Taste receptor cells encounter chemicals in foods and transmit this information to the gustatory neurons, which convey it further to the gustatory relay nuclei in the lower brainstem. Characterizing neurons involved in the transmission of gustatory information in the peripheral and central nervous systems helps us better understand how we perceive and discriminate tastes. However, it is difficult to anatomically identify them. Using cell-type-specific promoters/enhancers and a transneuronal tracer, we generated transgenic mice to visualize neurons in the gustatory neural pathways. We observed the tracer in the neurons of cranial sensory ganglia and the nucleus of the solitary tract in the medulla where gustatory neurons project. The tracer was also distributed in the reticular formation and several motor nuclei in the medulla that have not been recognized as gustatory ascending pathways. These transgenic mice revealed gustatory relay neurons in the known gustatory ascending pathway and an unexpected, thus presumably novel, neural circuit of gustatory system.

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Some carbohydrate-active enzymes display a modular structure in which catalytic modules that target an insoluble substrate are often attached to one or more noncatalytic carbohydrate-binding modules (CBMs) that assist enzymatic activity. CBMs have been classified into more than 60 families based on amino acid sequence similarities. CBM family 13 (CBM13) and family 42 (CBM42) possess a β-trefoil fold and are grouped into CBM fold family 2. The β-trefoil fold contains a sequence of approximately 45 amino acid residues that is repeated 3 times, resulting in three subdomains (α, β and γ) that fold into an overall globular structure. Each subdomain is composed of four β-strands that fold into a Y-shaped β-hairpin structure. CBM13 and CBM42 have multivalent sugar-binding ability. In this review, I describe the sugar-binding mechanisms of the CBM13 and CBM42 domains of a β-xylanase, a β-L-arabinopyranosidase, and an α-L-arabinofuranosidase.

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Mycotoxins are frequent contaminants of grains, and breweries need, therefore, to pay close attention to the risk of contamination in beer made from such grains as barley and corn. The fate of 14 types of mycotoxin (aflatoxins, fumonisins, ochratoxin A, patulin, trichothecenes, and zearalenone) during beer brewing was investigated in this study. Malt artificially spiked with each mycotoxin was put through the mashing, filtration, boiling and fermentation processes involved in brewing. After brewing, the levels of aflatoxins, ochratoxin A, patulin, and zearalenone were found to have decreased to less than 20% of their initial concentration. They had been adsorbed mainly to the spent grain and removed from the unhopped wort. Additionally, as zearalenone was known, patulin was metabolized to the less toxic compound during the fermentation process. The risk of carry-over to beer was therefore reduced for half of the mycotoxins studied. However, attention still needs to be paid to the risk of trichothecene contamination.

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A second isoprene unit biosynthetic pathway, via 2-C-methyl-D-erythritol 4-phosphate (MEP), was discovered in the 1990s. We screened and isolated the cyclic dipeptide, maculosin, which is a probable novel MEP pathway inhibitor, from the culture broth of Bacillus subtilis strain KN07. To identify the target enzyme of maculosin, we applied an avidin-biotin complex method using biotinylated maculosin and the lysates of seven Escherichia coli strains, each overexpressing one enzyme of the MEP pathway, and performed quartz crystal microbalance (QCM) experiments using maculosin and each enzyme. The results indicate that IspG, the sixth enzyme on the MEP pathway, was bound to maculosin.

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Two terphenyl quinones were synthesized for a structural study on a naturally occurring biologically active terphenyl quinone. 3-Methoxy-5,6-diphenylcyclohexa-3,5-dien-1,2-dione, a possible structure proposed by our analysis of the NMR spectra, was synthesized by Suzuki-Miyaura coupling and subsequent oxidation of the resulting substituted phenol, although not being identical to the natural product. Recently isolated 3-methoxy-2,5-diphenylcyclohexa-2,5-dien-1,4-dione was synthesized from a commercially available 2,5-diphenyl-1,4-benzoquinone in three steps in a good overall yield, and its NMR spectra were identical to those of the natural product.

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Aqueous extracts of Rhizopus oryzae (Aq-ROU) have a broad range of physiological activity. Here we identified a new physiological effect of Aq-ROU in rat hepatocyte cell line RLN-10. Aq-ROU induced the accumulation of nitrite, a stable metabolite nitric oxide (NO), in cell culture medium and induced potent diaminofluorescein-FM diacetate staining in the cells. Real-time reverse transcriptase (RT)-PCR analysis showed marked inducible NO synthase gene expression. Additionally, markedly enhanced expression of p22^phox and temporally increased expression of NADPH oxidase1 indicated that superoxide was produced. Nuclear translocation of nuclear factor-kappa (NF-κ) B p65 increased remarkably following Aq-ROU and following lipopolysaccharide treatment, a potent activator of NF-κB. Ammonium pyrrolidine-1-carbodithioate, an inhibitor of NF-κB, inhibited NO production following Aq-ROU treatment. Our data indicate that Aq-ROU induces NO production and potentially the production of superoxide, which may contribute to the broad range of physiological effects observed for Aq-ROU ingested by animals.

