In our previous studies,
B1/1, which was renamed
(
), was able to modulate the production of pro-inflammatory cytokines and other components of the innate immune response in vitro and in vivo. In ...this study, we evaluated the effect of
B1/1 in two concentrations (1 × 10
and 1 × 10
CFU) on the metabolic activity, adherence ability and relative gene expression of pro-inflammatory interleukins (IL-1β, IL-6, IL-8, IL-18), lumican and olfactomedin 4 produced by non-carcinogenic porcine-derived enterocytes (CLAB). CLAB cells were cultured in a 12-well cell culture plate at a concentration of 4 × 10
cells/well in DMEM medium in a controlled humidified atmosphere for 48 h. A 1 mL volume of each probiotic bacterial suspension was added to the CLAB cells. Plates were incubated for 2 h and 4 h. Our results revealed that
B1/1 was able to adhere to CLAB cells in sufficient numbers in both concentrations. In particular, the concentration of 10
B1/1 allowed to modulate the gene expression of pro-inflammatory cytokines, as well as to increase the metabolic activity of the cells. In addition, administration of
B1/1 in both concentrations significantly stimulated gene expression for both proteins in the CLAB cell line after 4 h of incubation.
Increasing antimicrobial resistance has caused a great interest in natural products as alternatives or potentiators of antibiotics. The objective of this study was to isolate individual tannins from ...crude chestnut extract as well as to determine the influence of both crude extracts (tannic acid extract, chestnut extract) and individual pure tannins (gallic acid, vescalin, vescalagin, castalin, castalagin) on the growth of Gram-positive Staphylococcus aureus bacteria. Their antibacterial activity was monitored by measuring the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) as well as the duration of the lag phase, growth rate and generation time. The effect of growth medium strength on the MIC of different tannins was also investigated. Bacterial growth was followed spectrophotometrically, and MIC values were determined by the microdilution method. The MIC values of various isolated compounds allowed us to determine the bioactive compounds and their contribution to antimicrobial activity. It was found that MIC values increase with increasing growth medium strength and that the lag phase lengthens with increasing tannin concentrations, while the growth rates decrease. Comparing the results of the two studies, the antimicrobial activity of tannins against S. aureus was not as pronounced as in the case of E. coli, which may indicate that a different mechanism of action is responsible for the antimicrobial effects of tannins on Gram-positive than on Gram-negative bacteria, or that a different mechanism is more pronounced.
The antioxidant activity (AA) of hop extracts obtained from different hop genotypes (n = 14) was studied. For comparison, the purified β-acids-rich fraction and α-acids-with-β-acids-rich fraction ...were also used to test the antioxidative potential. The AA of purified hydroacetonic hop extracts was investigated using the Ferric Reducing Ability of Plasma (FRAP), Oxygen Radical Absorption Capacity (ORAC) and Intracellular Antioxidant (IA) methods. The FRAP values in different hop genotypes ranged between 63.5 and 101.6 μmol Trolox equivalent (TE)/g dry weight (DW), the ORAC values ranged between 1069 and 1910 μmol TE/g DW and IA potential values ranged between 52.7 and 118.0 mmol TE/g DW. Significant differences in AA between hop genotypes were observed with all three methods. AAs were determined using three different methods, which did not highly correlate with each other. We also did not find significant correlations between AA and different chemical components, which applies both to AA determined using individual methods as well as the total AA. Based on this fact, we assume that the synergistic or antagonistic effects between hop compounds have a more pronounced effect on AA than the presence and quantity of individual hop compounds.
The antimicrobial activity of hop extracts obtained from different hop genotypes were investigated against
and
. In this study the pure xanthohumol, purified β-acids rich fraction, as well as α-acids ...with β-acids rich fraction were used to test antimicrobial activity against
and
; whereby, the antimicrobial activity of different hop extracts against
was studied for the first time. Microbial susceptibility to purified hydroacetonic extracts from different hop varieties was investigated by the broth microdilution assay to determine the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC). The hop hydroacetonic extracts were more effective against
than against
. Strong inverse correlations of MIC and MBC values were obtained with xanthohumol, cohumulone, n+adhumulone, colupulone and n+adlupulone contents, suggesting that the identified chemical hop compounds are directly responsible for antimicrobial effects. Moreover, the effect of the growth medium strength on the MIC values of hop extracts against
was systematically investigated for the first time. The current study also reveals the effect of different hop extracts on
, which responds to their presence by lag phase extension and generation time prolongation.
