CBP22, a Novel Bacteriocin Isolated from Clostridium butyricum ZJU-F1, Protects against LPS-Induced Intestinal Injury through Maintaining the Tight Junction Complex

A novel bacteriocin secreted by Clostridium butyricum ZJU-F1 was isolated using ammonium sulfate fractionation, cation exchange chromatography, affinity chromatography, and reverse-phase high-performance liquid chromatography (RP-HPLC). The bacteriocin, named CBP22, contained 22 amino acids with the sequence PSAWQITKCAGSIAWALGSGIF. Analysis of its structure and physicochemical properties indicated that CBP22 had a molecular weight of 2264.63 Da and a +1 net charge. CBP22 showed activity against E. col K88, E. coli ATCC25922, and S. aureus ATCC26923. The effects and potential mechanisms of bacteriocin CBP22 on the innate immune response were investigated with a lipopolysaccharide- (LPS-) induced mouse model. The results showed that pretreatment with CBP22 prevented LPS-induced impairment in epithelial tissues and significantly reduced serum levels of IgG, IgA, IgM, TNF-α, and sIgA. Moreover, CBP22 treatment increased the expression of the zonula occludens and reduced permeability as well as apoptosis in the jejunum in LPS-treated mice. In summary, CBP22 inhibits the intestinal injury and prevents the gut barrier dysfunction induced by LPS, suggesting the potential use of CBP22 for treating intestinal damage.

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Levansucrase from Bacillus amyloliquefaciens KK9 and Its Y237S Variant Producing the High Bioactive Levan-Type Fructooligosaccharides

Biomolecules ◽

10.3390/biom10050692 ◽

2020 ◽

Vol 10 (5) ◽

pp. 692

Author(s):

Pongsakorn Phengnoi ◽

Thanapon Charoenwongpaiboon ◽

Karan Wangpaiboon ◽

Methus Klaewkla ◽

Santhana Nakapong ◽

...

Keyword(s):

Bacillus Amyloliquefaciens ◽

High Performance ◽

Amperometric Detection ◽

Anion Exchange Chromatography ◽

Exchange Chromatography ◽

Wild Type ◽

E Coli ◽

Prebiotic Activity ◽

Food Biotechnology ◽

Variant Enzyme

Levan-typed fructooligosaccharide (LFOS), a β-2,6 linked oligofructose, displays the potential application as a prebiotic and therapeutic dietary supplement. In the present study, LFOS was synthesized using levansucrase from Bacillus amyloliquefaciens KK9 (LsKK9). The wild-type LsKK9 was cloned and expressed in E. coli, and purified by cation exchanger chromatography. Additionally, Y237S variant of LsKK9 was constructed based on sequence alignment and structural analysis to enhance the LFOS production. High-performance anion-exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD) analysis indicated that Y237S variant efficiently produced a higher amount of short-chain LFOS than wild type. Also, the concentration of enzyme and sucrose in the reactions was optimized. Finally, prebiotic activity assay demonstrated that LFOS produced by Y237S variant had higher prebiotic activity than that of the wild-type enzyme, making the variant enzyme attractive for food biotechnology.

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Tailored Enzymatic Synthesis of Chitooligosaccharides with Different Deacetylation Degrees and Their Anti-Inflammatory Activity

Catalysts ◽

10.3390/catal9050405 ◽

2019 ◽

Vol 9 (5) ◽

pp. 405 ◽

Cited By ~ 5

Author(s):

P. Santos-Moriano ◽

P. Kidibule ◽

N. Míguez ◽

L. Fernández-Arrojo ◽

A.O. Ballesteros ◽

...

Keyword(s):

Tumor Necrosis Factor ◽

High Performance ◽

Tumor Necrosis ◽

Amperometric Detection ◽

Anion Exchange Chromatography ◽

Exchange Chromatography ◽

Inflammatory Activity ◽

Tnf Α ◽

Anti Inflammatory ◽

Necrosis Factor

By controlled hydrolysis of chitosan or chitin with different enzymes, three types of chitooligosaccharides (COS) with MW between 0.2 and 1.2 kDa were obtained: fully deacetylated (fdCOS), partially acetylated (paCOS), and fully acetylated (faCOS). The chemical composition of the samples was analyzed by high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) and MALDI-TOF mass spectrometry. The synthesized fdCOS was basically formed by GlcN, (GlcN)2, (GlcN)3, and (GlcN)4. On the contrary, faCOS contained mostly GlcNAc, (GlcNAc)2 and (GlcNAc)3, while paCOS corresponded to a mixture of at least 11 oligosaccharides with different proportions of GlcNAc and GlcN. The anti-inflammatory activity of the three COS mixtures was studied by measuring their ability to reduce the level of TNF-α (tumor necrosis factor) in murine macrophages (RAW 264.7) after stimulation with a mixture of lipopolysaccharides (LPS). Only fdCOS and faCOS were able to significantly reduce the production of tumor necrosis factor (TNF)-α at 6 h after stimulation with lipopolysaccharides.

