Direct synthesis of conjugated tetraenes from 1,3-enynes with 1,3-dienes.

New direct access to conjugated tetraenes has been achieved. A Ru(0)-catalysed reaction of 1,3-enynes with 1,3-dienes gives 1,3,5,7-octatetraene derivatives by formal regioselective insertion of the alkynyl group of 1,3-enynes into the terminal C-H bond in 1,3-dienes. With a silyl substituent on the alkynyl side in 1,3-enynes, the reaction regioselectively proceeds to give the linear cross-dimerisation product having the silyl group at the internal position. Stoichiometric and DFT calculations support the oxidative coupling mechanism for the linear cross-dimerisation. Methyl (2E,4E,6E,8E)-10-hydroxy-2,4,6,8-decatetraenoate, a versatile polyene intermediate, is accessed by this method as a formal synthesis of biologically active compounds.

Stimulation of glucose uptake by theasinensins through the AMP-activated protein kinase pathway in rat skeletal muscle cells.

Theasinensins, dimeric catechins, have been reported to possess anti-hyperglycemic activity, but the underlying mechanism for this activity remains unknown. In this study, the effect of theasinensins A and B on glucose uptake into rat skeletal muscle cells (L6 myotubes) was investigated. A glucose uptake study using 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxyglucose (2-NBDG) indicated that both theasinensins A and B stimulated glucose uptake in a concentration-dependent manner and translocation of glucose transporter 4 (GLUT4) to the plasma membrane. In addition, inhibition studies measuring 2-NBDG uptake in L6 cells revealed that compound C (AMP-activated protein kinase inhibitor) suppressed theasinensin-stimulated glucose uptake, whereas genistein (insulin receptor tyrosine kinase inhibitor) and wortmannin (phosphatidylinositol 3-kinase inhibitor) were inactive. Subsequent experiments on GLUT4-related signaling pathways in L6 cells demonstrated that theasinensins promoted the phosphorylation of AMPK, but not that of Akt, and that the theasinensin-promoted glucose uptake was blocked in the presence of a CaMKK inhibitor. The promotion of AMPK phosphorylation by theasinensins was not blocked in LKB1-knockdown cells. Consequently, it was concluded that theasinensins A and B did in fact promote GLUT4 translocation to the plasma membrane in L6 myotubes through the CaMKK/AMPK signaling pathway, but not through the PI3K/Akt pathway.

Isolation of Cryptosporidium andersoni Kawatabi type in a slaughterhouse in the northern island of Japan.

Fecal samples were collected from 325 adult cattle and 108 pigs in a slaughterhouse in Hokkaido, the northern island of Japan. Five adult cattle were found to be positive for oocysts of Cryptopsoridium (1.5%). The oocysts were morphologically similar to those of Cryptosporidium andersoni. The partial sequence of the 18S rRNA gene of the isolate was 100% identical with that of the C. andersoni Kawatabi strain. SCID mice were infected after oral administration. Based on the morphology of the oocysts, the sequence of the 18S rRNA gene and the infectivity to SCID mice, the isolate was concluded to be of the same type as the C. andersoni Kawatabi strain that has been isolated in Honshu, the main island of Japan.

A novel enzymatic approach to the massproduction of L-galactose from L-sorbose.

Wild-type strain of Pseudomonas cichorii ST-24 was unable to grow on D -psicose and inductively produced D -tagatose 3-epimerase (D -TE) with D -tagatose as an inducer. We have isolated a constitutive mutant, designated strain Ka75, which had acquired a new ability to grow on a mineral salts medium containing D -psicose as a sole carbon source. The D -psicose-metabolizing mutant synthesized a high level of D -TE. When grown on the culture medium supplemented with Mn(2+), the mutant strain produced around 250-fold higher activity than did the parent strain. Enzymatic properties of the constitutive enzyme were similar to those of the wild-type. Using the immobilized D -TE and recombinant L-rhamnose isomerase (L-RhI) from Escherichia coli strain JM109, a two-step enzymatic reaction was performed for massproduction of a rare aldo-hexose monosaccharide, L-galactose, from a common one, L-sorbose. In the first step, L-sorbose was epimerized to L-tagatose in a yield of 28%. The L-tagatose obtained was utilized as a starting material for L-galactose preparation by the immobilized L-RhI. At equilibrium, approximately 30% L-tagatose was isomerized to L-galactose. Finally, 7.5 g of L-galactose was obtained from 100 g of L-sorbose, viz an overall yield of 7.5%. The product obtained was purified and identified to be L-galactose by specific optical rotation and high performance liquid chromatography (HPLC) analysis, and was ultimately confirmed by (13)C nuclear magnetic resonance ((13)C NMR) and IR spectra.