Search PubMed⌕ Search

Biomedical subjects

M Tagawa

Publications and source records attributed to M Tagawa.

At least 163 records · Page 9Linked to original sources

Changes in reticuloendothelial function in dogs with endotoxin-induced shock.

A shock model was experimentally produced by intravenous injection of a lethal dose (3 mg/kg) of endotoxin under general anesthesia induced by pentobarbital sodium using 7 beagles. The effect of this endotoxic shock on the reticuloendothelial function was investigated. The blood endotoxin concentration peaked immediately after administration and decreased subsequently. However, the value still remained on an increased level (1,051 pg/ml) even at 360 min after endotoxin treatment. The lipid emulsion test as an index of reticuloendothelial phagocytotic activity and the arterial ketone body ratio as an index of the energy charge in the liver decreased after endotoxin treatment and failed to recover during the experiment. Fibronectin, one of opsonic proteins, tended to decrease after injection of the endotoxin and was significantly (p < 0.01) low at 180 and 360 min compared with the value before injection of the endotoxin. These results suggested the depression of the reticuloendothelial function during endotoxin-induced shock.

Animals↗

Prophylactic effect of milbemycin oxime against Dirofilaria immitis infection in dogs: optimum dose and administration time.

To examine the prophylactic efficacy of milbemycin oxime against Dirofilaria immitis infection, 42 filaria-free beagles were inoculated with 100 infective larvae of the parasite. Milbemycin oxime was administered orally at doses of 0.05 to 1.0 mg/kg body weight one month after inoculation, and at a single dose of 0.25 mg/kg body weight on days 1 to 90 after inoculation. All dogs were euthanized and autopsied to evaluate the prophylactic effect of the drug against D. immitis infection 6 months after inoculation. It was confirmed that a dose of 0.25 mg/kg of milbemycin oxime completely-protected the infection 15 to 60 days after infection.

Animals↗

Evaluation of extravascular thermal volume in the lung in dogs with endotoxin-induced shock by double indicator dilution method using heat and sodium ions.

Accuracy assessment was undertaken under varying hemodynamic conditions for a lung water volume measurement device which is based on the principle of a double indicator dilution method using heat and sodium ions. Changes in extravascular thermal volume were investigated in dogs with endotoxin-induced shock. The isoproterenol- or propranolol-induced changes in hemodynamics had no effect on the measurement. This confirmed the high accuracy of this measuring method. The measurement revealed a tendency for the extravascular thermal volume to gradually increase (p < 0.05) during endotoxin shock. This confirmed the gradual progression of pulmonary edema during endotoxin shock.

Animals↗

Inhibition of steroid-induced prostatic hyperplasia in rats by treatment with anti-androgen (TZP-4238).

The effect of a synthetic steroidal anti-androgen, TZP-4238, on steroid-induced rat prostatic hyperplasia was investigated. Male Wistar rats were divided into four experimental groups. Group 1 consisted of intact controls. The other animals were castrated. The castrated animals were treated for 7 weeks with 1) testosterone 1 mg/head plus 17 beta-estradiol (E2) 0.01 mg/head (Group 2), 2) testosterone plus E2 + TZP-4238 8 mg/kg (Group 3) and 3) testosterone plus E2 + chlormadinone acetate (CMA) 20 mg/kg (Group 4). TZP-4238 and CMA were administered orally for 4 weeks after 3 weeks treatment with testosterone plus E2. In group 2, glandular hyperplasia of the prostate was clearly observed, and the number of bromo-deoxyuridine (BrdU)-positive cells showed a significant increase. In contrast, combined treatment with TZP-4238 (Group 3) or CMA (Group 4) produced marked atrophy of the glandular epithelium, and the number of BrdU-positive cells were remarkably decreased compared with Group 2. In addition, the localization of glutathione-peroxidase (GSH-PO) which effectively reduces the lipid peroxides in the glandular epithelial cells was markedly decreased. Furthermore, nuclear immunostaining of androgen receptor was remarkably decreased after combined treatment with TZP-4238 or CMA. Our data indicate that TZP-4238 is a potent steroidal androgen receptor antagonist for the prevention of rat prostatic growth in the steroid-induced prostatic hyperplasia model.