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Lonicera japonica is used in Chinese medicine as a source of antioxidants, primarily flavonoids, and a phenolic acid chlorogenic acid (CGA). Here we report the isolation and characterization of the full-length cDNA of LjC3H, a gene encoding p-coumaroyl ester 3-hydroxylase, an enzyme involved in CGA synthesis. Phylogenetic analysis indicated that is protein belongs to the CYP98A subfamily, and homology modeling revealed that its structure resembles that of other cytochrome P450 family proteins. Southern blot analysis indicated that more than one copy of sequences homologous to LjC3H is present in the L. japonica genome. Heterologous expression of LjC3H cDNA in Escherichia coli allowed an in vitro assay of LjC3H to be performed. This experiment revealed that the enzyme favors p-coumaroylshikimate over p-coumaroylquinate as substrate. LjC3H transcript abundance was increased both by treatment of the leaves with methyl jasmonate and by exposure to UV-B radiation. The CGA levels in the leaves of L. japonica were positively correlated with LjC3H transcript abundance.

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BtrC2 of the butirosin producer Bacillus circulans is a non-catalytic subunit of 2-deoxy-scyllo-inosose (DOI) synthase that is involved in butirosin biosynthesis, and also a homolog of glutamine amidotransferase subunit (PdxT) of pyridoxal 5'-phosphate (PLP) synthase of Bacillus subtilis. BtrC2 has been found to have functions in B. circulans both in primary and secondary metabolism. In this study, we investigated the properties of PdxT of B. subtilis in order to determine whether the property of enzyme stabilization is universal among PdxT homologs. Complementation with PdxT in the btrC2 disruptant of B. circulans restored the growth and short-term production of antibiotics, but long-term production of antibiotics cannot be restored. Additionally, PdxT did not bind physically with or stabilize BtrC. Our results indicate that the function of BtrC2 in secondary metabolism is specific properties, not universal among PdxT homologs.

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RNase Po1 is a guanylic acid-specific ribonuclease (a RNase T1 family RNase) from Pleurotus ostreatus. We determined the cDNA sequence encoding RNase Po1 and expressed RNase Po1 in Escherichia coli. A comparison of the enzymatic properties of RNase Po1 and RNase T1 indicated that the optimum temperature for RNase Po1 activity was 20 °C higher than that for RNase T1. An MTT assay indicated that RNase Po1 inhibits the proliferation of human neuroblastoma cells (IMR-32 and SK-N-SH) and human leukemia cells (Jurkat and HL-60). Furthermore, Hoechst 33342 staining showed morphological changes in HL-60 cells due to RNase Po1, and flow cytometry indicated the appearance of a sub-G1 cell population. The extent of these changes was dependent on the concentration of RNase Pol. We suggest that RNase Po1 induces apoptosis in tumor cells.

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Methylcitrate synthase (EC 2.3.3.5; MCS) is a key enzyme of the methylcitric acid cycle localized in the mitochondria of eukaryotic cells and related to propionic acid metabolism. In this study, cloning of the gene mcsA encoding MCS and heterologous expression of it in Escherichia coli were performed for functional analysis of the MCS of citric acid-producing Aspergillus niger WU-2223L. Only one copy of mcsA (1,495 bp) exists in the A. niger WU-2223L chromosome. It encodes a 51-kDa polypeptide consisting of 465 amino acids containing mitochondrial targeting signal peptides. Purified recombinant MCS showed not only MCS activity (27.6 U/mg) but also citrate synthase (EC 2.3.3.1; CS) activity (26.8 U/mg). For functional analysis of MCS, mcsA disruptant strain DMCS-1, derived from A. niger WU-2223L, was constructed. Although A. niger WU-2223L showed growth on propionate as sole carbon source, DMCS-1 showed no growth. These results suggest that MCS is an essential enzyme in propionic acid metabolism, and that the methylcitric acid cycle operates functionally in A. niger WU-2223L. To determine whether MCS makes a contribution to citric acid production, citric acid production tests on DMCS-1 were performed. The amount of citric acid produced from glucose consumed by DMCS-1 in citric acid production medium over 12 d of cultivation was on the same level to that by WU-2223L. Thus it was found that MCS made no contribution to citric acid production from glucose in A. niger WU-2223L, although MCS showed CS activity.

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In plant Mesembryanthemum crystallinum, which has the inducible crassulacean acid metabolism (CAM), isoforms of plastidic phosphate translocators (pPTs) are categorized into three subfamilies: the triose phosphate/phosphate translocator (McTPT1), the phosphoenolpyruvate/phosphate translocator (McPPT1), and the glucose 6-phosphate/phosphate translocator (McGPT1 and McGPT2). In order to elucidate the physiological roles of these pPTs in M. crystallinum, we determined the substrate specificity of each pPT isoform. The substrate specificities of McTPT1, McPPT1, and McGPT1 showed overall similarities to those of orthologs that have been characterized. In contrast, for glucose 6-phosphate, McGPT2 showed higher selectivity than McGPT1 and other GPT orthologs. Because the expression of McGTP2 is specific to CAM while that of McGTP1 is constitutively expressed in both the C₃- and the CAM-state in M. crystallinum, we propose that McGPT2 functions as a CAM system-specific GPT in this plant.

