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Biomedical subjects

H Hashiba

Publications and source records attributed to H Hashiba.

33 records · Page 2Linked to original sources
PubMed1986

Characterization of the genes coding for two major cell wall proteins from protein-producing Bacillus brevis 47: complete nucleotide sequence of the outer wall protein gene.

Bacillus brevis 47 contains two cell wall proteins termed the outer wall protein (OWP) and middle wall protein (MWP), each of which forms hexagonal arrays in the cell wall. A 6-kilobase BglII-BclI fragment of B. brevis 47 DNA cloned into Bacillus subtilis with a derivative of pHW1 as a vector directed the synthesis of a polypeptide, with almost the same molecular weight as the authentic OWP, as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, which was specifically recognized by the anti-OWP antibody. Nucleotide sequence analysis of the subfragment revealed that it contains two open reading frames in tandem. The upstream truncated open reading frame corresponds to the carboxy-terminal portion of the MWP, and the downstream open reading frame corresponds to the entire translational portion of the OWP. The latter encodes a secretory precursor of the OWP, consisting of a total of 1,004 amino acid residues with a signal peptide of 24 amino acid residues at its amino-terminal end. Futhermore, analysis of transcripts in B. brevis 47 suggests that the MWP and OWP genes, in that order, constitute a cotranscriptional unit and that the major promoter shared by the two genes is located upstream of the MWP gene.

Amino Acid Sequence↗
PubMed1986

Enhancement by 1 alpha,25-dihydroxyvitamin D3 of chemically induced transformation of BALB 3T3 cells without induction of ornithine decarboxylase or activation of protein kinase C1.

We reported previously that 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25(OH)2D3], a hormonally active form of vitamin D3, markedly enhanced methylcholanthrene-induced transformation of BALB 3T3 A31-1-1 cells. When the cells were treated with methylcholanthrene (1 microgram/ml) for 72 h and then with 1 alpha,25(OH)2D3 (5 ng/ml) for 2 wk, the transformation frequency was 1.95 +/- 0.73 (SD) foci/dish in 8 independent experiments, which was about 20 times that in cultures treated with methylcholanthrene only. Even at a physiological concentration in plasma, i.e., 0.05 ng/ml, 1 alpha,25(OH)2D3 enhanced the transformation frequency significantly (P less than 0.001). 1 alpha,25(OH)2D3 was not cytotoxic but slightly inhibited growth of the cells. Cells treated with 1 alpha,25(OH)2D3 were thin and became arranged in a meshwork with wide intercellular spaces. These morphological changes were reversible. 1 alpha,25(OH)2D3 induced DNA synthesis in quiescent BALB 3T3 cells dose and time dependently, but this effect was less than that of 12-O-tetradecanoylphorbol-13-acetate. Unlike 12-O-tetradecanoylphorbol-13-acetate, 1 alpha,25(OH)2D3 did not interfere with the binding of epidermal growth factor or phorbol dibutyrate. 1 alpha,25(OH)2D3 did not induce ornithine decarboxylase. Moreover, it did not activate protein kinase C in quiescent BALB 3T3 cells or this enzyme isolated from mouse brain. BALB 3T3 cells and their transformants contain a specific cytosol receptor for 1 alpha,25(OH)2D3, but the binding sites of the transformants were fewer and had lower affinity than those of untransformed BALB 3T3 cells. These effects of 1 alpha,25(OH)2D3 were specific, because other derivatives of vitamin D3 induced the same effects only at 200 times or more higher concentrations.

Animals↗
PubMed1986

Activation of protein kinase C and specific phosphorylation of a Mr 90,000 membrane protein of promotable BALB/3T3 and C3H/10T1/2 cells by tumor promoters.

