Biomedical subjects

H Hashiba

Publications and source records attributed to H Hashiba.

At least 19 recordsLinked to original sources

PubMed1994

Effect of Lactobacillus acidophilus on iron bioavailability in rats.

The effect of Lactobacillus acidophilus on iron bioavailability in rats was examined by the hemoglobin regeneration method. For hemoglobin depletion, female Wistar rats were fed an iron-deficient diet at 3 weeks of age for 13 days. Rats were then assigned to one of four groups, such that average blood hemoglobin value and average body weight were similar among the groups. For hemoglobin regeneration, they were fed one of two ferrous sulfate-supplemented diets that contained the following iron levels (mg/kg): 13.7 for two groups; 21.7 for the other two groups. In the two groups fed the same diet, one group additionally received oral administration of 2 ml of skim milk fortified with 0.3% yeast extract once or twice a day for 7 days, and other rats were administered with skim milk fermented by L. acidophilus SBT 2062 in the same manner. Hemoglobin regeneration efficiency (HRE) was significantly higher in the fermented product-given rats than in the skim milk-supplied rats. There was no significant interaction in HRE between the dietary iron group and the oral administration group. These results indicate that L. acidophilus SBT 2062 is effective for increasing of iron bioavailability in rats.

Animals↗

PubMed1994

Effects of skim milk and its fermented product by Lactobacillus acidophilus on plasma and liver lipid levels in diet-induced hypertriglyceridemic rats.

Effects of skim milk and its fermented product by Lactobacillus acidophilus on plasma and liver triglyceride and cholesterol levels were examined in diet-induced hypertriglyceridemic rats. Male Sprague-Dawley rats at 4 weeks of age were fed a hypertriglyceridemic diet that contained 20% coconut oil, 17.5% fructose, and 17.5% sucrose for 14 days. The test diet was supplemented with either 20% skim milk powder or 20% powder of skim milk fermented by L. acidophilus SBT 2062. Hypertriglyceridemia was observed in the control group, but plasma cholesterol levels were not increased. Skim milk suppressed the elevation of plasma triglyceride levels, while its fermented product had no significant effect. Both dairy products prevented the elevation of liver triglyceride and cholesterol levels, but had no effect on plasma cholesterol levels.

Animals↗

PubMed1993

Identification of the replication region of Streptococcus thermophilus No. 29 plasmid pST1.

The replication region of the 2.77-kilobases (kb) plasmid pST1 from Streptococcus thermophilus No. 29 was identified. Deletion derivatives of pST1 were introduced into plasmid-free S. thermophilus No. 29 and examined for their ability to replicate autonomously. The nucleotide sequence had one open reading frame encoded for a 315 amino acid protein (Rep). Comparisons with proteins encoded by other Gram-positive bacterial plasmids strongly suggest that the deduced protein (RepS) encoded by pST1 has a replicative role. pST1 also contains a DNA sequence similar to the origin nick sequences of pLP1 or pLAB1000, which initiate plasmid replication at the plus origin. In a maxicell system, E. coli CSR603 carrying pSTUC4 produced a protein that was considered to correspond to the product of the RepS gene. These results strongly suggest that pST1 is replicated following a rolling-circle mechanism via single-stranded DNA intermediates.

Amino Acid Sequence↗

PubMed1992

Establishment of a host-vector system in Lactobacillus helveticus with beta-galactosidase activity as a selection marker.

A host-vector system was established in Lactobacillus helveticus with beta-galactosidase activity as a selection marker. Plasmid pBG10 was constructed by joining the beta-galactosidase gene from L. bulgaricus, the promoter region of the erythromycin resistance gene from pAM beta 1, the replication region of pBR329, and the replication region of the L. helveticus cryptic plasmid pLJ1. L. helveticus SBT2195 (Lac- mutant), transformed with pBG10, was selected on skim milk plates. The structural gene of alpha-amylase (1536 bp) from Bacillus licheniformis, inserted downstream of the promoter region of the erythromycin resistance gene of pBG10, was expressed in L. helveticus SBT2195. Plasmid pBG10 is a food-grade and expression vector in L. helveticus.