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Kynurenic acid, a tryptophan metabolite, is involved in psychiatric disease. Our laboratory previously described its transport by rat/human organic anion transporters rOAT1, hOAT1, rOAT3 and hOAT3, which are involved in drug disposition. In this study, we performed an uptake experiment using Xenopus laevis oocytes to examine the transport of xanthurenic acid, a tryptophan catabolite and kynurenic acid analog, by various transporters. All the transporters tested stimulated the uptake of xanthurenic acid into oocytes. The transport activity of xanthurenic acid by hOAT1 was greater than that by rOAT1. In OAT3, the rat homolog showed efficient transport, compared with hOAT3. The apparent values of K_m and V_max for the transport by hOAT1 were 4.83 µM and 26.0 pmol/oocyte/h respectively. In rOAT3, the respective values were 6.87 µM and 21.7 pmol/oocyte/h. This is the first report on xanthurenic acid transport by OAT1 and OAT3.

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The facultative piezophile Shewanella violacea DSS12 is known to have respiratory components that alter under the influence of hydrostatic pressure during growth, suggesting that its respiratory system is adapted to high pressure. We analyzed the expression of the genes encoding terminal oxidases and some respiratory components of DSS12 under various growth conditions. The expression of some of the genes during growth was regulated by both the O₂ concentration and hydrostatic pressure. Additionally, the activities of cytochrome c oxidase and quinol oxidase of the membrane fraction of DSS12 grown under various conditions were measured under high pressure. The piezotolerance of cytochrome c oxidase activity was dependent on the O₂ concentration during growth, while that of quinol oxidase was influenced by pressure during growth. The activity of quinol oxidase was more piezotolerant than that of cytochrome c oxidase under all growth conditions. Even in the membranes of the non-piezophile Shewanella amazonensis, quinol oxidase was more piezotolerant than cytochrome c oxidase, although both were highly piezosensitive as compared to the activities in DSS12. By phylogenetic analysis, piezophile-specific cytochrome c oxidase, which is also found in the genome of DSS12, was identified in piezophilic Shewanella and related genera. Our observations suggest that DSS12 constitutively expresses piezotolerant respiratory terminal oxidases, and that lower O₂ concentrations and higher hydrostatic pressures induce higher piezotolerance in both types of terminal oxidases. Quinol oxidase might be the dominant terminal oxidase in high-pressure environments, while cytochrome c oxidase might also contribute. These features should contribute to adaptation of DSS12 in deep-sea environments.

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The Schizosaccharomyces pombe php2⁺ gene encodes a subunit of the CCAAT-binding factor complex. We found that disruption of the php2⁺ gene extended the chronological lifespan of the fission yeast. Moreover, the lifespan of the Δphp2 mutant was barely extended under calorie restricted (CR) conditions. Many other phenotypes of the Δphp2 mutant resembled those of wild-type cells grown under CR conditions, suggesting that the Δphp2 mutant might undergo CR. The mutant also showed low respiratory activity concomitant with decreased expression of the cyc1⁺ and rip1⁺ genes, both of which are involved in mitochondrial electron transport. On the basis of a chromatin immunoprecipitation assay, we determined that Php2 binds to a DNA region upstream of cyc1⁺ and rip1⁺ in S. pombe. Here we discuss the possible mechanisms by which the chronological lifespan of Δphp2 mutant is extended.

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Jasmonate plays key roles in plant growth and stress responses, as in defense against pathogen attack. Jasmonoyl-isoleucine (JA-Ile), a major active form of jasmonates, is thought to play a pivotal role in plant defense responses, but the involvement of JA-Ile in rice defense responses, including phytoalexin production, remains largely unknown. Here we found that OsJAR1 contributes mainly to stress-induced JA-Ile production by the use of an osjar1 Tos17 mutant. The osjar1 mutant was impaired in JA-induced expression of JA-responsive genes and phytoalexin production, and these defects were restored genetically. Endogenous JA-Ile was indispensable to the production of a flavonoid phytoalexin, sakuranetin, but not to that of diterpenoid phytoalexins in response to heavy metal stress and the rice blast fungus. The osjar1 mutant was also found to be more susceptible to the blast fungus than the parental wild type. These results suggest that JA-Ile production makes a contribution to rice defense responses with a great impact on stress-induced sakuranetin production.

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Small ubiquitin-related modifier (SUMO) research requires large amounts of SUMO-activating enzyme (SUMO-E1) for in vitro SUMOylation assays. In this study the expression and purification of a hexahistidine-tagged mouse Aos1-Uba2-fusion protein (His₆-mAU) in a baculovirus-insect cell system revealed that approximately 1.0 mg of highly-purified His₆-mAU was obtained from 100 mL of Sf9 cell culture. The recombinant protein had SUMO-E1 activity in multiple in vitro SUMOylation assays.