The activation of protein kinase C and protein phosphorylation by tumor promoters were examined using quiescent cultures of BALB/3T3 and C3H/10T1/2 cells, because in these cells tumor promoters enhance chemically induced transformation and also induce DNA synthesis and ornithine decarboxylase. The cytosol and membrane fractions were partially purified, and the activity of protein kinase C was assayed. In quiescent cells, protein kinase C activity was found only in the cytosol fraction. Treatment with 100 ng of 12-O-tetradecanoylphorbol-13-acetate or teleocidin B per ml caused rapid translocation of protein kinase C from the cytosol to the membrane fraction. The activity in the cytosol disappeared almost completely after 15 min when the activity in the membrane reached a peak. The membrane activity gradually decreased to the control level after 6 h, while no activity reappeared in the cytosol within 6 h. Under these circumstances, a membrane protein with a molecular weight of 90,000 and pl of 4.0-4.4 (termed p90) was specifically phosphorylated, possibly by the activated protein kinase C, in both cell-free and intact-cell systems. On treatment of quiescent BALB/3T3 cells with 100 ng of 12-O-tetradecanoylphorbol-13-acetate, p90 phosphorylation increased 2-fold in 1 min, reaching a peak in 15 min of 3.4-fold the initial value. The phosphorylation of p90 increased with increase in the concentrations of 12-O-tetradecanoylphorbol-13-acetate between 0.1 and 10 ng/ml and reached a plateau at 10 ng/ml. p90 phosphorylation also occurred on exposure of the cells to non-phorbol ester tumor promoters (mezerein and teleocidin B) and growth factors, such as platelet-derived growth factor and fibroblast growth factor. p90 was not immunoprecipitated by antibody against the insulin receptor. Phosphorylation of p90 occurred at a serine residue. The present study suggests that activation of protein kinase C and phosphorylation of p90 by it are early events leading to tumor promotion.

Animals↗
PubMed1985

Inhibition of tumor promotion in mouse skin by 1 alpha,25-dihydroxyvitamin D3.

1 alpha,25-Dihydroxyvitamin D3 [1 alpha,25(OH)2D3], a hormonally active form of vitamin D3, was found to inhibit the promotional phase of 7,12-dimethylbenz[a]anthracene-induced skin carcinogenesis in female Sencar mice. Topical application of 1 alpha,25-(OH)2D3 once a week at a dose of 1 micrograms or less, a tolerable dose from hypercalcemia, dose dependently inhibited tumor promotion by 12-O-tetradecanoylphorbol-13-acetate (TPA). When 1 micrograms of 1 alpha,25(OH)2D3 was applied 30 min before 5 micrograms of TPA, the times required for 50 and 100% tumor incidence were delayed about 2.5 and 7 weeks, respectively, and the number of tumors per mouse was decreased by 25-30%. This inhibitory effect was more pronounced when examined by a two-stage promotion protocol, in which a single application of 5 micrograms of TPA (Stage I) was followed by repeated applications of 5 micrograms of mezerein once a week for 19 weeks (Stage II). When 1 alpha,25(OH)2D3 at 1 micrograms was applied at Stage I + II or Stage II, tumor formation was markedly suppressed, resulting in decrease of about 70-80% in the incidence and 87-90% in the number of tumors per mouse. Application of 1 alpha,25(OH)2D3 at Stage I only did not inhibit tumor formation, indicating that 1 alpha,25(OH)2D3 specifically inhibited Stage II promotion. These results are in good agreement with the previous and present findings that 1 alpha,25(OH)2D3 inhibited induction of epidermal ornithine decarboxylase by TPA and mezerein. The possibility that 1 alpha,25(OH)2D3 suppressed tumor promotion by killing initiated cells rather than inhibiting promotion was ruled out by an experiment in which TPA was applied to the 1 alpha,25(OH)2D3 alone-treated animals.

Animals↗
PubMed1984

Inhibition by 1 alpha, 25-dihydroxyvitamin D3 of induction of epidermal ornithine decarboxylase caused by 12-O-tetradecanoylphorbol-13-acetate and teleocidin B.