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C/EBPδ protects from radiation-induced intestinal injury and sepsis by suppression of inflammatory and nitrosative stress

Scientific Reports ◽

10.1038/s41598-019-49437-x ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 2

Author(s):

Sudip Banerjee ◽

Qiang Fu ◽

Sumit K. Shah ◽

Stepan B Melnyk ◽

Esta Sterneck ◽

...

Keyword(s):

Nitrosative Stress ◽

Intestinal Barrier ◽

Intestinal Injury ◽

Nuclear Antigen ◽

Intestinal Damage ◽

Oxidative And Nitrosative Stress ◽

Tnf Α ◽

Radiation Induced ◽

Villous Height ◽

Proliferating Cell

Abstract Ionizing radiation (IR)-induced intestinal damage is characterized by a loss of intestinal crypt cells, intestinal barrier disruption and translocation of intestinal microflora resulting in sepsis-mediated lethality. We have shown that mice lacking C/EBPδ display IR-induced intestinal and hematopoietic injury and lethality. The purpose of this study was to investigate whether increased IR-induced inflammatory, oxidative and nitrosative stress promote intestinal injury and sepsis-mediated lethality in Cebpd−/− mice. We found that irradiated Cebpd−/− mice show decreased villous height, crypt depth, crypt to villi ratio and expression of the proliferation marker, proliferating cell nuclear antigen, indicative of intestinal injury. Cebpd−/− mice show increased expression of the pro-inflammatory cytokines (Il-6, Tnf-α) and chemokines (Cxcl1, Mcp-1, Mif-1α) and Nos2 in the intestinal tissues compared to Cebpd+/+ mice after exposure to TBI. Cebpd−/− mice show decreased GSH/GSSG ratio, increased S-nitrosoglutathione and 3-nitrotyrosine in the intestine indicative of basal oxidative and nitrosative stress, which was exacerbated by IR. Irradiated Cebpd-deficient mice showed upregulation of Claudin-2 that correlated with increased intestinal permeability, presence of plasma endotoxin and bacterial translocation to the liver. Overall these results uncover a novel role for C/EBPδ in protection against IR-induced intestinal injury by suppressing inflammation and nitrosative stress and underlying sepsis-induced lethality.

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Effect of Resin Acid and Zinc Oxide on Immune Status of Weaned Piglets Challenged With E. coli Lipopolysaccharide

Frontiers in Veterinary Science ◽

10.3389/fvets.2021.761742 ◽

2021 ◽

Vol 8 ◽

Author(s):

Xiaonan Guan ◽

Regiane R. Santos ◽

Hannele Kettunen ◽

Juhani Vuorenmaa ◽

Francesc Molist

Keyword(s):

Zinc Oxide ◽

Resin Acid ◽

Serum Levels ◽

Immune Defence ◽

Weaned Piglets ◽

Necrosis Factor Alpha ◽

E Coli ◽

Tnf Α ◽

Factor Alpha ◽

Necrosis Factor

With the ban of zinc oxide (ZnO) at high dosages in piglet diets in Europe by 2022, alternative nutritional solutions are being tested to support piglet immune defence during their weaning, the most critical and stressful moment of pig production. The present study evaluated the effect of zinc oxide (ZnO; 2,500 mg/kg diet) and resin acid concentrate (RAC; 200 mg/kg diet) on the immune defence of weaned piglets challenged with lipopolysaccharide (LPS). Piglets were challenged at days 7 and 21 post-weaning, and blood was sampled 1.5 and 3.0 h after each challenge to determine serum levels of pro- and anti-inflammatory cytokines. The levels of serum tumour necrosis factor alpha (TNF-α) and interleukin 8 (IL-8) increased at days 7 and 21, and those of IL-6 at day 21 when challenged piglets were fed a diet supplemented with ZnO. In challenged piglets fed with RAC, the serum levels of IL-1β, IL-6, IL-8, IL-10 and TNF-α were increased at days 7 and 21, except for that of IL-1β, which was not affected at day 21. The increased levels of these cytokines indicate the successful immune-modulatory effect of ZnO and RAC, which appears as a candidate to replace ZnO in weaned piglets' diets.

