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Biomedical subjects

A Ohara

Publications and source records attributed to A Ohara.

At least 73 records · Page 4Linked to original sources

Production of phytochelatins in Polygonum cuspidatum on exposure to copper but not to zinc.

We studied cellular resistance to copper of plant cells Polygonum cuspidatum. When callus of P. cuspidatum was incubated on medium containing 100 microM cupric sulfate, the callus grew as well as the control callus did. The copper content of the callus, however, was elevated to a similar level of the medium. When cell extracts of callus exposed to 100 microM cupric sulfate were fractionated by gel filtration chromatography, a specific copper peak was eluted at the region of molecular weights between 4000 and 1000. Since an appearance of the copper-containing materials was inhibited by buthionine sulfoximine and the partially purified copper-containing materials contained only three amino acids: glutamic acid, glycine and cystine, the materials were supposed to be gamma-glutamyl peptides phytochelatins. Callus of P. cuspidatum synthesized phytochelatins from 50 microM cupric sulfate and maximally at 100-150 microM cupric sulfate. When induction of phytochelatins by another heavy metal, zinc, was analyzed, the callus, however, did not synthesize phytochelatins on exposure to zinc sulfate up to 1 mM. These findings suggested that phytochelatins were required for resistance to copper but probably not to zinc in the plant cells.

Cells, Cultured↗

Characterization of cationic acid phosphatase isozyme from rat liver mitochondria.

Acid phosphatase isozyme was highly purified from rat liver mitochondrial fraction. The enzyme showed an isoelectric point value of above 9.5 on isoelectric focusing, and the apparent molecular weight was estimated to be 32000 by Sephadex G-100 gel filtration or 16000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme catalyzed the hydrolysis of adenosine 5'-triphosphate, adenosine 5'-diphosphate, thiamine pyrophosphate, inorganic pyrophosphate, and phosphoprotein such as casein and phosvitin, but not of several phosphomonoesters, except for p-nitrophenyl phosphate and o-phosphotyrosine. The enzyme was not inhibited by L-(+)-tartrate, and was significantly activated by Fe2+ and reducing agents such as ascorbic acid, L-cysteine,and dithiothreitol. The enzyme was found to be distributed in various rat tissues including liver, spleen, kidney, small intestine, lung, stomach, brain and heart, but not in skeletal muscle.

Acid Phosphatase↗

[Determination of acetone, methanol, and methyl ethyl ketone in urine using head-space gas chromatography (HS.GC)].

Using HS.GC, We have succeeded in simultaneous determination of Ac, MeOH and MEK in urine without any complicated pretreatment or correction by internal standard. Moreover, in order to lower the detection limits of these materials, study was made on the salting out effect using 14 kinds of salts. As pretreatment, 2.0 ml of urine, 3.0 g of sodium sulfate and small sized magnetic stirrer are put into vial, which is sealed by septum. This is then heated for 10 min in warm bath of 50 degrees C. In order to dissolve the added salts as much as possible, the specimen is stirred by the stirrer. After cooling the liquid to room temperature, the specimen is analysed by HS.GC. The results showed that sodium sulfate was excellent synthetically. 1) Using the urine of workers not exposed to organic solvents three kinds of urine having specific gravity of 1.010, 1.024 and 1.034 were prepared and mixed standard organic solvents (Ac, MeOH and MEK) were added. Recovery percentages and coefficients of variation were calculated. The results showed that recovery percentages ranged from 92.0 to 101.7% and coefficients of variation from 0.2 to 4.6%. 2) The regression equations of standard curves were satisfactory with y = 9053x - 200(r = 0.999, n = 12) for Ac, y = 801x - 400 (r = 0.999, n = 12) for MeOH, and y = 15488x - 277 (r = 0.999, n = 12) for MEK. 3) The detection limits calculated by IUPAC formula were 0.0092 mg/l for Ac, 0.11 mg/l for MeOH and 0.0063 mg/l for MEK. These results indicated that this method is superior to other methods because the pretreatment is very simple, specificity is excellent, analysis by standard curves is possible, and this method is not affected by specific gravity of the urine.

