Phenotypical characteristics of peritoneal macrophages of two rat strains during inflammatory response: the importance of gut microbiota representatives

Predmet istraživanja ove doktorske disertacije je ispitivanje promena u fenotipu i aktivnosti makrofaga peritonealne šupljine Albino Oxford (AO) i Dark Agouti (DA) pacova tokom razvoja inflamatornog odgovora koji je indukovan delovanjem bakterija mikrobiote creva direktno, intraperitonealnom (i.p.) primenom, ili indirektno, tokom razvoja kolitisa, kao i ispitivanje modulacije ovih promena primenom probiotske bakterije Lactobacillus rhamnosus (L. rhamnosus). Rezultati su pokazali da i.p. primena umrtvljenih E. coli ili Enterococcus spp. dovodi do izraženije inflamatorne polarizacije makrofaga peritonealne šupljine DA pacova u odnosu na AO soj, dok i.p. primena L. rhamnosus ima veći potencijal da spreči funkcionalne promene do kojih dovodi in vitro stimulacija komensalnim bakterijama u makrofagima pacova AO soja. Tokom razvoja kolitisa makrofagi DA pacova produkuju više IL-6 u odnosu na AO soj, a značajno manja smrtnost AO pacova je praćena intenzivnijom produkcijom IL-10 u makrofagima u odgovoru na dodatnu stimulaciju komensalnim bakterijama in vitro i višim nivoom antitela klase IgA specifičnih za E. coli u serumu. Rani postnatalni oralni tretman bakterijom L. rhamnosus u DA pacovima povećava raznovrsnost Bifidobacterium vrsta u fecesu, a nakon indukcije kolitisa u adultnom dobu smanjuje stepen ulceracije kolona, povećava zastupljenost rezidentnih makrofaga peritonealne šupljine i suprimira povećanja produkcije NO i IL-10 stimulisana komensalnim bakterijama in vitro. Nalazi ukazuju na značaj genetski determinisane predispozicije ka određenom obrascu aktivacije makrofaga za promene u fenotipu i aktivnosti tokom razvoja inflamatornog odgovora koji je indukovan delovanjem bakterija mikrobiote creva, kao i za modulaciju ovih promena primenom probiotske bakterije L. rhamnosus.The subject of this doctoral dissertation is the study of changes in the phenotype and activity of peritoneal cavity macrophages from Albino Oxford (AO) and Dark Agouti (DA) rats during the development of inflammatory response induced by intestinal microbiota directly, intraperitoneally (i.p.), or indirectly, through development of colitis, as well as study of the modulation of these changes using the probiotic bacterium Lactobacillus rhamnosus (L. rhamnosus). I.p. application of killed E. coli or Enterococcus spp. leads to a more pronounced inflammatory polarization of the peritoneal cavity macrophages from DA rats compared the AO strain, while i.p. application of L. rhamnosus has a greater potential to prevent functional changes caused by in vitro stimulation with commensal bacteria in AO rat macrophages. During the development of colitis, DA rat macrophages produce more IL-6 than the AO strain, and significantly lower AO rat mortality was accompanied by more intense IL-10 production in macrophages in response to additional stimulation with commensal bacteria in vitro and higher levels of IgA-class antibodies specific to E. coli in the serum. Early postnatal oral treatment with L. rhamnosus in DA rats increased the diversity of Bifidobacterium species in feces, and after induction of colitis in adulthood reduced the degree of colonic ulceration, increased the presence of resident peritoneal cavity macrophages and suppressed the increased production of NO and IL-10 stimulated by commensal bacteria . The findings indicate the importance of genetically determined predisposition to a certain pattern of macrophage activation for changes in phenotype and activity during the development of inflammatory response induced by intestinal microbiota, as well as for modulation of these changes using probiotic bacterium L. rhamnosus

Od ustanka do međunarodnog priznanja - vojska u stvaranju državnog identiteta Kneževine Srbije

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Field reversal DC machine braking with DCM drive

This study was supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia, and these results are parts of the Grant No. 451-03-68/2022-14/200132 with University of Kragujevac Faculty of Technical Sciences Čačak.This paper gives a detailed description of implementation of braking by field reversal at laboratory setup of DC machine with modern Siemens DCM drive. The main focus of the paper is implementation of braking procedure of the DC machine using field reversal through didactic approach suited to students of electrical engineering within the course of the electric drives at Faculty of Technical Sciences Čačak. The laboratory test bench allows students to upgrade parts of the setup and test different operation regimes of DC and AC machines, as well as gaining experience of tuning and optimization of the overall motor-load system.Publishe

