Adlercreutzia equolifaciens gen. nov., sp. nov., an equol-producing bacterium isolated from human faeces, and emended description of the genus Eggerthella Nine strains capable of metabolizing isoflavones to equol were isolated from human faeces. Four of the strains were characterized by determining phenotypic and biochemical features and their phylogenetic position based on 16S rRNA gene sequence analysis. These strains were related to Eggerthella sinensis HKU14 T with about 93 % 16S rRNA gene sequence similarity; they were asaccharolytic, obligately anaerobic, non-spore-forming, non-motile and Gram-positive coccobacilli. In enzyme activity tests, arginine dihydrolase, arginine and leucine arylamidases were positive but nitrate reduction, urease and b-glucosidase were negative. The major menaquinone was DMMK-6 (dimethylmenaquinone-6), while that of members of the genus Eggerthella was MMK-6 (methylmenaquinone-6). Furthermore, the cell-wall peptidoglycan type of these strains was A1c, while that of members of the genus Eggerthella was A4c. On the basis of these data, a new genus, Adlercreutzia gen. nov., is proposed with one species, Adlercreutzia equolifaciens sp. , 2006). We isolated nine strains capable of metabolizing isoflavones to equol from human faeces. Seven of these strains could metabolize daidzein via dihydrodaidzein to equol, while the other two isolates could metabolize only dihydrodaidzein to equol. These strains were divided into four groups by 16S rRNA gene sequence analysis. Representative strains selected from each group were asaccharolytic, obligately anaerobic, non-spore-forming, non-motile and Gram-positive coccobacilli. Although these strains were genetically related to the genus Eggerthella, they did not belong to the genera Eggerthella, Slackia or Denitrobacterium from 16S rRNA gene sequence analysis.The presence of dimethylmenaquinone-6 (DMMK-6) as the predominant menaquinone of these strains is unique. Furthermore, the cell-wall peptidoglycan type of these strains was A1c, while that of members of the genus Eggerthella was A4c.Strains FJC-A10, FJC-A161, FJC-B9 T , FJC-B12, FJC-B15, FJC-B19, FJC-B20, FJC-D47 and FJC-D53 were cultivated for 3 days at 37 u C on BL agar (Nissui) with 5 % (v/v) horse blood. Coriobacterium glomerans JCM 10262 T was cultivated for 2 days at 30 u C on GAM agar (Nissui). Detailed menaquinone and fatty acid profiles of representative strains and related type strains and minimum-evolution and maximumparsimony 16S rRNA gene sequence-based trees are available as supplementary material with the online version of this paper.
Autophagy is a cellular self-catabolic process wherein organelles, macromolecules, and invading microbes are sequestered in autophagosomes that fuse with lysosomes. In this study, we uncover the role of nitric oxide (NO) as a signaling molecule for autophagy induction via its downstream mediator, 8-nitroguanosine 3',5'-cyclic monophosphate (8-nitro-cGMP). We found that 8-nitro-cGMP-induced autophagy is mediated by Lys63-linked polyubiquitination and that endogenous 8-nitro-cGMP promotes autophagic exclusion of invading group A Streptococcus (GAS) from cells. 8-nitro-cGMP can modify Cys residues by S-guanylation of proteins. We showed that intracellular GAS is modified with S-guanylation extensively in autophagosomes-like vacuoles, suggesting the role of S-guanylation as a marker for selective autophagic degradation. This finding is supported by the fact that S-guanylated bacteria were selectively marked with polyubiquitin, a known molecular tag for selective transport to autophagosomes. These results collectively indicate that 8-nitro-cGMP plays a crucial role in cytoprotection during bacterial infections or inflammations via autophagy upregulation.
