Isamu Maeda

Novel transdermal drug delivery system with polyhydroxyalkanoate and starburst polyamidoamine dendrimer.

In search of an efficient transdermal drug delivery system (TDDS), a polyhydroxyalkanoate (PHA)-based system with a polyamidoamine dendrimer was examined. Tamsulosin was used as the model drug. The dendrimer was found to act as the weak enhancer. By adding the dendrimer, the dendrimer-containing PHA matrix achieved the clinically required amount of tamsulosin permeating through the skin model. This is also the first report of the application of PHA and dendrimer to the TDDS.

Mechanism of Enhancement Effect of Dendrimer on Transdermal Drug Permeation through Polyhydroxyalkanoate Matrix

The possible application of polyhydroxyalkanoate (PHA) in transdermal drug delivery systems (TDDSs) for tamsulosin was previously reported. PHAs containing the drugs, ketoprofen, clonidine and tamsulosin showed good adhesiveness to the skin model used, that is, shed snake skin, and dispersed well all model drugs tested. The model drugs hardly permeated through snake skin in solution form. However, these drugs permeated well through snake skin from the PHA matrix. It was previously reported that the addition of a dendrimer, a polymeric permeation enhancer, is effective for the TDDS for tamsulosin to establish an effective clinical TDDS. The effect of dendrimer addition was examined in TDDSs for ketoprofen and clonidine. The dendrimer added did not show an enhancement effect on the TDDSs for the two drugs. To investigate the mechanism of the enhancement effect of a dendrimer on the tamsulosin TDDS, X-ray analyses were performed. With dendrimer addition, drug crystallization in PHA was promoted. The crystal in PHA had highly ordered and changed its space group. These findings are very important for exploiting high-performance PHA-based TDDSs.

Stably sustained hydrogen production by biophotolysis in natural day/night cycle

Our photobiological hydrogen production system was successfully scaled up to a pilot plant scale in the photosynthetic starch accumulation and the fermentative production of organic compounds from starch by Chlamydomonas MGA 161 and the hydrogen photoproduction by Rhodovulum sulfidophilum W-1S from organic compounds produced by Chlamydomonas MGA 161 in the natural day/night cycle. The conversion yield of organic compounds from starch of Chlamydomonas MGA 161 was 80 to 100 % of the theoretical yield, but the molar yield of hydrogen photoproduction of Rv. sulfidophilum W-1S was approximately 40 %. One main cause for the low hydrogen yield of Rv. sulfidophilum W-1S was a competition between the accumulation of intracellular Poly (3-hydroxybutyric acid), PHB, and the hydrogen production under light anaerobic conditions. The PHB accumulation was repressed and the hydrogen photoproduction was promoted by enhancing the nitrogenase activity of Rv. sulfidophilum W-1S.

Excretion of glycerol by the marine Chlamydomonas sp. strain W-80 in high CO2 cultures☆

Abstract A Chlamydomonas sp. strain, which excretes glycerol as the major carbon compound, was isolated and designated as strain W-80. The glycerol production was much enhanced by a high concentration of CO2, and the final concentration of the product was measured at levels up to 24.9 mM (2.29 g/l).

Antibiotic production by the immobilized cyanobacterium, Scytonema sp. TISTR 8208, in a seaweed-type photobioreactor

A photobioreactor was constructed using either anchored polyurethane foam strips (1 × 1 × 40 cm, PU-strips) fixed on a stainless-steel ring to prevent flotation, or free-floating polyurethane foam blocks (1 × 1 × 1 cm, PU-blocks) as biomass supporting materials (BSM). The cyanobacterium,Scytonema sp. TISTR 8208, which produces an antibiotic, was immobilized onto PU-strips or -blocks. The free-floating PU-blocks could immobilize only about 70% of the total cells, while the anchored PU-strips could immobilize as much as 97%. PU-strips were chosen as the BSM and we named this type of reactor, seaweed-type bioreactor (STB). Optimal physical conditions for antibiotic production were determined in the STB. Inoculum density was 0.4 g l−1 and cells were sparged with air containing 5% CO2 circulated at the gas flow rate of 250 ml min−1 and illuminated at a light intensity of 200 μmol photon m−2 s−1. The production of antibiotic could be increased 3-fold.

Adenoviral transfection of hepatocytes with the thioredoxin gene confers protection against apoptosis and necrosis.