Androgen Antagonists↗

Immunolocalization of glutathione-peroxidase and androgen receptors in the rat dorsolateral prostate.

The effects of testosterone (T) and 17 beta-estradiol (E2) on the dorsolateral prostate of castrated rats were investigated by histopathological and immunohistochemical procedures. A significant increase in the prostatic weight of the castrated rats occurred after treatment with testosterone alone, or in combination with E2. Histologically, glandular hypertrophy was observed and fibromuscular stromal proliferation was also seen in the rats receiving the combination treatment. Immunohistochemical localization of glutathione-peroxidase (GSH-PO), an enzyme which effectively reduces the lipid peroxides, was clearly detected in the glandular epithelial of the dorsolateral prostate following testosterone alone or testosterone plus E2-treatment. Therefore, it appeared that GSH-PO protein synthesis in the glandular epithelium of the dorsolateral prostate can be enhanced (induced?) by sex hormones such as testosterone and E2. In the dorsolateral prostate of intact rats, positive staining for androgen receptors (AR) was observed in nuclei of the glandular epithelial cells. In addition, immunodetectable AR decreased within 2 days after castration, but returned to intact levels after administration of testosterone. These findings agree with previous work on the prostate. It is concluded that immunohistochemical analysis is very useful in studying the effects of androgen treatment on the prostate.

Animals↗

Immunohistochemical localization of tubulin and actin in rat prostate.

Immunohistochemical localization of the cytoskeleton components alpha tubulin and actin in the rat prostate was achieved. Formalin-fixed paraffin sections were prepared and examined by the avidin-biotin-peroxidase-complex method (ABC method). We observed that the cytoplasm of the glandular epithelial cells were stained by alpha tubulin and actin antibodies. The staining intensity for these proteins was decreased following castration, and recovered by testosterone plus 17 beta-estradiol-administration to the castrated rats. Based on these data, we suggest that alpha tubulin and actin in the rat prostate very likely participate in the secretory and metabolic activities of the glandular epithelial cells of the prostate.

Actins↗

Absorption, distribution and excretion of [carbonyl-14C]mosapride citrate after a single oral administration in rats, dogs and monkeys.

Absorption, distribution and excretion of mosapride citrate ((+/-)-4-amino-5-chloro-2-ethoxy-N-[[4-(4-fluorobenzyl)-2-morph oli nyl] methyl]benzamide citrate, AS-4370, CAS 112885-42-4), a novel gastric prokinetic drug, were studied with 14C-labeled drug in male and female rats mainly after a single oral administration. Plasma concentrations and excretion following oral administration of [14C]mosapride were also investigated in dogs and monkeys of both sexes. The main experimental dose was 10 mg/kg. After oral administration, [14C]mosapride radioactivity was rapidly absorbed through the intestinal tract. In male rats, concentration of plasma radioactivity reached the maximum (Cmax; 1410 ng eq./ml) 1 h after administration and decreased biphasically with half-lives of about 2 h in alpha-phase (t1/2 alpha) and in beta-phase (t1/2 beta) of about 8 h. t1/2 beta was virtually constant in the dose range from 1 mg/kg to 100 mg/kg, and the area under the plasma concentration-time curve (AUC) was proportional to the dose. In female rats, biphasic plasma concentration-time profile with similar half-lives was also observed, but Cmax (2070 ng eq./ml) and AUC were larger than those in male rats, suggesting the sex difference in pharmacokinetics. In dogs and monkeys, Cmaxs of plasma concentration were about 1000 ng eq./ml and 2000-3000 ng eq./ml, respectively, and sex difference was not observed. Plasma concentrations declined in a biphasic manner and t1/2 alpha and t1/2 beta were about 4 h and 15 h in dogs and about 3 h and 10 h in monkeys, respectively. The [14C]mosapride radioactivity was distributed to many tissues including the stomach and small intestine at the higher concentration, while to the brain and eye ball at the lower concentration than the plasma in male rats. Radioactivities in most tissues decreased essentially in parallel with those in plasma. In pregnant rats, concentrations of radioactivity in fetus were a little higher than those in the maternal plasma. In lactating rats, milk radioactivity concentrations were about 5 times higher than corresponding plasma concentrations, and both of them decreased with similar half-lives. Mosapride was bound to serum protein of various animal species, albumin and alpha 1-acid glycoprotein, in about 93-99%. After oral administration in rats, about 40% of dosed radioactivity was excreted into urine and about 60% into feces via bile. Neither dose dependency nor sex differences was observed in excretion. In dogs, about 20% of dosed radioactivity was recovered in urine and about 70% in feces.(ABSTRACT TRUNCATED AT 400 WORDS)

