the presence of BHb was evaluated after induced differentiation, by histochemical staining and by RT-PCR analysis of the expression of genes specific for chondrogenic, adipogenic and osteogenic lineages. The effects of BHb on the cell proliferation and motility were dependent on both, cell type and BHb concentration (0.1 mu M,1 mu M and 10 mu M). In the lowest concentration (0.1 mu M) BHb showed the least prominent effect on the cell proliferation and migration. In this concentration BHb reduced the differentiation capacity of all tested cells and its effect was dependent of composition of induction medium and the culture period. Obtained data suggest that BHb has the potential to be used as a component of cell culture media through maintaining proliferation and reducing differentiation capacity of mesenchymal stromal cells
Aronia juice consumption prior to half-marathon race can acutely affect platelet activation in recreational runners
Get PDFLong-distance running, especially in non-professional runners, can increase cardiac arrest risk by enhancing platelet activation and aggregation. Polyphenols can exert cardioprotective effects by positively influencing platelet function. This study aimed to examine the acute effects of polyphenol-rich aronia juice consumption, before simulation of a half-marathon race, on platelet activation and aggregation with leukocytes in recreational runners. In this acute crossover study,10 healthy male runners (age 30.8 +/- 2.3 years) consumed breakfast with 200 mL, of aronia juice or 200 mL of placebo. They warmed-up and ran a simulated half-marathon race (21.1 km). Blood was collected at baseline, and at 15 min, 1 h, and 24 h after the run. All variables were analyzed with 4 (time) x 2 (group)ANOVA with repeated measures on both factors. Results revealed a significant effect of group on platelet activation parameters: P-selectin and CPIIb-IIIa expressions significantly decreased in the aronia group compared with the placebo group (F-[1.(9]) = 10.282, p = 0.011 and F-[1.9] = 7.860, p = 0.021, respectively). The effect of rime was significant on both platelet aggregation markers: platelet-monocyte and platelet-neutrophil aggregates were significantly lower after the race (F-[(3).7] = 4.227, p = 0.014 and F-[3.7] = 70.065, p = 0.000, respectively), with changes more pronounced in the later. All effects remained when platelets were exposed to an agonist. These results suggest that aronia consumption could counteract the half-marathon race-induced changes in platelet function. Novelty Aronia juice consumption significantly decreased the expression of platelet activation markers but did not affect platelet aggregation. The race itself did significantly reduce platelet-neutrophil aggregation. Aronia juice may serve as a supplement beverage for recreational runners to alleviate enhanced platelet reactivity caused by prolonged running.This is the peer-reviewed version of the article: Stevanović, V. B.; Pantović, A.; Krga, I. S.; Zeković, M. T.; Šarac, I. R.; Glibetić, M. D.; Vidović, N. Đ. Aronia Juice Consumption Prior to Half-Marathon Race Can Acutely Affect Platelet Activation in Recreational Runners. Applied Physiology Nutrition & Metabolism 2020, 45 (4), 393–400. [https://doi.org/10.1139/apnm-2019-0267
Anaerobic digestion of sludge over a decade's work of the WWTP in Subotica
Get PDFOstaci iz prečišćavanja otpadnih voda predstavljaju ekološki rizik, iziskuju rešenje, uvođenje raznih tehnologija na liniji mulja. Anaerobno digestovanje je jedno od mogućnosti rešavanja ostataka iz tretmana gradskih komunalnih otpadnih voda. Linija mulja kao tehnološka celina na PPOV u Subotici beleži decenijski rad. U radu će se prikazati rezultati anaerobne digestije i proizvodnje biogasa u ovom decenijskom periodu u PPOV Subotica.Wastewater treatment residues present ecological risks, require solutions and utilization of various technologies on the sludge lines. Anaerobic digestion is one of the possibilities for solving the residues of municipal communal wastewater treatment. The sludge line as a technological unit of WWTP Subotica has been working for а decade. This paper will show the results of anaerobic digestion and biogas production in this decade in WWTP Subotica
Hydroxyurea-induced senescent peripheral blood mesenchymal stromal cells inhibit bystander cell proliferation of JAK2V617F-positive human erythroleukemia cells