Organic fertilizers, such as spent mushroom substrate (SMS), improve soil fertility, but studies comparing their effects on different agricultural soils are limited. In this study, the effects of ...standard, SMS and composed fertilizers on soils from conventional–integrated, organic and biodynamic farming were investigated. Soil samples were analyzed for microorganisms and the activity of β-glucosidase (β-GLU), β-1,4-N-acetylglucosaminidase (NAG), urease (URE), arylamidase (ARN), phosphatase (PHOS), acid phosphatase (PAC), alkaline phosphatase (PAH) and arylsulphatase (ARS). Biodynamic soil showed the highest microbial counts and enzyme activities, followed by organic and conventional soils. SMS significantly increased the number of microorganisms and enzyme activities, especially in biodynamic and organic soils. Seasonal variations affected all microorganisms and most enzymes in all soils, except NAG in conventional and organic soils. Biodynamic soil showed stable activity of enzymes and microorganisms throughout the year, indicating greater stability. This study concludes that soil microorganisms and enzyme activities respond differently to fertilization depending on the soil type, with SMS demonstrating beneficial effects in all tested soils.
The staining of intracellular antigens is a standard method in flow cytometry but still only occasionally used for immunologic studies in veterinary species. In order to improve information about ...suitable fixation/permeabilisation protocols in porcine lymphocytes we tested six fixation/permeabilisation variants for the staining of different intracellular antigens: CD79α, perforin, interferon-γ and the cell cycle associated protein Ki-67. For the fixation/permeabilisation procedure, four commercial kits from BD Biosciences, ADG Bio Research, Dako and AbD Serotec were tested in comparison to non-commercial fixation solutions of formaldehyde together with either saponin or Tween-20 for permeabilisation. Perforin and Ki-67 expressing cells were optimally stained only with permeabilisation reagents containing saponin, which includes the BD kit. In contrast, labelling of CD79α and IFN-γ was possible with all investigated fixation/permeabilisation variants; however, here saponin and Tween protocols induced a higher degree of unspecific binding. Also, scatter properties of cells treated with saponin were worse compared to samples prepared with the kits from ADG, Dako and Serotec. Simultaneous staining of cell surface antigens was not negatively affected by any of the fixation/permeabilisation variants. A universal recommendation for a single fixation/permeabilisation strategy could not be deduced from our data but our work provides a useful guideline for optimal staining of common intracellular antigens analyzed in swine lymphocytes.
The maintenance of a healthy status depends on the coexistence between the host organism and the microbiota. Early studies have already focused on the nutritional properties of probiotics, which may ...also contribute to the structural changes in the gut microbiota, thereby affecting host metabolism and homeostasis. Maintaining homeostasis in the body is therefore crucial and is reflected at all levels, including that of glucose, a simple sugar molecule that is an essential fuel for normal cellular function. Despite numerous clinical studies that have shown the effect of various probiotics on glucose and its homeostasis, knowledge about the exact function of their mechanism is still scarce. The aim of our review was to select in vivo and in vitro studies in English published in the last eleven years dealing with the effects of probiotics on glucose metabolism and its homeostasis. In this context, diverse probiotic effects at different organ levels were highlighted, summarizing their potential mechanisms to influence glucose metabolism and its homeostasis. Variations in results due to different methodological approaches were discussed, as well as limitations, especially in in vivo studies. Further studies on the interactions between probiotics, host microorganisms and their immunity are needed.
