Institute of Virology, Vaccines and Sera “Torlak”

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    983 research outputs found

    Aging diminishes the resistance of AO rats to EAE: putative role of enhanced generation of GM-CSF Expressing CD4+T cells in aged rats

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    Background: Aging influences immune response and susceptibility to EAE in a strain specific manner. The study was designed to examine influence of aging on EAE induction in Albino Oxford (AO) rats. Results: Differently from 3-month-old (young) rats, which were resistant to EAE induction, the majority of aged (24-26-month-old) rats developed mild chronic form of EAE. On 16th day post-immunization, when in aged rats the neurological deficit reached plateau, more mononuclear cells, including CD4+ T lymphocytes was retrieved from spinal cord of aged than young rats. The frequencies of IL-17+ and GM-CSF+ cells within spinal cord infiltrating CD4+ lymphocytes were greater in aged rats. To their increased frequency contributed the expansion of GM-CSF + IL-17 + IFN-gamma+ cells, which are highly pathogenic in mice. The expression of the cytokines (IL-1 beta and IL-23/p19) driving GM-CSF + IL-17 + IFN-gamma + cell differentiation in mice was also augmented in aged rat spinal cord mononuclear cells. Additionally, in aged rat spinal cord the expansion of GM-CSF + IL-17-IFN-gamma- CD4+ T lymphocytes was found. Consistently, the expression of mRNAs for IL-3, the cytokine exhibiting the same expression pattern as GM-CSF, and IL-7, the cytokine driving differentiation of GM-CSF + IL-17-IFN-gamma- CD4 + lymphocytes in mice, was upregulated in aged rat spinal cord mononuclear cells, and the tissue, respectively. This was in accordance with the enhanced generation of the brain antigen-specific GM-CSF+ CD4+ lymphocytes in aged rat draining lymph nodes, as suggested by (i) the higher frequency of GM-CSF+ cells (reflecting the expansion of IL-17-IFN-gamma- cells) within their CD4+ lymphocytes and (ii) the upregulated GM-CSF and IL-3 mRNA expression in fresh CD4+ lymphocytes and MBP-stimulated draining lymph node cells and IL-7 mRNA in lymph node tissue from aged rats. In agreement with the upregulated GM-CSF expression in aged rats, strikingly more CD11b + CD45(int) (activated microglia) and CD45(hi) (mainly proinflammatory dendritic cells and macrophages) cells was retrieved from aged than young rat spinal cord. Besides, expression of mRNA for SOCS1, a negative regulator of proinflammatory cytokine expression in innate immunity cells, was downregulated in aged rat spinal cord mononuclear cells. Conclusions: The study revealed that aging may overcome genetic resistance to EAE, and indicated the cellular and molecular mechanisms contributing to this phenomenon in AO rats

    Effectiveness of Wharton's jelly stem cells in gastroschisis repair using the inner surface of the umbilical cord as a patch: long-term results

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    Objective This retrospective study was designed to assess and compare the anatomical, functional, and esthetic appearance of the umbilical area in patients after repair of gastroschisis using the inner surface of the umbilical cord (UC) and in patients with omphalocele conservatively treated. Background Our procedure transformed gastroschisis into an 'artificial-surgical omphalocele' in which the prolapsed intestine was covered with an umbilical patch, the inner surface of which contained Wharton's jelly (WJ). Methods We have summarized an experience of 17 years in repairing gastroschisis using the inner surface of the UC. From 1986 to 2003, 21 infants with gastroschisis and one with a ruptured omphalocele were treated with this technique. We used Eagle's medium to prove the validity of the umbilical stump and the duration of its viability. The inner surface of the umbilical patch is a 'live' structure with WJ, which contains mucoid connective tissue and fibroblast-like cells-that is, stem cells producing cutis, adipose, and connective tissue. Results Using our method, early control assessment of 18 of 21 patients with gastroschisis, at intervals of 1-3 months, showed good functional and esthetic results. Clinical long-term results in terms of anatomical, clinical, and functional findings were excellent. Besides clinical testimony, we used high-frequency ultrasonography to make an appraisal of the effectiveness of WJ stem cells in the repair of gastroschisis, and compared our results with healthy volunteers and patients with omphalocele conservatively treated. Conclusion This paper describes the effect of the local application of WJ-that is, mesenchymal stromal cells derived from the inner surface of the umbilical stump - and its influence on the healing process of the birth defect and wound. (C) 2015 Annals of Pediatric Surgery

