Lanthionine as an essential constituent of cell wall peptidoglycan ofFusobacterium nucleatum

Lanthionine, a sulfur-containing diamino acid which had not previously been reported as one of the main amino acids of any bacterial cell wall peptidoglycan, was demonstrated inFusobacterium nucleatum peptidoglycan isolated by sodium dodecyl sulfate extraction and protease digestion. Lysine, diaminopimelic acid, and ornithine were absent. Lanthionine seems to be an essential dibasic amino acid, involved in cross-linkages betwen stem peptide subunits inF. nucleatum.     

Meg1/Grb10 overexpression causes postnatal growth retardation and insulin resistance via negative modulation of the IGF1R and IR cascades

The Meg1/Grb10 protein has been implicated as an adapter protein in the signaling pathways from insulin receptor (IR) and insulin-like growth factor 1 receptor (IGF1R) in vitro. To elucidate its in vivo function, four independent Meg1/Grb10 transgenic mouse lines were established, and the effects of excess Meg1/Grb10 on both postnatal growth and glucose metabolism were examined. All of the Meg1/Grb10 transgenic mice showed growth retardation after weaning (3-4 weeks), which indicates that ectopic overexpression of Meg1/Grb10 inhibits postnatal growth that is mediated by IGF1 via IGF1R. In addition, the mice became hyperinsulinemic owing to high levels of insulin resistance, which demonstrates that Meg1/Grb10 also modulates the insulin receptor cascade negatively in vivo. Type II diabetes arose frequently in the two transgenic lines, which also showed impaired glucose tolerance. In these mice, severe atrophy of the pancreatic acinus cells was associated with high-level production of Meg1/Grb10 in the pancreas. These results suggest that Meg1/Grb10 inhibits the function of both insulin and IGF1 receptors in these cells, since a similar phenotype has been reported for Ir and Igf1r double knockout mice. Taken together, these results indicate that Meg1/Grb10 interacts with both insulin and IGF1 receptors in vivo, and negatively regulates the IGF growth pathways via these receptors.     

Novel Daple-like protein positively regulates both the Wnt/beta-catenin pathway and the Wnt/JNK pathway in Xenopus

Wnt signaling pathways are essential in various developmental processes including differentiation, proliferation, cell migration, and cell polarity. Wnt proteins execute their multiple functions by activating distinct intracellular signaling cascades, although the mechanisms underlying this activation are not fully understood. We identified a novel Daple-like protein in Xenopus and named it xDal (Xenopus Daple-like). As with Daple, xDal contains several leucine zipper-like regions (LZLs) and a putative PDZ domain-binding motif, and can interact directly with the dishevelled protein. In contrast to mDaple, injection of xDal mRNA into the dorso-vegetal blastomere does not induce ventralization and acted synergistically with xdsh in secondary axis induction. XDal also induced expression of siamois and xnr-3, suggesting that XDal functions as a positive regulator of the Wnt/beta-catenin pathway. Injection of xDal mRNA into the dorso-animal blastomere, however, induced gastrulation-defective phenotypes in a dose-dependent manner. In addition, xDal inhibited activin-induced elongation of animal caps and enhanced c-jun phosphorylation. Based on these findings, xDal is also thought to function in the Wnt/JNK pathway. Moreover, functional domain analysis with several deletion mutants indicated that xDal requires both a putative PDZ domain-binding motif and at least one LZL for its activity. These findings with xDal will provide new information on the Wnt signaling pathways.     

