Effect of medium-chain triglycerides on the postprandial triglyceride concentration in healthy men

This study compared the serum lipid concentrations after a single dose of medium-chain triglycerides (MCT) or long-chain triglycerides (LCT) between individuals grouped according to the body mass index (BMI). Twenty-five males participated as volunteers, the test diet containing 10 g of MCT or LCT. Blood samples were collected up to 6 h after the intake of a test diets. The LCT diet resulted in significantly greater increases in areas under the curves (AUCs) for serum and chylomicron triglyceride in the BMI > or = 23 kg/m2 group than those in the BMI < 23 kg/m2 group. The magnitude of response after intake of the MCT diet by the BMI > or = 23 kg/m2 group was significantly lower than that after the LCT diet. These results suggest that, in subjects with BMI > or = 23 kg/m2, the intake of MCT is preferable to that of LCT for maintaining postprandial triglyceride at a low concentration.     

Activation of protein kinase C reverses capsaicin-induced calcium-dependent desensitization of TRPV1 ion channels

Ca2+ selective ion channels of vanilloid receptor subtype-1 (TRPV1) in capsaicin-sensitive dorsal root ganglion (DRG) neurons and TRPV1 transfected Chinese hamster ovarian (CHO) cells are desensitized following calcium-dependent tachyphylaxis induced by successive applications of 100 nM capsaicin. Tachyphylaxis of TRPV1 to 100 nM capsaicin stimuli was not observed in the absence of extracellular calcium. Capsaicin sensitivity of desensitized TRPV1 ion channels recovered on application of phorbol-12-myristate-13-acetate (PMA). PMA-induced recovery of desensitized TRPV1 was primarily due to influx of extracellular calcium observed during re-application of capsaicin following desensitization. Capsazepine blocked the re-sensitization to capsaicin by PMA. Protein kinase C (PKC) inhibitory peptide PKC fragment 19-36 also inhibited re-sensitization to capsaicin by PMA. Reversal of capsaicin-induced desensitization by PMA was prevented by a mutation of TRPV1 where phosphorylation sites serine502 and serine800 were replaced with alanine. This study provides evidence for a role of PKC in reversing capsaicin-induced calcium-dependent desensitization of TRPV1 ion channels.     

Tirucallane-type triterpenes from Juliania adstringens

Five tirucallane-type triterpenes were isolated along with nine known triterpenes from the bark of Juliania adstringens. The structures of the five triterpenes were determined by analysis of their 1H and 13C NMR and mass spectral data, and each compound exhibited growth inhibitory activity against leukemia cells (L-1210).     

Antagonist of secondary lymphoid-tissue chemokine (CCR ligand 21) prevents the development of chronic graft-versus-host disease in mice

The use of receptor antagonists for chemokines is an alternative approach to blocking chemokine actions and has the potential to provide novel therapeutics. We determined the receptor antagonist properties of murine N-terminally truncated secondary lymphoid tissue chemokine (SLC)/6Ckine/CCR ligand 21 analogs and evaluated the preventive effects of SLC antagonists on chronic graft-vs-host disease (GVHD) in a murine model by blocking the homing of donor CCR7-expressing T cells into the recipient's lymphoid organs. SLC analogs truncated >4 aa residues from the N terminus showed a loss of chemotaxis and Ca2+ influx of CCR7-expressing cells and also inhibited SLC-stimulated chemotaxis and SLC-induced Ca2+ influx completely. To determine whether SLC antagonist inhibits the development of chronic GVHD, chronic GVHD was induced by injecting DBA/2 spleen cells into (C57BL/6 x DBA/2) F1 mice. Total numbers of spleen cells and host B cells, serum levels of IgE, and of total IgG and IgG1 of anti-DNA Abs in SLC antagonist-treated GVHD mice were significantly lower than those in control PBS-treated GVHD mice. This was due to a reduction in the levels of activated donor CD4+ T cells and a decrease in IL-4 production, resulting in a reduction in the numbers of activated host B cells. Therefore, our results suggest that SLC antagonist has beneficial effects for the prevention of chronic GVHD.     

