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Lactobacillus salivarius BGHO1 and Lactobacillus reuteri BGGO6-55 modify nutritive profile of Artemia franciscana nauplii in a strain ratio, dose and application timing-dependent manner
No full textThis study aimed to analyze an impact of Lactobacillus salivarius BGHO1 and Lactobacillus reuteri BGGO6-55 supplemented to Anemia franciscana cultivation medium on biochemical profile of hatched nauplii. Impacts of different BGHO1:BGGO6-55 ratios (75:25, 50:50 and 25:75) and their total concentrations (250 and 500 g/kg of Anemia cysts) at different timings of bacteria application (pre- and post-hatching) were examined. The effects were evaluated by quantification of naupliar peptide, soluble protein, phospho-, neutral and unsaturated lipid content. Uni and multifactorial analysis of variance (ANOVA) were used to estimate the effects of treatments relative to control and to model factor interactions, respectively Statistical analysis indicated that posthatching application of 75:25 strain ratio at high dose was associated with an increase of neutral lipid amount. Furthermore, factor interaction profiling identified positive correlation of lacto-bacilli concentration with the level of free amino acids/short peptides, phospho-, neutral and unsaturated lipids, but only at 50:50 strain ratio combination. Application of lower lactobacilli dose at 75:25 strain ratio caused an increase of soluble protein and phospholipid amount. Hypothetically, graded response of Artemia nauplii to lactobacilli supplementation was induced with different BGHO1 doses. It assumedly ranged from stress-response protein synthesis at lower doses to membrane permeability alterations and triglyceride-mediated defense mechanism activation at higher BGHO1 doses. In contrast to lactobacilli supplementation after Anemia hatching, pre-hatching application of lactobacilli was not associated with an increase of Anemia nutritive profile, though again there was a positive association of lactobacilli concentration with nutrient amount. This implicates interference of lactobacilli with the hatching process. Results presented here facilitate the design of future studies aiming to modify Artemia nutritive profile in accordance with nutritional demands of cultivated fish species
Role of the End-Point Mediators of Sympathoadrenal and Sympathoneural Stress Axes in the Pathogenesis of Experimental Autoimmune Encephalomyelitis and Multiple Sclerosis
Get PDFThe role of stress effector systems in the initiation and progression of multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE), the most commonly used experimental model of MS, has strongly been suggested. To corroborate this notion, alterations in activity of the sympathoadrenal and sympathoneural axes of sympathoadrenal system (a major communication pathway between the central nervous system and the immune system), mirrored in altered release of their end-point mediators (adrenaline and noradrenaline, respectively), are shown to precede (in MS) and/or occur during development of MS and EAE in response to immune cell activation (in early phase of disease) and disease-related damage of sympathoadrenal system neurons and their projections (in late phase of disease). To add to the complexity, innate immunity cells and T-lymphocytes synthesize noradrenaline that may be implicated in a local autocrine/paracrine self-amplifying feed-forward loop to enhance myeloid-cell synthesis of proinflammatory cytokines and inflammatory injury. Furthermore, experimental manipulations targeting noradrenaline/adrenaline action are shown to influence clinical outcome of EAE, in a disease phase-specific manner. This is partly related to the fact that virtually all types of cells involved in the instigation and progression of autoimmune inflammation and target tissue damage in EAE/MS express functional adrenoceptors. Although catecholamines exert majority of immunomodulatory effects through beta(2)-adrenoceptor, a role for alpha-adrenoceptors in EAE pathogenesis has also been indicated. In this review, we summarize all aforementioned aspects of immunopathogenetic action of catecholamines in EAE/MS as possibly important for designing new strategies targeting their action to prevent/mitigate autoimmune neuroinflammation and tissue damage
Supplementary information for the article: Dimitrijević, M.; Arsenović-Ranin, N.; Bufan, B.; Nacka-Aleksić, M.; Kosec, D.; Pilipović, I.; Kotur-Stevuljević, J.; Simić, L.; Sopta, J.; Leposavić, G. Sex-Based Differences in Monocytic Lineage Cells Contribute to More Severe Collagen-Induced Arthritis in Female Rats Compared with Male Rats. Inflammation 2020, 43 (6), 2312–2331. https://doi.org/10.1007/s10753-020-01302-0.