Acetone↗

Reduction of adverse effects of indomethacin by anti-allergic drugs in rat stomachs.

This study was designed to elucidate the role of leukotrienes (LT) in indomethacin-induced gastric ulcers in relation to effects of clinically available anti-allergic drugs, azelastine (Azeptin, CAS 58581-89-8) and oxatomide (CAS 60607-34-3). Azelastine (2 mg/kg) or oxatomide (30 mg/kg) was administered intragastrically once 15 min before intragastrical administration of 12 mg/kg of indomethacin. Mucosal prostaglandins (PGs) and LTs were measured by high-performance liquid chromatography (HPLC). In the control rats, 4 kinds of PGs, i.e., 6-keto-PGF1 alpha, PGF2 alpha, PGE2 and PGD2, were detected in gastric mucosa, but no LT was detected. Administration of indomethacin caused severe gastric ulcers (lesion scores; 31.2 +/- 11.1 mm), and all prostaglandins were diminished completely. In contrast, LTC4 and LTD4 (peptide-LTs) were detected and the sum of their levels was 18.5 +/- 7.1 ng/g tissue. Irrespective of indomethacin administration, lesion scores were remarkably reduced in rats treated with azelastine or oxatomide (17.0 +/- 8.1 and 16.0 +/- 8.0, respectively). Azelastine and oxatomide treatment did not improve mucosal PG levels, however, both drugs reduced significantly increases in peptide-LT level, 9.3 +/- 4.6 and 8.6 +/- 4.4, respectively. These results suggest that increases in mucosal LT levels are also involved in the formation of indomethacin-induced gastric ulcers. Combined therapy using anti-allergic drugs and non-steroidal anti-inflammatory drugs might be recommended.

Animals↗

[Hematopoietic failure of a healthy child with human parvovirus B19 infection--studies on the hematopoiesis].

An 8 year-old boy, who had been healthy since his birth, presented fever, ecchymosis and nasal bleeding. His peripheral blood picture showed pancytopenia and bone marrow showed erythroblastopenia, but these findings recovered spontaneously in two weeks. The diagnosis of human parvovirus B19 (HPVB19) infection was made by the presence of anti HPVB19 IgM antibodies in the sera. In order to investigate the mechanisms of erythroblastopenia and pancytopenia, we performed in vitro hematopoietic cell culture in both acute phase and the convalescent phase (3 months after the onset). In the acute phase, the formation of CFU-E colony was inhibited but that of BFU-E, CFU-G, GEMM colony was not inhibited. Unexpectedly, HPVB19 DNA was detected in CFU-GEMM colony cells with the PCR technique. It is suggested that HPVB19 infected not only to erythroid progenitor cells but also to other progenitors.

Cells, Cultured↗

Diagnosis of paraaortic and pelvic lymph node metastasis of gynecologic malignant tumors by ultrasound-guided percutaneous fine-needle aspiration biopsy.

Transabdominal ultrasonography was done in patients with gynecologic malignancies to detect paraaortic and pelvic lymphadenopathy. Of the 78 patients who underwent ultrasonic examination of pelvic lymph nodes, pelvic lymphadenopathy was detected in 24, of whom ultrasound-guided percutaneous fine-needle aspiration biopsies (FNAB) were done in 16. Of these 16 patients, the results of 13 biopsies were positive. Of the 41 patients who underwent ultrasonographic examination of the paraaortic lymph nodes, 27 had paraaortic lymphadenopathy. Ultrasound-guided percutaneous FNAB were done in 26 of these 27 patients, with positive findings in 21 of the 26. It was concluded that ultrasound-guided percutaneous FNAB is useful in detecting metastatic lymph nodes in gynecologic malignancies.

Abdominal Neoplasms↗

Cyclic GMP-activated channel activity in renal epithelial cells (A6).

We studied the effects of guanosine 3',5'-cyclic monophosphate (cGMP) and nitroprusside on ion channels in the apical membrane of confluent A6 cells (a distal nephron cell line) cultured on permeable supports for 10-14 days using patch clamp techniques. In cell-attached patches without any detectable channel activity, activity of a non-selective cation channel with a single-channel conductance of 1 pS was observed after adding nitroprusside. After adding cGMP to the cytosolic surface of inside-out patches with no detectable channel activity, we observed single channel activity similar to the channel observed after adding nitroprusside. These observations imply that nitroprusside activates a non-selective cation channel with small single channel conductance (1 pS) via an increase in cGMP which activates the channel.

