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Biomedical subjects

A Fujishima

Publications and source records attributed to A Fujishima.

At least 55 records · Page 3Linked to original sources

Photokilling of T-24 human bladder cancer cells with titanium dioxide.

A photoexcited titanium dioxide surface has a strong ability to decompose water into hydrogen and oxygen. We have studied this effect in order to use it to kill cancer cells in vitro and in vivo. A distinct cell killing effect was observed on cultured T-24 human bladder cancer cells treated with titanium dioxide particles and 300-400 nm UV light irradiation. Titanium dioxide plus UV light also dramatically suppressed the tumour growth of T-24 cells that were implanted in nude mice. Cells cultured on the titanium dioxide electrode were also killed under UV irradiation when the electrode was anodically polarised, suggesting that photogenerated holes are involved in the cell killing. The cell killing effect caused by titanium dioxide particles plus UV light irradiation was significantly hampered in the presence of L-cysteine and catalase, scavengers of hydroxyl radicals and hydrogen peroxide respectively. Transmission electron microscopic observations showed the titanium dioxide particles to be distributed on the cell surface and inside the cells. These results suggest that titanium dioxide particles under UV light irradiation produced photogenerated holes on the surface yielding hydroxyl radicals and hydrogen peroxide inside or outside the cells and the cells were then killed by the action of these highly oxidising molecules. The possible application of photoexcited titanium dioxide particles to cancer treatment as a new anti-cancer modality is discussed.

Catalase↗

Human parathyroid hormone: efficient synthesis in Escherichia coli using a synthetic gene, purification and characterization.

Human parathyroid hormone is a peptide hormone consisting of 84 amino acid residues. Production of small proteins by direct expression in Escherichia coli is often unsuccessful owing to susceptibility of the mRNA and/or the product to endogenous enzymes. In this study, direct expression of the hormone at an excellent level (over 100 mg/L) has been achieved by using a suitably designed synthetic gene under the control of the T7 promoter. The protein produced in bacteria was extracted and easily purified in a good yield of 27 mg/L. The purified product was physico-chemically identified as intact human parathyroid hormone from the results of amino acid analysis, N-terminal sequencing, and peptide mapping using fast atom bombardment mass spectrometry. In biological assays the purified product stimulated adenylate cyclase in vitro, promoted bone growth and increased the serum osteocalcin in rats to the same extent as the authentic hormone.

Amino Acid Sequence↗

Induction of cytotoxicity by photoexcited TiO2 particles.

Photoexcited TiO2 particles can drive various chemical reactions due to their strong oxidizing and reducing ability. To investigate the possible use of this effect for cancer treatment, the antitumor activity of photoexcited TiO2 particles was studied in vitro and in vivo. HeLa cells cultured in vitro were completely killed in the presence of TiO2 (50 micrograms/ml) with 10-min UV irradiation by a 500-W-Hg lamp. In contrast, very little cell death was observed from TiO2 treatment without UV irradiation. Photoexcited TiO2 particles also significantly suppressed the growth of HeLa cells implanted in nude mice, compared with those receiving TiO2 alone or UV irradiation alone. The cell death caused by photoexcited TiO2 particles was significantly protected in the presence of L-tryptophan and catalase. These molecules are quenchers of hydroxyl radicals and scavengers of hydrogen peroxide, respectively, suggesting that the cells were killed by the OH. and H2O2 produced from photoexcited TiO2 particles.

Animals↗

Crystal structure of basic fibroblast growth factor at 1.6 A resolution.

We have determined the crystal structures of two types of human basic fibroblast growth factor, the serine analogue and the wild-type, at 1.6 and 2.5 A resolution, respectively. Two good heavy atom derivatives were found and used for multiple isomorphous replacement phasing. The atomic coordinates were refined using the Hendrickson & Konnert program for stereochemically restrained refinement against structure factors. The crystallographic R factors were reduced to 15.3% for the serine analogue structure and 16.0% for the wild-type structure. The serine analogue and wild-type structures have been found to be almost identical, the root-mean-square deviation between the corresponding C alpha atoms being 0.11 A. Their structures are composed of twelve beta-strands forming a barrel and three loops. Their molecules have an approximate threefold internal symmetry and are similar in architecture to that of interleukin-1 beta. A possible heparin-binding site, which comprises five basic residues, Lys119, Arg120, Lys125, Lys129, and Lys135, has been revealed by calculating the electrostatic potential energy.

Amino Acid Sequence↗

[The direct tensile test of composite resins using the small specimen--effect of the preparation of specimen, the size of specimen and the testing condition on the tensile properties].

