Rhodobacter azotoformans LPS (RAP99-LPS) Is a TLR4 Agonist That Inhibits Lung Metastasis and Enhances TLR3-Mediated Chemokine Expression.

The lipopolysaccharides (LPSs) of Rhodobacter are reported to be TLR4 antagonists. Accordingly, the extract of Rhodobacter azotoformans (RAP99) is used as a health supplement for humans and animals in Japan to regulate immune responses in vivo. We previously analyzed the LPS structure of RAP99 (RAP99-LPS) and found it is different from that of E. coli-LPS but similar to lipid A from Rhodobacter sphaeroides (RSLA), a known antagonist of TLR4, with both having three C14 fatty acyl groups, two C10 fatty acyl groups, and two phosphates. Here we show that RAP99-LPS has an immune stimulatory activity and acts as a TLR4 agonist. Pretreatment of RAP99-LPS suppressed E. coli-LPS-mediated weight loss, suggesting it is an antagonist against E. coli-LPS like other LPS isolated from Rhodobacter. However, injections of RAP99-LPS caused splenomegaly and increased immune cell numbers in C57BL/6 mice but not in C3H/HeJ mice, suggesting that RAP99-LPS stimulates immune cells via TLR4. Consistently, RAP99-LPS suppressed the lung metastasis of B16F1 tumor cells and enhanced the expression of TLR3-mediated chemokines. These results suggest that RAP99-LPS is a TLR4 agonist that enhances the activation status of the immune system to promote anti-viral and anti-tumor activity in vivo.

Identification of a Point Mutation in the Granule-bound Starch Synthase I Gene (GBSSI) in a waxy Diploid Wheat Mutant and Design of Molecular Markers for Backcrossing.

In cereals, granule-bound starch synthase I (GBSSI)-deficient mutants accumulate glutinous (amylose-free) starch in their storage tissues. The amylose-free starch produced by waxy (wx) mutants of hexaploid bread wheat (Triticum aestivum L.) is used in cakes and breads. However, wx mutants of diploid wheat (T. monococcum L.) have so far no commercial applications. In this study, we identified a mutation in exon 6 of GBSSI in a diploid wheat wx mutant that resulted in the replacement of Trp355 with a stop codon. Molecular markers were developed for the rapid screening of the mutation, which should allow the selection of heterozygous and homozygous plants during backcrossing. This will facilitate the improvement of the agricultural traits of the wx mutant and the generation of new amylose-free wx lines.

Video-endoscopic comparison of swallowing waxy rice mochi and waxy wheat mochi: improvement of a traditional Japanese food that presents a choking hazard.

Mochi is highly cohesive and adhesive, and easy to choke on. Many of the fatal suffocation accidents with mochi occur in the elderly aged 65 years or older. These circumstances prompted us to investigate a special property of waxy wheat which is similar in texture to waxy rice, but is less cohesive and adhesive. We compared the differences in chewing and swallowing movements associated with eating waxy rice mochi and waxy wheat mochi between healthy adults and healthy elderly. Healthy elderly chewed mochi more and longer than healthy adults. Although there was no difference in the number of chewing cycles or total duration of chewing between the two types of mochi, waxy wheat mochi was easier to chew and left less pharyngeal residue. These findings lead us to suggest that waxy wheat mochi is promising as a food that is easy to swallow and difficult to choke on.

Effect of lignin-derived lignophenols on hepatic lipid metabolism in rats fed a high-fat diet.

The effect of lignin-derived lignophenols on lipid metabolism in the livers of rats fed a high-fat diet was investigated. Rats fed a diet providing 45% of energy from fat were divided into 2 groups, namely 0% and 0.5% lignophenols-containing diets. The controls were fed a diet providing 10% of energy from fat. Plasma blood parameters, protein expression of acetyl-CoA carboxylase (ACC) and sterol regulatory element-binding protein (SREBP)-1, and SREBP-1c mRNA expression in the livers were examined. The plasma triglyceride levels in the rats fed lignophenols-containing diets were decreased. SREBP-1c mRNA expression in the rats fed lignophenols-containing diets was significantly reduced compared with the rats fed high-fat diets, and phosphorylated ACC protein in the rats fed lignophenols-containing diets was significantly increased. Our results suggested that lignophenols suppress the expression of SREBP-1c mRNA and the phosphorylation of ACC in the liver, and may lead to a decrease in plasma triglyceride levels.

Effect of lignin-derived lignophenols on vascular oxidative stress and inflammation in streptozotocin-induced diabetic rats.