Administration, Oral↗

Metabolism of [carbonyl-14C]mosapride citrate after a single oral administration in rats, dogs and monkeys.

The metabolism of mosapride citrate ((+-)-4-amino-5-chloro-2-ethoxy-N-[[4-(4-fluorobenzyl)-2-morpho li nyl] methyl]benzamide citrate, AS-4370, CAS 112885-42-4) was investigated in rats, dogs and monkeys after a single oral administration. Four metabolites, M-1 to M-4, were isolated from rat urine. Two of them were identified as des-p-fluorobenzyl (M-1) and 5'-oxo-des-p-fluorobenzyl mosapride (M-2) and the other two were considered to be as 3-hydroxy des-p-fluorobenzyl (M-3) and 3-hydroxy 5'-oxo-des-p-fluorobenzyl mosapride (M-4) by mass spectrometry and/or nuclear magnetic resonance spectrometry. Metabolites in biological specimens such as plasma and urine after oral administration of [14C]mosapride in rats, dogs and monkeys were analyzed by thin layer chromatography (TLC). Unchanged mosapride, M-1, M-2 and some other metabolites were observed in the plasma of male rats. On the other hand, female rat plasma contained mosapride as the major radioactive component with a small amount of M-1 and little other metabolite(s). In rat urine, irrespective of sex, M-1, M-2, M-3, M-4 and a few other metabolites were observed but the unchanged drug was hardly detected. These facts suggested that the metabolic rate of mosapride was different between sexes in rats. In male rat bile, mosapride, M-1 and polar metabolite(s) were observed. In dogs, mosapride and M-1 were observed as major radioactive components in plasma, and M-1 was seen also as the major metabolite in urine but unchanged drug was hardly detected. Similar results were obtained in monkeys.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗

Absorption, distribution, metabolism and excretion of [carbonyl-14C]mosapride citrate after repeated oral administration in rats.

Absorption, distribution, metabolism and excretion of mosapride citrate ((+/-)-4-amino-5-chloro-2-ethoxy-N [[4-(4-fluorobenzyl)-2-morpholinyl]methyl]benzamide citrate, AS-4370, CAS 112885-42-4), a novel gastrokinetic agent, were studied with 14C-labeled compound in male rats during and after 21 consecutive daily oral administration at a dose of 10 mg/kg/d. After the first administration, plasma radioactivity concentrations were essentially equal to those in the single dose experiment, including the maximum concentration (Cmax; 1130 ng eq./ml) at 1 h. Plasma concentrations at 1 h after each administration were virtually constant in the range of 770-1350 ng eq./ml for 21 days. On the other hand, the plasma concentration at 24 h gradually increased for 6-7 days to about 120 ng eq./ml and became substantially constant, suggesting that apparent steady state was achieved by around 7 consecutive daily administration. Plasma concentration reached Cmax of 1230 ng eq./ml at 0.5 h after the 21st administration and decreased biphasically with half-lives of 3.4 h (t1/2 alpha) and 14.9 h (t1/2 beta). t1/2 alpha was virtually similar but t1/2 beta was about 2 times longer compared with single dose experiment. About 40% and 55% of dose radioactivity were excreted in urine and feces, respectively, for 21 days and radioactivity was almost completely excreted within 168 h after the last administration. The analysis by thin layer chromatography elucidated that composition of radioactive metabolites in plasma and urine after repeated administration was similar to that in the single dose study.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗

Involvement of protein kinase C in proliferative response of osteoblastic cell lines stimulated with prostaglandin E2.