In our previous studies, Lactobacillus reuteri B1/1, which was renamed Limosilactobacillus reuteri (L. reuteri), was able to modulate the production of pro-inflammatory cytokines and other components ...of the innate immune response in vitro and in vivo. In this study, we evaluated the effect of Lactobacillus reuteri B1/1 in two concentrations (1 × 10sup.7 and 1 × 10sup.9 CFU) on the metabolic activity, adherence ability and relative gene expression of pro-inflammatory interleukins (IL-1β, IL-6, IL-8, IL-18), lumican and olfactomedin 4 produced by non-carcinogenic porcine-derived enterocytes (CLAB). CLAB cells were cultured in a 12-well cell culture plate at a concentration of 4 × 10sup.5 cells/well in DMEM medium in a controlled humidified atmosphere for 48 h. A 1 mL volume of each probiotic bacterial suspension was added to the CLAB cells. Plates were incubated for 2 h and 4 h. Our results revealed that L. reuteri B1/1 was able to adhere to CLAB cells in sufficient numbers in both concentrations. In particular, the concentration of 10sup.9 L. reuteri B1/1 allowed to modulate the gene expression of pro-inflammatory cytokines, as well as to increase the metabolic activity of the cells. In addition, administration of L. reuteri B1/1 in both concentrations significantly stimulated gene expression for both proteins in the CLAB cell line after 4 h of incubation.
Increasing antimicrobial resistance has caused a great interest in natural products as alternatives or potentiators of antibiotics. The objective of this study was to isolate individual tannins from ...crude chestnut extract as well as to determine the influence of both crude extracts (tannic acid extract, chestnut extract) and individual pure tannins (gallic acid, vescalin, vescalagin, castalin, castalagin) on the growth of Gram-positive Staphylococcus aureus bacteria. Their antibacterial activity was monitored by measuring the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) as well as the duration of the lag phase, growth rate and generation time. The effect of growth medium strength on the MIC of different tannins was also investigated. Bacterial growth was followed spectrophotometrically, and MIC values were determined by the microdilution method. The MIC values of various isolated compounds allowed us to determine the bioactive compounds and their contribution to antimicrobial activity. It was found that MIC values increase with increasing growth medium strength and that the lag phase lengthens with increasing tannin concentrations, while the growth rates decrease. Comparing the results of the two studies, the antimicrobial activity of tannins against S. aureus was not as pronounced as in the case of E. coli, which may indicate that a different mechanism of action is responsible for the antimicrobial effects of tannins on Gram-positive than on Gram-negative bacteria, or that a different mechanism is more pronounced.
Natural killer (NK) cells are one of the main cellular components of the innate immune system. They play an important role in the immune response against infections as well as tumour cells and ...therefore have two major properties: production of immune regulatory cytokines and chemokines as well as cytolytic destruction of particular target cells.
The existence of NK cells in swine is well known as well as the phenotype of resting NK cells, but their response following activation by cytokines is still poorly understood. Therefore, we tested the influence of the immune regulatory cytokines IL-2, IL-12 and IL-18 on cytolytic activity, phenotype, IFN-γ production and the accumulation of perforin in cytoplasm of peripheral blood mononuclear cells (PBMC) as well as purified NK cells. NK cells were enriched from PBMC using a magnetic cell separation (MACS) strategy with monoclonal antibodies against CD3, CD21 and SWC3, thereby removing T-, B- and myeloid cells. Respective fractions were used in flow cytometry (FCM) based cytolytic assays with the human tumour cell line K562 as target. After stimulation with the cytokines described above, the NK cell enriched CD3
−CD21
−SWC3
− fraction showed an evident increase in the cytolytic activity compared to PBMC. This enhanced cytolytic activity was accompanied by a strong enrichment of IFN-γ producing cells when a combination of all three cytokines (IL-2/IL-12/IL-18) was used; as determined in ELISPOT assays and intracellular staining of IFN-γ in FCM. Also, the combination of these three cytokines led to an accumulation of perforin in the cytoplasm and an up-regulation of CD25 compared to control cultures incubated in medium without cytokines.
The experiments performed clearly indicate a stimulatory role and strong synergistic effects of the investigated cytokines in the activation of porcine NK cells in vitro, inducing IFN-γ, perforin production and cytotoxicity against target cells.
