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K Mashima

Publications and source records attributed to K Mashima.

36 records · Page 2Linked to original sources
PubMed1991

Purification and properties of growth inhibitor from normal rabbit serum.

It was previously found that rabbit serum contains a growth-inhibitory substance for a tumorigenic rat liver cell line RSV-BRL. In the present study, the growth inhibitor was purified from normal rabbit serum to show a homogeneous protein band with a molecular weight (Mr) of 56 k on SDS-polyacrylamide gel electrophoresis under non-reducing conditions. The purified growth inhibitor, tentatively named rabbit serum-derived growth inhibitor (RSGI), potently inhibited the growth of RSV-BRL and nine kinds of other cell lines including three human tumor cell lines at a concentration of 20 ng/ml or higher. The growth-inhibitory effect of RSGI was reversible and appeared to be cytostatic rather than cytotoxic. RSGI was stable to heating at 56 degrees C for 30 min or treatment with 0.1 M 2-mercaptoethanol, but labile to heating at 100 degrees C for 3 min or treatment with 1 M acetic acid (pH 2.3), 6 M urea, 50% (v/v) 1-propanol, or 0.1% (w/v) trypsin. These properties of RSGI suggested that it was different from type beta transforming growth factors, tumor necrosis factor-alpha, and other known growth-regulatory factors.

Animals↗
PubMed1989

Structure and expression of elongation factor 2 gene during development of Dictyostelium discoideum.

A cDNA library constructed from poly(A)+ RNA isolated from Dictyostelium discoideum cells at 12 h of development was screened with the hamster elongation factor 2 (EF-2) cDNA. Several different cDNA clones which hybridized were isolated after a second screening. A cDNA clone representing the 5'-end of the mRNA was obtained by primer extension. By comparing the amino acid sequence deduced from the nucleotide sequences of these clones with that of hamster EF-2, we found enough homology between them to conclude that the isolated clones were complementary to the mRNA of D. discoideum EF-2. The N terminus which is the GTP-binding domain and the C-terminal half where it interacts with a ribosome showed a high degree of homology. The amino acid sequence of the carboxyl half includes that it contain a site of ADP-ribosylation by diphtheria toxin. From the Northern blotting analysis, the size of the mRNA was estimated to be 2.6 kilobases. The expression of the mRNA was high in vegetative cells, became maximal at the aggregation stage, and decreased thereafter through development. Upon differentiation of prespore and prestalk cells, the mRNA was highly enriched in the former over the latter. ADP-ribosylation assay of EF-2 protein by diphtheria toxin showed nearly the same developmental changes for the protein as the mRNA. However, prestalk cells were found to contain the same amount of the protein as prespore cells. The Southern blot analyses indicated that the gene encoding EF-2 is unique.

Amino Acid Sequence↗
PubMed1989

A study on the toxicity of antineoplastic drugs (bleomycin, peplomycin and cis-diamminedichloroplatinum) by simultaneous administration (Part 1).

In order to clarify the toxicity of the antineoplastic drugs, bleomycin (BLM), peplomycin (PEP) and cis-diamminedichloroplatinum (CDDP), which are commonly used to treat head and neck cancer by simultaneous administration, the semiacute toxicity of each of these drugs in rats was studied as an initial step. Body-weight change, general behavior, red blood cell count (RBC), white blood cell count (WBC), serum biochemistry (s-GOT, s-GPT, BUN, GLU, ALB, TP), A/G ratio, relative organ weight and histopathological features were determined. BLM and PEP given by ip administration once a week produced severe diarrhea, decreased diet consumption, emaciation, piloerection and loss of hair, enhancements of RBC and WBC, several changes in serum biochemistry, proliferation and mitosis of epithelial cells in the forestomach and occasional granular and vacuolar degeneration in the liver. CDDP administered in the same manner produced a significant decrease of WBC, several changes in serum biochemistry parameters especially BUN, an increased focal-segmental mesangial matrix in the glomeruli and vacuolar degeneration of epithelial cells of the convoluted tubule of the kidney.

Animals↗
PubMed1988

Multiple forms of growth inhibitors secreted from cultured rat liver cells: purification and characterization.

It was found that a non-tumorigenic epithelial cell line from the liver of a Buffalo-strain rat (BRL) secreted into the culture medium various inhibitors of the growth of BRL and RSV-BRL (tumorigenic BRL transformed by infection of Rous sarcoma virus). The secreted inhibitors were classified into two types: one inhibited the growth of BRL to a greater extent than that of RSV-BRL (non-tumorigenic BRL growth inhibitor, NGI), and the other, vice versa (tumorigenic BRL growth inhibitor, TGI). Two NGI (NGI-I and NGI-II) and two TGI (TGI-I and TGI-II) were highly purified from the serum-free conditioned medium. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis without 2-mercaptoethanol, NGI-I and II gave protein bands with molecular weights (Mr) of 56,000 and 21,000, respectively. TGI-I and II gave a band that migrated faster than bromophenol blue marker dye, but they did not pass through an ultrafiltration membrane with an Mr cutoff of 5,000. In the presence of a reducing reagent, only NGI-II showed a decrease of Mr, from 21,000 to 11,000. NGI and TGI showed 50% growth inhibition with BRL and RSV-BRL, respectively, at 5-15 ng/ml in the medium containing 10% fetal calf serum. NGI and TGI all were stable to 1 M acetic acid (pH 2.3) and 6 M urea, but labile to 5 mM dithiothreitol or trypsin. Of the eight cell lines tested, NGI-I was most effective on BRL, NGI-II on BRL and HSC-3 (human tongue squamous carcinoma), and both TGI-I and II on RSV-BRL.