A recombinant adenovirus vector containing the human thioredoxin (TRX) gene was constructed using the Cre-loxP recombination system and used to transfect rat hepatocytes with very high efficiency. The TRX gene was expressed in a dose-dependent manner and significantly modulated rat cellular functions. The TRX gene conferred resistance to oxidative stress, such as hydrogen peroxide treatment, on the host hepatocytes. FACS analysis of DNA fragmentation showed that the TRX gene suppressed hepatocyte apoptosis. It also significantly extended the life span of hepatocytes cultured conventionally on polystyrene plates. Liver-specific functions were maintained in the viability-modulated hepatocytes. Moreover, TRX expression did not affect hepatocyte spheroid formation and it extensively suppressed necrosis in the internal cells. Thus, the transfection of hepatocytes with the TRX gene successfully confers global maintenance of liver functions. These findings provide important information for the development of bioartificial liver support systems and gene therapy for liver diseases.

Influence of Sulfate-Reducing Bacteria on Outdoor Hydrogen Production by Photosynthetic Bacterium with Seawater

Abstract. The application of seawater for bacterial fermentative production is a cost-effective technology. Hydrogen production by marine photosynthetic bacterium with seawater failed to continue after more than 10 days, and was accompanied by the formation of hydrogen sulfide and a change in culture color from red to black. However, substrate consumption in the blackish culture was comparable to that in a hydrogen-producing culture. A decrease in hydrogen production occurred upon the addition of sodium sulfide at concentrations of 1.5 mM or higher. PCR analysis targeted at the 16S rDNA sequence selective for sulfate-reducing bacteria revealed the existence of sulfate-reducing bacteria in inoculation cultures of the phototrophic bacterium and medium for hydrogen production. Hence, the high sulfate concentration of seawater, the low oxidation-reduction potential under hydrogen-producing conditions, and the presence of electron donors such as acetate might promote the metabolic activities of sulfate-reducing bacteria, resulting in the deterioration of hydrogen production with seawater.

Broad Spectrum and Mode of Action of an Antibiotic Produced by Scytonema sp. TISTR 8208 in a Seaweed-Type Bioreactor

A photobioreactor was constructed using anchored polyurethane foam strips (1 x 1 x 40 cm) fixed onto a stainless-steel ring to prevent flotation, as a biomass support material (BSM). This type of reactor was named a seaweed-type bioreactor. A filamentous cyanobacterium, Scytonema sp. TISTR 8208, which produces a novel cyclic dodecapeptide antibiotic, was immobilized in seaweed-type photobioreactor and cultivated with air containing 5% CO2 sparged at a gas flow rate of 250 mL/min under illumination at a light intensity of 200 mmol photon m-2 s-1. The antibiotic produced in the seaweed-type photobioreactor was purified by HPLC and examined regarding its spectrum and mode of action. The antibiotic effectively inhibited the growth of Gram-positive bacteria, pathogenic yeasts, and filamentous fungi, but it had only a weak effect on Gram-negative bacteria. Scanning electron micrograph analysis showed that the most characteristic change was swelling of the cells after exposure to the antibiotic. The antibiotic seems to alter the conformation of the microbial cell membrane, thereby changing its permeability, leading to osmotic shock.

Unusual Accumulation of Demethylspheroidene in Anaerobic-Phototrophic Growth of crtA-Deleted Mutants of Rhodovulum sulfidophilum

Rhodovulum sulfidophilum produces carotenoids in the spheroidene pathway. Spheroidene monooxygenase, CrtA, catalyzes the conversion of spheroidene to spheroidenone. crtA-deleted mutants of R. sulfidophilum did not produce spheroidenone and demethylspheroidenone. In these mutants, the ratio of demethylspheroidene to spheroidene increased with exposure to light. One mutant exhibiting a spheroidene-predominant phenotype did not grow under anaerobic-light conditions and was devoid of bacteriochlorophyll a, even under semiaerobic-light conditions There was no difference in the growth of the mutants under aerobic-dark conditions. These data suggest that demethylspheroidene is important for photosynthesis in R. sulfidophilum.

Polyethyleneimine/chitosan hexamer-mediated gene transfection into intestinal epithelial cell cultured in serum-containing medium

In search of an efficient nonviral vector, polyethyleneimine (PEI)-based vectors were examined. In general, the transfection efficiency of nonviral vectors is suppressed by serum. Here we show that PEI based vectors, particularly, the chitosan hexamer-PEI vector, could perform efficient gene transfection into intestinal epithelial cells (IEC-6) in the presence of serum. The conjugation order of the two polymers with a plasmid (first, chitosan hexamer; second, PEI) was found to be an important factor in enhancing transfection efficiency.