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Dose-dependent effects of adalimumab in neonatal rats with hypoxia/reoxygenation-induced intestinal damage

Bosnian Journal of Basic Medical Sciences ◽

10.17305/bjbms.2020.4823 ◽

2020 ◽

Author(s):

Halil Kocamaz ◽

Özmert MA Özdemir ◽

Nilay Şen Türk ◽

Yaşar Enli ◽

Barbaros Şahin ◽

...

Keyword(s):

Low Dose ◽

Treated Group ◽

Intestinal Injury ◽

Control Group ◽

High Dose ◽

Intestinal Damage ◽

Neonatal Rats ◽

Tnf Α ◽

Dose Dependent ◽

Hypoxia Reoxygenation

Tumor necrosis factor-alpha (TNF-α) has an important role in hypoxia/reoxygenation (H/R)-induced intestinal damage. It was shown that blocking TNF-α with infliximab has beneficial effects on experimental necrotizing enterocolitis and hypoxic intestinal injury. However, there is no data about the effect of adalimumab on H/R-induced intestinal damage. Therefore, we aimed to determine potential dose-dependent benefits of adalimumab in such damage in neonatal rats. Wistar albino rat pups were assigned to one of the four groups: control group, hypoxia group, low-dose adalimumab (5 mg/kg/day) treated group (LDAT), and high-dose adalimumab (50 mg/kg/day) treated group (HDAT). On the fourth day of the experiment, all rats except for the control group were exposed to H/R followed by euthanasia. Malondialdehyde (MDA), myeloperoxidase (MPO), TNF-α, total antioxidant capacity (TAC), and total oxidant capacity (TOC) were measured in intestinal tissue. TAC and TOC values were used to calculate the oxidative stress index (OSI). Histopathological injury scores (HIS) were also evaluated in the tissue samples. MDA levels were significantly lower in the LDAT and HDAT groups (p < 0.001). TNF-α levels were significantly lower in the LDAT group (p < 0.001). OSI was significantly higher in the H/R group than in the control and LDAT groups (p < 0.001). Mean HIS values in the LDAT group were significantly lower than those in the H/R and HDAT groups (p < 0.001). This experimental study showed that low-dose adalimumab appears to have a beneficial effect on intestinal injury induced with H/R in neonatal rats.

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PSVII-3 Effects of oral administration of essential oils on anti-immune stress, antimicrobial and repairing the intestinal damage in broilers challenged by lipopolysaccharide

Journal of Animal Science ◽

10.1093/jas/skz122.302 ◽

2019 ◽

Vol 97 (Supplement_2) ◽

pp. 171-171

Author(s):

Hanjin Oh ◽

Shudong Liu ◽

Won Yun ◽

Jihwan Lee ◽

Jiseon An ◽

...

Keyword(s):

Oral Administration ◽

Essential Oils ◽

Basal Diet ◽

Stress Reaction ◽

Intestinal Damage ◽

E Coli ◽

Abnormal Growth ◽

Immune Stress ◽

Tnf Α

Abstract The aim of the present study was to evaluate the effects of oral administration of essential oils (Carvacrol, Thyme and Oregano essential oils) on anti-immune stress, antimicrobial and repairing the intestinal damage, caused by Salmonella enterica Lipopolysaccharide, in vivo of broilers. A total of 100 ROSS 308 broilers (21-day-old; 1.10 ± 0.11kg) were randomly allocated to 5 groups, T1: basal diet + Saline; T2: basal diet + LPS; T3: basal diet+ 200μL carvacrol oils+ LPS; T4: basal diet+ 200μL thyme oils+ LPS; T5: basal diet+ 200μL oregano oils+ LPS, with 20 replicates every group, and 1 chicken per replicate per cage. The challenged by LPS resulted in an immune stress, which manifests as the abnormal growth (P < 0.05) in immune organs, the content of IgG (P < 0.05) and TNF-α(P < 0.05) and the rectum temperature (P < 0.05) increased compared with other groups; the oral administration of essential oils controlled the immune stress to a certain extent. The essential oils could withhold maleficent bacteria, as E. coli (P < 0.05) and Salmonella enumeration (P < 0.05), in vivo of the broiler. Meanwhile, the essential oils repaired the intestinal damage which shown a reduction in the villi height (P < 0.05) and goblet cell (P < 0.05) caused by LPS. In conclusion, the essential oils (Carvacrol, Thyme and Oregano essential oils) controlled the stress reaction and maintain intestinal health to a certain extent. http://www.conferenceharvester.com/

