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Biomedical subjects

F Wada

Publications and source records attributed to F Wada.

At least 37 records · Page 2Linked to original sources
PubMed1990

Purification and identification of a growth factor produced by Paramecium tetraurelia.

We previously reported that the jumyo mutant of a cilate protozoan Paramecium tetraurelia excretes into the medium a factor which promotes its own cell division. Here, the factor was purified to electrophoretic homogeneity through a series of liquid chromatographic procedures. This substance is a protein with a molecular weight of 17,000 which at concentrations of 1 X 10(-9) M (17 ng/ml) or more results in the recovery of the cell division rate of the jumyo mutant to the level of the wild type. The factor is therefore considered to be a growth factor and was named Paramecium growth factor (ParGF). This is the first report of direct proof for the production of a growth factor in an organism other than vertebrates.

Animals↗
PubMed1990

Differences in growth requirements between epithelial and stromal cells derived from rat ventral prostate in serum-free primary culture.

Different procedures of enzymatic digestion of rat prostatic tissue and unique sets of mitogenic factors made it possible to culture practically pure populations of epithelial and stromal cells without previous separation of the two types of cells. Keratin-positive epithelial cells dissociated by trypsin and collagenase from adult rat ventral prostate proliferated in medium WAJC 404 supplemented with epidermal growth factor, insulin, cholera toxin, and bovine pituitary extract. Proliferation of epithelial cells was completely inhibited by dexamethasone as low as 30 nM. On the other hand, fibroblast-like stromal cells released by trypsin digestion required a plastic substratum coated with calf serum or fibronectin, and proliferated in Eagle's minimum essential medium supplemented with cholera toxin, bovine pituitary extract, dexamethasone, and bovine serum albumin. Epidermal growth factor and insulin had negligible effect on proliferation of stromal cells. Physiological concentrations of dihydrotestosterone and estradiol showed no effect on proliferation of both types of cells.

Animals↗
PubMed1989

Promotion of calcification by imidazole and its suppression by diltiazem in the growth cartilage of rats with HEBP induced rickets.

The object of our experiments was to determine the effect of imidazole on the growth cartilage of rats with HEBP induced rickets. When HEBP (1-hydroxyethylidene-1, 1-bisphophonic acid) was given to young rats in large doses over a short period, rickets was consistently produced. We found that imidazole had a calcification promoting action in the growth plate cartilage where there had been an increase in thickness due to the inhibition of calcification. In an attempt to clarify the mechanism of accelerated calcification due to imidazole, the effects of diltiazem, a calcium antagonist, were observed; it was found to suppress the accelerated calcification. If diltiazem inhibits the entry of calcium ions into the cells of the growth cartilage, as it does in smooth muscle and myocardial cells, then our results indicate that intracellular concentrations of calcium may play an important role in the accelerated calcification due to imidazole.

Animals↗
PubMed1988

Comparative analysis of growth factors in normal and pathologic human prostates.

Growth factors, as detected by DNA synthesis stimulating activity for BALB/c 3T3 cells, in normal, benign hypertrophic and cancerous human prostates were analyzed. The total content (units per gram of tissue) in benign hypertrophic prostate was two to four times higher than those in normal and cancerous prostates. In all the three groups, heparin-binding growth factor, growth factor adsorbed to heparin-Sepharose in the presence of 0.5 NaCl. accounted for about 80-95% of the total growth factor content. Qualitative analysis using a heparin-Sepharose column revealed two types of heparin-binding growth factor in the prostates, one eluted from the column at 1.2-1.3 M NaCl and the other at 1.5-1.8 M NaCl. The latter was the predominant type in all groups. In addition to the growth factors detected with BALB/c 3T3, a growth factor with specific action upon MC3T3-E1 mouse osteoblasts was found in prostatic cancer, but not in normal and benign hypertrophic prostates.

Animals↗
PubMed1988

Partial purification of a major type of rat prostatic growth factor: characterization as an epidermal growth factor-related mitogen.

The dorsolateral prostate of rats contains a mitogen that shares several properties with epidermal growth factor (EGF), which was designated as prostatic EGF-related mitogen (PEM). PEM was purified about 2,100-fold using molecular-sieve and ion-exchange chromatography. Final preparation stimulated DNA synthesis in BALB/c 3T3 cells at a concentration as low as 1.5 ng/ml and competed with 125I-EGF for binding to cell surface receptors. PEM had a molecular weight of about 14,000 and an isoelectric point of about 4.5, being heat- and acid-stable but inactivated by dithiothreitol. The primary cultured rat dorsolateral prostate epithelial cells required EGF for maximum growth. Partially purified PEM fully substituted for EGF in the primary culture system at a concentration as low as 90 ng/ml. However, the activity of PEM was hardly suppressed by antimouse EGF antiserum. These findings suggest that PEM is a member of the EGF family but has a higher molecular weight (high molecular weight EGF).

Animals↗
PubMed1988

Production of IGF-II-related peptide by an anaplastic cell line (AT-3) established from the Dunning prostatic carcinoma of rats.