The proliferative action of prostaglandin E2 in a rat osteoblastic cell line was studied. Prostaglandin E2 has a crucial effect on bone remodeling process and is shown to stimulate protein and DNA synthesis in serum depleted condition. This reaction is completely inhibited by protein kinase C inhibitors, suggesting that protein kinase C activation induced by prostaglandin E2 is involved in the proliferative response of osteoblast cells.

Animals↗

Nucleotide sequence of prothrombin gene in abnormal prothrombin-producing hepatocellular carcinoma cell lines.

A protein induced by vitamin K absence or antagonist II, PIVKA-II is synthesized in the liver and possesses a structure similar to prothrombin except that ten glutamic acid residues in amino-terminal Gla domain are not completely gamma-carboxylated and are functionally inactive. This protein can be detected in the plasma of patients with hepatocellular carcinoma (HCC) and used as a new tumor marker. To analyze the mechanism of PIVKA-II production in HCC tissue, the prothrombin gene of PIVKA-II-secreting HCC cell lines was sequenced to detect the mutation in the Gla domain and carboxylase recognition site of leader sequence located on exons I and II that may cause the inhibition of carboxylation. Exons I and II and donor and acceptor site of intron I of the prothrombin gene in two HCC cell lines, PLC/PRF/5 and huH-2, were analyzed by polymerase chain reaction (PCR), and the product was sequenced directly. In addition, RNA samples of these cell lines were used for complementary DNA synthesis, followed by PCR and sequencing. The nucleotide sequences of the Gla domain in both HCC cell lines were conserved. One nucleotide change was detected at nt.554 (adenine to guanine), but this did not influence the amino acid sequence. Splicing sites between exons I and II, the leader sequence of the precursor prothrombin, and protease target sites also were conserved as the reported prothrombin gene, and mutations reported for other des-gamma-carboxy coagulation factors were not detected. These results also were confirmed by DNA analysis of seven human fresh-frozen samples (three PIVKA-II-positive HCC samples and four control specimens). The mechanism of PIVKA-II production in HCC is still unclear, but it is not caused by mutation in the prothrombin gene.

Amino Acid Sequence↗

Neuropeptide signal transduction in lung cancer: clinical implications of bradykinin sensitivity and overall heterogeneity.

To define the role of neuropeptides in lung cancer biology, we evaluated the effect of seven peptide classes on signal transduction and growth in human lung and breast cancer cell lines. Flow cytometric methods were used to quantitate the calcium response in individual cells produced by these peptides alone or in combination. The effects on growth were assessed by [3H]thymidine, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, and soft agarose colony assays. All lung cancer cells demonstrated calcium responses to one or more peptides with classic small cell lines displaying the greatest responsiveness, followed by variant small cell lines and non-small cell lines. Breast cancer cell lines demonstrated little or no response. There was great variability in the magnitude of calcium response and pattern of response between lung cancer cell lines to individual neuropeptides. Bradykinin was the most potent peptide and produced responses in the highest fraction of lung cancer cell lines. Combinations of peptides produced greater intracellular calcium release than the single peptides, although in less than a quantitatively additive manner. Each peptide produced a refractory period which was peptide class specific. The growth stimulating effects of these neuropeptides were absent or small in magnitude and did not correlate with calcium signal transduction. These results imply that lung cancer cells display a wide sensitivity to neuropeptides but in a very heterogeneous manner. Knowledge of this heterogeneity should be incorporated into the design of antitumor strategies based on this autocrine pathway.