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Protection against Legionnaire's Disease: Recombinant Flagellin A of Legionella pneumophila Can Induce Protective Immunity against Bacteremia in a BALB/c Murine Model

Journal of Molecular Microbiology and Biotechnology ◽

10.1159/000460295 ◽

2017 ◽

Vol 27 (2) ◽

pp. 110-116 ◽

Cited By ~ 4

Author(s):

Ashraf Mohabati Mobarez ◽

Roya Ahmadrajabi ◽

Nima Khoramabadi ◽

Ali-Hatef Salmanian

Keyword(s):

Legionella Pneumophila ◽

Serum Antibody ◽

Exchange Chromatography ◽

Sublethal Dose ◽

Cell Mediated Immunity ◽

Lethal Challenge ◽

E Coli ◽

Tnf Α ◽

Recombinant Type

To investigate the immunoprotective effects of the recombinant type A flagellin (FLA), the flaA gene of Legionella pneumophila serogroup 1 strain Paris was cloned into pET28a(+). Recombinant FLA (rFLA) was overexpressed in E. coli BL21 (DE3) and purified by Ni2+ exchange chromatography. Female BALB/c aged 6-8 weeks were immunized with 20 μg of rFLA. Nonimmunized mice along with mice inoculated with a sublethal dose of live L. pneumophila intravenously were considered as negative and positive controls, respectively. A significant serum antibody response was observed in female BALB/c mice immunized with rFLA. Production of IFN-γ and IL-12, and TNF-α in the serum and the splenocyte cultures, and antigen-specific splenocyte proliferation suggested a strong innate and adaptive cell-mediated immunity response in rFLA-immunized mice. Intravenous lethal challenge with L. pneumophila serogroup 1 (strain Paris) showed that 60% of mice immunized with rFLA survived in a 10-day follow-up survey. These results show that rFLA from L. pneumophila can elicit strong innate and adaptive immune responses and suggest the possibility of a long-term immunity against lethal challenge with L. pneumophila.

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Mitochondrial DNA: A Biomarker of Disease Severity in Necrotizing Enterocolitis

European Journal of Pediatric Surgery ◽

10.1055/s-0039-1697910 ◽

2019 ◽

Vol 30 (01) ◽

pp. 085-089

Author(s):

Edoardo Bindi ◽

Bo Li ◽

Haitao Zhou ◽

Maarten Janssen Lok ◽

Mashriq Alganabi ◽

...

Keyword(s):

Necrotizing Enterocolitis ◽

Intestinal Ischemia ◽

Organ Damage ◽

Intestinal Injury ◽

Intestinal Damage ◽

Tnf Α ◽

Polymerase Chain ◽

Factor Α ◽

Mtdna Markers ◽

Necrosis Factor

Abstract Introduction There is a need to develop sensitive markers to diagnose or monitor the severity of intestinal damage in necrotizing enterocolitis (NEC). Mitochondrial deoxyribonucleic acid (mtDNA) is increased in the intestine and blood of adults in response to intestinal ischemia and can trigger secondary organ damage. We hypothesize that mtDNA is increased during experimental NEC and that mtDNA levels are correlated to the degree of intestinal injury. Materials and Methods NEC was induced in C57BL/6 mice (n = 18) (approval: 44032) by gavage feeding with hyperosmolar formula, hypoxia, and lipopolysaccharide administration from postnatal day (P) 5 to 9. Breastfed pups served as control (n = 15). Blood was collected by cardiac puncture and terminal ileum was harvested on P9. Reverse transcription quatitative polymerase chain reaction was used to measure mtDNA (markers COX3, CYTB, ND1) and inflammatory cytokines (interleukin 6 [IL-6] and tumor necrosis factor-α[TNF-α]) in blood and ileum. Intestinal injury was scored blindly by four investigators and classified as no/minor injury (score 0 or 1) or NEC (score ≥2). Results mtDNA is significantly increased in gut and blood of NEC mice (p < 0.05). Furthermore, mtDNA increases in intestine and blood proportionally to the degree of intestinal injury as indicated by a positive correlation with histological scoring and inflammation (r = 0.6; p < 0.05) (expression of IL-6 and TNF-α). Conclusion Following NEC intestinal injury, mtDNA is released from the intestine into circulation. The blood level of mtDNA is related to the degree of intestinal injury. mtDNA can be a novel marker of intestinal injury and can be useful for monitoring the progression of NEC.