AT-3 cells, one of anaplastic cell lines established from the Dunning prostatic carcinoma of rats, were able to grow under serum-free conditions in a state of suspension detached from a substratum. Radioimmunoassay using monoclonal antibody against rat insulin-like growth factor II (IGF-II) revealed the presence of IGF-II-related peptide in acid-ethanol extracts of lyophilized serum-free media conditioned by AT-3 cells. The peptide contents in the culture media increased with increase in cell number; 71 ng at 3.0 X 10(6) cells and 449 ng at 4.6 X 10(7) cells. IGF-II-related peptide was hardly detectable in acid-ethanol extracts of AT-3 cells harvested after 13-days culture. These results indicate that AT-3 cells produce IGF-II-related peptide and may release it into the culture media.

Animals↗
PubMed1988

Proliferation of epithelial cells derived from rat dorsolateral prostate in serum-free primary cell culture and their response to androgen.

Primary cultured epithelial cells derived from the rat dorsolateral prostate proliferated in serum-free nutrient medium WAJC 404 supplemented with mitogens: insulin (650 nM), cholera toxin (120 pM), epidermal growth factor (EGF) (2.5 nM), dexamethasone (300 nM), and bovine pituitary extract (25 micrograms/ml). The culture consisted of two types of epithelial cell colonies: one originated from single cells or small cell aggregates and the other was epithelial cell outgrowth from small tissue fragments attached to a substratum. There were differences in requirements for the mitogens between the two types of colonies. Requirements for cholera toxin, bovine pituitary extract, and dexamethasone were higher in the former type of colonies, and those for EGF were higher in the latter type of colonies. Proliferation of the epithelial cells in either type of colony was suppressed more than 50% by 1 nM dihydrotestosterone. This suppressive effect was not mediated by stromal component in the tissue fragments, and was counteracted by cyproterone acetate, indicating specific and direct action of the androgen on prostate epithelial cells. The results suggest that there is discrete participation of polypeptide growth factors and androgen in proliferation and differentiation, respectively, of prostate epithelial cells in vivo.

Androgens↗
PubMed1988

Stabilization of fibroblast growth factors by a non-cytotoxic zwitterionic detergent, 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS).

The potential usefulness of a zwitterionic detergent, 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHPAS), in the stabilization of acidic and basic fibroblast growth factors (FGFs) was examined. Among several detergents, CHAPS was found to be not only non-cytotoxic but also most useful in handling the diluted preparations of FGFs. The advantages are as follows: 1) at lower concentrations than 0.01% CHAPS did not affect growth factor activity of calf serum (CS) and the growth rate of BALB/c 3T3 cells. The primary culture of rat prostate epithelium and colony formation of NRK-49F cells were hardly influenced by CHAPS lower than 0.003%; 2) the loss of FGFs that usually occurs due to their adherence to the surface of storage containers was effectively prevented by inclusion of 0.1% CHAPS; 3) the recovery of FGFs after storage or dialysis was significantly enhanced by inclusion of 0.1% CHAPS; 4) CHAPS at lower concentrations than 0.1% does not interfere with amino acid analysis, except that Thr may be misled only when the ratio of protein/CHAPS is low; 5) amino acid sequence analysis was hardly disturbed by CHAPS up to 0.5%. These results indicate that CHAPS is useful as a stabilizing agent for various kinds of polypeptides capable of showing biological activity at a low concentration.

Amino Acids↗
PubMed1988

[New infusion port for intermittent infusion chemotherapy].

We devised a new infusion port for the intermittent infusion chemotherapy to the hepatic artery. This device has the following advantages: 1. A large needle (18 G) may be used to infuse the medicine because of the special urethane gum parts of the port. 2. One need only exchange the port for a new one or to the tube for continuous infusion chemotherapy under local anesthesia. 3. This port can be employed for arterial embolization because of its large inner diameter. In this report we introduce the structure and advantages of this new port device.

Antineoplastic Agents↗
PubMed1987

Heparin binding affinity of rat prostatic growth factor in normal and cancerous prostates: partial purification and characterization of rat prostatic growth factor in the Dunning tumor.

The rat prostate contains two types of growth factors capable of stimulating DNA synthesis in BALB/3T3 cells. These rat prostatic growth factors (RPGF) were separable by a different affinity for heparin: low affinity type RPGF and high affinity (HiA) type RPGF. About 80% of the RPGF in the cytosol from normal prostates was low affinity type, whereas more than 80% in the cytosol from the Dunning tumors was HiA type. Elution profile of HiA-RPGF showed two peaks of activity eluted from the heparin-Sepharose column, one at 1.3-1.4 M NaCl (HiA1-RPGF) and the other at 1.6-1.7 M NaCl (HiA2-RPGF). HiA2-RPGF could be purified 1100-fold from the Dunning tumor (AT-3 subline) in about 20% recovery by heparin-Sepharose chromatography. The partially purified HiA2-RPGF in the Dunning tumor has a molecular weight of about 19,000 and isoelectric point of about 3.8, and stimulated DNA synthesis at a concentration of about 0.25 nM. The activity was lost by heat treatment at 70 degrees C for 5 min and by acid treatment, whereas it was stimulated by incubating with dithiothreitol. The HiA2-RPGF did not have transforming growth factor activity at a concentration of 250 ng/ml or lower in the presence of epidermal growth factor.