Base Sequence↗

PubMed1991

Localization of metallothionein in hair follicles of normal skin and the basal cell layer of hyperplastic epidermis: possible association with cell proliferation.

Metallothionein is a low-molecular-weight metal-binding protein. Although it is inducible by a variety of agents and ubiquitously present in many tissues, its physiologic functions are still not clear. The present study was undertaken to determine the possible functions of metallothionein in both the proliferation and differentiation of epidermal keratinocytes. Metallothionein was detected immunohistochemically in hair matrix cells of the bulb and cells of the outer root sheath of anagen hair follicles, but not in dermal papillae in normal skin in the back of mice. In hyperplastic epidermal tissue, induced by either a phorbol ester tumor promoter or cholera toxin, the basal cells of the interfollicular epidermis stained strongly for metallothionein. Elevated expression of mRNA of the metallothionein gene was also demonstrated when the skin was stimulated by agents that induced hyperplasia. Papillomas produced by two-stage carcinogenesis protocols also stained for metallothionein. These observations suggest that metallothionein is involved in the proliferation of epidermal keratinocytes.

Animals↗

PubMed1991

[Interleukin-2 receptor expression in pulmonary granulomatous diseases].

Interleukin-2 receptor expression (IL-2R) on monocytes and alveolar macrophages (AM) was determined in patients with sarcoidosis and pulmonary tuberculosis. In sarcoidosis and tuberculosis, IL-2R on monocytes was detectable, while it was undetectable in healthy controls. IL-2R on AM in sarcoidosis and tuberculosis was significantly increased as compared to healthy controls. IFN-gamma, which has been shown to be increased in sarcoidosis and tuberculosis as compared to healthy controls, induced IL-2R on monocytes in healthy controls, suggesting that IFN-gamma is at least in part responsible for the induction or enhancement of IL-2R on monocytes or AM in sarcoidosis and tuberculosis. Phorbol myristate acetate which is known to be protein kinase C (PKC) activator induced IL-2R on monocytes, and PKC inhibitor, H7, inhibited IFN-gamma-induced IL-2R on monocytes in healthy controls. Calcium ionophore, A23187, induced IL-2R on monocytes and calmodulin antagonist, W7, inhibited IFN-gamma-induced IL-2R on monocytes. Based on these results, it seems that not only the PKC pathway but also the calcium-calmodulin pathway is involved in IFN-gamma-induced IL-2R.

Adult↗

PubMed1991

[Serum soluble IL-2 receptor level in patients with sarcoidosis].

Serum levels of soluble IL-2 receptors (sIL-2R) by an ELISA method in 28 patients with sarcoidosis and 16 healthy controls were studied, and the source of sIL-2R was further examined. sIL-2R in serum was significantly higher in sarcoidosis than in controls. In sarcoidosis sIL-2R in serum significantly correlated with serum ACE level, and was significantly higher in stage II or III patients than in stage O patients. sIL-2R in supernatants of cultured monocytes (Mo) and alveolar macrophages (AM) was significantly higher in sarcoidosis than in controls. sIL-2R in supernatants of cultured T lymphocytes obtained from peripheral blood or BALF was barely detectable in sarcoidosis, while it was undetectable in controls. Furthermore, sIL-2R in serum was significantly correlated with sIL-2R in supernatants of cultured Mo and AM. These results indicate that sIL-2R in serum is an useful index of the disease activity of sarcoidosis, and may be mainly derived from IL-2R on Mo and AM.

Adult↗

PubMed1990

Elevated expression of secondary, but not early, responding genes to phorbol ester tumor promoters in papillomas and carcinomas of mouse skin.