Get PDFHydroxyurea (HU) is a nonalkylating antineoplastic agent used in the treatment of hematological malignancies. HU is a DNA replication stress inducer, and as such, it may induce a premature senescence-like cell phenotype; however, its repercussion on bystander cell proliferation has not been revealed so far. Our results indicate that HU strongly inhibited peripheral blood mesenchymal stromal cells (PBMSC) proliferation by cell cycle arrest in S phase, and that, consequently, PBMSC acquire senescence-related phenotypical changes. HU-treated PBMSC display increased senescence-associated beta-galactosidase levels and p16(INK4) expression, as well as DNA damage response and genotoxic effects, evidenced by expression of gamma H2A.X and micronuclei. Moreover, HU-induced PBMSC senescence is mediated by increased reactive oxygen species (ROS) levels, as demonstrated by the inhibition of senescence markers in the presence of ROS scavenger N-acetylcysteine and NADPH oxidase inhibitor Apocynin. To determine the HU-induced bystander effect, we used the JAK2V617F-positive human erythroleukemia 92.1.7 (HEL) cells. Co-culture with HU-induced senescent PBMSC (HU-S-PBMSC) strongly inhibited bystander HEL cell proliferation, and this effect is mediated by both ROS and transforming growth factor (TGF)-beta expression. Besides induction of premature senescence, HU educates PBMSC toward an inhibitory phenotype of HEL cell proliferation. Finally, our study contributes to the understanding of the role of HU-induced PBMSC senescence as a potential adjuvant in hematological malignancy therapies
Candidatus neoehrlichia sp. (fu98) and borrelia burgdorferi sensu lato in red foxes (vulpes vulpes) from Serbia
Get PDFHuman activities such as deforestation, urbanization, and environmental pollution lead to a reduction in the spatial boundary between wild animals, domestic animals and humans. These activities increase the risk for the emergence of pathogens from the sylvatic cycle in the population of domestic animals and humans. Foxes are recognized as potential reservoirs for a number of bacterial pathogens of medical and public health concern. The aim of the present study was to investigate the prevalence and spatial distribution of bacterial tick-borne pathogens from the Anaplasmataceae family, Borrelia burgdorferi sensu lato (s.l.), Rickettsia spp., Coxiella burnetii, Francisella tularensis, Bartonella spp., in the red fox population from Serbia and to discuss the obtained results from the epidemiological point of view. Legally hunted red foxes (Vulpes vulpes) from 14 localities in Serbia were included in the study and spleen samples from 129 animals were tested with conventional PCR assays for the presence of bacterial tick-borne pathogens. DNA of Candidatus Neoehrlichia sp. (FU98), Borrelia burgdorferi sensu stricto, Borrelia lusitaniae, and Borrelia garinii was detected in 6 (4.7%), 1 (0.8%), 2 (1.6%) and 1 (0.8%) animals, respectively. Co-infection by Candidatus Neoehrlichia sp. (FU98) and B. garinii was detected in one animal. All samples were negative for other tested bacterial tick-borne pathogens. The results of the present study indicate the potential role of foxes in natural cycles of Candidatus Neoehrlichia sp. (FU98) and causative agents of Lyme borreliosis in the investigated areas. Further research is required to elucidate the role of foxes in the epidemiology of these and other tick-borne zoonotic pathogens in the Republic of Serbia
Improving the reporting quality of intervention trials addressing the inter-individual variability in response to the consumption of plant bioactives: quality index and recommendations
Get PDFKey messages The reporting quality of human studies on inter-individual variation in response to plant bioactives is generally low and should be significantly improved. There is no specific guidance for reporting studies on inter-individual variation in response to plant bioactives. The assessment of reporting quality using a scale approach is considered a valuable tool in assessing compliance with the recommendations in the submission phase or during the reviewing process. It also provides a quantitative measure of the quality of studies to be used in meta-analysis. Eleven reporting criteria were developed and supported by detailed definitions and guidance for their scoring. The POSITIVe quality index was tested and demonstrated to be valid, reliable, and responsive. The use of the quality index and its supporting explanatory material (dictionary) as a guide for researchers, peer-reviewers, and journal editors will foster further complete and transparent reporting of data on inter-individual variability. The criteria used in the quality index can serve as additional guidance to inform the design and conduction of further studies on inter-individual variations in response to plant bioactives. Better reporting is expected to lead to a