Heat‐killed Lactobacillus reuteri and cell‐free culture supernatant have similar effects to viable probiotics during interaction with Porphyromonas gingivalis

2020 ◽  
Vol 55 (2) ◽  
pp. 215-220 ◽  
Author(s):  
Barbara M. C. Geraldo ◽  
Marianna N. Batalha ◽  
Noala V. M. Milhan ◽  
Rodnei D. Rossoni ◽  
Liliana Scorzoni ◽  
...  
Keyword(s):  
Porphyromonas Gingivalis ◽  
Culture Supernatant ◽  
Lactobacillus Reuteri ◽  
Cell Free Culture Supernatant

scholarly journals Production of biologically active human interleukin-10 by Bifidobacterium bifidum BGN4

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Nayoun Hong ◽  
Seockmo Ku ◽  
Kyungjin Yuk ◽  
Tony V. Johnston ◽  
Geun Eog Ji ◽  
...  
Keyword(s):  
Culture Supernatant ◽  
Shuttle Vector ◽  
Sodium Dodecyl ◽  
Interleukin 10 ◽  
Biologically Active ◽  
Enzyme Linked Immunosorbent Assay ◽  
Functional Evaluation ◽  
Ht 29 ◽  
Cell Free Culture Supernatant

Abstract Background Bifidobacterium spp. are representative probiotics that play an important role in the health of their hosts. Among various Bifidobacterium spp., B. bifidum BGN4 exhibits relatively high cell adhesion to colonic cells and has been reported to have various in vivo and in vitro bio functionalities (e.g., anti-allergic effect, anti-cancer effect, and modulatory effects on immune cells). Interleukin-10 (IL-10) has emerged as a major suppressor of immune response in macrophages and other antigen presenting cells and plays an essential role in the regulation and resolution of inflammation. In this study, recombinant B. bifidum BGN4 [pBESIL10] was developed to deliver human IL-10 effectively to the intestines. Results The vector pBESIL10 was constructed by cloning the human IL-10 gene under a gap promoter and signal peptide from Bifidobacterium spp. into the E. coli-Bifidobacterium shuttle vector pBES2. The secreted human IL-10 from B. bifidum BGN4 [pBESIL10] was analyzed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), Western Blotting, and enzyme-linked immunosorbent assay (ELISA). More than 1,473 ± 300 ng/mL (n = 4) of human IL-10 was obtained in the cell free culture supernatant of B. bifidum BGN4 [pBESIL10]. This productivity is significantly higher than other previously reported human IL-10 level from food grade bacteria. In vitro functional evaluation of the cell free culture supernatant of B. bifidum BGN4 [pBESIL10] revealed significantly inhibited interleukin-6 (IL-6) production in lipopolysaccharide (LPS)-induced Raw 264.7 cells (n = 6, p < 0.0001) and interleukin-8 (IL-8) production in LPS-induced HT-29 cells (n = 6, p < 0.01) or TNFα-induced HT-29 cells (n = 6, p < 0.001). Conclusion B. bifidum BGN4 [pBESIL10] efficiently produces and secretes significant amounts of biologically active human IL-10. The human IL-10 production level in this study is the highest of all human IL-10 production reported to date. Further research should be pursued to evaluate B. bifidum BGN4 [pBESIL10] producing IL-10 as a treatment for various inflammation-related diseases, including inflammatory bowel disease, rheumatoid arthritis, allergic asthma, and cancer immunotherapy.

Anti-tumor potential of cell free culture supernatant of Lactobacillus rhamnosus strains isolated from human breast milk

2019 ◽  
Vol 123 ◽  
pp. 286-297 ◽  
Author(s):  
Muhammad Shahid Riaz Rajoka ◽  
Haobin Zhao ◽  
Hafiza Mahreen Mehwish ◽  
Na Li ◽  
Yao Lu ◽  
...  
Keyword(s):  
Breast Milk ◽  
Culture Supernatant ◽  
Lactobacillus Rhamnosus ◽  
Human Breast Milk ◽  
Human Breast ◽  
Cell Free Culture Supernatant

Changes in Heat Resistance Resulting from pH and Nutritional Shifts of Acid-Adapted and Non–Acid-Adapted Listeria monocytogenes Scott A†