Cell Membrane↗

Activation of blood coagulation and fibrinolysis in vibration syndrome.

The pathophysiology of peripheral circulatory disturbance in patients presenting with vibration syndrome was studied from the viewpoint of blood coagulation. Plasma levels of fibronectin (FN), vitronectin (VN), thrombin-antithrombin III complex (TAT), and alpha 2-plasmin inhibitor-plasmin complex (PIC) were measured in 23 subjects who showed no evidence of vibration-induced white finger [VWF(-) group] and in 24 patients who presented with VWF [VWF(+) group]. In the VWF(-) group, plasma FN concentrations were elevated but plasma TAT and PIC levels were within the normal ranges. In the VWF(+) group, plasma FN concentrations were normal but plasma TAT and PIC levels were significantly elevated. In both groups, plasma VN concentrations were similar to those in normal controls. For purposes of comparison, 32 patients presenting with diabetes mellitus were also studied. They were divided into 2 groups, 13 subjects who showed no evidence of angiopathy [complication(-) group] and 19 patients who presented with angiopathy [complication(+) group]. In the complication(+) group, plasma TAT and PIC concentrations were significantly elevated, as in the VWF(+) group. These results suggest that in vibration syndrome, vibration, cold stimulus, or other factors first injure the vascular endothelium, resulting in a rise in plasma FN, and that in the VWF(+) group, augmentation of coagulation and fibrinolysis induces a state of compensated disseminated intravascular coagulation (DIC).

Adult↗

Generation of B lymphocytes from a single hemopoietic progenitor cell in vitro.

The first stages of the pathway by which lymphocytes differentiate from hemopoietic stem cells were studied at a clonal level. When 211 interleukin 3 (IL-3)-induced blast colonies shown to be capable of differentiating into a variety of hemopoietic cells were individually transferred into wells containing a monolayer of stromal cells, growth in granulocyte, macrophage, megakaryocyte, or mast cell lineages was observed in 192 wells. In seven of these 192 wells, lymphoid cell growth also was seen. The lymphoid cells were proved to be B lymphocytes by phenotype and immunoglobulin gene rearrangement analyses and by demonstration of surface expression of IgM. The clonal origin of myeloid and B lymphocyte lineage cells was further confirmed by the generation of both myeloid and B lymphoid cells in the same well following FACS clone-sorting of IL-3 induced blast cells. These results provide in vitro evidence that cells of B lymphoid and myeloid lineage can originate clonally from single primitive hemopoietic stem cells.

Animals↗

Serotonin-induced decrease of duodenal mucosal blood flow plus acid load produces duodenal mucosal lesion in rats.

To clarify the role of duodenal mucosal blood flow in the genesis of duodenal mucosal lesions, a single dose of serotonin, 20 mg/kg, was administered subcutaneously to rats. Serotonin significantly decreased both the duodenal mucosal blood flow and the output of gastric acid and pepsin. However, mild duodenal erosions were observed in only 2 of the 9 rats receiving that agent. The repeated intragastric administration of 1 ml of 0.1 N HCl hourly for 6 h failed to induce duodenal mucosal lesions. On the other hand, the administration of both serotonin and repeated doses of HCl produced marked duodenal mucosal lesions in all rats. It is concluded, therefore, that one of the factors responsible for the genesis of duodenal mucosal lesions is a decrease of duodenal mucosal blood flow combined with an acid load against the duodenal mucosa.

Animals↗

[The effect of copper on o- and m-tyrosine content in the serum of guinea pigs].