better understanding of the mechanisms and factors involved and thus better study designs with greater impact on policies and practice. Purpose The quality of the study design and data reporting in human trials dealing with the inter-individual variability in response to the consumption of plant bioactives is, in general, low. There is a lack of recommendations supporting the scientific community on this topic. This study aimed at developing a quality index to assist the assessment of the reporting quality of intervention trials addressing the inter-individual variability in response to plant bioactive consumption. Recommendations for better designing and reporting studies were discussed. Methods The selection of the parameters used for the development of the quality index was carried out in agreement with the scientific community through a survey. Parameters were defined, grouped into categories, and scored for different quality levels. The applicability of the scoring system was tested in terms of consistency and effort, and its validity was assessed by comparison with a simultaneous evaluation by experts' criteria. Results The "POSITIVe quality index" included 11 reporting criteria grouped into four categories (Statistics, Reporting, Data presentation, and Individual data availability). It was supported by detailed definitions and guidance for their scoring. The quality index score was tested, and the index demonstrated to be valid, reliable, and responsive. Conclusions The evaluation of the reporting quality of studies addressing inter-individual variability in response to plant bioactives highlighted the aspects requiring major improvements. Specific tools and recommendations favoring a complete and transparent reporting on inter-individual variability have been provided to support the scientific community on this field
Diversity of Lyme borreliosis spirochetes isolated from ticks in Serbia
No full textSpirochetes from the Borrelia burgdorferi sensu lato (s.l.). (Spirochaetales: Spirochaetaceae) species complex, including the causative agents of Lyme borreliosis, have been isolated from ticks, vertebrate reservoirs and humans. Previous analyses based on direct molecular detection in ticks indicated a considerable diversity of B. burgdorferi s.l. complex in Serbia. The present study aimed (a) to isolate borrelia strains from Serbia; (b) to determine their genotypic characteristics; and (c) to establish a collection of viable B. burgdorferi s.l. strains for further biological, ecological and genetic studies. For the present study, 231 adult Ixodes ricinus (Ixodida: Ixodidae) ticks from 16 ecologically different localities in Serbia were individually processed to cultivate B. burgdorferi s.l. This led to the isolation of 36 strains. A hbb gene quantitative real-time polymerase chain reaction (PCR) based on melting temperature determination and ospA gene sequencing were used to genotype the isolated spirochetes. The species identified based on the hbb gene real-time PCR were: Borrelia lusitaniae (44.4%), Borrelia afzelii (36.1%), Borrelia garinii (13.9%) and Borrelia valaisiana (5.6%), whereas the ospA sequence analysis revealed the occurrence of Borrelia bavariensis. This is the first report of the isolation of B. lusitaniae, B. garinii, B. bavariensis and B. valaisiana strains in Serbia
Trichinella spp. In wild mesocarnivores in an endemic setting
Get PDFHuman trichinellosis and Trichinella infection in pigs are both still endemic in the Balkans, including Serbia. Because of the flow between the sylvatic and the domestic cycle of Trichinella spp., monitoring wildlife has been recommended for the risk assessment of Trichinella spp. infection in swine. We have previously shown the presence of Trichinella infection in wild carnivores including the wolf and the golden jackal, and here we report on Trichinella infection in several other mesocarnivore species. From a total of 469 animals collected between 1994 and 2013, Trichinella larvae were detected in 29 (6.2%, 95% CI = 4.0-8.4) animals, including 14 red foxes (4.7%), 7 wild cats (35%), 5 beech martens (4.8%), 2 pine martens (16.7%), and 1 European badger (6.25%). No Trichinella larvae were detected in the examined specimens of European polecats, steppe polecats and European otters. Species identification of the Trichinella larvae performed for 18 positive samples revealed T. spiralis in 77.8% and T. britovi in 22.2% of the isolates. Both species were detected in red foxes and wild cats. The predominance of T. spiralis in wildlife in Serbia indicates the (past or present) spillover of this pathogen from domestic to wild animals
