Role of Rotavirus Enterotoxin NSP4 in the Inflammatory Response in Murine Macrophage RAW 264.7 Cells

The rotavirus nonstructural glycoprotein, NSP4, has been identified as the first viral enterotoxin capable of inducing diarrhea. To investigate the biological function of NSP4 in the inflammatory process, a cDNA from human rotavirus (Wa strain) RNA segment 10 was amplified by RT-PCR, cloned into TA vector, and subsequently subcloned into pET23b expression plasmid. The expression of NSP4 protein was determined by SDS-PAGE and Western blotting, then, the protein was purified by affinity chromatography on Ni-NTA-agarose column. The inflammatory effects of NSP4, namely, production of nitric oxide (NO), pro-inflammatory cytokines (IL-1β, IL-6, IL-10, and TNF-α), and prostaglandin E2 (PGE₂), was evaluated using NSP4-stimulated RAW 264.7 murine macrophages and compared with those observed after stimulation with lipopolysaccharide (LPS). The levels of IL-1β, IL-6, and TNF-α were significantly increased, and those of NO and PGE₂ also increased in NSP4-stimulated RAW 264.7 cells. These findings indicate that NSP4 plays an important role in the inflammatory response observed during rotavirus infection.

Anti-Inflammatory and Anti-Superbacterial Activity of Polyphenols Isolated from Black Raspberry

The fruit of the black raspberry (Rubus coreanus Miquel) has been employed in traditional medicine, and recent studies have demonstrated its measureable biological activities. However, the root of the black raspberry has not been studied. Therefore, in this study, we evaluated the anti-inflammatory and antibacterial properties of the root and unripe fruit polyphenols of the black raspberry. Both polyphenols proved to have anti-inflammatory activity as evidenced by the decreased nitric oxide (NO), cytokines (IL-1beta , IL-6, and IL-10) and prostaglandin E2 (PGE2) levels in lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophages. However, root polyphenols showed stronger anti-inflammatory activity than fruit polyphenols. LPS-induced mRNA and protein expressions of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 levels were also decreased, confirming the anti-inflammatory activity. Root polyphenols showed lethal activity against methicillin-resistant Staphylococcus aureus (MRSA), carbapenem-resistant Acinetobacter baumannii (CRAB), and Bacillus anthracis. In contrast, the black raspberry fruit did not demonstrate these properties. These data provide the first demonstration that black raspberry root has potential anti-inflammatory and anti-superbacterial properties that can be exploited as alternatives for use in the food and cosmetic industries and/or as pharmaceuticals.

Analysis of Oropharyngeal Microbiota between the Patients with Bronchial Asthma and the Non-Asthmatic Persons

Bronchial asthma can be triggered by microbial agents in the oropharynx. This study was designed to identify the differences in microbiota of oropharynx of bronchial asthmatic patients in contrast to normal controls. In order to resolve the qualitative and quantitative diversity of the 16S rRNA gene present in the oropharynx microbiota of 4 patients and 4 controls, we compared microbial communities using Sanger sequencing and 376 sequences of 16S rRNA gene were analyzed. Of the total microbial diversity detected in the oropharynx in asthmatic patients 45.6% comprised members of the Firmicutes. In contrast, Proteobacteria (44.0%) dominated the oropharyngeal microbiota in the normal control group. Members of the Bacteroidetes, Fusobacteria, Actinobacteria, TM7, Cyanobacteria and unclassified bacteria were present in both groups. In conclusion, the difference in the microbiota of the oropharynx between patients and normal individuals could trigger symptomatic attacks in bronchial asthma.

A Metaviromic Analysis of Viral Communities in the Feces of Unexplained Acute Gastroenteritis

Although viruses are the most common cause of acute gastroenteritis (AGE) in humans, details about the causative viruses in AGE are largely unknown because many causative viruses are unable to be cultured by current culture techniques. In our study, fecal samples from 10 children under five years of age with unexplained AGE and 10 healthy children were investigated for RNA viruses using random priming (RP)-mediated sequence-independent single primer amplification (SISPA). The causative viruses in cases of cryptogenic diarrhea were then assessed for their potential diagnostic value. Of the 1,129 viral clones identified, rotavirus was most commonly associated with AGE (125 sequences, 22.4%). In contrast, bacteriophage was most common (43 sequences, 13.6%) in healthy children. The remaining 515 viral clones were unidentifiable. These findings suggest that investigation of cases or outbreaks of unexplained diarrhea using a metaviromic strategy is a new avenue for diagnosis.