No full textSupplementary Fig. 1 Sex differences in the clinical and histological presentation of CIA. (a) A line graph indicates daily arthritic score (mean ± SEM) from the 12th to the 39th day post-immunization (d.p.i.) in male (n = 9) and female (n = 10) CIA rats. Mann–Whitney U test: * p ≤ 0.05, from the 17th to the 39th d.p.i. (b) Line graph indicates daily arthritic score (mean ± SEM) from the 13th to the 21st day post-immunization (d.p.i.) in male and female CIA rats. Mann–Whitney U test: n = 8 rats/sex. * p ≤ 0.05. Photographs show representative arthritic joints (arrows) of hind paws from male and female CIA rats. (c) Photomicrographs of HE-stained sections of paraffin-embedded joints from male and female CIA rats show replacement of the normal bone marrow cell populations by inflammatory cells. In females, numerous multinuclear giant cells (red arrows) are present as opposed to male CIA rats. Original magnification × 400. The bar indicates 100 μm (PNG 2723 kb) Supplementary Fig. 2 Fluorescence minus one controls for flow cytometry analysis of CD11b/CCR2/CX3CR1 staining of splenocytes. For setting cutoff boundaries, gates were controlled using fluorescence minus one (FMO) controls obtained by omitting a single antibody from the labeling antibody cocktail. Flow cytometry dot plots represent FMO controls without anti-CX3CR1 or anti-CCR2 Abs within CD11b+ splenocytes (gated as shown in Fig. 3) isolated from CIA rats on the 21st day post-immunization (PNG 98 kb). Supplementary Fig. 3 Fluorescence minus one controls for flow cytometry analysis of CD11b/CD43/CCR2/CX3CR1 staining of peripheral blood cells. For setting cutoff boundaries, gates were controlled using fluorescence minus one (FMO) controls obtained by omitting a single antibody from the labeling antibody cocktail. Flow cytometry dot plots represent FMO controls without (upper) CD43 mAb within CD11b+ peripheral blood cells (gated as shown in Fig. 4a) and (lower) anti-CX3CR1 or anti-CCR2 Abs within CD11b+CD43+ peripheral blood cells isolated from CIA rats on the 21st day post-immunization (PNG 157 kb). Supplementary Fig. 4 Sex differences in the activation of Th cells, Th17 cell function, and frequency of CD40+CD11b+ antigen presenting cells in draining lymph nodes from CIA rats, popliteal draining lymph nodes (DLNs) were retrieved from male and female CIA rats on the 21st day post-immunization. (a) Scatter plots with bar indicate the frequencies of activated Th cells (CD25+Foxp3-CD4+) and Th17 cells (IL-17+CD4+TCRαβ+) in DLNs from CIA rats and the concentration of IL-17 in supernatants of collagen type II-stimulated and unstimulated (medium) DLN cell cultures from male and female rats (see MATERIAL AND METHODS). Linear graph shows the correlation between the frequency of activated Th cells (CD25+Foxp3-CD4+) and the frequency of Th17 cells (IL-17+CD4+TCRαβ+) in DLNs from CIA rats. Pearson’s r value is shown in the graph. (b) Representative flow cytometry dot plots show (upper) CD11b staining and (lower) CD40 vs CD11b staining of DLN cells from male and female rats. Number indicates percent in the region. Scatter plots with bar indicate the frequency and the number of (upper) CD11b+ cells and (lower) CD40+CD11b+ cells in DLNs of male and female rats. The Number indicates percent in the region. Results are expressed as mean ± SEM. (c) The linear graph shows the correlation between the frequency of activated Th cells (CD25+Foxp3-CD4+) and the frequency of CD40+CD11b+ cells in DLNs from CIA rats. Pearson’s r value is shown in the graph. n = 8 rats/sex. * p ≤ 0.05, ** p ≤ 0.01, and *** p ≤ 0.001 (PNG 590 kb). Supplementary Fig. 5 Gating strategy for activated Th cells and Th17 cells, popliteal draining lymph nodes (DLNs) were retrieved from CIA rats on the 21st day post-immunization. Flow cytometry dot plots show gating strategy for (a) activated The cells (CD25+Foxp3-CD4+) and (b) Th17 cells (IL-17+CD4+TCRαβ+) (PNG 101 kb).Supplementary material for: [https://doi.org/10.1007/s10753-020-01302-0]Related to the published version: [https://intor.torlakinstitut.com/handle/123456789/543
Benzimidazole-based dual dipeptidyl peptidase-4 and xanthine oxidase inhibitors