2004 ◽  
Vol 67 (2) ◽  
pp. 316-321 ◽  
Author(s):  
DARRELL O. BAYLES
Keyword(s):  
Listeria Monocytogenes ◽  
Heat Resistance ◽  
Frequency Distribution ◽  
Culture Supernatant ◽  
Physiological State ◽  
Distribution Data ◽  
Treatment Conditions ◽  
Lower Heat ◽  
Full Factorial ◽  
Cell Free Culture Supernatant

Stationary-phase Listeria monocytogenes cells that were either pH dependent acid adapted or not acid adapted were heat challenged at 60°C in a two-level full factorial design for three variables. The three variables and the levels consisted of tryptic soy broth (TSB) and sterile cell-free culture supernatant (sterile TSB), the presence and absence of 1% added glucose, and pH 4.8 and pH 7. Non–acid-adapted cells were most heat resistant when challenged in TSB (mean decimal reduction times at 60°C: D60 = 1.16 min). In the absence of added glucose, non–acid-adapted cells had similar D60-values for inactivations at pH 4.8 and pH 7; however, the presence of glucose caused non–acid-adapted cells challenged at pH 4.8 to be more heat sensitive (D60 = 0.65 min) than those inactivated at pH 7 (D60 = 1.03 min), indicating an interaction between glucose and pH. Overall, the significantly decreased heat resistance of the acid-adapted cells was due to the presence of glucose (D60 = 0.78 min without glucose, D60 = 0.59 min with glucose). Acid-adapted cells heat challenged in TSB had similar D60-values for inactivations at pH 4.8 and pH 7; however, acid-adapted cells in sterile TSB challenged at pH 4.8 (D60 = 0.52 min) had significantly lower heat resistance than did cells challenged at pH 7 (D60 = 0.76 min), indicating an interaction between the medium and pH. The L. monocytogenes survivor data were modeled to extract information on the frequency distribution of heat resistance within heat-challenged populations, and the frequency distribution characteristics of mean, mode, and variance were compared among treatment conditions. Significant differences in the frequency distribution data were compared with the D60-values. These data indicated that the presence and level of cross-protection is highly dependent on the physiological state of the cells and nutrient availability at the time of heat challenge. Such conditions should be considered to ensure that stressed pathogens in foods are destroyed or inactivated.

scholarly journals Altered Gingipain Maturation in vimA- and vimE-Defective Isogenic Mutants of Porphyromonas gingivalis

2005 ◽  
Vol 73 (3) ◽  
pp. 1357-1366 ◽  
Author(s):  
Elaine Vanterpool ◽  
Francis Roy ◽  
Lawrence Sandberg ◽  
Hansel M. Fletcher
Keyword(s):  
Cell Surface ◽  
Porphyromonas Gingivalis ◽  
Culture Supernatant ◽  
Specific Activity ◽  
Phenotypic Effect ◽  
Western Blots ◽  
Defective Mutant ◽  
Surface Polysaccharide ◽  
Isogenic Mutants ◽  
Specific Sugar

ABSTRACT We have previously shown that gingipain activity in Porphyromonas gingivalis is modulated by the unique vimA and vimE genes. To determine if these genes had a similar phenotypic effect on protease maturation and activation, isogenic mutants defective in those genes were further characterized. Western blot analyses with antigingipain antibodies showed RgpA-, RgpB-, and Kgp-immunoreactive bands in membrane fractions as well as the culture supernatant of both P. gingivalis W83 and FLL93, the vimE-defective mutant. In contrast, the membrane of P. gingivalis FLL92, the vimA-defective mutant, demonstrated immunoreactivity only with RgpB antibodies. With mass spectrometry or Western blots, full-length RgpA and RgpB were identified from extracellular fractions. In similar extracellular fractions from P. gingivalis FLL92 and FLL93, purified RgpB activated only arginine-specific activity. In addition, the lipopolysaccharide profiles of the vimA and vimE mutants were truncated in comparison to that of W83. While glycosylated proteins were detected in the membrane and extracellular fractions from the vimA- and vimE-defective mutants, a monoclonal antibody (1B5) that reacts with specific sugar moieties of the P. gingivalis cell surface polysaccharide and membrane-associated Rgp gingipain showed no immunoreactivity with these fractions. Taken together, these results indicate a possible defect in sugar biogenesis in both the vimA- and vimE-defective mutants. These modulating genes play a role in the secretion, processing, and/or anchorage of gingipains on the cell surface.

scholarly journals Lactobacillus reuteri ZJ617 Culture Supernatant Attenuates Acute Liver Injury Induced in Mice by Lipopolysaccharide

