Antifreeze Emulsions Produced from Linoleic Acid and Water Using Enzymatically Modified Gelatin

An enzymatically modified gelatin with covalently attached leucine dodecyl ester, referred to as EMG-12, was used as a surfactant to prepare emulsions with different properties by changing the surfactant concentration, oil volume fraction, and pH in the water phase. The emulsions generally resisted the freezing of their constituent bulk water at approximately ?10°C, but similar emulsions produced with soy protein isolate, casein, or Tween-80 as control agents were less resistant. The freezing (or unfreezing) of the bulk water in these emulsions depended on the kind of agent used, not on the emulsion properties such as average area of the oil/water interface, stability against coalescence, and stability against creaming. The emulsion produced with EMG-12, like that produced with polyglycerol stearate, tended to maintain its unfrozen state even in the presence of silver iodide crystals added as heterogeneous ice-nuclei. The significance of producing such an antifreeze emulsion is discussed from the standpoint of cryopreservation of cold-sensitive food and biological systems.     

Structural Influence of Sugar and Aglycon Moieties on the Acetolysis of Some Glycosylamines

The acetolysis was applied to a novel cleavage of N-p-tolyl-β-d-glycosylamines in glucogalacto- and manno-configurations and their 2-acetamido-2-deoxy sugars. The results were compared with the acetolysis of furanosyl- and pyranosylnucleosides. N-Acetyl-p-toluidine was isolated as crystals in up to 92% yield in the acetolysis of N-P-tolyl-β-d-glycosylamines of neutral sugars and in 1.5~2.0% yields in that of 2-acetamido-2-deoxy sugars. The stability of glycosylamine linkages against the acetolysis was found in the following order: pyranosylnucleosides (the most stable) > N-P-tolyl-β-d-glycosylamine of 2-acetamido-2-deoxy sugars > furanosylnucleosides > N-P-tolyl-β-d-glycosylamines of neutral sugars (the most unstable).     

Nutritive Value of 1,2-Propanediol Di-laurate,Di-lauryl Succinate and 1,3-Butanediol by Calves

Procedure of bioassay for chicks was applied to ruminants. Calves of about 8-week-old were fed the same but restricted amount of basal diet and graded levels of corn starch or test materials. The feeding conditions was so adjusted that body weight gain of calves from 1st to 6th week was in proportion to dietary energy supply. Digestive ability of the calves was not disturbed and growth was smooth in the feeding conditions. All of the 3 samples tested were suggested to be available by calves, although palatability of di-lauryl succinate was low. Reliability of the estimate was discussed.     

Enzymatic synthesis of capsaicin 4-O-β-xylooligosaccharides by β-xylosidase from Aspergillus sp

Capsaicin 4-O-β-xylooligosaccharides were synthesized by a biocatalytic xylosylation using Aspergillus sp. β-xylosidase. Capsaicin was converted into three new capsaicin glycosides, i.e. capsaicin 4-O-β-xyloside, capsaicin 4-O-β-xylobioside, and capsaicin 4-O-β-xylotrioside in 15, 12 and 10% yield, respectively. All products were isolated from the reaction mixtures by preparative HPLC. The structures of the products were determined by NMR spectroscopic method.     

A multifaceted genomics approach allows the isolation of the rice Pia-blast resistance gene consisting of two adjacent NBS-LRR protein genes

The Oryza sativa (rice) resistance gene Pia confers resistance to the blast fungus Magnaporthe oryzae carrying the AVR-Pia avirulence gene. To clone Pia, we employed a multifaceted genomics approach. First, we selected 12 R-gene analog (RGA) genes encoding nucleotide binding site-leucine rich repeats (NBS-LRRs) proteins from a region on chromosome 11 that shows linkage to Pia. By using seven rice accessions, we examined the association between Pia phenotypes and DNA polymorphisms in the 10 genes, which revealed three genes (Os11gRGA3-Os11gRGA5) exhibiting a perfect association with the Pia phenotypes. We also screened ethyl methane sulfonate (EMS)-treated mutant lines of the rice cultivar 'Sasanishiki' harboring Pia, and isolated two mutants that lost the Pia phenotype. DNA sequencing of Os11gRGA3-Os11gRGA5 from the two mutant lines identified independent mutations of major effects in Os11gRGA4. The wild-type 'Sasanishiki' allele of Os11gRGA4 (SasRGA4) complemented Pia function in both mutants, suggesting that SasRGA4 is necessary for Pia function. However, when the rice cultivar 'Himenomochi' lacking Pia was transfected with SasRGA4, the Pia phenotype was not recovered. An additional complementation study revealed that the two NBS-LRR-type R genes, SasRGA4 and SasRGA5, that are located next to each other and oriented in the opposite direction are necessary for Pia function. A population genetics analysis of SasRGA4 and SasRGA5 suggests that the two genes are under long-term balancing selection.     