Military cliques in the Serbian strategic culture of the 20th century

Throughout the entire 20th century, the Serbian strategic culture was "torn" by various discontinuities, from the state system, through foreign policy and ideological, to strategic and doctrinal orientation, and it can be said that discontinuity is almost its only constant. Nevertheless, the existence of military cliques can be considered to be one of the constants of the Serbian strategic culture. They followed the Serbian Army and influenced the political life of the Kingdom of Serbia. The dynastic coup of May 1903 can be taken as an example of their actions. After that, younger conspirators established the so-called Black Hand, whose leader was Colonel Apis, who was shot after the Thessaloniki process in 1917. The interwar period was marked by the action of the so-called White Hand, under the leadership of General Petar Živković, which was active until the assassination of King Alexander Karađorđević in Marseilles in 1934. The Second Yugoslavia even had a formalized "military clique" represented in the League of Communists, which was an integral part of the League of Communists of Yugoslavia at federal level, together with the republican leagues. Thus, the influence of the party on the military organization was secured, and the influence of the army in the society was also "secured" by its representation in the party forum at federal level. Consequently, the Serbian political elite welcomed the dissolution of the SFRY with perhaps the only constant in its strategic culture, and that is the reliance on the military in resolving political crises, which can be vividly presented by the statement of Slobodan Milosević: "We do not know how to work, but we know how to fight"

Organized crime in Western Balkans: Case Serbia

This paper discusses the common heritage of the Western Balkans in connection with organized crime, which, it can be claimed, did not significantly manifest under socialism. The disintegration of the former Yugoslavia led to war, destruction, and overall increase in crime, including organized crime. The factors that especially contributed to that, besides the war, are: the transition process, the increase in the level of corruption, inefficient functioning of the state apparatus, economic decline, and trade sanctions imposed by the United Nations which resulted in a sort of semi-legal cooperation between governmental authorities and organized criminal groups in the Western Balkans. Local wars in the former Yugoslavia enabled the expansion of illegal arms trafficking, theft and smuggling of motor vehicles, and smuggling of all kinds of excise goods, especially oil and cigarettes. The trafficking of narcotics became a very lucrative and widespread activity of criminal groups, with the geo-strategic position of the Western Balkans countries being one of the contributing factors. The beginning of democratic changes in Serbia marked the commencement of the fight against organized crime. It was also the start of the creation of the legal regulatory system to counter organized crime, which involves interstate cooperation at both the regional and global level, as well as the adoption of international conventions and standards. Consequently, a special police unit for combating organized crime was established within the Serbian Ministry of Interior. The Republic of Serbia, like most countries in the region, has adopted a series of legislative measures to combat organized crime, which come from experiences of other countries that have shown good results. Specific aspects of the investigative procedure and collecting evidence of organized crime cannot be achieved using conventional methods of investigation, such as home, other premises and persons’ search warrants, temporary confiscation of items, questioning of the accused, witnesses’ hearing, etc. In these efforts the basic dilemma is the level of proportionality between security and freedom, because the methods of an undercover investigation are special methods that are used to temporarily restrict the fundamental rights and freedoms of men, for the purposes of combating complex forms of crime

Lactobacillus rhamnosus Affects Rat Peritoneal Cavity Cell Response to Stimulation with Gut Microbiota: Focus on the Host Innate Immunity

Gut microbiota contribute to shaping the immune repertoire of the host, whereas probiotics may exert beneficial effects by modulating immune responses. Having in mind the differences in both the composition of gut microbiota and the immune response between rats of Albino Oxford (AO) and Dark Agouti (DA) rat strains, we investigated if intraperitoneal (i.p.) injection of live Lactobacillus rhamnosus (LB) may influence peri-toneal cavity cell response to invitro treatments with selected microbiota in the rat strain-dependent manner. Peritoneal cavity cells from AO and DA rats were lavaged two (d2) and seven days (d7) following i.p. injection with LB and tested for NO, urea, and H2O2 release basally, or upon invitro stimulation with autologous E.coli and Enterococcus spp. Whereas the single i.p. injection of LB nearly depleted resident macrophages and increased the proportion of small inflammatory macrophages and monocytes on d2 in both rat strains, greater proportion of MHCIIhiCD163− and CCR7+ cells and increased NO/diminished H2O2 release in DA compared with AO rats suggest a more intense inflammatory prim-ing by LB in this rat strain. Even though E.coli- and/or Enterococcus spp.-induced rise in H2O2 release invitro was abrogated by LB in cells from both rat strains, LB prevented microbiota-induced increase in NO/urea ratio only in cells from AO and augmented it in cells from DA rats. Thus, the immunomodulatory properties may not be constant for particular probiotic bacteria, but shaped by innate immunity of the host.https://rdcu.be/cxyn

17β-Estradiol and genistein affect macrophage inflammatory cytokine production during aging in sex-specific manner