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In a previous study, heat-treated noradrenaline induced flowering of the short-day plant Lemna paucicostata Hegelmaier 151. In the present study, we found that heat-treated noradrenaline also had flower-inducing activity in short-day L. paucicostata strains 441 and 6746 and in long-day L. gibba strain G3. The flower-inducing activity in these plants was enhanced by water homogenates of eggplant (Solanum melongena L.).

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Here we describe the generation of potato plants that constitutively overexpressed, UDP-N-acetylglucosamine:dolichol phosphate-N-acetylglucosamine-phosphotransferase (GPT). Such transgenic plants can be formed in a medium with tunicamycin at 9.8 ± 0.28% efficiency, similar to the 9.4 ± 1.10 for the bialaphos resistance gene (Bar) gene. This study indicated that GPT transformation was very stable with high reproducibility, and that growth and tuber production in the GPT-transformed plants were stronger than in the wild-type plants.

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In this study, we examined the effects of Jeju dwarf bamboo (Sasa quelpaertensis Nakai) extract (JBE) and p-coumaric acid (CA) on oleic acid (OA)-induced lipid accumulation in HepG2 cells. JBE and CA increased the phosphorylation of AMP-activated protein kinase (AMPK), and acetyl-CoA carboxylase (ACC) and the expression of carnitine palmitoyl transferase 1a (CPT1a) in OA-treated HepG2 cells. Additionally, these compounds decreased sterol regulatory element-binding protein-1c (SREBP-1c), fatty acid synthase (FAS), and OA-induced lipid accumulation, suggesting that JBE and CA modulate lipid metabolism in HepG2 cells via the AMPK activation pathway.

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We have developed a time- and cost-effective method for isolating low molecular weight (LMW) RNA from plants. In our protocol, the isolation procedure can be completed within 3 h. Polyethylene glycol (PEG) and absolute ethanol are used to isolate LMW RNA, and the LMW RNA yields were >80 µg/g of fresh-weight tissues for several of the plant species tested.

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Acetylation of α-tubulin on Lys⁴⁰ is thought to be a modification that regulates the dynamic instability of microtubules, but little is known about the occurrence of α-tubulin acetylation in plants. Here we report on a growth stage-dependent change in levels of α-tubulin acetylation and the organ distribution of acetylated α-tubulin in Arabidopsis thaliana plants. Widespread occurrence of α-tubulin acetylation in the leaves of 15 species (20 cultivars) of angiosperms was also confirmed. Our data indicate that acetylated α-tubulin is widespread in many angiosperms, but levels can differ, sometimes considerably, among different organs and developmental stages.

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We investigated 10 lactic acid bacteria strains with probiotic potential prepared from Mongolian dairy products for their ability to induce T helper type-1 (Th1) cytokine production in mouse immune cells in vitro and in vivo. Among these strains, the Lactobacillus plantarum 06CC2 strain was effective in elevating the level of interleukin (IL)-12p40 in co-culture with J774.1 cells and the levels of IL-12 and interferon (IFN)-γ in co-culture with mouse spleen cells in vitro. Oral administration of this strain augmented the gene expression of IFN-γ and IL-12p40 and enlarged the population of CD4⁺, CD25⁺, and CD49b⁺ cells in the spleens of normal mice. It also significantly elevated the gene expression of IL-12 receptor β2 as well as IL-12p40 and IFN-γ in Peyer's patches. Thus oral administration of strain 06CC2 was effective in inducing Th1 cytokine production activating the Th1 immune response associated with intestinal immunity in normal mice.

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The objective of the present study was to investigate the cholesterol-reducing effect of medium-chain fatty acids (MCFAs) completed by elevated excretion of fecal neutral steroids and/or bile acids. Blood and liver lipid profiles, fecal neutral steroids, bile acids, and mRNA and protein expression of the genes relevant to cholesterol homeostasis were measured and analyzed in C57BL/6J mice fed a cholesterol-rich diet with 2% caprylic acid or capric acid for 12 weeks. Blood total cholesterol and low-density lipoprotein cholesterol (LDL-c) levels were reduced significantly as compared to diet with palmitic acid or stearic acid. Caprylic acid promoted the excretion of fecal neutral steroids, especially cholesterol. The excretion of fecal bile acids, mainly in the form of cholic acid was enhanced and accompanied by elevated expression of mRNA and the protein of hepatic cholesterol 7α-hydroxylase (CYP7A1). These results indicate that MCFAs can reduce blood cholesterol by promoting the excretion of fecal cholesterol and cholic acid.