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Degradation of Pectins with Different Degrees of Esterification by Bacteroides thetaiotaomicron Isolated from Human Gut Flora

Applied and Environmental Microbiology ◽

10.1128/aem.66.4.1321-1327.2000 ◽

2000 ◽

Vol 66 (4) ◽

pp. 1321-1327 ◽

Cited By ~ 75

Author(s):

Gerhard Dongowski ◽

Angelika Lorenz ◽

Horst Anger

Keyword(s):

High Performance ◽

Culture Media ◽

Short Chain Fatty Acids ◽

Anion Exchange Chromatography ◽

Exchange Chromatography ◽

Fecal Flora ◽

Bacteroides Thetaiotaomicron ◽

E Coli ◽

Pure Cultures

ABSTRACT A complete human fecal flora and cultures of defined species obtained from fecal flora were investigated in vitro to determine their ability to ferment the dietary fiber pectin. Bacteroides thetaiotaomicron was tested as a pectin-degrading microorganism alone and in coculture with Escherichia coli. Macromolecular pectins with different degrees of esterification were used as substrates in microbial degradation studies. The levels of oligogalacturonic acids formed in batch cultures were estimated during a 24- or 48-h incubation period by using high-performance thin-layer chromatography and high-performance anion-exchange chromatography. The spectrum and the amount of unsaturated oligogalacturonic acids formed as intermediate products of pectin fermentation changed permanently in the culture media during incubation with the complete fecal flora. After 24 h, no oligogalacturonic acids were detected. The pectin-degrading activities of pure cultures of B. thetaiotaomicron were lower than the pectin-degrading activity of a complete fecal flora. Cocultures of B. thetaiotaomicronand E. coli exhibited intermediate levels of degradation activity. In pure cultures of E. coli no pectin-degrading activity was found. Additionally, the rate of pectin degradation was affected by the degree of esterification of the substrate. Saturated oligogalacturonic acids were not found during pectin fermentation. The disappearance of oligogalacturonic acids in the later stages of fermentation with both the complete fecal flora and B. thetaiotaomicron was accompanied by increased formation of short-chain fatty acids.

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Generation of Mature Nα-Terminal Acetylated Thymosinα1 by Cleavage of Recombinant Prothymosinα

The Scientific World JOURNAL ◽

10.1155/2013/387282 ◽

2013 ◽

Vol 2013 ◽

pp. 1-8 ◽

Cited By ~ 1

Author(s):

Bo Liu ◽

Xin Gong ◽

Shaohong Chang ◽

Peng Sun ◽

Jun Wu

Keyword(s):

Mass Spectrometry ◽

High Performance Liquid Chromatography ◽

High Performance ◽

Expression System ◽

Exchange Chromatography ◽

Cation Exchange Chromatography ◽

Reverse Phase ◽

E Coli ◽

Amino Acid Peptide

Nα-terminal acetylation of peptides plays an important biological role but is rarely observed in prokaryotes. Nα-terminal acetylated thymosinα1 (Tα1), a 28-amino-acid peptide, is an immune modifier that has been used in the clinic to treat hepatitis B and C virus (HBV/HCV) infections. We previously documented Nα-terminal acetylation of recombinant prothymosinα(ProTα) inE. coli. Here we present a method for production of Nα-acetylated Tα1 from recombinant ProTα. The recombinant ProTαwas cleaved by human legumain expressed inPichia pastoristo release Tα1in vitro. The Nα-acetylated Tα1 peptide was subsequently purified by reverse phase and cation exchange chromatography. Mass spectrometry indicated that the molecular mass of recombinant Nα-acetylated Tα1 was 3108.79 in, which is identical to the mass of Nα-acetylated Tα1 produced by total chemical synthesis. This mass corresponded to the nonacetylated Tα1 mass with a 42 Da increment. The retention time of recombinant Nα-acetylated Tα1 and chemosynthetic Nα-acetylated Tα1 were both 15.4 min in RP-high performance liquid chromatography (HPLC). These data support the use of anE. coliexpression system for the production of recombinant human Nα-acetylated Tα1 and also will provide the basis for the preparation of recombinant acetylated peptides inE. coli.

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