No full textMultiple-targeting compounds might reduce complex polypharmacy of multifactorial diseases, such as diabetes, and contribute to the greater therapeutic success. Targeting reactive oxygen species-producing enzymes, as xanthine oxidase (XO), might suppress progression of diabetes-associated vascular complications. In this study a small series of benzimidazole derivatives (1-9) was evaluated for inhibitory activity against dipeptidyl peptidase-4 (DPP-4) and XO. One 1,3-disubstituted-benzimidazole-2-imine (5) and 1,3-thiazolo [3,2-a] benzimidazolone derivative (8) were shown as effective dual DPP-4 and XO inhibitors, with IC50 values lower than 200 mu M, and predicted binding modes with both target enzymes. Both selected dual inhibitors (compounds 5 and 8) did not show cytotoxicity to a greater extent on Caco-2 cells even at concentration of 250 mu M. These structures represent new non-purine scaffolds bearing two therapeutic functionalities, being DPP-4 and XO inhibitors, more favorable in comparison to DPP-4 inhibitors with DPP-4 as a single target due to pleiotropic effects of XO inhibition.Peer-reviewed version: [https://intor.torlakinstitut.com/handle/123456789/699
Genomic Characteristics of Colistin-Resistant Salmonella enterica subsp. enterica Serovar Infantis from Poultry Farms in the Republic of Serbia
Get PDFThe antimicrobial susceptibility testing was conducted on 174 single isolates from poultry farms in Serbia and it was determined that seven Salmonella spp. were multidrug resistant. Sixteen serotypes were detected, but only serotype Infantis confirmed reduced susceptibility to colistin. Seven colistin resistant Salmonella Infantis were studied in detail using the WGS approach. Three sequence types were identified corresponding to different epizootiology region. The isolate from the Province of Vojvodina 3842 and isolates from Jagodina (92 and 821) are represented by the sequence type ST413 and ST11, respectively. Four isolates from Kraljevo are ST32, a common S. Infantis sequence type in humans, poultry and food. The fosfomycin resistance gene fosA7 in isolate 3842 and the vgaA gene in isolate 8418/2948 encoding resistance to pleuromutilins were reported for the first time in serovar Infantis. The changes in relative expression of the phoP/Q, mgrB and pmrA/B genes were detected. Single nucleotide polymorphisms of the pmrB gene, including transitions Val164Gly or Val164Met, and Arg92Pro are described. Analyses of quinolone resistance determining region revealed substitutions Ser83Tyr in GyrA protein and Thr57Ser and Ser80Arg in ParC protein. Based on WGS data, there are two major clusters among analyzed Salmonella Infantis isolates from central Serbia
Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia
Get PDFVaccines can have heterologous effects on the immune system, i.e., effects other than triggering an immune response against the disease targeted by the vaccine. We investigated whether monoclonal antibodies (mAbs) specific for tetanus could cross-react with Chlamydia and confer heterologous protection against chlamydial infection. The capability of two tetanus-specific mAbs, namely mAb26 and mAb51, to prevent chlamydial infection has been assessed: (i) in vitro, by performing a neutralization assay using human conjunctival epithelial (HCjE) cells infected with Chlamydia trachomatis serovar B, and (ii) in vivo, by using a guinea pig model of Chlamydia caviae-induced inclusion conjunctivitis. The mAb26 has been superior in comparison with mAb51 in the prevention of chlamydial infection in HCjE cells. The mAb26 has conferred approximate to 40% inhibition of the infection, compared to less than 5% inhibition in the presence of the mAb51. In vivo, mAb26 significantly diminished ocular pathology intensity in guinea pigs infected with C. caviae compared to either the mAb51-treated or sham-treated guinea pigs. Our data provide insights that tetanus immunization generates antibodies which induce heterologous chlamydial immunity and promote protection beyond the intended target pathogen.Supplementary information: [https://hdl.handle.net/21.15107/rcub_intor_643
The effect of live and inert feed treatment with lactobacilli on weaning success in intensively reared pike-perch larvae