2019 ◽  
Vol 149 (11) ◽  
pp. 2046-2055 ◽  
Author(s):  
Yanjun Cui ◽  
Sirui Qi ◽  
Wenming Zhang ◽  
Jiangdi Mao ◽  
Renlong Tang ◽  
...  
Keyword(s):  
Liver Injury ◽  
Intraperitoneal Injection ◽  
Molecular Mechanisms ◽  
Culture Supernatant ◽  
Lactobacillus Reuteri ◽  
Acute Liver Injury ◽  
Saline Control ◽  
Control Group ◽  
Intestinal Integrity ◽  
Oral Inoculation

ABSTRACT Background Lactobacillus rhamnosus GG culture supernatant (LGGs) promotes intestinal integrity and ameliorates acute liver injury induced by alcohol in mice. Objectives The aim of this study was to investigate the protective effects and molecular mechanisms of Lactobacillus reuteri ZJ617 culture supernatant (ZJ617s) on acute liver injury induced by lipopolysaccharide (LPS) in mice. Methods Male C57BL/6 mice (20 ± 2 g, 8 wk old) were randomly divided into 4 groups (6 mice/group): oral inoculation with phosphate-buffered saline (control), intraperitoneal injection of LPS (10 mg/kg body weight) (LPS), oral inoculation with ZJ617s 2 wk before intraperitoneal injection of LPS (ZJ617s + LPS), or oral inoculation with LGGs 2 wk before intraperitoneal injection of LPS (LGGs + LPS). Systemic inflammation, intestinal integrity, biomarkers of hepatic function, autophagy, and apoptosis signals in the liver were determined. Results Twenty-four hours after LPS injection, the activities of serum alanine transaminase and aspartate transaminase were 32.2% and 30.3% lower in the ZJ617s + LPS group compared with the LPS group, respectively (P < 0.05). The ZJ617s + LPS group exhibited higher intestinal expression of claudin 3 (62.5%), occludin (60.1%), and zonula occludens 1 (60.5%) compared with the LPS group (P < 0.05). The concentrations of hepatic interleukin-6 and tumor necrosis factor-α were 21.4% and 27.3% lower in the ZJ617s + LPS group compared with the LPS group, respectively (P < 0.05). However, the concentration of interleukin-10 was 22.2% higher in the ZJ617s + LPS group. LPS increased the expression of Toll-like receptor 4 (TLR4; by 50.5%), phosphorylation p38 mitogen-activated protein kinase (p38MAPK; by 57.1%), extracellular signal-regulated kinase (by 77.8%), c-Jun N-terminal kinase (by 42.9%), and nuclear factor-κB (NF-κB; by 36.0%) compared with the control group. Supplementation with ZJ617s or LGGs ameliorated these effects (P < 0.05). Moreover, the hepatic expression of active caspase-3 and microtubule-associated protein 1 light chain 3 II was 23.8% and 28.6% lower in the ZJ617s + LPS group compared with the LPS group, respectively (P < 0.05). Conclusions ZJ617s exerts beneficial effects on the mouse liver through suppression of hepatic TLR4/MAPK/NF-κB activation, apoptosis, and autophagy. This trial was registered at Zhejiang University (http://www.lac.zju.edu.cn) as NO.ZJU20170529.

Growth and hemolysin production by Haemophilus pleuropneumoniae cultivated in a chemically defined medium

1986 ◽  
Vol 32 (10) ◽  
pp. 801-805 ◽  
Author(s):  
James R. Maudsley ◽  
Solomon Kadis
Keyword(s):  
Growth Rate ◽  
Yeast Extract ◽  
Hemolytic Activity ◽  
Culture Supernatant ◽  
Iron Concentration ◽  
Defined Medium ◽  
Chemically Defined Medium ◽  
Sterile Cell ◽  
Stages Of Growth ◽  
Cell Free Culture Supernatant