Pharmacological properties of strychnine-sensitive and -insensitive glycine receptors have been investigated in rat suprachiasmatic nucleus (SCN) neurons. Because the SCN neurons were too small for stable intracellular recordings by the glass-microelectrode technique, a conventional whole cell mode patch-clamp technique was employed on the acutely dissociated SCN neurons. Dissociated SCN neurons were morphologically heterogeneous and could be distinguished into several types. All cells responded to glycine in a concentration-dependent manner. The glycine-induced current was primarily Cl- sensitive and competitively blocked by strychnine. The SCN neurons also responded to excitatory amino acids: glutamate, quisqualate, kainate, and N-methyl-D-aspartate (NMDA). Responses to glutamate and aspartate, which are endogenous neurotransmitter candidates, were enhanced by adding glycine. Glycine especially augmented the maximum response to NMDA in a full concentration range. 6-Cyano-7-nitroquinoxaline-2,3-dione (CNQX) did not suppress the strychnine-sensitive glycine response but did suppress the strychnine-insensitive NMDA response in a competitive manner for glycine. The results suggest that glycine influences neural activity in the SCN as a classical inhibitory neurotransmitter and an excitatory neuromodulator.
Experiments were conducted to determine the effects of different types of paddy rice (dehulled, crushed and untreated whole-grain forms) on growth performance in broiler chickens. In Experiment 1, a control diet containing 41.6% corn (control), and three experimental diets containing 40.7% dehulled, 43% crushed or 43% whole-grain paddy rice, were formulated to be iso-caloric (3,100 kcal/kg ME) and iso-nitrogenous (20% crude protein), but to contain different levels of fat (6%, 5.6%, 10.7%, 10.7%, respectively). Groups of 0-day-old chickens were the fed experimental diets ad libitum for 28 d. The average final body weight of groups fed the crushed and untreated wholegrain paddy rice was significantly lower (P<0.05) than that of the control group, but that of dehulled rice-fed group was slightly higher than the control group. This finding implies that the growth retardation seen with crushed or untreated whole rice could be caused by the feed intake-reduction due to the fat addition to the experimental diets. In Experiment 2, the control diet containing corn and the experimental diets containing dehulled and untreated wholegrain paddy rice were formulated to contain similar levels of fat (5.6-6%) and to be iso-nitrogenous (20%), but not to be iso-caloric. As a result, the average body weight gain of the whole-grain rice-fed chickens showed a considerable increase compared with the control diet-fed chickens even though the energy content of the former diet was only about 90% that of the control diet. It is concluded that whole-grain paddy rice could serve as a valuable constituent of broiler chicken feed, but that excess fat in the diet to maintain the ME content (3,100 kcal/g) could have a negative effect on growth performance.
This study showed a low prevalence of EBF at 6 months in the studied area in Lao PDR. Some of the factors that had a strong impact on EBF at 6 months included: location of residence, ethnicity, father's involvement, early breastfeeding plan, Mother's Card in antenatal care and television advertisement. There may be opportunities for government to review a range of policies relating to paternal involvement, antenatal care and formula advertising that could help to improve EBF rate.
Autophagy plays a critical role in tumorigenesis, but how autophagy contributes to cancer cells’ responses to chemotherapeutics remains controversial. To investigate the roles of autophagy in malignant gliomas, we used CRISPR/CAS9 to knock out the ATG5 gene, which is essential for autophagosome formation, in tumor cells derived from patients with glioblastoma. While ATG5 disruption inhibited autophagy, it did not change the phenotypes of glioma cells and did not alter their sensitivity to temozolomide, an agent used for glioblastoma patient therapy. Screening of an anticancer drug library identified compounds that showed greater efficacy to ATG5‐knockout glioma cells compared to control. While several selected compounds, including nigericin and salinomycin, remarkably induced autophagy, potent autophagy inducers by mTOR inhibition did not exhibit the ATG5‐dependent cytoprotective effects. Nigericin in combination with ATG5 deficiency synergistically suppressed spheroid formation by glioma cells in a manner mitigated by Ca2+ chelation or CaMKK inhibition, indicating that, in combination with autophagy inhibition, calcium‐mobilizing compounds contribute to efficient anticancer therapeutics. ATG5‐knockout cells treated with nigericin showed increased mitochondria‐derived reactive oxygen species and apoptosis compared to controls, indicating that autophagy protects glioma cells from mitochondrial reactive oxygen species‐mediated damage. Finally, using a patient‐derived xenograft model, we demonstrated that chloroquine, a pharmacological autophagy inhibitor, dramatically enhanced the efficacy of compounds selected in this study. Our findings propose a novel therapeutic strategy in which calcium‐mobilizing compounds are combined with autophagy inhibitors to treat patients with glioblastoma.
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