Bombesin↗

Inhibitory influence of a new steroidal anti-androgen, TZP-4238, on prostatic hyperplasia in the beagle dog.

The effect of a synthetic steroidal anti-androgen, TZP-4238, on spontaneous benign prostatic hyperplasia (BPH) in dogs was investigated. Old male beagle dogs (5-9 years old) were divided into three experimental groups. Group 1 consisted of BPH controls. Groups 2 and 3 received TZP-4238 0.1 mg/kg/day and chlormadinone acetate (CMA) 0.3 mg/kg/day p.o., respectively, for 5 months. In group 1, glandular hyperplasia of the prostate was clearly detected. In contrast, TZP-4238 (Group 2) or CMA (Group 3) produced marked atrophy of the glandular epithelium. In addition, a histopathological study showed that TZP-4238 or CMA medication for 5 months exerted no effect on the testes and the pituitary luteinizing hormone (LH) cells. Therefore, it is suggested that TZP-4238 (0.1 mg/kg) or CMA (0.3 mg/kg) causes regression of spontaneous canine BPH without any histopathological effects on the testes and pituitary LH cells. However, slightly decreased serum testosterone levels were found in TZP-4238-treated animals, due apparently to a direct and/or indirect effect on the testes. Thus, it is suggested that a marginal antigonadotrophic effect cannot be excluded. It is concluded that TZP-4238 is a potent anti-androgen for the treatment of spontaneous canine BPH, without any negative influence on the function of the testes and the pituitary LH cells.

Animals↗

The effects of new steroidal anti-androgen, TZP-4238, and chlormadinone acetate on the pituitary, prostate and adrenal gland of the rat: histopathological and immunocytochemical studies.

The atrophic effects of a synthetic steroidal anti-androgen, TZP-4238, on the pituitary, prostate and adrenal gland of rats were investigated. Male Sprague-Dawley rats were divided into three experimental groups. Group 1 consisted of controls. Groups 2 and 3 received chlormadinone acetate (CMA) 50 mg/kg/day and TZP-4238 10 mg/kg/day p.o., respectively, for 3 weeks. CMA (Group 2) produced marked atrophy of the prostate. Furthermore, CMA caused marked atrophy of the adrenal gland. Histopathologically, the remarkable atrophy was observed in the adrenal cortical cells of zonae fasciculata and reticularis. The most striking ultrastructural alterations were noted in the mitochondria. In addition, intramitochondrial localization of glutathione-peroxidase (GSH-PO) which effectively reduces the lipid peroxides, was less than that in the controls. In the anterior pituitary gland, CMA induced a reduction in the size of ACTH cells. TZP-4238 (Group 3) produced marked atrophy of the prostate. However, TZP-4238 exerted no effect on the adrenal gland or anterior pituitary ACTH cells. In addition, the present histopathological study showed that TZP-4238 or CMA exerted no effect on the testes or anterior pituitary LH cells. Therefore, it is suggested that TZP-4238 causes atrophy of the prostate without any significant histopathological changes in the adrenal glands or anterior pituitary ACTH cells under the present experimental conditions. We further speculated that TZP-4238 had a more potent anti-prostatic effect than CMA and TZP-4238 had a less inhibitory influence than CMA on the pituitary-adrenal axis.

Adrenal Glands↗

[Improvement of pituitary homograft under the kidney capsule in mice].

The model of hyperprolactinemia induced by pituitary homografts under the kidney capsule has been used mainly in the field of reproductive physiology. The authors report an improved method for pituitary grafting in mice. The procedure was as follows: 1. The male pituitary glands with normal saline were aspirated into a polyethylene tube. 2. Two incisions were made in the kidney capsule. 3. The polyethylene tube with pituitary glands was inserted via a large incision. 4. Blowing air into the tube, the pituitary glands were left under the kidney capsule and normal saline streamed out of a small incision. Using this method, all pituitary grafted mice became pseudopregnant.

Animals↗