    Lactobacillus rhamnosus LA68 and Lactobacillus plantarum WCFS1 differently influence metabolic and immunological parameters in high fat diet-induced hypercholesterolemia and hepatic steatosis

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    In this study, two Lactobacillus strains (L. rhamnosus LA68 and L. plantarum WCFS1) were evaluated for their effects on high fat diet induced pathology in mice. The aim was to determine whether the administration of lactic acid bacteria had beneficial effects on ameliorating pathology. C57BL/6 mice fed a high fat diet were orally administered with the Lactobacillus strains. Both the metabolic and immunological parameters were analyzed. The administration of both of the strains had beneficial effects on mouse weight, serum cholesterol, TNF-alpha levels and liver histology. LA68 lowered the total cholesterol and HDL levels more prominently, whereas WCFS1 was more potent in lowering the TG and LDL levels. Leptin and adiponectin levels were increased in all experimental groups to different extents. The administration of L. plantarum WCFS1 led to a marked increase in leptin levels, as well as an increase in CD3+CD4+ and CD3+CD8+ cells, and a decrease of CD25+ cells, and had a lowering effect on IL-6 production and cell metabolic activity. In conclusion, active administration of both Lactobacillus strains had a positive effect on HFD-induced pathology. Although both of the tested strains had beneficial effects, oral administration of WCFS1 increased leptin levels and had a more prominent immunomodulatory effect, which should be taken into consideration in case of humane usage

    Aged rat macrophages exhibit ipaired response to in vitro stimulation with IL-4

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    The influence of aging on phagocytic, antigen presenting and secretory capacity of resident Albino Oxford rat peritoneal macrophages cultured for 24 hours in medium alone and in the presence of either LPS or IL-4 was examined. The frequencies of CD68+, CD169+ and CCR7+ cells were comparable among fresh peritoneal cells from young (3-months-old) and aged (i8-months-old) rats, whereas those of CD163+, MHCII+ and CD86+ cells were lower within the same cell population in aged relative to young rats. Congruent with the latter findings, aged rat macrophages cultured in medium alone exhibited impaired allostimulatory properties. Additionally, their zymosan phagocytizing ability was reduced. Differently from young rat macrophages, aged rat macrophages changed neither allostimulatory nor zymosan phagocytizing ability upon in vitro treatment with either LPS or IL-4. In the presence of LPS, NO production comparably increased in macrophages from young and aged rats. However, in the presence of either LPS or IL-4 urea production increased only in macrophages from aged rats. Differently from LPS, which increased the prototypic proinflamrnatory cytokine production in macrophages from rats of both ages, the diminishing effect of IL-4 on their production was evident only in macrophages from young rats. Collectively, our results suggest that aging, besides diminishing influence on macrophage allostimulatory and zymosan phagocytizing ability, may impair their responsiveness to IL-4 in vitro. Consequently, an altered efficacy of inflammatory/immune responses in aged rats may be expected

    Antimicrobial activity of oregano (Origanum vulgare L.): And basil (Ocimum basilicum L.): Extracts

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    Komercijalni uzorci nadzemnih delova Origanum vulgare L. i Ocimum basilicum L. (Lamiaceae) su korišćeni za određivanje antimikrobne aktivnosti. Aktivnost ekstrakata različite polarnosti je testirana protiv izabranih mikroorganizama, uključujući laboratorijski soj Helicobacter pilori NCTC 12868. Testirani ekstrakti su imali umerenu aktivnost. Ekstrakti O. vulgare su bili aktivniji protiv bakterija, posebno Gram pozitivnih bakterija sa minimalnim inhibitornim koncentracijama (MIK) između 62,5 i 125 μg/mL, od ekstrakata O. basilicum koji su pokazali aktivnost protiv Candida albicans (MIK 125 μg/mL). Cikloheksanski ekstrakt O. vulgare nije pokazao antimikrobnu aktivnost na testirani H. pylori, dok su ostali testirani ekstrakti bili aktivni sa MIK vrednostima između 250 μg/mL i 500 μg/mL. Identifikovana i kvantifikovana ruzmarinska kiselina i druga polarna jedinjenja mogu predstavljati aktivna jedinjenja sa antibakterijskom aktivnošću u ovim začinima.The commercial samples of aerial parts of Origanum vulgare L. and Ocimum basilicum L. (Lamiaceae) were tested for antimicrobial activity. The activity of the extracts with different polarity was tested against a panel of microorganisms, including laboratory strain Helicobacter pylori NCTC 12868. The tested extracts showed a moderate activity. The extracts of O. vulgare were more active against bacteria, especially against Gram positive bacteria with the minimal inhibitory concentration (MIC) between 62.5 and 125 μg/mL, than the extracts of O. basilicum which were active against Candida albicans (MIC 125 μg/mL). Cyclohexane extract of O. vulgare did not show any activity against tested H. pylori, while all other tested extracts were active with MICs between 250 μg/mL and 500 μg/mL. Identified and quantified rosmarinic acid and other polar compounds could be active antibacterial compounds in these spices