3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (trp-P-1) is incorporated into rat splenocytes,thymocytes, and hepatocytes through monoamine transporters and induces apoptosis

3-Amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1), which is a tryptophan pyrolysate formed during cooking, induces apoptosis in rat splenocytes, thymocytes, and hepatocytes. In this study, we investigated whether Trp-P-1 is transported into these cells and causes apoptosis. Trp-P-1 was immediately incorporated into rat splenocytes, thymocytes, and hepatocytes in a dose- and time-dependent manner. Dopamine and serotonin significantly competed with the uptake of Trp-P-1 into these cells, and nomifensine and indatraline, which are inhibitors of dopamine- and serotonin-transporters, respectively, markedly suppressed the uptake of Trp-P-1. On the other hand, amino acids including tryptophan did not compete with Trp-P-1. Inhibition of monoamine transporters using nomifensine and indatraline partially suppressed Trp-P-1-induced cell death in these cells. In hepatocytes, the inhibition of transporters prevented Trp-P-1-induced morphological changes and activation of caspase-3. These results demonstrated that Trp-P-1 is incorporated into the cells through monoamine transporters and induces apoptosis.     

The liver-enriched transcription factor CREBH is nutritionally regulated and activated by fatty acids and PPARα

To elucidate the physiological role of CREBH, the hepatic mRNA and protein levels of CREBH were estimated in various feeding states of wild and obesity mice. In the fast state, the expression of CREBH mRNA and nuclear protein were high and profoundly suppressed by refeeding in the wild-type mice. In ob/ob mice, the refeeding suppression was impaired. The diet studies suggested that CREBH expression was activated by fatty acids. CREBH mRNA levels in the mouse primary hepatocytes were elevated by addition of the palmitate, oleate and eicosapenonate. It was also induced by PPARα agonist and repressed by PPARα antagonist. Luciferase reporter gene assays indicated that the CREBH promoter activity was induced by fatty acids and co-expression of PPARα. Deletion studies identified the PPRE for PPARα activation. Electrophoretic mobility shift assay and chromatin immunoprecipitation (ChIP) assay confirmed that PPARα directly binds to the PPRE. Activation of CREBH at fasting through fatty acids and PPARα suggest that CREBH is involved in nutritional regulation.     

Some properties of an immobilized glycolysis system of yeast in fermentative phosphorylation of nucleotides

Summary Immobilized dried yeast cells, which contain glycolytic and some other emzymes, required NAD but not ATP for the (e.g. choline kinase, pyrophosphorylase) fermentative production of CDP-choline, when washed and reused. The immobilized system was more resistant to heat than dried cells, which had previously been used for the same purpose. However, when too many cells were immobilized, leakage of the enzymes from the resin lattice was observed during repeated use. To prevent this leakage, the ratio of cells to resin should be considered. Present name: Yoshinori Wakai     

TeleOphta: Machine learning and image processing methods for teleophthalmology

A complete prototype for the automatic detection of normal examinations on a teleophthalmology network for diabetic retinopathy screening is presented. The system combines pathological pattern mining methods, with specific lesion detection methods, to extract information from the images. This information, plus patient and other contextual data, is used by a classifier to compute an abnormality risk. Such a system should reduce the burden on readers on teleophthalmology networks.     

Studies on Chrysanthemic Acid

Synthesis of (±)-trans-chrysanthemic acid from (±)-1′-hydroxydihydro-trans-chrysanthemic acid by the dehydration with p-toluene-sulfonic acid was attempted. However, the attempt was found to be unsuccessful giving a compound believed to be methyl methyl 2,6 dimethylhepta-3.6-diene-5-carboxylate upon dehydration.

A cleavage upon cyclopropane ring was confirmed by deriving the acid obtained by the hydrolysis of the above ester to already known 2,6-dimethyl-heptane-5-carboxylic acid.

Analogous mode of dehydration and cleavage upon the ester of (±)-2,2-dimethyl-3-trans-hydroxylbenzyl-cyclopropane-l-carboxylic acid was also observed to give 1-phenyl-4-methyl-penta-1,3-diene-3-carboxylic acid. On the other hand, (±)-trans-caronic acid being derived to (±)-1′-oxo-2′-hydroxy-dihydro-trans-chrysanthemic acid, the synthesis of (±)-trans-chrysanthemic acid from (±)-trans-caronic acid became possible using (±)-1′-oxo-2′-hydroxy-dihydro-trans-chrysanthemic acid as a relay substance.