Animals↗
PubMed1987

Growth-inhibitory protein present in rabbit serum, which is more effective on tumorigenic rat liver epithelial cells than on non-tumorigenic ones: its species, and mode of existence.

We have previously reported that in culture, rabbit serum inhibits the growth of the epithelial cell line from Buffalo rat liver (BRL) lower than that of the tumorigenic one transformed by Rous sarcoma virus (RSV-BRL). Here, the serum was fractionated by several different methods. The findings are: 1) the growth inhibitor present (GI) existed as large complexes with non-inhibitory proteins; 2) the complexes were dissociated by 1 M NaCl plus 6 M urea; 3) the dissociated GI did not pass through membrane filter with Mr cutoff 10k; 4) it was stable in 8.5 M urea and 1 M acetic acid (pH 2.5), but labile against either dithiothreitol and trypsin; 5) it was separable into two species with pI 7.5 and 9.5; 6) both species were more effective on RSV-BRL than on BRL.

Animals↗
PubMed1987

Growth inhibitors in serum, platelets, and normal and malignant tissues.

The non-malignant epithelial cell line from Buffalo rat liver (BRL) and its malignant transformant obtained by Rous sarcoma virus infection (RSV-BRL) were used as the indicators. A survey was made in animals for growth inhibitors which could inhibit the growth of BRL, but not the growth of RSV-BRL in culture, under the consideration that the loss of the sensitivity to such inhibitors might be the most dominant cause for acquisition of tumorigenicity. From human platelets, a growth inhibitor was extracted with acid-ethanol and prepared to a homogeneous purity, separated from the major TGF-beta activity, which stimulated the anchorage-independent growth of NRK cells in the presence of EGF. The purified inhibitor (Mr 25-27k; pI 8.7-9.1) showed 50% inhibition on the growth of BRL at 20 pg/ml (0.7 pM), but hardly influenced the growth of RSV-BRL. Its inhibition was specific to epithelial cell lines, thus called epithelial growth inhibitor (EGI). It consisted of two subunits with practically identical Mr, which were cross-linked with disulfide bonds. Its sequence of N-terminal 10 amino acids was consistent with that of TGF-beta. In rats, serum contained a growth inhibitor with Mm 220k. The inhibitor, when treated with high concentrations of urea or guanidine-HCl, liberated an active component with Mr 30k and pI 5.2. When the growth inhibitor with Mm 220k was treated with ethanol-acetone or 1 M acetic acid, its activity increased more than 10-fold. Growth inhibitors were extracted with acid-ethanol from various tissues of rats. The activities of the extracts to inhibit the growth of BRL cells were high in spleen, lung and tumors with relatively high mitotic activities, but low in muscle, heart and brain. Acid-ethanol extract from BRL cells showed growth inhibitory activity toward BRL cells themselves significantly higher than that from RSV-BRL cells. The serum-free media conditioned with BRL and RSV-BRL cells showed a little growth inhibitory activity toward BRL cells. On treatment with acetic acid, their activities increased significantly, comparable for both types of cells. The media thus activated inhibited the growth of BRL cells, but not that of RSV-BRL cells. All the growth inhibitors described above including human platelet inhibitor, whether present in extracts or purified, resembled one another in stability against acid, heat and reducing reagents.

Animals↗
PubMed1987

Transformation of rat liver cell line by Rous sarcoma virus causes loss of cell surface fibronectin, accompanied with secretion of metallo-proteinase that preferentially digests the fibronectin.

An epithelial cell line derived from the liver of a normal Buffalo rat (BRL) was transformed by Rous sarcoma virus (RSV). The RSV-transformed cells were separated into five clones (RSV-BRL1 through 5), which were morphologically different. RSV-BRL cells exhibited the following characteristics distinct from those of BRL cells: tumorigenicity, irregular cell arrangement, loose intercellular junction, growth in soft agar (anchorage-independent growth) except for RSV-BRL3 and 5, and loss of cell surface fibronectin. When BRL cells were cultured in the standard medium supplemented with the serum-free conditioned medium of RSV-BRL cells, the amount of the cell surface fibronectin decreased significantly. It was found that RSV-BRL cells secreted a proteinase capable of hydrolyzing the fibronectin, whereas BRL cells secreted hardly any of this proteinase. The fibronectin-hydrolyzing proteinase (FNase) could also hydrolyze plasma fibronectin added as an exogenous substrate. The hydrolysis of plasma fibronectin was inhibited by ethylenediamine tetraacetate, but stimulated by rho-chloromercuribenzoate and calcium ion. This indicates that FNase is a metallo-enzyme, but not a serine or thiol enzyme. In addition to the proteinase, RSV-BRL cells secreted plasminogen activator and a proteinase inhibitor which inhibited the activity of plasmin but not FNase.