Animals↗
PubMed1987

Isolation of a 41 kilodalton cytosol protein from the Dunning rat prostatic adenocarcinoma: characterization as depolymerized actin isomers.

Except for albumin, a 41,000-dalton protein (41K) in the cytosol of the Dunning R-3327 rat prostatic adenocarcinoma was found to be the most abundant soluble protein. This protein was purified in nearly homogeneous state by conventional chromatographies. After the first chromatography, because of the adhesive nature of the protein, 0.5% SDS and 2 M urea were necessary for subsequent steps of purification. The amino acid composition of the purified 41K was similar to that of actin isolated from rabbit skeletal muscle. Alternatively, 41K could be extracted from the Dunning tumor in the presence of ATP and dithiothreitol, under conditions in which actin molecules are depolymerized, and could be purified by the same method as cytoskeletal actin. The purified protein showed properties similar to rat skeletal muscle actin in amino acid composition and antigenicity. Both 41K proteins were found to be composed of four components having different isoelectric points. These results indicate that most actin exists in a depolymerized form as a cytosol protein of 41,000 daltons in the Dunning tumor and is composed of at least four isomers.

Actins↗
PubMed1987

A comparative study of the calcification-promoting action of 1,25 (OH)2D3 and calcitonin on the growth cartilage of rats with 1-hydroxyethylidene-1, 1-biphosphonic acid (HEBP)-induced rickets.

When HEBP (1-hydroxyethylidene-1, 1-biphosphonic acid) was administered to young rats in large doses over a short period rickets was consistently produced. When HEBP was administered concomitantly with 1,25 (OH)2D3 or calcitonin (CT), calcification appeared in the growth-plate cartilage where there had been an increase in thickness due to the inhibition of calcification. This experiment was done in an attempt to clarify differences in the calcification-promoting mechanisms of 1,25 (OH)2D3 and CT. The serum alkaline phosphatase level was reduced in rats with an accelerated calcification following the administration of 1,25 (OH)2D3, but there was no reduction in the serum alkaline phosphatase level in rats in which the calcification was accelerated by the administration of CT. The mode of appearance of calcification in the growth-plate cartilage by 1,25 (OH)2D3 or CT differed, depending on the time of administration. These results suggest that mechanisms involved in the enhancement of calcification by 1,25 (OH)2D3 and CT differ in cases where rickets are induced by HEBP.

Alkaline Phosphatase↗
PubMed1987

Protein profiles of benign hypertrophic prostate: stroma-abundant distribution of BPH-associated nonhistone proteins.

Mechanically isolated epithelium and stroma from benign hypertrophic prostates were highly pure on the basis of histochemical and biochemical criteria. By electrophoretic analyses, whole cellular and nuclear proteins were compared among whole tissues, epithelium, and stroma. The characteristic protein profiles of benign hypertrophic prostates were reflected in the electrophoretic patterns of the stroma. Two-dimensional gel electrophoretic patterns of the epithelium were different from those of the stroma with exception of about 18 major spots that were common to both fractions. Of the protein species separated, 35K/6.7 (molecular weight/pI) and 36K, which was composed of two species with pI of 4.4 and 4.6, were abundant in the epithelium and stroma, respectively. Nuclei prepared from whole tissues of benign hypertrophic prostates contained three kinds of nonhistone proteins (NHP) closely associated with benign prostatic hypertrophy (BPH); 42 K-NHP, 55 K-NHP and 190 K-NHP. Electrophoretic analysis of the nuclear proteins revealed that all the BPH-associated nonhistone proteins were abundantly distributed in the nuclei of the stroma.

DNA↗
PubMed1987

Growth factors in the prostate.

Certain local tissue factors, such as growth factor, in addition to androgens, are involved in the prostate growth. The prostate contains two types of growth factors capable of stimulating DNA synthesis in BALB/3T3 cells. They were divided into low affinity (LoA) type and high affinity (HiA) type by a different affinity for heparin-Sepharose. HiA-type growth factor is further classified into acidic HiA and basic HiA types. Acidic HiA type could be purified from the Dunning tumor (R 3327), a rat prostatic adenocarcinoma, and has a molecular weight of about 19,000 and a pI of about 3.8. Basic HiA type could be isolated from the tissues of human benign prostatic hypertrophy and has a molecular weight of about 12,000 and a pI of about 10.5. They are inactivated by heat and acid treatments. Acidic HiA type appears to be involved in growth of the rat prostate epithelium, and LoA type growth factor is possibly relevant to reproductive physiology because of its coexistence with "probasin," a major secretory protein in the dorsolateral prostate having a strong affinity for spermatozoa.

Aging↗