A chemically defined medium (CDM) has been developed which supports both growth and hemolysin production by Haemophilus pleuropneumoniae. Although the growth rate in stationary cultures was substantially slower in CDM than in trypticase soy broth plus 0.6% yeast extract (TSBYE) and slightly slower than in heart infusion broth (HIB), extracellular hemolysin activity in CDM was slightly higher than in HIB and 16-fold greater than in TSBYE. Maximum hemolytic activity was produced in CDM in early to mid log phase of growth. Hemolytic activity in sterile, cell-free culture supernatant fluids persisted for over 10 days at 4 °C and 3–5 days at 37 °C, but was completely destroyed at 56 °C after 30 min. Total hemolysin inactivation was also achieved in the presence of trypsin or pronase (10 units/mL), but no decrease in hemolytic activity was noted in the presence of DNase or RNase. Iron had little effect on the hemolytic activity in the early stages of growth. However, in the later stages of growth, iron had a pronounced effect with hemolyic activity decreasing as the iron concentration increased from 1 to 500 μM. None of these iron concentrations had any effect on the hemolytic activity when added directly to prepared cell-free culture supernatant fluids. The extracellur hemolysin produced by H. pleuropneumoniae in CDM appears to be a heat-labile protein the activity of which is influenced by iron at certain phases of growth.

scholarly journals Characterization of Reutericyclin Produced by Lactobacillus reuteri LTH2584

2000 ◽  
Vol 66 (10) ◽  
pp. 4325-4333 ◽  
Author(s):  
Michael G. Gänzle ◽  
Alexandra Höltzel ◽  
Jens Walter ◽  
Günther Jung ◽  
Walter P. Hammes
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Activity ◽  
Culture Supernatant ◽  
Lactobacillus Reuteri ◽  
Gel Filtration ◽  
Reversed Phase ◽  
Exchange Chromatography ◽  
Growth Media ◽  
Dependent Manner ◽  
Cell Extracts

ABSTRACT Lactobacillus reuteri LTH2584 exhibits antimicrobial activity that can be attributed neither to bacteriocins nor to the production of reuterin or organic acids. We have purified the active compound, named reutericyclin, to homogeneity and characterized its antimicrobial activity. Reutericyclin exhibited a broad inhibitory spectrum including Lactobacillus spp., Bacillus subtilis, B. cereus, Enterococcus faecalis, Staphylococcus aureus, and Listeria innocua. It did not affect the growth of gram-negative bacteria; however, the growth of lipopolysaccharide mutant strains ofEscherichia coli was inhibited. Reutericyclin exhibited a bactericidal mode of action against Lactobacillus sanfranciscensis, Staphylococcus aureus, and B. subtilis and triggered the lysis of cells of L. sanfranciscensis in a dose-dependent manner. Germination of spores of B. subtilis was inhibited, but the spores remained unaffected under conditions that do not permit germination. The fatty acid supply of the growth media had a strong effect on reutericyclin production and its distribution between producer cells and the culture supernatant. Reutericyclin was purified from cell extracts and culture supernatant of L. reuteri LTH2584 cultures grown in mMRS by solvent extraction, gel filtration, RP-C8 chromatography, and anion-exchange chromatography, followed by rechromatography by reversed-phase high-pressure liquid chromatography. Reutericyclin was characterized as a negatively charged, highly hydrophobic molecule with a molecular mass of 349 Da. Structural characterization (A. Höltzel, M. G. Gänzle, G. J. Nicholson, W. P. Hammes, and G. Jung, Angew. Chem. Int. Ed. 39:2766–2768, 2000) revealed that reutericyclin is a novel tetramic acid derivative. The inhibitory activity of culture supernatant of L. reuteri LTH2584 corresponded to that of purified as well as synthetic reutericyclin.

In vitro evaluation of indole-3-carboxaldehyde on Vibrio parahaemolyticus biofilms

Biologia ◽  
2016 ◽  
Vol 71 (3) ◽  
Author(s):  
Murugan Rajalaxmi ◽  
Vivekanandham Amsa Devi ◽  
Shunmugiah Karutha Pandian
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Culture Supernatant ◽  
In Vitro Evaluation ◽  
Palk Bay ◽  
Antibiofilm Agents ◽  
Cell Free Culture Supernatant

AbstractThis study was aimed to explore antibiofilm agents from the hitherto underexplored Palk Bay seawater bacteria. The cell free culture supernatant of the isolate

Gingipains in the culture supernatant of Porphyromonas gingivalis cleave CD4 and CD8 on human T cells

2002 ◽  
Vol 37 (6) ◽  
pp. 464-468 ◽  
Author(s):  
Y. Kitamura ◽  
M. Yoneda ◽  
T. Imamura ◽  
S. Matono ◽  
Y. Aida ◽  
...  
Keyword(s):  
T Cells ◽  
Porphyromonas Gingivalis ◽  
Culture Supernatant ◽  
Human T Cells
Sign in / Sign up

Export Citation Format

Share Document