The interplay between copper(II), human serum albumin, fatty acids, and carbonylating agent interferes with Cys 34 thiol reactivity and copper binding
No full textCys34 thiol group of human serum albumin (HSA) represents major plasma antioxidant. Its reactivity is influenced by multiple factors. The influence of fatty acids (FA; saturated, mono, and poly unsaturated acids from fish oil) binding to HSA, on copper(II) binding affinity and Cys34 thiol group accessibility/reactivity, in the presence of carbonylation agent (methylglyoxal, MG) was examined. HSA-copper(II) content, thiol group reactivity, and HSA carbonylation level were monitored spectrophotometrically. Changes in HSA were followed by fluorescence spectroscopy and native PAG electrophoresis. FA/HSA molar ratio was screened by GC. Together, binding of copper(II) ions and FA to HSA increase the reactivity of Cys34 thiol group (depending on the type of FA), with constant contribution of copper(II) ions of one-third. Carbonylation of FA-HSA-Cu(II) complexes caused a decrease in the Cys34 thiol group content, accompanied by a decrease in the content of HSA-bound copper. The carbonylation level of guanidine groups was not affected by FAs and copper(II) binding. Fluorescent emission spectra of FA-HSA-Cu(II)-MG complexes showed conformational changes in HSA molecule. Although binding of fatty acids and copper ions caused a significant increase in the thiol group reactivity, Cys34 thiol from FA-HSA-Cu(II) complexes reacted with MG in smaller extent than expected, probably as a consequence of conformational changes introduced by carbonylation. Increase in the percentage of reacted-free thiol groups with MG (due to FA and copper binding) may not seem to be very significant, but it is very important in complex biological systems, where catalytic metal is present. [GRAPHICS]
Epizootiološko-epidemiološki značaj zlatnog šakala (Canis aureus) u održavanju vektorima prenosivih zoonoza na teritoriji Republike Srbije
Get PDFepizootiological and epidemiological role of the golden jackals (Canis aureus) as potential reservoirs for zoonotic vector-borne pathogens, for the first time in Serbia. Jackal spleens and collected ticks were tested by molecular methods (qPCR, conventional PCR, RFLP, sequencing) for the presence of vector-borne pathogens from the genera: Babesia spp., Rickettsia spp., Anaplasma spp., Ehrlichia spp., Francisella spp., Bartonella spp., Leishmania spp., and species from Borrelia burgdorferi sensu lato complex and Coxiella burneti. A total of 216 jackals originating from 10 localities throughout the territory of the Republic of Serbia were analyzed. Beside that, 118 ticks collected from jackals were also tested. The collected ticks belonged to one of three species: Ixodes ricinus (54,2%), Dermacentor reticulatus (39,8%) i Haemaphysalis concinna (5,9%). The presence of DNA of Babesia canis with the prevalence of 4.2% (9/216), Anaplasma phagocytophilum (0.9%, 2/216) and Leishmania spp. (6.9%, 15/216) was confirmed by molecular methods in the spleen of the tested jackals. In ticks collected from jackals, the presence of DNA from two Babesia spp. (Babesia canis, Babesia microti), Anaplasma phagocytophilum and three species from the Borrelia burgdorferi s.l. complex (Borrelia garinii, Borrelia lusitaniae, Borrelia valaisiana) was confirmed...Istraživanjem sprovedenim u okviru ove doktorske disertacije, po prvi put u Srbiji, analiziran je epizootiološko-epidemiološki značaj šakala (Canis aureus) kao potencijalnih rezervoara za uzročnike zoonoza koje se prenose vektorima. Molekularnim metodama (qPCR, konvencionalni PCR, RFLP, sekvenciranje) u slezinama šakala i krpeljima sakupljenim sa šakala, dokazivano je prisustvo vektorima prenosivih patogena iz rodova: Babesia spp., Rickettsia spp., Anaplasma spp., Ehrlichia spp., Francisella spp., Bartonella spp., Leishmania spp., i vrsta u okviru Borrelia burgdorferi sensu lato kompleksa i Coxiella burneti. Ukupno je analizirano 216 šakala poreklom sa 10 lokaliteta širom teritorije Republike Srbije. Na prisustvo patogena ispitano je i 118 sakupljenih krpelja koji su pripadali jednoj od tri vrste: Ixodes ricinus (54,2%), Dermacentor reticulatus (39,8%) i Haemaphysalis concinna (5,9%). Molekularnim metodama u slezinama šakala potvrđeno je prisustvo DNK Babesia canis sa prevalencijom od 4,2% (9/216), Anaplasma phagocytophilum (0,9%, 2/216) i Leishmania spp. (6,9%, 15/216). U krpeljima sakupljenim sa šakala potvrđeno je prisustvo DNK dve vrste Babesia (Babesia canis, Babesia microti), Anaplasma phagocytophilum i tri vrste Borrelia burgdorferi s.l. kompleksa (Borrelia garinii, Borrelia lusitaniae, Borrelia valaisiana)..