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α-Amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD) plays a key role in the regulation of NAD biosynthesis or the production of quinolinate from tryptophan (Trp). We investigated in this study the effect of phytol, a phytochemical known as a peroxisome proliferator-activated receptor α (PPARα) ligand, on NAD synthesis and ACMSD expression in rats. Male Sprague-Dawley rats were fed a diet containing 0.5%, 1%, or 2% phytol for 7 d. Phytol decreased the ACMSD activity and its mRNA expression in a dose-dependent manner in the liver. Phytol similarly and significantly suppressed ACMSD mRNA expression in primary rat hepatocytes. However, the mRNA expression of ACO (a known PPARα target gene) was higher in the low-phytol groups than in the high-phytol group in vivo and in vitro. Phytol increased the blood NAD level by suppressing ACMSD mRNA expression in the liver of the rats. It is possible that this mechanism occurred by the activation of PPARα and also of other transcriptional factors.

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Recent reports have proposed possible anti-obesity mechanisms for antioxidants involving increased energy expenditure, pre-adipocyte differentiation and proliferation, decreased lipogenesis and increased lipolysis, and fat oxidation. The aim of this study was to examine and to confirm the anti-obesity effect of the oligomeric and polymeric procyanidin fractions from defatted grape seeds. The lipid metabolism-related mRNA level in the mouse preadipocytes, 3T3-L1 cells, was determined to evaluate the anti-obesity effect of the phenolic fractions from a grape seed meal. Lipid accumulation was reduced by 19% of the control level by the procyanidin fraction originating from the grape seed meal. Emerging from the effect of the treatment on HSL and LPL mRNA expression, lipolytic enzyme activity was not involved in the anti-obesity effects of CPE and FPP from the defatted grape seed meal. We tested and confirmed in this study the effect of the biological activities of oligomeric and polymeric procyanidins from the defatted grape seed meal. It is suggested from the results of this brief study that further studies would be desirable to focus on the anti-obesity effect of the purified extracts of a defatted grape seed meal.

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Cirsium setidens is a perennial medicinal herb that is rich in flavonoids. We investigated in this study the effect of a C. setidens ethanol extract (CSE) on the development of nonalcoholic fatty liver in mice fed a high-fat diet (HF). C57BL/6J mice were fed either a control diet (CON) or HF for 8 weeks, and then fed CON, HF, or HF with 100 mg/kg of BW CSE (HF+CSE) for an additional 7 weeks. The final body weight and adipose tissue weight of the mice in the HF+CSE group were significantly lower than those in the HF group. CSE also markedly diminished both the lipid droplets in the liver tissues and decreased the hepatic and serum triglycerides (TG) concentrations. CSE strongly increased the hepatic mRNA levels of carnitine palmitoyltransferase (CPT1) and medium-chain acyl-CoA dehydrogenase (MCAD), the fatty acid β-oxidation enzymes. The hepatic levels of phosphorylated-AMP-activated protein kinase (AMPK) were significantly higher in the HF+CSF group than in the HF group. These results suggest that CSE inhibited hepatic fat accumulation by up-regulating the expression of the fatty acid β-oxidation genes.

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L-Cysteine works as a precursor of the antioxidant, glutathione. We investigated the effects of L-cysteine (1% and 2%) on lipid metabolism and the antioxidative system in rats fed a normal diet. Administering L-cysteine dependently decreased the food intake, fat mass weight and body weight dose. Dietary L-cysteine also decreased the triglyceride levels in the serum and liver. However, there were no significant differences in the hepatic TBARS and glutathione (GSH) levels among the groups. The activities of catalase and glutathione reductase in the rats receiving 2% L-cysteine were significantly higher (p<0.05) than in the control rats. These results suggest that dietary L-cysteine dose-dependently affected the antioxidative enzyme activities, and the lipid levels in the serum and liver which might be related to the reduced food intake.

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waxy/amylose-extender (wx/ae) double-mutant japonica rice (Oryza sativa L.) produces resistant starch (RS) and a large amount of γ-oryzanol. Our previous study has shown the hypolipidemic effect of wx/ae brown rice on mice. To identify the functional constituents of the hypolipidemic activity in wx/ae rice, we prepared pure wx/ae starch and γ-oryzanol from wx/ae rice and investigated their effect on the lipid metabolism in BALB/c.KOR/Stm Slc-Apoe^shl mice. The mice were fed for 3 weeks a diet containing non-mutant rice starch, non-mutant rice starch plus γ-oryzanol, wx/ae starch, or wx/ae starch plus γ-oryzanol. γ-Oryzanol by itself had no effect on the lipid metabolism, and wx/ae starch prevented an accumulation of triacylglycerol (TAG) in the liver. Interestingly, the combination of wx/ae starch plus γ-oryzanol not only prevented a TAG accumulation in the liver, but also partially suppressed the rise in plasma TAG concentration, indicating that wx/ae starch and γ-oryzanol could have a synergistic effect on the lipid metabolism.

