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Surimi gel is a commonly found gelled product in Japan. Disintegration of the surimi gel is mainly caused by proteolytic degradation of the myosin heavy chain (MHC) under an inappropriate heating process. Many studies have reported the decrease in MHC in the disintegrated surimi gel but the mechanistic details of this degradation remain unclear. This study employed peptidomic analysis of disintegrated surimi gels from deep-sea bonefish Pterothrissus gissu to reveal the MHC cleavage causing gel disintegration. More peptides derived from an MHC rod were found in the disintegrated P. gissu surimi gels than in the integrated gel. Most MHC peptides were derived from the Src homology 3 domain or near the skip residues. The results of the terminome analysis suggest that the catalytic type of the proteases is responsible for light meromyosin cleavage activated at ∼35 °C. These results showed the temperature-dependent cleavage of the MHC rod, causing disintegration of the P. gissu surimi gel.

The O 2 consumption rate of embryos has been attracting much attention as a key indicator of cell metabolisms and development. In this study, we propose an on-chip device that enables the accurate, easy, and noninvasive measurement of O 2 consumption rates of single embryos. Pt electrodes and micropits for embryo settlement were fabricated on Si chips via microfabrication techniques. The configuration of the device enables the detection of O 2 concentration profiles surrounding the embryos by settling embryos into the pits with a mouth pipet. Moreover, as the detection is based on an electrochemical method, the influence of O 2 consumption on the electrodes was also considered. By using a simulator (COMSOL Multiphysics), we estimated the O 2 concentration profiles in the device with and without the effects of the electrodes. Based on the simulation results, we developed a normalization process to calculate the precise O 2 consumption rate of the sample. Finally, using both the measurement system and the algorithm for the analysis, the respiratory activities of mouse embryos were successfully measured.

γ-Oryzanol (γ-ORZ) is a mixture of phytosteryl ferulates purified from rice bran oil. In this study, we examined whether γ-ORZ represents a suppressive effect on the lipopolysaccharide (LPS)-induced adhesion molecule expression on vascular endothelium. Treatment with LPS elevated the mRNA expression of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and E-selectin in bovine aortic endothelial cells (BAECs). Pretreatment with γ-ORZ dose-dependently decreased the LPS-mediated expression of these genes. Western blotting also revealed that pretreatment with γ-ORZ dose-dependently inhibited LPS-induced VCAM-1 expression in human umbilical vein endothelial cells. Consistently, pretreatment with γ-ORZ dose-dependently reduced LPS-induced U937 monocyte adhesion to BAECs. In immunofluorescence, LPS caused nuclear factor-κB (NF-κB) nuclear translocation in 40% of BAECs, which indicates NF-κB activation. Pretreatment with γ-ORZ, as well as its components (cycloartenyl ferulate, ferulic acid, or cycloartenol), dose-dependently inhibited LPS-mediated NF-κB activation. Collectively, our results suggested that γ-ORZ reduced LPS-mediated adhesion molecule expression through NF-κB inhibition in vascular endothelium.

The antimelanosic and antioxidative properties of a hot water extract prepared from the fruiting body of the edible mushroom (Flammulina velutipes) were evaluated by dietary supplementation in Kuruma shrimp (Marsupenaeus japonicus) for possible aquaculture application. The extract contained ergothioneine (ERT) at a level of 2.05 mg/mL. A commercial standard of l-ergothioneine (l-ERT) and the mushroom extract showed inhibitory activity against mushroom polyphenoloxidase (PPO). Feeding of the extract had no adverse effects on the immune systems of the shrimp under the present experimental conditions. Supplementation of the extract in the diet significantly suppressed PPO activities in the hemolymphs of the shrimp. Expression of the prophenoloxidase (proPO) gene decreased in the hemocyte of the Kuruma shrimp fed with the mushroom extract. Consequently, development of melanosis was significantly suppressed in the supplement fed shrimp during ice storage. Lipid oxidation was also effectively controlled in the supplement fed group throughout the storage period. In vitro experiments showed that l-ERT effectively inhibited the activation of proPO in the hemocyte lysate supernatant (HLS). The transcript of the proPO gene in the hemocyte showed lower expression in the l-ERT-treated HLS. It was concluded that dietary supplementation of the mushroom extract in shrimp could be a promising approach to control post mortem development of melanosis and lipid oxidation in shrimp muscles.

The antioxidative property of a hydrophilic extract prepared from the fruiting body of edible mushroom ( Flammulina velutipes) was evaluated. The mushroom extract contained ergothioneine (ERT) at a level of 3.03 +/- 0.07 mg/mL, showed higher 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, and suppressed lipid oxidation of bigeye tuna meat more effectively than authentic L-ERT added at the same concentration. The authentic L-ERT had stronger total reducing power than the mushroom extract and inhibited the formation of metmyoglobin (metMb) more significantly in bigeye tuna meat. Lipid oxidation in beef and fish meats to which the mushroom extract had been added was "virtually" controlled during storage on ice. Ground beef and bigeye tuna meat with the extract added kept their natural colors unchanged for longer than 12 and 7 days of ice storage, respectively. Contrary to this, browning in meat color was observed in the control samples without the extract after 6 and 2 days of storage, respectively, when stored under similar conditions. There was significant correlation between meat color and chemical parameters, including total lipid hydroperoxides, thiobarbituric acid reactive substances, and metMb. However, there was no significant correlation between pH value and meat discoloration. These results suggest that ERT in the hydrophilic extract of F. velutipes plays an important role as a color stabilizer of meats.

Fluorometric and fluorescent image analysis methods with 3-perylene diphenylphosphine (3-PeDPP) were developed to measure lipid hydroperoxides in refined tuna oil. The fluorescence intensity of 3-perylene diphenylphosphineoxide (3-PeDPPO, the corresponding peroxidation product of 3-PeDPP) has been used for assessing lipid hydroperoxides in refined tuna oil models, and the results are compared to those of a classical method using ferric-xylenol orange complex. There were good correlations between the lipid hydroperoxide contents determined by the 3-PeDPPO fluorescence and the xylenol orange methods (R(2) = 0.984). Moreover, the novel probe 3-PeDPP and the fluorescent image analysis enabled us to evaluate two-dimensionally the lipid hydroperoxide contents in tuna oil droplets. These results suggest that the fluorometric and fluorescent image analyses using 3-PeDPP would be suitable for batch and two-dimensional determinations of lipid hydroperoxides in oil and oil-containing materials.