Severe acute cholestatic hepatitis of unknown etiology successfully treated with the Chinese herbal medicine Inchinko-to (TJ-135)

Severe acute hepatitis of unknown etiology is difficult to treat and often progresses to subacute fulminant hepatitis or late-onset hepatic failure. A 45-year-old wellnourished, healthy man had progressive fatigue and his liver function tests showed severe liver dysfunction. The etiology of sever acute cholestatic hepatitis was unknown. The liver function tests normalized gradually, which excluded high persistent total bilirubin after starting on predonine. A liver biopsy showed chronic active hepatitis with mild f ibrosis (A2, F1). Oral Inchinko-to, a Chinese herbal medicine, at 7.5 g daily was prescribed. The treatment was effective with no adverse effects. We present a successfully treated case and discuss hepatoprotective and choleretic effects of Inchinko-to.

Antimicrobial Oil-in-Water Nanoemulsions： Synergistic Effect of Nisin and Carvacrol against Bacillus subtilis

The aim of this study was to elucidate the mode of action of two antimicrobial compounds, nisin, which is hydrophilic, and the lipophilic compound carvacrol. For that purpose, both antimicrobials were loaded into Oil-in-/Water nanoemulsions, and their antimicrobial activity against Bacillus subtilis was investigated. The experimental results indicated that the O/W nanoemulsions loaded with both, nisin and carvacrol, having Sauter mean diameter （d3,2） of around 120 nm, were considerably stable under the conditions investigated （up to 1 week storage at room temperature）. The interfacial tension between nisin aqueous solutions and soybean oil could be reduced up to 12 mN/m, as compared to that of pure water and soybean oil （21 mN/m）, so that demonstrating the potential of nisin to be used as emulsifier. The results obtained for O/W nanoemulsions loaded with both antimicrobial compounds indicated a reduction of nearly two log cycles （around 100 times） on B. subtilis population upon incubation for 24 h, compared to the blank （10 mM phosphate buffer）.

Size Reduction of Hydrophobically Modified Starch on Beads Mill： Effect of Dispersing Medium and Milling Time

The primary interest to this study was to investigate the effect of milling parameters on the size of hydrophobically modified starch particles, aiming to produce small, uniformly sized modified starch microspheres. Octie, a commercial product originated from cornstarch modified using Octenyl Succinate Anhydride （OSA）, was dispersed （3 wt%） using different media （water or ethanol） and subsequently wet-milled using a beads mill with zirconium beads at a rotation of 6,000 rpm up to 30 min. It was found that milling Octie in water dispersion for 3 min resulted in the smallest mean particle size （2.04 i 0.91 ktm）, compared to unmilled modified starch granules （15.2 ~ 6.0 lam）. Granular size and morphology changed considerably with further milling. For instance, very dense clusters with variable particle sizes （20.6 ~ 10.0 lam） were obtained after 30 min milling. As depicted by Scanning Electronic Microscopy, a large number of particles were apparently flattened during the milling process rather than broken, forming aggregates. Ultimately, within the range of experimental conditions tested, production of sub-micron modified starch particles was not possible.

Rescue of white egg 1 mutant by introduction of the wildtype Bombyx kynurenine 3-monooxygenase gene

In silkworms, the white egg 1 （w-1） mutant, which is characterized by white eyes and white eggs, is deficient in Bombyx kynurenine 3-monooxygenase （KMO） activity. To investigate whether the w-1 mutant phenotype is rescued by introducing the wild-type KMO gene, we constructed transgenic silkworms with the wild-type Bombyx KMO gene under the control of either the cytoplasmic actin gene promoter （A3KMO） or the native KMO gene promoter （KKMO）. We created two transgenic lines with A3KMO and one line with KKMO constructs. The eyes of adults in these lines were brown, and the eggs laid by the transgenic females were also brown. Reverse transcription-polymerase chain reaction（RT-PCR） analysis showed that the A3KMO silkworm lines expressed the transcript in the mid-gut, fat bodies, and Malpighian tubules. The KKMO line expressed the transcript only in the fat bodies and Malpighian tubules. The intensity of eye and egg color in the transgenic lines was proportional to the KMO expression level. Interestingly, transgenic larvae with the A3KMO construct had a light brown larval cuticle, but the KKMO line did not. These results indicate that the wild-type KMO gene can be used as a marker gene for visually screening transgenic silkworms.

Expression of the Japanese oak silkworm Antheraea yamamai fibroin gene in the domesticated silkworm Bombyx mori

To understand the evolutionary conservation ofthe gene expression mechanism and secretion machinery between Antheraea and Bombyx fibroins, we introduced the genomic A. yamamai fibroin gene into the domesticated silkworm, B. mori. The spliced A. yamamai fibroin mRNA appeared only in the posterior region of the silk gland of the transgenic silkworm, suggesting that the functions of the fibroin promoter region and the splicing machinery are conserved between these two species. The A. yamarnai fibroin protein was detected in the lumen of the silk gland of the transgenic silkworm, albeit at lower levels compared with the B. mori-type fibroin. We found a strong degeneration of the posterior region of the silk gland of the transgenic silkworm. As a result, the cocoon shell weight was much lower in the transgenic silkworm than in the non-transgenic line. These results indicate that the promoter function and splicing machinery are well conserved between A. yamamai and B. mori but that the secretion mechanism of fibroin is diversified between the two.