A single topical treatment of mouse skin with the potent tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) results in transient inductions of a variety of genes. Based on the time courses of their inductions, these genes can be classified into two main groups: "early" response genes whose mRNA expression reaches a maximum 0.5-2 h after TPA treatment and "secondary" response genes whose mRNA expression is maximal 4 h or more after treatment. The nuclear oncogenes c-fos, c-myc, and c-jun belong to the early response group, whereas the metallothionein, osteopontin, and urokinase genes belong to the secondary response group. The steady-state expressions of these early and secondary response genes are all very low in normal skin, except that of c-jun, which is relatively high. Steady-state levels of expression and inducibility of these genes by TPA were not altered in initiated skin or in apparently normal skin during tumor promotion. We examined the expressions of these genes in papillomas and carcinomas produced by two-stage (initiator-promoter) and three-stage (initiator-promoter-initiator) protocols in mouse skin. Steady-state expression of the early responding nuclear oncogenes in papillomas and carcinomas was found to remain at the same low level as in normal skin. However, all the secondary responding genes were found to be expressed constitutively at high levels in these tumors. Elevated expressions of the genes for transforming growth factor alpha and beta were also observed in papillomas and to varying extents in carcinomas. These observations suggest that the regulatory machinery for transcription by the protein kinase C-mediated pathway through nuclear oncogenes is altered during the processes of tumor promotion and progression. The genes whose expression is elevated may be associated directly or indirectly with tumor promotion and progression.

9,10-Dimethyl-1,2-benzanthracene↗

PubMed1990

Normal human chromosome 1 carries suppressor activity for various phenotypes of a Kirsten murine sarcoma virus-transformed NIH/3T3 cell line.

In order to identify chromosomes that carry putative tumor-suppressor genes for the various phenotypes of Kirsten sarcoma virus-transformed NIH/3T3 (DT) cells, we performed microcell-mediated chromosome transfer into DT cells. We first isolated mouse A9 clones, containing a single human chromosome 1, 11 or 12 tagged with pSV2-neo plasmid DNA. Then, chromosome 1, 11 or 12 was transferred from the A9 clones into DT cells by microcell fusion. The growth rate, colony-forming ability in soft agar and tumorigenicity of the DT cells were controlled by chromosome 1, but not by chromosome 11 or 12, indicating that normal human chromosome 1 carries a putative tumor-suppressor gene(s) that affects various transformed phenotypes of DT cells.

Animals↗

PubMed1990

Transformation of Lactobacillus helveticus subsp. jugurti with plasmid pLHR by electroporation.

Lactobacillus helveticus subsp. jugurti was transformed with plasmid, pLHR (8.5 kilobases), by electroporation. The plasmid, pLHR, consists of a cryptic plasmid, pLJ1, from L. helveticus subsp. jugurti, the Escherichia coli vector pBR329, and the erythromycin resistance gene of pAM beta 1 from Enterococcus faecalis. Maximum transformation efficiency of 1.3 x 10(4) transformants per microgram of DNA was obtained by exposure to a pulse with an exponential decay waveform at 4 kV/cm with 25 microF capacitance. The presence of glycine in the growth medium was essential for transformation. Plasmid DNA isolated from transformants had not undergone detectable rearrangements or deletions. In addition, it was found that L. helveticus subsp. jugurti has a restriction and modification system.

Culture Media↗

PubMed1990

Suggestive evidence for functionally distinct, tumor-suppressor genes on chromosomes 1 and 11 for a human fibrosarcoma cell line, HT1080.

One approach for identifying chromosomes which carry putative tumor-suppressor genes is the introduction of specific chromosomes into the tumor cells of interest. We examined the ability of human chromosomes derived from normal fibroblasts to suppress or modulate tumorigenicity in nude mice and the in vitro properties of HT1080, a human fibrosarcoma cell line. We first isolated mouse A9 cells containing a single human chromosome (1, 2, 7, 11, or 12) integrated with pSV2neo plasmid DNA. Following fusion of microcells from these A9 cells with the HT1080 cells, clones that were resistant to G418 were isolated and karyotypically analysed. Three of 4 microcell-hybrids with an introduced chromosome 1 were non-tumorigenic (#1-7, -8 and -13), whereas the parental HT1080 cells were highly tumorigenic. The other microcell-hybrid clone (#1-1) formed tumors, the cells of which had lost one copy of chromosome 1. Two clones from the #1-1 cells were isolated; one contained an extra copy of chromosome 1, and the other one did not. The former was non-tumorigenic and the latter was tumorigenic. The introduction of chromosome 11 also suppressed the tumorigenicity of HT1080 cells, while the introduction of other chromosomes, i.e., 2, 7, or 12, had minimal or no effect on the tumorigenicity of these cells. Cells from tumors formed by microcell-hybrids with the introduction of chromosome 2, 7, or 12 still contained the introduced chromosome. Interestingly, only the microcell-hybrids with an introduced chromosome 1 had an alteration in cellular morphology and modulation of in vitro transformed properties, i.e., cell-growth and saturation density in a medium containing 10% calf serum and cell-growth in soft-agar. Thus, the results indicate the presence of putative tumor-suppressor genes for HT1080 cells on chromosomes 1 and 11, and further suggest that the genes on these chromosomes control different neoplastic phenotypes.

