Effects of ß-Glucan on the Release of Nitric Oxide by Macrophages Stimulated with Lipopolysaccharide.

This research analyzed the effect of ß-glucan that is expected to alleviate the production of the inflammatory mediator in macrophagocytes, which are processed by the lipopolysaccharide (LPS) of Escherichia. The incubated layer was used for a nitric oxide (NO) analysis. The DNA-binding activation of the small unit of nuclear factor-κB was measured using the enzyme-linked immunosorbent assay-based kit. In the RAW264.7 cells that were vitalized by Escherichia coli (E. coli) LPS, the ß-glucan inhibited both the combatant and rendering phases of the inducible NO synthase (iNOS)-derived NO. ß-Glucan increased the expression of the heme oxygenase-1 (HO-1) in the cells that were stimulated by E. coli LPS, and the HO-1 activation was inhibited by the tin protoporphyrin IX (SnPP). This shows that the NO production induced by LPS is related to the inhibition effect of ß-glucan. The phosphorylation of c-Jun N-terminal kinases (JNK) and the p38 induced by the LPS were not influenced by the ß-glucan, and the inhibitory κB-α (IκB-α) decomposition was not influenced either. Instead, ß-glucan remarkably inhibited the phosphorylation of the signal transducer and activator of transcription-1 (STAT1) that was induced by the E. coli LPS. Overall, the ß-glucan inhibited the production of NO in macrophagocytes that was vitalized by the E .coli LPS through the HO-1 induction and the STAT1 pathways inhibition in this research. As the host immune response control by ß-glucan weakens the progress of the inflammatory disease, ß-glucan can be used as an effective immunomodulator.

Effect of caffeic acid phenethyl ester on Prevotella intermedia lipopolysaccharide-induced production of proinflammatory mediators in murine macrophages.

BACKGROUND AND OBJECTIVE: Caffeic acid phenethyl ester (CAPE) has numerous potentially beneficial properties, including antioxidant, immunomodulatory and anti-inflammatory activities. However, the effect of CAPE on periodontal disease has not been studied before. This study was designed to investigate the efficacy of CAPE in ameliorating the production of proinflammatory mediators in macrophages activated by lipopolysaccharide (LPS) from Prevotella intermedia, a pathogen implicated in periodontal disease. MATERIAL AND METHODS: LPS from P. intermedia ATCC 25611 was isolated by using the standard hot phenol-water method. Culture supernatants were assayed for nitric oxide (NO), interleukin (IL)-1ß and IL-6. We used real-time polymerase chain reaction to quantify inducible NO synthase, IL-1ß, IL-6, heme oxygenase (HO)-1 and suppressors of cytokine signaling (SOCS) 1 mRNA expression. HO-1 protein expression and levels of signaling proteins were assessed by immunoblot analysis. DNA-binding activities of NF-κB subunits were analyzed by using the enzyme-linked immunosorbent assay-based kits. RESULTS: CAPE exerted significant inhibitory effects on P. intermedia LPS-induced production of NO, IL-1ß and IL-6 as well as their mRNA expression in RAW264.7 cells. CAPE-induced HO-1 expression in cells activated with P. intermedia LPS, and selective inhibition of HO-1 activity by tin protoporphyrin IX attenuated the inhibitory effect of CAPE on LPS-induced NO production. CAPE did not interfere with IκB-α degradation induced by P. intermedia LPS. Instead, CAPE decreased nuclear translocation of NF-κB p65 and p50 subunits induced with LPS, and lessened LPS-induced p50 binding activity. Further, CAPE showed strong inhibitory effects on LPS-induced signal transducer and activator of transcription 1 and 3 phosphorylation. Besides, CAPE significantly elevated SOCS1 mRNA expression in P. intermedia LPS-stimulated cells. CONCLUSION: Modulation of host response by CAPE may represent an attractive strategy towards the treatment of periodontal disease. In vivo studies are required to appraise the potential of CAPE further as an immunomodulator in the treatment of periodontal disease.

Prevalence, phenotypic traits and molecular characterization of emetic toxin-producing Bacillus cereus strains isolated from human stools in Korea.

