OM-X®, a Fermented Vegetables Extract, Facilitates Muscle Endurance Capacity in Swimming Exercise Mice.

The anti-fatigue effect was investigated of the probiotic supplement, OM-X®, on forced swimming capacity in mice. Mice were administered either vehicle (distilled water; DW) or OM-X® (85 mg/kg body weight) by gavage for 4 weeks. Forced swimming tests were conducted weekly using the Ishihara-modified Matsumoto swimming pool. The endurance swimming time of the final forced swimming exercise in mice fed with OM-X® group showed an approximately 2-fold increase compared with the vehicle control group. Biomedical parameters, including blood lactate, blood superoxide dismutase (SOD) activity, serum triacylglycerol (TG), hepatic total lipids (TL), TG and phospholipid (PL) were significantly lower in mice fed-with OM-X® than those in the vehicle control group. Furthermore, the mRNA expression levels of carbamoyl phosphate synthetase 1 (Cpsl) and arginase I (Argl), in the urea cycle, were increased by OM-X® feeding. Thus, our findings suggest promotion of lipid metabolism and up-regulation of the urea cycle, at least in part, for the anti-fatigue effect mediated by OM-X®.

OM-X®, Fermented Vegetables Extract Suppresses Antigen-Stimulated Degranulation in Rat Basophilic Leukemia RBL-2H3 Cells and Passive Cutaneous Anaphylaxis Reaction in Mice.

OM-X® is a hand-made and naturally manufactured probiotic supplement. This fermented food product is made from vegetables, fruits, seaweeds and mushrooms, using 12 strains of lactic acid bacteria and bifidobacteria. OM-X® is also known to have beneficial health properties, and some of its components show effects on antigen (Ag)-stimulated degranulation activity, indicating that OM-X® may be useful in the treatment of allergy responses and symptoms. In this study, we evaluated the inhibitory effects of OM-X® on Ag-stimulated degranulation in rat basophilic leukemia RBL-2H3 cells, clarified the underlying mechanisms, and determined the active compounds in OM-X® for suppression of degranulation. Treatment with OM-X® gradually suppressed Ag-stimulated degranulation throughout the maturation period. OM-X® also gradually produced melanoidins by lactic acid bacterial fermentation during the maturation process. There was a high correlation between the suppression levels of Ag-stimulated degranulation and the browning of OM-X®. Furthermore, the inhibition of Ag-stimulated degranulation by OM-X® was found to be partially due to the direct inactivation of NADPH oxidase. To elucidate the in vivo effects of OM- X®, type I allergy model mice were orally administered with OM-X®, and the passive cutaneous anaphylaxis (PCA) reaction was measured. OM-X® intake remarkably suppressed the PCA reaction. Taken together, our findings suggest that OMX® could be a beneficial food to ameliorate allergic reactions.

Complete sequence analysis of two cryptic plasmids from Bifidobacterium kashiwanohense†JCM 15439 (type strain) isolated from healthy infant feces.

Bifidobacterial plasmids reported so far are derived from a limited number of strains and plasmids of bifidobacterial type strains isolated from humans are unknown. We found that Bifidobacterium kashiwanohense JCM 15439 (type strain) isolated from a healthy infant contained two cryptic plasmids, designated pBBKW-1 and pBBKW-2. We determined and analyzed the complete sequences of both plasmids. pBBKW-1 (7716 bp) was predicted to replicate by a rolling-circle mechanism and encode six protein-coding genes, two of which are putative replication proteins. pBBKW-1 seems to be a cointegrate plasmid containing two copies of the plasmid pMG1 from Bifidobacterium longum. pBBKW-2 (2920 bp) was predicted to encode six protein-coding genes and be a theta-type replicating plasmid, which has been reported to be more stable than a rolling circle-type replicating plasmid frequently found in bifidobacteria. Our finding will provide new insights into safe recombinant plasmid constructions for humans.

Genomic adaptation of the Lactobacillus casei group.

Lactobacillus casei, L. paracasei, and L. rhamnosus form a closely related taxonomic group (Lactobacillus casei group) within the facultatively heterofermentative lactobacilli. Here, we report the complete genome sequences of L. paracasei JCM 8130 and L. casei ATCC 393, and the draft genome sequence of L. paracasei COM0101, all of which were isolated from daily products. Furthermore, we re-annotated the genome of L. rhamnosus ATCC 53103 (also known as L. rhamnosus GG), which we have previously reported. We confirmed that ATCC 393 is distinct from other strains previously described as L. paracasei. The core genome of 10 completely sequenced strains of the L. casei group comprised 1,682 protein-coding genes. Although extensive genome-wide synteny was found among the L. casei group, the genomes of ATCC 53103, JCM 8130, and ATCC 393 contained genomic islands compared with L. paracasei ATCC 334. Several genomic islands, including carbohydrate utilization gene clusters, were found at the same loci in the chromosomes of the L. casei group. The spaCBA pilus gene cluster, which was first identified in GG, was also found in other strains of the L. casei group, but several L. paracasei strains including COM0101 contained truncated spaC gene. ATCC 53103 encoded a higher number of proteins involved in carbohydrate utilization compared with intestinal lactobacilli, and extracellular adhesion proteins, several of which are absent in other strains of the L. casei group. In addition to previously fully sequenced L. rhamnosus and L. paracasei strains, the complete genome sequences of L. casei will provide valuable insights into the evolution of the L. casei group.