Lignophenols (LP) are the derivatives of native lignin, which is an abundant organic polymer in the plant kingdom. This study investigated whether LP can attenuate vascular oxidative stress and inflammation in streptozotocin (STZ)-induced diabetic rats. The diabetic rats induced by a single intravenous injection of STZ were randomly divided into two groups fed either 0 or 1.0% LP-containing diet. After 5 weeks of treatment, the superoxide (O(2)(-)) production, mRNA expression levels of nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) oxidase subunits, monocyte chemoattractant protein-1 (MCP-1) and its receptor C-C chemokine receptor 2 (CCR2), and protein expression level of inducible nitric oxide synthase (iNOS) were examined in the aorta of vehicle-injected control and diabetic rats treated with or without LP. The increased O(2)(-) production and mRNA expression levels of NAD(P)H oxidase subunits Nox4 and p47phox were found to be significantly reduced in the aorta of diabetic rats treated with LP. The mRNA expression of MCP-1 and CCR2, and the protein expression of iNOS were found to be increased in the aorta of untreated diabetic rats, whereas these levels were significantly lower in the LP-treated group. These findings suggest that LP could attenuate vascular oxidative stress and/or inflammation via inhibition of NAD(P)H oxidase. This may lead to an improvement in the vascular impairment of diabetes.

Lignophenols decrease oleate-induced apolipoprotein-B secretion in HepG2 cells.

Lignin is one of the components in the plant cell wall, after cellulose, making up 20-30% of the global plant biomass. Lignophenols (LPs) are derivatives of lignin with high phenolic function and antioxidant properties. However, their medicinal property is not well characterised. Apolipoprotein-B (apo-B) is an essential component in very low-density lipoprotein, and high levels of serum apolipoprotein-B (apo-B) are a major factor for coronary heart disease. In this study, we examined the effect of lignophenols on apo-B secretion in HepG2 cells. HepG2 cells were treated with varying concentrations of LPs and 0.8 mm sodium oleate. LPs decreased oleate-induced apo-B secretion in a dose-dependent manner. LPs also decreased oleate-induced microsomal triglyceride transfer protein (MTTP) mRNA expression and cellular total cholesterol, suggesting that lipid bioavailability and lipidation of lipoprotein are likely involved in the decreased secretion of apo-B. Furthermore, LPs decreased oleate-induced mature sterol regulatory element binding protein 2 (SREBP-2), a transcription factor that activates cholesterol biosynthesis. This is the first study to show that LPs can decrease oleate-induced apo-B secretion in HepG2 cells. The modulations of MTTP mRNA expression, cellular total cholesterol metabolism and mature SREBP-2 expression may be important factors in the regulation of apo-B secretion by LPs.

Lignin-derived lignophenols attenuate oxidative and inflammatory damage to the kidney in streptozotocin-induced diabetic rats.

This study investigated the effects of lignin-derived lignophenols (LPs) on the oxidative stress and infiltration of macrophages in the kidney of streptozotocin (STZ)-induced diabetic rats. The diabetic rats were divided into four groups with 0%, 0.11%, 0.33% and 1.0% LP diets. The vehicle-injected controls were given a commercial diet. At 5 weeks, superoxide (O(2)(-)) production, macrophage kinetics, the degree of fibrosis in glomeruli and mRNA expression for monocyte chemoattractant protein-1 (MCP-1) were examined. The NADPH-stimulated O(2)(-) levels in the kidney of the diabetic rats treated with 1.0% LP were significantly lower than those in untreated diabetic rats. The number of macrophages, levels of MCP-1 mRNA expression and degree of glomerular fibrosis increased in untreated LP and these levels were significantly lower in 1.0%LP-treated rats. The results suggested that LPs suppress the excess oxidative stress, the infiltration and activation of macrophages and the glomerular expansion in STZ-induced diabetic kidneys.

Protective effect of lignophenol derivative from beech (Fagus crenata Blume) on copper- and zinc-mediated cell death in PC12 cells.

Lignophenol, prepared using a phase-separation system, is a derivative of lignin, which is one of the components in the plant cell wall, and possesses high phenolic function, high stability and antioxidant properties. However, little is known about the beneficial effect of lignophenol. In this study, we investigated the protective effect of lignophenol from the beech tree (Fagus crenata Blume) on copper- and zinc-mediated apoptosis in PC12 cells by using DNA fragmentation and TUNEL assays. In DNA fragmentation assays, the DNA ladder patterns in the PC12 cells treated with 200 microM Cu and 200 microM Zn were enhanced, whereas the DNA ladder pattern was hardly observed in these cells treated with 20 mM lignophenol. In the TUNEL assay, TUNEL signals increased significantly in the untreated PC12 cells exposed to 200 microM Cu compared with the control. In contrast, the degree of apoptosis in the 20 mM lignophenol-treated cells was significantly lower than in the untreated cells, indicating that lignophenol inhibited Cu-induced apoptotic cell death in PC 12 cells. In the 200 microM Zn-exposed group, the degree of apoptosis in the 20 mM lignophenol-treated cells was also low compared with the untreated cells. In conclusion, these results suggest that lignophenol plays a role in protecting against Cu- and Zn-mediated PC12 apoptotic cell death.