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Although pu-erh tea has been shown to suppress hyperlipidemia, it is unclear how it modulates fatty acid synthase expression in mice fed on a high-fat diet. We investigated the effects of a pu-erh tea extract (PTE) on diet-induced body fat accumulation. C57BL/6J mice were fed a control diet, a high-fat diet (HFD), and HFD supplemented with 0.225% or 0.45% PTE for 70 d. Supplementation with PTE reduced the body weight gain, and the abdominal and liver fat accumulation. A significant difference in the triglyceride level were observed between the HFD control and HFD+0.45% PTE groups. A PTE intake tended to decrease sterol regulatory element-binding protein (SREBP)-1c and fatty acid synthase (FAS) mRNA expression in the liver of the mice. These findings indicate that PTE reduced lipogenesis by down-regulating SREBP-1c and related molecules, leading to the suppression of body fat accumulation.

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Dipeptidyl peptidase IV (DPPIV) degrades some peptide hormones and cytokines, resulting in homeostatic modulation. However, the role of DPPIV in inflammatory bowel diseases remains controversial. To determine the role of DPPIV in colitis, we used F344/DuCrlCrlj (F344/Du) rats as a DPPIV-deficient model. The serum DPPIV activity was much lower in the F344/Du rats than in F344/Jcl rats which were used as a DPPIV-positive model. Interestingly, the disease activity index (DAI) was different in the early phase of 2% dextran sulfate sodium (DSS)-induced colitis, as judged by the mucosal myeloperoxidase activity, colonic weight, and cecal fermentation. Similarly, retarded DAI was apparent in the DPPIV-deficient rats with 1% DSS-induced colitis. These findings suggest that a low level of DPPIV activity contributed to maintaining intestinal homeostasis by suppressing the cleavage of cytokines and growth hormones in DSS-induced colitis, especially in the early phase of colitis and with moderate inflammation.

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An investigation by using an aroma extract dilution analysis (AEDA) of the aroma concentrates made from freshly roasted in-shell peanuts and stored peanuts revealed a total of 43 key aroma compounds, including 8 newly identified compounds in peanuts. Among them, 2-isobutyl-3-methoxypyrazine, exhibiting an earthy note, and 4-hydroxy-2,5-dimethyl-3(2H)-furanone, exhibiting a caramel-like note, were detected with the highest flavor dilution (FD) factor of 4096 in the fresh peanuts, followed by 3,5-dimethyl-2-ethylpyrazine, exhibiting a nutty note, as having the next highest FD factor of 1024. A quantitative analysis of the key aroma compounds having high FD factors in the fresh peanuts and stored peanuts revealed that 2-methyl-3-furanthiol, 2-acetyl-1-pyrroline, 2-propionyl-1-pyrroline, and 3,5-dimethyl-2-vinylpyrazine significantly decreased during storage, while methyl 2-methyl-3-furyl disulfide, 4-hydroxy-2,5-dimethyl-3(2H)-furanone, and 2-methoxy-4-vinylphenol significantly increased. The sensory experiments revealed that the fresh peanuts presented strong roasty/meaty, popcorn-like, and nutty notes, as well as moderate spicy/burnt and caramel-like notes, whereas the stored peanuts presented significantly weak roasty/meaty and popcorn-like notes and a significantly strong spicy/burnt note. Based on the comparative AEDAs, the quantitative analysis, and the sensory analysis, it was concluded that the freshly roasted peanut aroma comprised the significant contributions of 2-methyl-3-furanthiol exhibiting a roasty/meaty note, and of 2-acetyl-1-pyrroline and 2-propionyl-1-pyrroline exhibiting a popcorn-like note, and the lesser contribution of 2-methoxy-4-vinylphenol exhibiting a spicy/burnt note. In particular, 2-methyl-3-furanthiol, which was only detected in the freshly roasted peanut aroma concentrate, might be an essential component describing the freshness of the roasted peanut aroma by its diffusive roasty/meaty note.

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Moisture sorption isotherms were measured at 25 °C for untreated, dry-heated and pre-gelatinized durum wheat flour samples. The isotherms could be expressed by the Guggenheim-Anderson-de Boer equation. The amount of water sorbed to the untreated flour was highest for low water activity, with water sorbed to the pre-gelatinized and dry-heated flour samples following. The dry-heated and pregelatinized flour samples exhibited the same dependence of the moisture content on the partial molar volume of water at 25 °C as the untreated flour. The partial molar volume of water was ca. 9 cm³/mol at a moisture content of 0.03 kg-H₂O/kg-d.m. The volume increased with increasing moisture content, and reached a constant value of ca. 17.5 cm³/mol at a moisture content of 0.2 kg-H₂O/kg-d.m. or higher.

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The extract from bulbs of Scilla scilloides exhibited inhibitory effects in lipoxygenase and hyaluronidase assays and various oxidation models in vitro. Incubating the cells in the presence of this extract ameliorated t-butyl hydroperoxide-induced cytotoxicity from 27% to 57% in a macrophage model. The results may indicate the potential role of S. scilloides for its anti-inflammatory and antioxidative effects.

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The effects of dietary coenzyme Q10 (CoQ10) on cholesterol metabolism in laying hens were investigated. Dietary CoQ10 significantly reduced egg yolk cholesterol content and suppressed hepatic hydroxymethylglutaryl-CoA reductase (HMGR) activity. It is therefore likely that CoQ10 acts as an HMGR inhibitor in the livers of laying hens, which in turn results in a reduction in egg-yolk cholesterol.