Animals↗

PubMed1990

[A case of miliary tuberculosis with prolonged high fever for more than 2 months under antituberculous therapy].

A 28 year-old male was admitted to our hospital with persistent cough and high fever. He was diagnosed to have miliary tuberculosis by the transbronchial lung biopsy specimen and tuberculous choroidal lesions in the ocular fundus. Antituberculosis therapy was immediately started. In spite of the fact that the bacilli were sensitive to the antituberculosis drugs used and he had no other complications, high fever persisted and lasted for more than 2 months. When tuberculosis is suspected, and antituberculosis treatment is tried to observe its clinical response, the presence of similar cases mentioned above should be taken into consideration.

Adult↗

PubMed1989

Induction of metallothionein mRNA by tumor promoters in mouse skin and its constitutive expression in papillomas.

A single topical application of 12-O-tetradecanoylphorbol-13-acetate (TPA) was found to induce mRNA of a metallothionein (MT) gene or genes in the skin of Sencar mice, and papillomas produced by repeated applications of TPA were shown to have elevated levels of MT mRNA. Induction of MT mRNA was maximal 4-8 h after application of TPA and returned to the control level 24 h later. A dose-dependent increase of MT mRNA was observed with doses of TPA of 1-5 micrograms. Of the other promoters tested, phorbol-12, 13-didecanoate, mezerein, and the ionophore A23187 also induced MT mRNA, but 4-O-methyl-TPA and benzoyl peroxide did not. Phorbol and 4 alpha-phorbol-12,13-didecanoate, which are not promoters, also did not induce MT mRNA. Retinoic acid and 1 alpha, 25-dihydroxyvitamin D3, inhibitors of tumor promotion, did not induce MT mRNA themselves or inhibit the induction of MT mRNA by TPA. In C57BL/6 promotion-resistant mice, TPA caused only slight induction of MT mRNA. These data suggest a correlation between induction of MT mRNA and epidermal hyperplasia. The constitutive elevation of MT mRNA levels in papillomas may be due to the loss, during the process of tumor promotion, of some mechanism regulating MT gene expression.

Administration, Topical↗

PubMed1989

Complete nucleotide sequence and characterization of a cryptic plasmid from Lactobacillus helveticus subsp. jugurti.

A small cryptic plasmid, pLJ1, was isolated from Lactobacillus helveticus subsp. jugurti and was cloned into Escherichia coli HB101 by using pBR329 as a vector. Plasmid pLJ1 was 3,292 base pairs long and had single restriction endonuclease sites for PvuII, KpnI, AvaII, Acci, HindIII, and EcoRI. In a maxicell system, pLJ1 produced a protein of about 41 kilodaltons.

Amino Acid Sequence↗

PubMed1989

Multiple chromosomes carrying tumor suppressor activity, via microcell-mediated chromosome transfer, for various tumor cell lines.

The ability of normal human fibroblast-derived chromosomes to suppress tumorigenicity in nude mice and in vitro growth properties of various tumor cell lines was examined. Normal human chromosomes tagged with pSV2neo gene by DNA transfection were transferred to the following human tumor cell lines by microcell-fusion: SiHa (uterine cervical carcinoma), A204 (rhabdomyosarcoma), SK-NEP-1 (Wilms' tumor), HHUA (uterine endometrial carcinoma), SK-N-MC (neuroblastoma), YCR (renal cell carcinoma), HT1080 (fibrosarcoma), and CC1 (chorionic carcinoma). The results indicate the presence of a putative tumor-suppressor gene(s) in multiple chromosomes, and suggest that multiple genes may normally be involved in suppressing the transformed phenotypes at different stages in some tumors. Thus, the microcell transfer of chromosomes to specific tumor cell lines is a useful technique to demonstrate the presence of tumor-suppressor genes on individual chromosomes, and may also be useful in cloning of tumor-suppressor genes as well as elucidating their function in cell-growth and differentiation.