The effect of copper on the concentrations of o- and m-tyrosines in the serum of guinea pigs was studied in vivo. When guinea pigs were fed the normal diet and 0.1% CuSO4 solution as a drinking water for 13 d, the concentrations of o- and m-tyrosines in the serum increased more significantly than that of guinea pigs fed normal diet and water without copper. Phenylalanine hydroxylase in the liver and kidney, and tyrosine hydroxylase in the brain and adrenal were not activated by the administration of copper to guinea pigs. The administration of copper caused an abnormal accumulation of copper in the liver, but not in the kidney, adrenal and brain, and significantly depressed the ascorbic acid content in various organs including the liver, kidney, brain and adrenal. The results obtained suggest that o- and m-tyrosines may be also formed nonenzymatically in vivo, in addition to the formation by the participation of enzymes such as phenylalanine and tyrosine hydroxylases.

Animals↗

[Determination of trichloroacetic acid and trichloroethanol by head-space gas chromatography (HS.GC)].

Simultaneous determination of trichloroacetic acid (TCA) and trichloroethanol (TCE) in urine was made using head-space gas chromatography (HSGC). TCA was analyzed after methyl esterification by methanol, and TCE was measured with decomposition of conjugation adding sulfuric acid. (1) As preliminary treatment, 0.1 ml of urine and 0.6 ml of esterizer (pure water: sulfuric acid: methanol = 6:5:1, V/V/V) were mixed in a sample vial, which was sealed a septum. This was analyzed in HSGC. (2) By this method, the recovery, standard deviation and coefficient of variation of TCA were 95.7-104.3%, 0.001-0.783 mg/l and 0.8-4.0%, respectively, while those of TCE were 98.6-102.5%, 0.024-1.603 mg/l and 0.8-4.0%, respectively. (3) Calibration curves were linear up to 30 mg/l for TCA (y = 1.838x + 0.023, r = 0.999, n = 8) and 60 mg/l for TCE (y = 0.963x + 0.072, r = 0.999, n = 8). (4) A high correlation between HSGC and alkaline pyridine spectrophotometry was found for both TTC (TCA + TCE = TTC), (y = 0.917x - 3.08, r = 0.980, n = 100, p less than 0.001) and TCA (y = 0.891x - 2.36, r = 0.928, n = 100, p less than 0.001). The values for TCA and TCE obtained with HSGC were lower than those obtained with spectrophotometry. (5) The limits of the detection obtained with this method were 0.002 mg/l for TCA and 0.005 mg/l for TCE according to the formula recommended by International Union of Pure and Applied Chemistry (IUPAC). These results indicate that this simple method is accurate and useful in simultaneous detection of TCA and TCE.

Chromatography, Gas↗

Effects of two benzimidazoles as proton pump inhibitors on water immersion stress-induced gastric ulcers in rats.

This study was designed to clarify effects of proton pump inhibitors, E-3810(2-([4-(3-methoxypropoxy)-3methylpyridine-2-yl]methyl- sulfinyl)-1H-benzimidazol sodium salt) and omeprazole (CAS 73590-58-6), on water immersion stress-induced gastric ulcers in relation to mucosal prostaglandins (PGs) and leukotrienes (LTs). Mucosal PGs were measured by high performance liquid chromatography (HPLC). In untreated rats, 4 kinds of PGs, i.e., 6-keto-PGF1 alpha, PGF2 alpha, PGE2 and PGD2 were detected in gastric mucosa, but no LTs were detected. Water immersion stress for 6 h caused severe hemorrhagic lesions in the fundic portion and, concomitantly, significant decreases in mucosal PG levels. On the other hand, LTC4 and LTD4 were detected after 6 h stress treatment, though LTB4 and LTE4 were not detected throughout the experiments. Peptide-LT (the sum of LTC4 and LTD4) levels were 23.5 +/- 3.2 ng/g tissue 6 h after water immersion stress. Administration of 20 mg/kg of E-3810 or 20 mg/kg of omeprazole mitigated gastric lesions and increases in the mucosal peptide-LT levels, but dit not improve the decrease in mucosal PGs.

2-Pyridinylmethylsulfinylbenzimidazoles↗

[Requirements of diagnostic criteria for aplastic anemia in children].