No full textWeaning to inert diet in intensively reared pike-perch larvae is confronted with significant fish loss, which prevents successful commercialization of pike-perch production. Achievement of satisfactory feed quality and effective assimilation of nutrients by larval fish is the major challenge in larval production process. Aim of this study was to evaluate whether treatment of live and inert feed with lactobacilli could alleviate growth retardation associated with early weaning of pike-perch reared in recirculating system. Weaning started on 18th day post-hatching (DPH) either as sudden weaning (SW) or by co-administration of Anemia for six days (gradual weaning, GW). Prior to administration to fish, Anemia was treated with Lactobacillus salivarius BGHO1/Lb. reuteri BGGO6-55, while inert feed was treated with Lb. paracasei subsp. paracasei BGHN14/Lb. rhamnosus BGT10. Treatment with lactobacilli slightly raised neutral lipid level in Anemia nauplii, but significantly reduced their content in dry feed. Fish were sampled on the 24th DPH. Survival, morphometric indices, skeleton differentiation, digestive enzyme activity and opportunistic pathogenic bacteria level were assessed in whole fish specimens. GW fish were presented with better survival, body growth and phospholipase A2 (PLA2) activity. Alongside, Vibrio spp. growth was suppressed in these fish and skeleton development was improved, according to Alizarin Red staining and ColIA1/Sparc mRNA expression data. Lactobacilli application in GW fish correlated with an increase of survival, condition factor and growth rate, according to trypsin and chymotrypsin activities, indicating better utilization of dietary proteins for muscle building. In SW fish, lactobacilli elevated chymotrypsin activity, PLA2 to lipase activity ratio and improved survival and ossification, as evident from Alizarin Red staining and ColIA1/Sparc mRNA expression. This indicated improved fatty acid absorption and control of metamorphosis process. Furthermore, lactobacilli suppressed Vibrio spp. growth in SW fish. Aside from demonstrating the ability of lactobacilli to aid weaning in pike-perch larvae, this study indicated that different types of food treatment may direct fish growth in a predictable manner, allowing further cost-effective improvements of larval pike-perch rearing in intensive system
Supplementary information for the article: Stojanović, M.; Lukić, I.; Marinković, E.; Kovačević, A.; Miljković, R.; Tobias, J.; Schabussova, I.; Zlatović, M.; Barisani-Asenbauer, T.; Wiedermann, U.; Inić-Kanada, A. Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia. Vaccines 2020, 8 (4), 719. https://doi.org/10.3390/vaccines8040719.
Get PDFTable S1: Characteristics of selected anti-tetanus mAbs [35,36].Related to the published version:[https://intor.torlakinstitut.com/handle/123456789/552]Supplementary material for: [https://doi.org/10.3390/vaccines8040719
Benzimidazole-based dual dipeptidyl peptidase-4 and xanthine oxidase inhibitors
Get PDFMultiple-targeting compounds might reduce complex polypharmacy of multifactorial diseases, such as diabetes, and contribute to the greater therapeutic success. Targeting reactive oxygen species-producing enzymes, as xanthine oxidase (XO), might suppress progression of diabetes-associated vascular complications. In this study a small series of benzimidazole derivatives (1-9) was evaluated for inhibitory activity against dipeptidyl peptidase-4 (DPP-4) and XO. One 1,3-disubstituted-benzimidazole-2-imine (5) and 1,3-thiazolo [3,2-a] benzimidazolone derivative (8) were shown as effective dual DPP-4 and XO inhibitors, with IC50 values lower than 200 mu M, and predicted binding modes with both target enzymes. Both selected dual inhibitors (compounds 5 and 8) did not show cytotoxicity to a greater extent on Caco-2 cells even at concentration of 250 mu M. These structures represent new non-purine scaffolds bearing two therapeutic functionalities, being DPP-4 and XO inhibitors, more favorable in comparison to DPP-4 inhibitors with DPP-4 as a single target due to pleiotropic effects of XO inhibition.This is the peer-reviewed version of the article: Tomović, K.; Ilić, B. S.; Smelcerović, Z.; Miljković, M.; Yancheva, D.; Kojić, M.; Mavrova, A. T.; Kocić, G.; Smelcerović, A. Benzimidazole-Based Dual Dipeptidyl Peptidase-4 and Xanthine Oxidase Inhibitors. Chemico-Biological Interactions 2020, 315. [https://doi.org/10.1016/j.cbi.2019.108873]