AIMS: To investigate the prevalence and genotypic/phenotypic characters of emetic toxin-producing Bacillus cereus strains isolated from sporadic food poisoning cases in Korea. METHODS AND RESULTS: The prevalence of emetic B. cereus was determined in 56 899 stool samples from sporadic food poisoning cases in Korea between 2004 and 2006. We assessed toxin profiles, phenotypic traits and antibiotic resistance. The molecular subtyping was ascertained using an automated repetitive sequence-based PCR (rep-PCR) system, DiversiLab™, with these emetic strains isolated from sporadic food poisoning cases and other emetic strains isolated from an outbreak and food samples. Emetic B. cereus was present in 0·012% of sporadic food poisoning cases. The prevalence of nheABC, hblCDA, cytK and entFM enterotoxin genes among emetic strains was 100, 14·3, 14·3 and 100%, respectively. Most emetic strains were negative for salicin hydrolysis (100%), starch fermentation (85·7%) and haemolysis (85·7%). One emetic isolate, VK7, exhibited several unique traits, such as harbouring the hbl gene and ability to hydrolyse starch. All isolated strains were highly resistant to ß-lactam antibiotics. All emetic strains except VK7 exhibited an identical rep-PCR banding pattern, while nonemetic strains were classified into various pulsotypes. CONCLUSIONS: Most emetic strains except one isolate exhibited similar genotypic/phenotypic traits and subtyping pattern. Automatic rep-PCR (DiversiLab™) may be used to discriminate emetic strains from nonemetic strains, although we could not distinguish between most emetic strains using that. SIGNIFICANCE AND IMPACT OF THE STUDY: Result of this study may contribute an extended database on the prevalence and toxigenic traits of emetic B. cereus strains isolated from Korea.

Comparison of 2 types of broths and 3 selective agars for the detection of Campylobacter species in whole-chicken carcass-rinse samples.

In this study, we compared the effectiveness of 2 types of Bolton broths and 3 selective media for isolating Campylobacter spp. from naturally contaminated whole-chicken carcass-rinse samples. One hundred chickens were rinsed with buffered peptone water, and the rinses were added to 2× Bolton broth (with or without blood supplementation). The samples were incubated and then streaked onto Preston agar, modified cefoperazone charcoal deoxycholate agar (mCCDA), and Campy-Cefex agar, which was followed by incubation under microaerobic conditions. No statistical differences were observed (P > 0.05) in isolation rate and selectivity between the 2 types of Bolton broths. Among the 3 selective agars, Preston agar yielded a significantly (P < 0.05) better isolation rate and selectivity. The Campy-Cefex agar, which is recommended by many food authorities for its high quantitative detection ability, showed extensive contamination with competing microorganisms and exhibited the lowest isolation rate and selectivity.

Evaluation of selective enrichment broths and chromogenic media for Salmonella detection in highly contaminated chicken carcasses.

We evaluated the effectiveness of 2 selective enrichment broths, Rappaport-Vassiliadis Soy (RVS) and Muller-Kauffmann tetrathionate with novobiocin (MKTTn), for the isolation of Salmonella from chicken carcasses obtained from 3 different types of retail markets. We also compared a chromogenic agar, chromID Salmonella agar (SM-ID 2), with a classic plating medium, xylose lysine deoxycholate agar (XLD). Salmonella were isolated from 118 of the 180 samples (65.5%). Salmonella were detected in 105 samples (88%) plated on XLD and 111 samples (94%) plated on SM-ID 2 when RVS broth was used for enrichment, and 43 samples (36.4%) plated on XLD and 67 samples (56.8%) plated on SM-ID 2 when the MKTTn broth was used. The highest sensitivity was found in the RVS-XLD combination (0.99), followed by RVS-SM-ID 2 (0.97). The specificity of the RVS-SM-ID 2 combination was the highest (0.89), but that of the MKTTn-XLD combination was zero. The results of this study indicate that the selective enrichment broths had a great effect on the sensitivity and specificity of plating media, and our study confirms that the RVS broth is the most suitable enrichment for the investigation of Salmonella in chicken carcasses. This observation suggests that use of RVS broth for selective enrichment and SM-ID 2 for selective isolation may be the best combination to determine the presence of Salmonella in chicken carcasses.