Topical application of 1 alpha, 25-dihydroxyvitamin D3 [1 alpha, 25-(OH)2D3], an active form of vitamin D3, markedly inhibited induction of ornithine decarboxylase caused by 12-O-tetradecanoylphorbol-13-acetate (TPA) and teleocidin B in mouse skin. The degree of inhibition was dependent on the dose and time of application of 1 alpha, 25(OH)2D3. Application of 1 micrograms 1 alpha, 25(OH)2D3 within 30 min before or after treatment with 10 micrograms TPA caused about 72% inhibition of ODC induction at 4 hr. Similar degrees of inhibition were obtained with dose ratios of 1 alpha, 25(OH)2D3 to TPA of 1:3, 1:10, and 1:30. The dose required for 50% inhibition was 0.063 micrograms, or 0.15 nmol, which is about one-half that of retinoic acid, a known inhibitor of induction of ODC activity by TPA. 1 alpha, 25(OH)2D3 had a specific inhibitory effect, in which 100 times higher doses or more of other derivatives of vitamin D3, such as 1 alpha-hydroxyvitamin D3, 25-hydroxyvitamin D3, 24R,25-dihydroxyvitamin D3, and vitamin D3, were required to inhibit ODC induction by TPA. 1 alpha, 25(OH)2D3 did not inhibit epidermal hyperplasia induced by TPA.

Alkaloids↗
PubMed1979

A study of the multiple pinhole coded aperture and the application of the minicomputer in image decoding.

Research has been done on optically reconstructed imaging employing the Multiple Pinhole Coded Aperture (hereafter abreviated as MPCA) in radioisotope tomographic imaging. However, problems remain in the optically reconstructed image method. Therefore, we employed a minicomputer (hereafter abbreviated as CPU) and developed the software for decoding and managing the radioisotope tomographic image. Combining the MPCA and the CPU system, we were able to decode and manage the radioisotope tomographic image. 1) In comparison to the optically decoded MPCA image, various input commands are possibly in the CPU method according to the dialogue between the CPU and the on line typewriter. In addition to this, decoded tomographic images of unrestricted depth are readily attainable. 2) In the CPU method noise elimination and other aspects of image management can be easily performed.

Computers↗
PubMed1979

[Clinical studies on PC-904 in biliary tract diseases (author's transl)].

The clinical effects of PC-904, a new semisynthetic penicillin, were studied in patients with biliary tract diseases, and the results were as follows: 1) PC-904 showed an average peak serum level of 40.7 +/- 11.6 microgram/ml 2 hours after an intravenous drip infusion of 1 g of the agent. The biliary level showed a peak value of 126.5 +/- 85.4 microgram/ml 2 hours to 3 hours after the infusion. 2) Isolated organisms from bile before the treatment were E. coli, Klebsiella, Proteus, Pseudomonas, Enterococcus and Bacteroides. MIC of PC-904 on 18 strains of isolated organisms was almost 6.25 microgram/ml or less. All isolated organisms except one strain of Klebsiella oxytoca disappeared after the treatment. 3) Six patients with cholelithiasis were medicated with PC-904 to prevent post-operative infections. The clinical effects were good in 4, poor in 1 and unknown in 1 case. 4) As to side effects no adverse reactions and allergic reactions were noted. Also no significant abnormalities of laboratory findings were observed.

Ampicillin↗
PubMed1978

A method of tomographic imaging using a multiple pinhole-coded aperture.

High energy collimators in use today are limited with respect to resolution and efficiency. The multiple pinhole coded aperture (hereafter abbreviated as MPCA) is a method being considered to improve the above. In our study we conducted basic experiments with an MPCA using an Anger-type gamma-camera. Using this method, we were able to perform radioisotope imaging and radioisotope tomographic effect. We have studied the resolution, detection efficiency, and tomographic effects of the MPCA. We found that, in the MPCA, detection efficiency is proportional to the number of pinholes and resolution is dependent upon the pinhole diameter. This tomographic effect is of the same principle as the stretching-type tomography, images on both side of the desired image appearing as noise images, affecting the depth resolution. Using an MPCA with a pinhole diameter of 3 mm and 3 pinholes positioned in an equilateral triangular pattern measuring 7.8 cm on each side, we attained a depth resolution of 3 cm. Because the MPCA method involves complicated image processing, the resulting image contains a factor of 2n-2 (where n is the number of pinhole) of noise, affecting the depth resolution.

Radionuclide Imaging↗