Animals↗

PubMed1987

Systemic inhibition of tumor promoter-induced ornithine decarboxylase in 1 alpha-hydroxyvitamin D3-treated animals.

Topical application of 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25(OH)2D3], an active form of vitamin D3, was previously shown to inhibit the induction of ornithine decarboxylase (ODC) and tumor promotion by tumor promoters in mouse skin. In the present study, this observation in skin was extended to other tissues, such as the stomach, colon, and liver, using 1 alpha-hydroxyvitamin D3 [1 alpha (OH)D3], which is converted to 1 alpha,25(OH)2D3 in the liver without hormonal control and thus evokes the systemic effects, if any, of 1 alpha,25(OH)2D3. When mice were given 1 alpha (OH)D3 at a dose of 5 micrograms by gastric tube, their plasma level of 1 alpha,25(OH)2D3 increased to a peak of about 18-fold the normal level after 12 h, followed by hypercalcemia (about 14 mg/dl), which reached a peak on Days 2 to 3. In 1 alpha (OH)D3-treated mice, induction of epidermal ODC by 12-O-tetradecanoylphorbol-13-acetate was markedly inhibited, the inhibition being maximal 2 to 4 days after 1 alpha (OH)D3 administration. ODC induction in the glandular stomach mucosa of rats by NaCl, a tumor promoter in stomach carcinogenesis, was also inhibited dose and time dependently by 1 alpha (OH)D3. Similarly, 1 alpha (OH)D3 treatment of rats markedly inhibited the induction of ODC in the colon mucosa by deoxycholate, a tumor promoter of colon carcinogenesis, and of ODC in the liver by phenobarbital, a promoter of liver carcinogenesis. These results suggest that an active form of vitamin D3 has a systemic inhibitory effect on induction of ODC activity by tumor promoters.

Animals↗

PubMed1987

Regulation of metallothionein gene expression by 1 alpha,25-dihydroxyvitamin D3 in cultured cells and in mice.

1 alpha,25-Dihydroxyvitamin D3 [1 alpha,25(OH)2D3], a hormonally active form of vitamin D3, has been shown to modulate cell differentiation and tumor promotion. This report demonstrates that mRNA of the metallothionein (MT) gene was induced by 1 alpha,25(OH)2D3 in cultured epidermal keratinocytes and also in liver, kidney, and skin tissues when 1 alpha-hydroxyvitamin D3, a synthetic precursor of 1 alpha,25(OH)2D3, was applied in vivo. Exposure of FRSK cells, a cell line derived from fetal rat skin keratinocytes, to 1 alpha,25(OH)2D3 at 5 ng/ml (12 nM) increased MT mRNA to almost the same extent as that induced by 10 microM dexamethasone or 1 microM CdCl2. This increase in the level of MT mRNA was evident within 2 hr and was maximal 12-24 hr after the addition of 1 alpha,25(OH)2D3. The induction was dose-dependent with concentrations of 1 alpha,25(OH)2D3 from 0.05 to 5.0 ng/ml. Amounts of MT increased with the increase of MT mRNA induced by 1 alpha,25(OH)2D3. Of the derivatives of vitamin D3 tested, only 1 alpha,25(OH)2D3 caused marked induction. Treatment with cycloheximide did not inhibit MT mRNA induction by 1 alpha,25(OH)2D3. 1 alpha,25(OH)2D3 induced MT mRNA in primary cultures of mouse epidermal keratinocytes but not in IAR-20, a liver cell line. 1 alpha,25(OH)2D3 had a similar effect in vivo: oral administration of 1 alpha-hydroxyvitamin D3 to mice resulted in increased levels of MT mRNA in the liver, kidney, and skin 24 hr later. Increase in the level of MT mRNA may be relevant to some biological actions of 1 alpha,25(OH)2D3.

Administration, Oral↗

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↗