The direct tensile test of composite resins using the specimen with the gauge length 10 mm has been developed by authors. In this study smaller specimens with the gauge length 5 mm and 2 mm were also investigated. As the gauge length became smaller, tensile properties such as the proportional limit, the proof stress, and the tensile strength showed the tendency to become higher. The effect of strain rate on the tensile properties appeared clearly when using the specimen with the gauge length 2 mm. The small specimen was found to have many advantages for the preparation, the cost of material and the handling during the tensile test.

Composite Resins↗

[Chemical durability of composite resins].

To evaluate their chemical durability, samples of posterior composite resins were stored in NaOH (0.1 mol, 1 mol), HCl (0.1 mol, 1 mol), acetone (99%), and distilled water at 37 degrees C for a week and subjected to the direct tensile test, measurement of surface roughness and SEM observation. NaOH caused the degradation of composite resins, decrease of tensile strength and increase of surface roughness. The SEM photograph of fractured surface of tensile specimens revealed the degradation zone under the surface. It was considered that NaOH caused the hydrolysis of silan coupling of composite resins. HCl also caused the degradation of composite resins, but some resins showed the durability in this condition. More examination is needed to clarify the mechanism of degradation. Acetone caused the degradation of matrix resin and some composite resins. It was considered to dissolve the low degree of polymerized matrix resin of composite resins. Immersion in chemicals of composite resins was thought to be useful to evaluate the chemical durability and to accelerate the degradation in water.

Composite Resins↗

Crystallographic characterization of recombinant human interleukin 2.

Recombinant human interleukin 2 produced by Escherichia coli was purified to homogeneity and crystallized after being separated from methionyl interleukin 2. Crystals suitable for structural studies have been obtained by the seed enlargement technique, using the method of vapor diffusion with ammonium sulfate as the precipitant at pH 4.6. The space group is P2(1)2(1)2 with cell dimensions a = 49.2 A, b = 87.6 A and c = 32.4 A. The asymmetric unit contains one molecule of the protein. From preliminary results, the crystals are moderately stable to X-rays and produce measurable reflections to a resolution of about 2.2 A. The diffraction data for the native crystals have been collected on a diffractometer at 2.4 A resolution.

Crystallography↗

Importance of disulfide linkage for constructing the biologically active human interleukin-2.

Recombinant human interleukin-2 (rIL-2) produced in Escherichia coli possesses a free thiol group at Cys-125 and a disulfide linkage between Cys-58 and Cys-105, as in the case for natural human interleukin-2. Treatment of rIL-2 with 200 mM dithiothreitol resulted in the cleavage of the Cys-58-Cys-105 disulfide bond. The reduced form of rIL-2 thus obtained retained only 10% of the in vitro biological activity of the native form, as measured by the ability to stimulate the growth of an IL-2-dependent mouse natural killer cell line, NKC3. Far-uv circular dichroism studies indicated that the cleavage of the disulfide bond results in a decrease of alpha-helix content. Near-uv circular dichroism studies suggested that the native molecule is folded into a rigid tertiary structure, while the reduced form showed a spectrum similar to that of rIL-2 denatured in the presence of 6 M guanidine.HCl. The once-reduced molecule was readily reoxidized in the presence of 10 microM Cu2+ to form the native molecule with full biological activity. These results strongly demonstrate that the Cys-58-Cys-105 disulfide linkage in the IL-2 molecule is essential for constructing a rigid and biologically active form of IL-2.

Amino Acids↗

Time-resolved x-ray diffraction studies on the intensity changes of the 5.9 and 5.1 nm actin layer lines from frog skeletal muscle during an isometric tetanus using synchrotron radiation.

Time-resolved x-ray diffraction studies have been made on the 5.9- and 5.1-nm actin layer lines from frog skeletal muscles during an isometric tetanus at 6 degrees C, using synchrotron radiation. The integrated intensities of these actin layer lines were found to increase during a tetanus by 30-50% for the 5.9-nm reflection and approximately 70% for the 5.1-nm reflection of the resting values. The intensity increase of both reflections was greater than that taking place in the transition from rest to rigor state. The intensity change of the 5.9-nm reflection preceded those of the myosin 42.9-nm off-meridional reflection and of the equatorial reflections, as well as the isometric tension development. The intensity profile of the 5.9-nm layer line during contraction was found to be different from that observed in the rigor state.

Actins↗

[NK activities in cancer patients--their modification by an immunopotentiator].

NK activity of patients with cancer against gastric carcinoma cell line KATO-III, lung carcinoma cell line A-549 as well as K-562 was significantly lower than that in the control group. We investigated in vitro effects of immunopotentiator, OK-432, on the effector cells of both healthy donors and cancer patients. NK activity of healthy donors was clearly augmented after treatment of effector cells with OK-432. NK activity of some patients with cancer, however, showed no augmentation after OK-432 treatment. These results might be helpful for monitoring cancer patients from immunological view point.

Adjuvants, Immunologic↗