As a general rule, diagnostic criteria of aplastic anemia in children are the same as adult criteria. However, blood counts of normal children show wide age-related variation, therefore we must establish a system of adjustment for diagnosis of aplastic anemia in children. The data of children with aplastic anemia visiting our institutes from 1966 to 1990 were evaluated for this study. RBC below 350 x 10(4)/microliters, WBC below 4,000/microliters or neutrophils below 1,500/microliters, platelets below 8 x 10(4)/microliters, reticulocytes below 4 x 10(4)/microliters and lymphocytes over 60% were seemed to satisfy the diagnostic criteria of aplastic anemia proposed by the Study Group of hemopoietic Disorders sponsored by the Ministry of Health and Welfare of Japan. Fifteen children (4.6%) did not meet these criteria and as such were diagnosed as atypical aplastic anemia. Thirteen of them were in a pre-aplastic state and developed typical aplastic anemia within 6 months to 8 years after the initial diagnosis. Clinical findings of these patients showed the decrease in number of megakaryocytes and committed stem cells in bone marrow. Three of these patients developed acute non-lymphocytic leukemia, and 2 of them were diagnosed as Fanconi's anemias.

Adolescent↗

Effect of cimetidine on the mucosal hydroxyproline content of rat gastric lesions induced by compound 48/80.

Extensive gastric mucosal lesions developed in rats after the administration of compound 48/80 at a dose of 0.75 mg/kg daily for 3 days. The hydroxyproline content of gastric mucosa was significantly increased 4 h after the last injection of compound 48/80, and subsequently decreased as the lesions healed. Treatment with cimetidine (CAS 51481-61-9) at a dose of 50 mg/kg (twice a day for 3 days) did not affect either the development or the healing of the compound 48/80-induced gastric lesions. However, the increase in mucosal hydroxyproline content was significantly reduced by treatment with cimetidine. This fact suggests that cimetidine might have an inhibitory effect on the regeneration of collagen in gastric mucosa.

Animals↗

Role of leukotrienes in the genesis and healing of water immersion stress-induced gastric ulcers in rats.

We investigated the role of leukotrienes (LTs) in the genesis of water immersion stress-induced gastric ulcers. Peptide LTs were detected after 4 h stress (3.7 +/- 0.5 ng/g tissue), although they were not detected after 2 h stress, and considerable amounts (20.3 +/- 2.3 ng/g tissue) were detected after 6 h stress. In contrast, AA-861 (100 mg/kg, p.o.), a 5-lipoxygenase inhibitor, reduced ulcer indices significantly after 6 h stress, although no significant changes were observed after 2 or 4 h stress compared with the control group. Peptide LTs were not detected after 4 h and those detected after 6 h stress were remarkably reduced by AA-861 treatment. The role of LTs in the healing of water immersion stress-induced gastric ulcers was also investigated. Significant ulcer healing was not observed within 24 h after stress but was significantly recovered after 48 h. Peptide LTs decreased time-dependently and 48 h after treatment they were not detected. In the rats treated with AA-861, ulcer indices and peptide LTs levels were remarkably reduced after 12 h, concomitantly. These results suggest that the increase in mucosal peptide LTs might be an inhibitory factor to ulcer healing.

Animals↗

[Treatment and prognosis of pT2 endometrial carcinoma].

Between January, 1966 and December, 1988, 66 patients with surgical-pathological stage II adenocarcinoma of the endometrium who were treated with abdominal simple or radical hysterectomy at the Osaka National Hospital were studied retrospectively. The 5-year survival rate for all patients was 75.7%. In 33 patients adenocarcinoma had invaded the cervical stroma. These patients demonstrated a significantly lower survival (60.0%) than those patients whose involvement was limited to the endocervical gland (91.7%) (p less than 0.05). For all patients, both with endocervical glandular involvement and cervical stromal invasion, radical hysterectomy improved survival rates more than did simple hysterectomy, but the difference was not statistically significant. The factor found to have a significant influence on survival was the maximal depth of invasion of the myometrium (p less than 0.05). The histologic grade, on the other hand, did not appear to influence survival. It is suggested that patients with minimal involvement of the cervix can be satisfactorily treated with simple hysterectomy, while patients with gross cervical spread should be treated with radical hysterectomy.

Adenocarcinoma↗