Aging differently affects the expression of estrogen receptors alpha and beta (ERα and ERβ) and Toll-like receptors (TLR4) on peritoneal cavity cells of male and female rats. We explored the involvement of ERα and ERβ in the in vitro treatment of LPS-stimulated peritoneal macrophages with 17β-estradiol (which stimulates both receptors) or genistein (which is predominantly an ERβ agonist) on inflammatory cytokine secretion from young (3 months old) and middle-aged (16 months old) female and male AO rats. Aging diminished the proportion of TLR4+ cells and secretion of IL-1β and IL-6 in macrophages from female rats while the effect on male rat macrophages was opposite. 17βestradiol increased IL-1β secretion by middle-aged females’ macrophages via ERα, and suppressed it in cells from young females via ERβ. Genistein-induced decrease of IL-1β in macrophages from all experimental groups was probably mediated by ERβ. 17β-estradiol augmented IL-6 secretion by cells from all experimental groups via ERα while genistein diminished it in all females’ and in middle-aged male rats’ macrophages by activating ERβ. However, genistein increased IL-6 secretion from macrophages of young male rats via ERα. Although 17β-estradiol and genistein stimulated secretion of macrophage inflammatory cytokines via ERα and suppressed it probably via ERβ, their modulatory actions were determined by aging-induced changes in macrophage ERs expression and possible ERα / ERβ interactions (Supported by Ministry of Education, Science and Technological development, Republic of Serbia, Grant No 175050)

Potential impact of early-life probiotic supplementation on peritoneal macrophage function

Clinical and animal trials show that early life probiotic consumption provides health benefits in adult life by modulating the immune response. We tested the effects of early life oral consumption of the probiotic Lactobacillus rhamnosus on the function and phenotype of rat peritoneal cavity cells in a model of induced colitis. For the first month of their lives, rats were either fed with an aqueous probiotic bacteria suspension (LB group) or tap water (control group). When the rats grew to 3 months old, we studied the response of their peritoneal macrophages to autologous fecal bacteria stimulation in vitro, both before and after colitis induction (TNBS 40mg/kg of body mass in 50% ethanol). Compared to the controls, the peritoneal cavity cells of the LB group produced less nitric oxide (NO) and had an increased proportion of CD163+ cells. The rats in the LB group have shown milder symptoms of colitis (shorter length of colon under necrosis, less severe submucosal infiltration, lesser degree of colonic wall thickening), along with a diminished increase of peritoneal proinflammatory CCR7+ cells and blunted NO production in response to stimulation by autologous fecal bacteria. Our results may indicate that early oral probiotic administration attenuates macrophage responses to fecal bacteria, which are the primary cause of tissue inflammation and necrosis in chemically induced colitis models, and that this attenuation may be involved in improving the health of colitic rats

NMDA Receptor Antagonist Memantine Ameliorates Experimental Autoimmune Encephalomyelitis in Aged Rats

Aging is closely related to the main aspects of multiple sclerosis (MS). The average age of the MS population is increasing and the number of elderly MS patients is expected to increase. In addition to neurons, N-methyl-D-aspartate receptors (NMDARs) are also expressed on non-neuronal cells, such as immune cells. The aim of this study was to investigate the role of NMDARs in experimental autoimmune encephalomyelitis (EAE) in young and aged rats. Memantine, a non-competitive NMDAR antagonist, was administered to young and aged Dark Agouti rats from day 7 after immunization. Antagonizing NMDARs had a more favourable effect on clinical disease, reactivation, and apoptosis of CD4+ T cells in the target organ of aged EAE rats. The expression of the fractalkine receptor CX3CR1 was increased in memantine-treated rats, but to a greater extent in aged rats. Additionally, memantine increased Nrf2 and Nrf2-regulated enzymes’ mRNA expression in brain tissue. The concentrations of superoxide anion radicals, malondialdehyde, and advanced oxidation protein products in brain tissue were consistent with previous results. Overall, our results suggest that NMDARs play a more important role in the pathogenesis of EAE in aged than in young rats

Morfo-fiziološka proučavanja izolata Alternaria spp. poreklom sa celera

Genus Alternaria represents one of the most important pathogens of Apiaceae family. Alternariose symptoms may develop on foliage causing leaf spot, necrosis and eventually drying and black root. Celery plants with characteristic symptoms were collected during 2012 and 2013 in Serbia. The preliminary identification was done by analysis of morphological characteristics of conidia. After identification cultural characteristics, growth rate and spore morphology were examined and compared based on influence of ecological factors such as nutrient medium, temperature, pH value of the medium and light quality.Vrste roda Alternaria predstavljaju jedne od najznačajnijih gljiva patogena familije Apiaceae. Promene koje izazivaju na biljkama domaćinima su mrka trulež korena, pegavost lišća, sušenje listova i lisnih preteljki. Tokom 2012. i 2013. godine na teritoriji Srbije, prikupljeni su uzorci celera sa karakterističnim simptomima pegavosti lišća i sušenja listova. Identifikacija izolata je izvršena analizom morfoloških karakteristika spora gljive. Nakon identifikacije izvršena su uporedna proučavanja uticaja različitih hranljivih podloga, temperatura, pH vrednosti podloga i kvaliteta svetlosti na morfološke karakteristike izolata, porast kolonija i morfologiju spora