Effects of a plum (Prunus mume Siebold and Zucc.) ethanol extract on the immune system in vivo and in vitro

The effect of a plum ethanol extract (PEE) on immunity was analyzed. An oral administration of PEE increased the interleukin (IL)-12p40 concentration in the serum and T-cell ratio in the spleen. In vitro studies revealed that PEE stimulated IL-12p70 production in peritoneal macrophages and natural killer activity. These findings suggest that PEE enhanced the immune function by stimulating innate immune cells.     

Coordinated movements of the head and body during orienting behaviour in the praying mantis Tenodera aridifolia

The visual orienting behaviour towards prey in the free-moving mantis was investigated with a high-speed camera. The orienting behaviour consisted of head, prothorax, and abdomen rotations. Coordinated movements of these body parts in the horizontal plane were analysed frame-by-frame. Rotations of these body parts were initiated with no or slight (≤40 ms) differences in timing. The initiation timing of prothorax-abdomen rotation was affected by its initial angle before the onset of orienting. There were positive correlations in amplitude among head-prothorax, prothorax-abdomen, and abdomen rotations. The ratio of these rotations to total gaze rotation was affected by the initial prothorax-abdomen angle before the onset of orienting. Our data suggest that coordinated movements of the head, prothorax, and abdomen during orienting are ballistic events and are pre-determined according to visual and proprioceptive information before the onset of orienting.     

Construction of strong mammalian promoters by random cis-acting element elongation

Synthetic oligonucleotides containing one of four kinds of cis-acting elements, binding sites for activating protein-1 (AP-1), nuclear factor kappaB (NF-kappaB), CArG binding factor A (CBF-A), and nuclear factor Y (NF-Y), were randomly ligated to construct DNA fragments. These fragments were inserted into the SalI site of a promoter probe vector; pGL3-TATASal, which is located immediately upstream of the TATA box sequence of the human heme oxygenase 1 gene and linked to the luciferase gene to construct 11 plasmid vectors. When these vectors were introduced into PC-3 cells of human prostate cancer, 6 out of the 11 transfectants showed a significantly higher luciferase activity than pGL3-TATASal. The two strongest promoters (clone 6 and clone 11) were investigated further Clone 6 turned out to be the strongest, showing a 3.0- and 8.4-fold activity in comparison to the two frequently used promoters--the cytomegalovirus (CMV) immediate early promoter and the simian virus 40 (SV40) early promoter respectively. Clone 11 was less active than clone 6, but still showed higher activity than the two promoters. When the plasmids were introduced into nine other cell lines, their activities varied but were still comparable to the two promoters. These results indicate that the method used here is simple and efficient for constructing strong promoters that are potentially useful for vectors in either gene therapy or recombinant vaccine.     

Thrombin regulates the function of human blood dendritic cells

Thrombin is the key enzyme in the coagulation cascade and activates endothelial cells, neutrophils and monocytes via protease-activated receptors (PARs). At the inflammatory site, immune cells have an opportunity to encounter thrombin. However little is known about the effect of thrombin for dendritic cells (DC), which are efficient antigen-presenting cells and play important roles in initiating and regulating immune responses. The present study revealed that thrombin has the ability to stimulate blood DC. Plasmacytoid DC (PDC) and myeloid DC (MDC) isolated from PBMC expressed PAR-1 and released MCP-1, IL-10, and IL-12 after thrombin stimulation. Unlike blood DC, monocyte-derived DC (MoDC), differentiated in vitro did not express PAR-1 and were unresponsive to thrombin. Effects of thrombin on blood DC were significantly diminished by the addition of anti-PAR-1 Ab or hirudin, serine protease inhibitor. Moreover, thrombin induced HLA-DR and CD86 expression on DC and the thrombin-treated DC induced allogenic T cell proliferation. These findings indicate that thrombin plays a role in the regulation of blood DC functions.     

Right-elevated expression of charon is regulated by fluid flow in medaka Kupffer's vesicle

Recent studies have revealed that a cilium-generated liquid flow in the node has a crucial role in the establishment of the left-right (LR) axis in the mouse. In fish, Kupffer's vesicle (KV), a teleost-specific spherical organ attached to the tail region, is known to have an equivalent role to the mouse node during LR axis formation. However, at present, there has been no report of an asymmetric gene expressed in KV under the control of fluid flow. Here we report the earliest asymmetric gene in teleost KV, medaka charon, and its regulation. Charon is a member of the Cerberus/DAN family of proteins, first identified in zebrafish. Although zebrafish charon was reported to be symmetrically expressed in KV, medaka charon displays asymmetric expression with more intense expression on the right side. This asymmetric expression was found to be regulated by KV flow because symmetric and up-regulated charon expression was observed in flow-defective embryos with immotile cilia or disrupted KV. Taken together, medaka charon is a reliable gene marker for LR asymmetry in KV and thus, will be useful for the analysis of the early steps downstream of the fluid flow.