    Genetic detection of Dobrava-Belgrade hantavirus in the edible dormouse (Glis glis) in central Serbia

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    Hantaviruses are endemic in the Balkans, particularly in Serbia, where sporadic cases and/or outbreaks of hantaviral human disease have been reported repeatedly, and evidenced serologically. Here, we present genetic detection of Dobrava-Belgrade virus (DOBV) hantaviral sequences in wild rodents trapped in central Serbia. All the animals were pre-screened serologically by indirect immunofluorescence (IF) test and only those with a positive finding of hantaviral antigens were further tested by polymerase chain reaction. Of the total of 104 trapped animals, 20 were found to be IF positive and of those three were positive for hantaviral RNA: one Microtus arvalis for Tula virus, and one each of Apodemus agrarius and Glis glis for DOBV. Phylogenetic analysis of the obtained sequences implies putative DOBV spillover infection of A. agrarius and G. glis from Apodemus flavicollis. However, future investigations should help to identify the most common natural host and geographical distribution of DOBV in its reservoir hosts in Serbia

    Brain and liver fatty acid composition changes upon consumption of Lactobacillus rhamnosus LA68

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    Recent reports suggest that the metabolic activity of the enteric microbiota may influence the fatty acid composition of the host tissue. There are many studies dealing with the influence of lactobacilli on various pathological conditions, and some of the effects are strain-specific. This study was designed to test the effects of a particular Lactobacillus strain, Lactobacillus rhamnosus LA68 on fatty acid composition of the liver and the brain of C57BL/6 mice in the absence of an underlying pathological condition. Female mice were supplemented with live L. rhamnosus LA68 bacteria for the duration of 1 month. Serum biochemistry was analyzed and liver and brain fatty acid composition was assessed by gas-liquid chromatography. Significant changes in liver and brain fatty acid composition were detected. In the liver tissue we detected an increase in palmitoleic acid (p = 0.038), while in the brain compartment we found an increase in palmitic (p = 0.042), stearic (p = 0.017), arachidonic acid (p = 0.009) and docosahexaenoic acid (p = 0.004) for control versus experimental group. These results show discrete changes caused by LA68 strain consumption. Even short duration of administration of LA68 influences the fatty acid composition of the host which adds to the existing knowledge about Lactobacillus host interaction, and adds to the growing knowledge of metabolic intervention possibilities

    Sexual dimorphism in the aged rat CD4+T lymphocyte-mediated immune response elicited by inoculation with spinal cord homogenate

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    Considering the crucial pathogenic role of CD4+ T cells in experimental autoimmune encephalomyelitis (EAE) and the opposite direction of the sexual dimorphism in the severity of the disease in 22-24-and 3-month-old dark agouti rats, sex differences in CD4+ T-cell-mediated immune response in aged rats immunized for EAE were examined and compared with those in young animals. In the inductive phase of EAE, fewer activated CD4+ lymphocytes were-retrieved from draining lymph nodes of male (developing less severe disease) compared with female rats, due, at least partly, to their lesser expansion. The former reflected a greater suppressive capacity of CD4+CD25+Foxp3+ cells. Consequently, CD4+ lymphocyte infiltration into the spinal cord of aged male rats was diminished. At the peak of EAE, the frequency of reactivated cells was lower, whereas that of the regulatory CD4+ cells was higher in male rat spinal cord. Consistently, microglial activation and the expression of proinflammatory/damaging cytokines in male rat spinal cord mononuclear cells were diminished. Additionally, the frequency of the highly pathogenic IL-17+IFN-gamma+ T lymphocytes infiltrating their spinal cord was lower. Together, these results point to (i) an age-specificity in CD4+ cell-mediated immune response and (ii) mechanisms underlying the sex differences in this response in aged rats. (C) 2015 Elsevier Ireland Ltd. All rights reserved

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