Immunomodulatory effects of heat-killed Enterococcus faecalis TH10 on murine macrophage cells.

The objective of this study was to investigate the immunomodulatory effects of heat-killed Enterococcus faecalis TH10 (hk-TH10) and its signal transduction on murine macrophage RAW264 cells. RAW264 cells produced nitric oxide (NO) following hk-TH10 treatment. In order to investigate the mechanisms underlying hk-TH10-stimulated NO production, we further measured NO production in RAW264 cells treated with Toll-like receptor (TLR) 4 inhibitor peptide, NF-κB inhibitor, TLR1-siRNA, TLR2-siRNA, and TLR-6 siRNA. Furthermore, the activation of TLR2-TLR1/6 pathway molecules was analyzed by Western blotting. The result of this study showed that hk-TH10 stimulates NO in RAW264 cells through the activation of the TLR2-TLR1/6 pathway. From our findings, we can conclude that hk-TH10 isolated from a traditional side-dish fermented food (tempeh) may facilitate host immunomodulation.

Selenite assimilation into formate dehydrogenase H depends on thioredoxin reductase in Escherichia coli.

Escherichia coli growing under anaerobic conditions produce H(2) and CO(2) by the enzymatic cleavage of formate that is produced from pyruvate at the end of glycolysis. Selenium is an integral part of formate dehydrogenase H (FDH H), which catalyses the first step in the formate hydrogen lyase (FHL) system. The genes of FHL system are transcribed only under anaerobic conditions, in the presence of a sigma 54-dependent transcriptional activator FhlA that binds formate as an effector molecule. Although the formate addition to the nutrient media has been an established procedure for inducing high FDH H activity, we have identified a low-salt nutrient medium containing <0.1% NaCl enabled constitutive, high expression of FDH H even without formate and d-glucose added to the medium. The novel conditions allowed us to study the effects of disrupting genes like trxB (thioredoxin reductase) or gor (glutathione reductase) on the production of FDH H activity and also reductive assimilation of selenite ( SeO 3(2-)) into the selenoprotein. Despite the widely accepted hypothesis that selenite is reduced by glutathione reductase-dependent system, it was demonstrated that trxB gene was essential for FDH H production and for labelling the FDH H polypeptide with 75Se-selenite. Our present study reports for the first time the physiological involvement of thioredoxin reductase in the reductive assimilation of selenite in E. coli.

Selenophosphate synthetase genes from lung adenocarcinoma cells: Sps1 for recycling L-selenocysteine and Sps2 for selenite assimilation.

A labile selenium donor compound monoselenophosphate is synthesized from selenide and ATP by selenophosphate synthetase (SPS). In the present study, Sps1 and Sps2 were cloned from a cDNA library prepared from human lung adenocarcinoma cells (NCIH441). The human lung Sps1 has been cloned as an ORF of 1,179 bp, identical in sequence to that of the recently revised human liver Sps1. The in-frame TGA codon of the lung Sps2 was genetically altered to TGT (Cys) to obtain the Sps2Cys gene. Expression of the recombinant plasmids containing Sps1 or Sps2Cys was highly toxic to Escherichia coli host cells grown aerobically. Accordingly, the human lung Sps homologs were characterized by an in vivo complementation assay using a selD mutant strain. An added selenium source and a low salt concentration (0.1-0.25% NaCl) in the medium were required for reproducible and sensitive in vivo complementation. Sps2Cys effectively complemented the selD mutant, and the resulting formate dehydrogenase H activity was as high as that of WT E. coli MC4100. In contrast, only a weak complementation of the selD mutant by the Sps1 gene was observed when cells were grown in selenite media. Better complementation with added l-selenocysteine suggested involvement of a selenocysteine lyase for mobilization of selenium. Based on this apparent substrate specificity of the Sps1 and Sps2 gene products we suggest that the Sps1-encoded enzyme depends on a selenium salvage system that recycles l-selenocysteine, whereas the Sps2 enzyme can function with a selenite assimilation system.