Truncation of CDK5 activator p35 induces intensive phosphorylation of Ser202/Thr205 of human tau

Hyperphosphorylated tau is a major component of neurofibrillary tangles, one of the hallmarks of Alzheimer's disease. CDK5 is a kinase that phosphorylates the tau protein, and its endogenous activator, p35, regulates its activity. Recently, calpain was found to digest p35 to its truncated product, p25. Several lines of evidence suggest that p25-CDK5 has much more powerful kinase activity and that it may cause abnormal hyperphosphorylation of tau. In this study, we have examined the kinetic characteristics of in vitro phosphorylation of the longest isoform of human tau by CDK5 and its activators using recombinant proteins. Although the kinase activity of CDK5 in phosphorylating tau was significantly higher in the presence of p25, the affinity of CDK5 for tau was not different. Phosphopeptide mapping revealed enhanced phosphorylation of Ser(202)/Thr(205) residues by p25-CDK5 (amino acid residues of tau are numbered according to the longest isoform of human tau). These results suggest that cleavage of p35 to p25 greatly enhances the kinase activity of CDK5 and increases the phosphorylation of Ser(202)/Thr(205). Considering the fact that phosphorylation of Ser(202)/Thr(205) antagonizes the tau-mediated nucleation of tubulin, p25-CDK5 may play a pivotal role in neuronal cell death in Alzheimer's disease.     

A new technique to prevent self-ligation of DNA

The most widely used technique for preventing self-ligation (self-circularization and concatenation) of DNA is dephosphorylation of the 5'-end, which stops DNA ligase from catalyzing the formation of phosphodiester bonds between the 3'-hydroxyl and 5'-phosphate residues at the DNA ends. The 5'-dephosphorylation technique cannot be applied to both DNA species to be ligated and thus, the untreated DNA species remains capable of self-ligation. To prevent this self-ligation, we replaced the 2'-deoxyribose at the 3'-end of the untreated DNA species with a 2',3'-dideoxyribose. Self-ligation was prevented at the replaced 3'-end, while the 5'-phosphate remaining at the 5'-end permitted ligation with the 3'-hydroxyl end of the 5'-dephosphorylated DNA strand. We successfully applied this 3'-replacement technique to gene cloning, adapter-mediated polymerase chain reaction and messenger RNA fingerprinting. The 3'-replacement technique is simple and not restricted by sequence or conformation of the DNA termini and is thus applicable to a wide variety of methods involving ligation.     

Sesquiterpene pyridine alkaloids from Hippocratea excelsa

Nineteen sesquiterpene pyridine alkaloids including 17 new compounds have been isolated from the 70% aq. EtOH extract of stem barks of Hippocratea excelsa. The structures of these compounds were elucidated by various spectroscopic means.     

Restriction fragments homologous to mitochondrial plasmid-like DNAs are located within limited chromosomal regions on the rice nuclear genome

Summary The shifted multiplicative model (SHMM) is used with a cluster method to identify subsets of sites in an international maize (Zea mays L.) trial without genotypic rank-change. For cluster analysis, distance between two sites is defined as the residual sum of squares after fitting SHMM with one multiplicative term (SHMM₁) if SHMM₁ does not show genotypic rank-change. However, if SHMM₁ does show genotypic rank-change, the distance between two sites is defined as the smaller of the sums of squares owing to genotypes within each of the two sites. Calculation of distance between two sites is facilitated by using the site regression model with one multiplicative term (SREG₁), which can be reparameterized as SHMM₁ when only two sites are considered. The dichotomous splitting procedure, used on the dendrogram obtained from cluster analysis, will first perform SHMM analyses on each of the last two cluster groups to join (end of the dendrogram). If SHMM₁ does not give an adequate fit, the next step is to move down the branches of the tree until groups of sites (clusters) are found to which SHMM₁ provides an adequate fit and primary effects of sites are all of the same sign. Five final groups of sites to which SHMM₁ provides an adequate fit and primary effects of sites are all of the same sign were obtained. The procedure appears to be useful in identifying subsets of sites in which genotypic rank-change interactions are negligible.Research reported in this paper (Journal article no. 91-3-218) is part of a project of the Kentucky Agricultural Experiment Station, published with the approval of the Director     

Lanthanide- and aromatic solvent-induced shift effects on proton resonances in C-4-methylated steroids and tetracyclic triterpenoids

Lanthanide-induced shifts (LIS) with Eu (dpm)₃ and aromatic solvent induced shifts (ASIS) with C₄H₄, C₅H₅N₁ and C₆F₆ of PMR signals were examined for a series of C-4-methylated steroids and tetracyclic triterpenoids having a hydroxyl, carbonyl or acetoxyl group at position C-3. The magnitude and/or direction of the LIS (or ASIS) of corresponding protons were extensively influenced by the nature of the C-3-functional groups. The possible geometries of Eu (dpm)₃-substrate complexes were also discussed on the basis of the LIS data. The above two techniques in the PMR spectroscopy provided the confirmatory evidence for the structural and stereochemical determination of steroids and triterpenoids.