Animals↗
PubMed1987

Acoustic screening for laryngeal cancer.

An acoustic screening method for indicating the possible presence of laryngeal cancer was investigated. Three acoustic parameters, comprising perturbations in pitch-period and peak-amplitude sequences, and vocal noise, were measured from a sustained vowel, e, spoken by the subjects taking part in the investigation. Experiments to discriminate between normal and cancer groups were performed with voice samples taken from 64 normal control subjects and 57 patients with laryngeal cancer. Experiments were also carried out to test the perceptual significance of the three parameters. From the results, we have been able to conclude that their combined use will enable us to build an acoustic screening system for laryngeal cancer.

Aged↗
PubMed1986

Purification and properties of epithelial growth inhibitor (EGI) from human platelets: its separation from type beta transforming growth factor (TGF-beta).

We previously reported that sera from various kinds of animals contain a protein(s) capable of inhibiting the growth of the non-malignant epithelial cell line derived from Buffalo rat liver (BRL). In the present study, a similar epithelial cell-specific growth inhibitor (EGI) was purified to homogeneity from an acid-ethanol extract of human platelets. During purification, EGI was separated from the major component of type beta transforming growth factor (TGF-beta), which can stimulate the colony formation of the non-malignant fibroblastic cell line derived from rat kidney (NRK) in soft agar in the presence of epidermal growth factor (EGF). The purified EGI had an Mr of 27,000, and was composed of two subunits identical in Mr. It significantly inhibited the growth in monolayer cultures of three non-malignant epithelial cell lines, BRL, MDCK (from Madin-Darby canine kidney) and BSC-1 (from African green monkey kidney), at doses lower than 40 pg/ml in medium containing 10% fetal calf serum. Its inhibitory activity was stable against heating at 90 degrees C for 3 min, but not against treatment with 50 mM dithiothreitol. In addition, TGF-beta was also partially purified from the same extract. The purified TGF-beta did not show any inhibitory activity toward the growth of BRL, MDCK, BSC-1, or NRK.

Blood Platelets↗
PubMed1986

Normalized noise energy as an acoustic measure to evaluate pathologic voice.

In order to evaluate noise components included in pathologic voice signals, a novel acoustic measure, normalized noise energy (NNE), is proposed and its effectiveness for the detection of laryngeal pathologies is investigated with 250 vowel samples spoken by 64 control (normal) subjects and 186 patients with various laryngeal diseases. The NNE is automatically computed from the voice signals using an adaptive comb filtering method performed in the frequency domain. Experiments with the voice samples show that the NNE is especially effective for detecting glottic cancer, recurrent nerve paralysis, and vocal cord nodules. Specifically, when glottic cancer is represented in terms of the T classification adopted by the UICC (Union Internationale Contre le Cancer), glottic T2-T4 cancer can be perfectly discriminated from normal samples, but 22.6% of patients with glottic T1 cancer are incorrectly classified as normal, with an error rate of 9.4% for normal subjects.

Glottis↗
PubMed1985

Characterization of a growth-inhibiting protein present in rat serum that exerts a differential effect on in vitro growth of nonmalignant rat liver cells when compared with Rous sarcoma virus-transformed rat liver cells.

We have previously reported the transformation by Rous sarcoma virus of a cloned epithelial cell line (BRL) established from Buffalo rat liver by H. Coon. The nontransformed (BRL) and transformed (RSV-BRL) cells grew at comparable rates in culture, whereas only the transformed cells were tumorigenic in vivo. We report here on the existence in rat and mouse sera of a growth inhibitor for the nontransformed BRL cells. The transformed BRL cells (RSV-BRL) were insensitive to this inhibitor. The inhibitory activity was not prominent in sera from other species of animals tested except for rabbit; this serum inhibited the growth of RSV-BRL cells more strongly than that of BRL cells. The growth inhibitor was partially purified from rat serum. It is a protein free of lipid and has a molecular weight of about 220 000. The inhibitor could be separated into three components of pI 4.6, 5.2 (major) and 5.6 by isoelectric electrophoresis.

Animals↗
PubMed1984

Oral carcinoma of the upper jaw. Results of radiation treatment.

Thirty-eight patients with hard palate carcinoma and 82 with upper gum carcinoma received primary therapy consisting of radiation therapy alone or in combination with surgery. Fifty-three per cent of the primary lesions were successfully controlled by initial radiation therapy. Radiation therapy did not seem to give inferior results compared with surgery in the treatment of the primary lesion. The most important determining factors in the prognosis were the presence of cervical metastases, and bone involvement.

Adenocarcinoma↗