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Two nicotinic acid-related compounds were found to induce erythroid differentiation in K562 cells. We investigated the changes in nicotinamide adenine dinucleotide (NAD) content induced by nicotinic acid-related compounds during differentiation. The NAD content was reduced by a treatment with nicotinic acid and isonicotinic acid. These results provide important clues toward elucidating the erythroid differentiation mechanism.

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It is well known that large dose of leucine reduces the food intake and causes growth retardation in experimental animals when leucine is given with a low-protein diet. However, the mechanism for the anorectic effect of leucine has not yet been clarified. We demonstrate here that the anorectic effect of leucine was significantly reduced in a vagotomized rat.

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Chinese black tea extract (CBTE) fermented with Aspergillus sp. significantly promoted hair growth after 2 weeks of topical application in shaved 6 week-old male C3H/He mice. The hair growth-promoting effect of CBTE was potentiated synergistically by capsaicin, which has no effect on hair growth by itself. CBTE displayed an affinity for estrogen receptor (ER)α, with an IC₅₀ value of 74.8 µg/mL. This effect of CBTE might be mediated by the ERs, since a similar effect induced by orally administered soy isoflavone, a mixture of ERs ligands, has been reported to be synergistically potentiated by capsaicin.

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Sugarcane bagasse was pretreated by soaking it in aqueous ammonia (SAA) and methanolic aqueous ammonia (SMAA) at 70 °C for 12 h. Then the pretreated as well as untreated bagasse was subjected to enzymatic hydrolysis at 50 °C for 72 h by 15 FPU cellulase and 30 CBU cellobiase per g of substrate. The hydrolysis of SAA-pretreated bagasse with a solid to liquid (S:L) ratio of 1:10 resulted in 95.9% of the maximum theoretical yield. The production yield for SMAA at an S:L ratio of 1:6 with 15% methanol was 88.6%, while it was only 21.3% for the untreated bagasse. Ethanol production by simultaneous saccharification and fermentation was conducted at 37 °C for 72 h. The results revealed that the ethanol production yield was improved from 12.7% for the untreated bagasse to 92.45% and 90.8% for the SAA and the SMAA pretreated bagasse, respectively. The compositional and chemical structural analysis suggested that lignin removal and crystallinity reduction were responsible for the hydrolysis and SSF improvements.

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Waste money bills (WMB) is a by-product of the money making process that consists of rich-cellulosic material for many biotechnological applications. This waste money bills is unusable and usually exhausted. Saccharification was improved using various concentrations of sodium hydroxide, NaOH (0.0, 0.5, 1.0, 2.0, 2.5, and 3.0% v/v) and various reaction times (20, 30, and 40 min) during pretreatment at 121 °C. Prior to ethanol fermentation, the highest glucose yield (62.2 mg/mL) was found by pretreatment consisting of 30 min at 2.0% NaOH, and it increased 33.8% as compared to an untreated sample. The highest amount of ethanol was obtained (26.1 mg/mL) during fermentation, and this was increased 95.3 and 22.5% as compared to aerobic and anaerobic conditions respectively during pretreatment with 2.0% NaOH for 30 min. Under anaerobic conditions, ethanol fermentation was enhanced by adding 0.4 mmol benzoic acid. Production of ethanol from waste money bills would cut waste management costs and make profitable.

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We have reported that a recombinant Candida utilis strain expressing a Candida shehatae xylose reductase K275R/N277D, a C. shehatae xylitol dehydrogenase, and xylulokinase from Pichia stipitis produced ethanol from xylose, but its productivity was low. In the present study, metabolomic (CE-TOF MS) and transcriptomic (microarray) analyses were performed to characterize xylose metabolism by engineered C. utilis and to identify key genetic changes contributing to efficient xylose utilization. The metabolomic analysis revealed that the xylose-fermenting strain accumulated more pentose phosphate pathway intermediates, more NADH, and more glycolytic intermediates upstream of glyceraldehyde 3-phosphate than the wild-type. Transcriptomic analysis of the strain grown on xylose indicated a significant increase in expression of the genes encoding tricarboxylic acid cycle enzymes, respiratory enzymes, and enzymes involved in ethanol oxidation. To decrease the NADH/NAD⁺ ratio and increase the ethanol yield of the fermentation of xylose, ADH1 encoding NADH-dependent alcohol dehydrogenase was overexpressed. The resulting strain exhibited a 17% increase in ethanol production and a 22% decrease in xylitol accumulation relative to control.

