4-Hydroxy hexenal derived from dietary n-3 polyunsaturated fatty acids induces anti-oxidative enzyme heme oxygenase-1 in multiple organs.

It has recently been reported that expression of heme oxygenase-1 (HO-1) plays a protective role against many diseases. Furthermore, n-3 polyunsaturated fatty acids (PUFAs) were shown to induce HO-1 expression in several cells in vitro, and in a few cases also in vivo. However, very few reports have demonstrated that n-3 PUFAs induce HO-1 in vivo. In this study, we examined the effect of fish-oil dietary supplementation on the distribution of fatty acids and their peroxidative metabolites and on the expression of HO-1 in multiple tissues (liver, kidney, heart, lung, spleen, intestine, skeletal muscle, white adipose, brown adipose, brain, aorta, and plasma) of C57BL/6 mice. Mice were divided into 4 groups, and fed a control, safflower-oil, and fish-oil diet for 3 weeks. One group was fed a fish-oil diet for just 1 week. The concentration of fatty acids, 4-hydroxy hexenal (4-HHE), and 4-hydroxy nonenal (4-HNE), and the expression of HO-1 mRNA were measured in the same tissues. We found that the concentration of 4-HHE (a product of n-3 PUFAs peroxidation) and expression of HO-1 mRNA were significantly increased after fish-oil treatment in most tissues. In addition, these increases were paralleled by an increase in the level of docosahexaenoic acid (DHA) but not eicosapentaenoic acid (EPA) in each tissue. These results are consistent with our previous results showing that DHA induces HO-1 expression through 4-HHE in vascular endothelial cells. In conclusion, we hypothesize that the HO-1-mediated protective effect of the fish oil diet may be through production of 4-HHE from DHA but not EPA in various tissues.

Heterotopic ossification of the distal portion of biceps femoris: case report and review of the literature.

Heterotopic ossification is the formation of mature lamellar bone in soft tissue. We report a very unusual case of heterotopic ossification arising in the distal portion of the biceps femoris muscle. A 29-year-old woman presented with knee pain after playing golf. She had had no apparent history of trauma. Radiography of the knee showed a calcified mass adjacent to the lateral side of her knee joint. The radiological diagnosis was calcification of the lateral collateral ligament. The pain soon disappeared without treatment. At the age of 45 years, she complained of a growing mass in her left knee that was occasionally painful. Computed tomography and magnetic resonance imaging demonstrated a well matured ossified mass in the lateral side of her left knee. Radiologically and macroscopically, it was found to be in continuity with the distal part of biceps femoris. The mass was excised en bloc. Microscopically, the lesion was mainly composed of well-matured lamellar bone with bone marrow and islands of cartilage showing enchondral ossification. No apparent zoning was found. Cellular atypia was not observed. Extraskeletal osteosarcoma was ruled out. The mass was diagnosed as a heterotopic ossification arising from the distal part of biceps femoris.

Osteolipoma arising adjacent to the sternoclavicular joint. A case report.

A 45-year-old woman presented with one-year history of a mass on her chest. Computed tomography and magnetic resonance imaging demonstrated a tumour near the sternoclavicular joint. The tumour was diagnosed as osteolipoma histologically after resection. Osteolipoma is a rare tumour and this may be the first report of osteolipoma arising adjacent to the sternoclavicular joint.

Effect of head size in head-tail-type polycations on their in vitro performances as nonviral gene vectors.

Three kinds of head-tail-type block copolymers composed of polyamidoamine (PAMAM) dendron heads and poly(L-lysine) (PLL) tail blocks (PAMAM dendron-PLL), having PAMAM dendrons with different generations (G2.5-PLL, G3.5-PLL and G4.5-PLL) were synthesized. Some of the dendron heads were located at polyplex surface, and G2.5-PLL and G3.5-PLL could form small polyplexes (less than 150 nm in size). G2.5-PLL and G3.5-PLL polyplexes were taken up into the cells more effectively. PAMAM dendron-PLL that had a larger dendron head could show a more-effective buffering effect. The in vitro performance of the PAMAM dendron-PLL polyplexes was controlled by the balance of cellular uptake and endosomal escape by a buffering effect.

Synthesis and characterization of a head-tail type polycation block copolymer as a nonviral gene vector.

A head-tail type polycation block copolymer, which is composed of the polyamidoamine (PAMAM) dendron and poly(L-lysine) (PLL) blocks, was newly designed as a nonviral gene vector in this study. This block copolymer (PAMAM dendron-PLL) was successfully synthesized in two steps: the synthesis of the PAMAM dendron block and the polymerization of the PLL block from the PAMAM dendron block. PAMAM dendron and PLL blocks in block copolymer showed independent deprotonation behavior, and their pK(a) were determined to be 6.8 and 9.0, respectively. The complexation with pDNA was evaluated by gel retardation assay and dye exclusion assay, and both assays indicated that pDNA was selectively complexed with PLL block of block copolymer. Also, the PAMAM dendron-PLL poplyplexes showed 10(2) fold higher transfection efficiency to HeLa cells as that for PLL polyplexes. This might be due to the buffering effect of the PAMAM dendron block. This block copolymer could produce a function share in each block, i.e., tail block complexed with pDNA and head block showed a buffering effect. This molecular design of the head-tail type block copolymer might provide a new approach for realizing in vivo gene therapy.

Role of Thr(66) in porcine NADH-cytochrome b5 reductase in catalysis and control of the rate-limiting step in electron transfer.

Site-directed mutagenesis of Thr(66) in porcine liver NADH-cytochrome b(5) reductase demonstrated that this residue modulates the semiquinone form of FAD and the rate-limiting step in the catalytic sequence of electron transfer. The absorption spectrum of the T66V mutant showed a typical neutral blue semiquinone intermediate during turnover in the electron transfer from NADH to ferricyanide but showed an anionic red semiquinone form during anaerobic photoreduction. The apparent k(cat) values of this mutant were approximately 10% of that of the wild type enzyme (WT). These data suggest that the T66V mutation stabilizes the neutral blue semiquinone and that the conversion of the neutral blue to the anionic red semiquinone form is the rate-limiting step. In the WT, the value of the rate constant of FAD reduction (k(red)) was consistent with the k(cat) values, and the oxidized enzyme-NADH complex was observed during the turnover with ferricyanide. This indicates that the reduction of FAD by NADH in the WT-NADH complex is the rate-limiting step. In the T66A mutant, the k(red) value was larger than the k(cat) values, but the k(red) value in the presence of NAD(+) was consistent with the k(cat) values. The spectral shape of this mutant observed during turnover was similar to that during the reduction with NADH in the presence of NAD(+). These data suggest that the oxidized T66A-NADH-NAD(+) ternary complex is a major intermediate in the turnover and that the release of NAD(+) from this complex is the rate-limiting step. These results substantiate the important role of Thr(66) in the one-electron transfer reaction catalyzed by this enzyme. On the basis of these data, we present a new kinetic scheme to explain the mechanism of electron transfer from NADH to one-electron acceptors including cytochrome b(5).

Design and synthesis of a selective EP4-receptor agonist. Part 4: practical synthesis and biological evaluation of a novel highly selective EP4-receptor agonist.

A practical method of synthesizing a highly selective EP4-receptor agonist 1 using Corey lactone 2 as a key intermediate was developed. Selective methanesulfonylation of the primary alcohol of the diol 8 under the newly devised conditions followed by the protection of the remaining secondary alcohol are key reactions in this new method. Further biological evaluation of 1a-b is also reported.