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A set of 300 bacterial strains isolated from various extreme environments were screened for the presence of cellulase activity on CMC agar plates. Phylogenetic analysis of the positive strain, based on 16S rRNA gene sequences indicated that the isolates were clustered within Firmicutes and Actinobacteria. A majority (17) of the isolates were identified as Bacillus, Paenibacillus, and Lysinibacillus sp., and the remaining three were identified as Arthobacter, Rhodococcus, and Bhargavaea cecembensis. Among the 20 positive isolates, 6 were evaluated for the production of cellulases on five different cellulosic substrates. Two isolates, B. cecembensis and Bacillus sp., based on maximum enzyme production on all cellulosic substrates, especially CMC and rice straw, were evaluated in terms of enzyme properties and kinetics. The enzymes of these two isolates are found to be active over broad range of pH and temperature. Such thermostable enzymes facilitate the development of efficient and cost-effective forms of the simultaneous saccharification and fermentation process converting lignocellulosic biomass into biofuels and value-added products.

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Mucosal immunity is critical in preventing infectious bronchitis virus (IBV) infection. To deliver viral antigens to the mucosal immune system of chickens safely and effectively, we constructed a Lactococcus lactis strain carrying IBV multi-epitope gene EpiC fused with the gene of the cell-wall anchoring domain of Staphylococcus aureus protein A. SDS–PAGE and Western blot results indicated that the fused peptide was located partially on the cell surface. Oral and nasal inoculation with the recombinant L. lactis of chickens elicited significantly high humoral and mucosal immune responses, especially in the nasally immunized group. Eighty percent chickens of the nasally immunized group with recombinant L. lactis did not show any clinical signs after a lethal dose challenge with IBV SAIBk strain, while all the non-recombinant L. lactis immunized chickens exhibited obvious and typical symptoms. These results indicate that needle-free recombinant lactococci anchoring the IBV antigen makes a promising vaccine candidate against the spread of IB.

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The yeast Kluyveromyces marxianus is considered as a potential alternative to Saccharomyces cerevisiae in producing ethanol as a biofuel. In this study, we investigated the ethanol fermentation properties of novel K. marxianus strain DMB1, isolated from bagasse hydrolysates. This strain utilized sorbitol as well as various pentoses and hexoses as single carbon sources under aerobic conditions and produced ethanol from glucose in hydrolysates of the Japanese cedar at 42 °C. Reference strains K. marxianus NBRC1777 and S. cerevisiae BY4743 did not assimilate sorbitol or ferment lignocellulosic hydrolysates to ethanol at this temperature. Thus strain DMB1 appears to be optimal for producing bioethanol at high temperatures, and might provide a valuable means of increasing the efficiency of ethanol fermentation.

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Pink-pigmented facultative methylotrophs (PPFMs) are major inhabitants of the phyllosphere. In a preceding study, we found that perilla plants harbor a dominant population of PPFMs on their leaves and seeds, and that the closest relative of PPFMs (Methylobacterium sp. strain OR01 as representative strain) isolated from red perilla seeds was M. fujisawaense DSM5686^T. In the present study, the specific interaction between red perilla and Methylobacterium species was investigated. All the PPFMs isolated from red perilla seeds harvested in the Ohara area of Kyoto, Japan in 2009, 2010, and 2011 and the PPFMs isolated from red perilla leaves planted at four geographically different places in Japan had 16S rRNA sequences identical to that of strain OR01. Direct transmission of PPFMs from seeds to leaves and the competitiveness of strain OR01 were confirmed. This report is the first step toward understanding the species-level specificity of the interaction between perilla plants and Methylobacterium species.

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Roxithromycin (RXM), active against prokaryotes, has beneficial side effects such as anti-cancer activities on mammalian cells, but the mechanisms underlying these effects remain unclear. We found that RXM inhibited the cellular differentiation of the rice blast fungus Magnaporthe oryzae. Hence, we screened the targets of RXM by the T7 phage display method with fungal genomic DNA, and identified MoCDC27 (M. oryzae Cell Division Cycle 27) as a candidate. We generated mocdc27 knockdown mutants that the appressoria formation was less affected by RXM. A complemented mutant restored sensitivity against RXM to the level of the wild type. These results suggest that MoCDC27 was involved in the inhibition of appressorium formation by RXM, and that the complex of RXM-MoCDC27 affected another molecule involved in appressorium formation. The T7 phage display method with fungal genomic DNA can be a useful tool in the quest for drug target.

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We prepared eight recombinant Saccharomyces cerevisae strains, including three strains generated in this study that were produced by chromosomal integration of xylose utilization pathway enzymes genes. Among these strains, MA-R4 was the most efficient at producing ethanol from rice straw enzymatic hydrolysate, indicating that it is a superior strain for bioethanol production.

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The targets of rapamycin (Tor) kinases play central roles in the integrated regulation of cellular activities. Although the molecular mechanisms of Tor-mediated signaling pathways have been studied extensively in yeast, the relationship between kinase activity and the redox maintenance system remains obscure. In this study, we established a quantitative extraction and determination method for glutathione-related compounds in Saccharomyces cerevisiae utilizing liquid chromatography-tandem mass spectrometry (LC-MS/MS). We found decreases in the levels of glutathione and its precursors resulting from the introduction of a Tor1 hyper-active mutation. In line with this finding, the mutant was more sensitive to several heavy metal ions, indicating a physiological defect arising from a failure to regulate the kinase activity.

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