Search PubMed⌕ Search

Biomedical subjects

H Kawashima

Publications and source records attributed to H Kawashima.

At least 217 records · Page 12Linked to original sources

Suppression of inflammatory responses to 12-O-tetradecanoyl-phorbol-13-acetate and carrageenin by YM-26734, a selective inhibitor of extracellular group II phospholipase A2.

1. YM-26734 [4-(3,5-didodecanoyl-2,4,6-trihydroxyphenyl)-7-hydroxy-2-(4-hydroxyph eny l) chroman] dose-dependently inhibited the activities of extracellular phospholipase A2 (PLA2): rabbit platelet-derived group II and porcine pancreas-derived group I PLA2, with IC50 values of 0.085 (0.056-0.129, n = 5) and 6.8 (5.0-9.6, n = 5) microM, respectively. 2. In contrast, YM-26734 did not reduce the activity of intracellular PLA2 prepared from mouse macrophages, which preferentially hydrolyzed arachidonoyl phospholipids at concentrations up to 50 microM. YM-26734 also showed no effect against either sheep seminal vesicle cyclo-oxygenase or rat leukocyte 5-lipoxygenase. 3. Linewater-Burk analysis showed that YM-26567-1 behaved as a competitive inhibitor of group II PLA2 derived from rabbit platelets, with a Ki value of 48 nM. 4. In mice, YM-26734 inhibited 12-O-tetradecanoylphorbol-13-acetate (TPA, 1 microgram/ear)-induced ear oedema in a dose-dependent manner, with ED50 values of 45 (30-67) micrograms/ear (n = 5) and 11 (4-32) mg kg-1, i.v. (n = 5), but did not decrease arachidonic acid (4 mg/ear)-induced ear oedema at 1 mg/ear and 30 mg kg-1, i.v. 5. In rats, the accumulation of exudate fluids and leukocytes in the pleural cavity in response to carrageenin injection (2 mg) was significantly less in a group treated with YM-26734 (20 mg kg-1, i.v.) than in the control group (0.43 +/- 0.02 vs 0.59 +/- 0.03 g per cavity and 3.8 +/- 0.2 vs 4.9 +/- 0.3 x 10(7) cells per cavity, respectively; n = 5). 6. These results suggest that YM-26734 is a potent and competitive inhibitor of extracellular PLA2 with selectivity for group II PLA2, and that the inhibition of group II enzymes activity may cause the suppression of inflammatory responses to TPA and carrageenin.

Animals↗

[Acute mixed lineage leukemia showing resistance to AML and ALL therapy].

A 51-year-old female was who admitted complaining of cough and slight fever and lower limb petechia. The laboratory examinations revealed leukocytosis (49,400/microliters) with blasts (71%) in the peripheral blood. The NCC was 30 x 10(4)/microliters with 84.8% blasts in the bone marrow. Myeloperoxidase staining was positive for 6% of blasts. Auer rods were not seen in some blasts. Thus, acute myeloblastic leukemia (M1) was diagnosed according to FAB classification. In the peripheral blood, 43.3% of blasts expressed CD19, 10.3% of blasts expressed CD20 and 55.6% of blasts expressed CD33 on admission. Though she received two courses of DCMP according to the DCMP-85 protocol, and one combined course of mitoxantrone, etoposide, and Ara-C. The NCC was 20.0 x 10(4)/microliters with 70% blasts in the bone marrow. CD19 was expressed by 72.4% of blasts and 35.0% expressed CD20. The ALL-90 protocol was started, but remission was not achieved. Thus this case was considered to be acute mixed lineage leukemia.

Antigens, CD↗

Establishment of HIV-1-producing cells from peripheral mononuclear cells cultured with normal human serum.

Normal human serum (NHS) contributed to the establishment of cells producing HIV-1 under the conditions of coculture of peripheral mononuclear cells (PMC) from HIV-1 seropositive patients and of PHA-prestimulated or -non-stimulated PMC from seronegative healthy donors. No addition of IL-2 and Polybrene was necessary. Since, in the case 90101, the mitochondrial displacement-loop DNA showed identical sequences in the established cells and the HIV-1 seropositive patient's cells, it can be asserted that the HIV-1-producing cells originated from the patient. These cells are still releasing HIV-1-virion more than one year after their establishment.

Adult↗

[A case of renal cell carcinoma associated with von Hippel-Lindau disease].

We report a 27-year-old male with renal cell carcinoma associated with von Hippel-Lindau disease. The patient was admitted to the department of internal medicine because of pneumothorax. At that time liver and pancreatic cyst, and renal tumor were found incidentally on the abdominal computerized tomography. He had a history of cerebellar hemangioblastoma, which had been removed in 1988. Angiography revealed multiple hypervascular tumors in the right kidney. We performed right radical nephrectomy, and histopathology showed clear cell type of renal cell carcinoma.

Adult↗

[Ureteritis cystica diagnosed by biopsy under a ureteroscope: a case report].

We report a case diagnosed as ureteritis cystica by ureteroscopic examination. A 65-year-old female was admitted to our hospital for a thorough examination of anasarca and reduction of renal function. Computerized tomography showed atrophy in the right kidney and hydronephrosis in the left kidney. Nephrostomy was performed and antegrade pyelography showed ureteral stenosis. For the purpose of accurate diagnosis of ureteral stenosis, ureteroscopy and biopsy were performed. Histological examination revealed ureteritis cystica. Fifty-five cases including this case, reported in Japan, are herein reviewed.

Aged↗

Lack of blood pressure increase with age in long-term hospitalised patients with the sequelae of acute carbon monoxide poisoning.

We investigated retrospectively age-related changes in BP and body composition in 44 men who had been hospitalised for up to 26 yrs as a result of carbon monoxide poisoning. The average age of the patients was 42.5 +/- 9.1 (SD) yrs on the first admission. The average period of hospitalisation was 19.5 +/- 5.9 yrs. The patients received mild salt restriction and daily exercise in the hospital. During the study period, nine patients developed hypertension and received antihypertensive treatment. The prevalence of hypertension in our study population was 205 per 1,000 and the incidence of hypertension (DBP > 95 mmHg) was 9.1 per 100 person years. Both prevalence and incidence appear to be lower than those in the general population. In the other 35 patients BP significantly decreased from 127.5 +/- 10.0 mmHg (SBP)/79.8 +/- 8.1 mmHg (DBP) at the time of the first examination to 120.8 +/- 16.3 mmHg/71.9 +/- 10.3 mmHg on the last examination. Body mass index did not change significantly during the study period. Hypertensive cardiovascular death was not observed throughout the study period. In conclusion, controlled diet, salt intake, physical exercise and isolation from the social stimuli achieved by a long-term hospitalisation may prevent increased BP and cardiovascular death.

Acute Disease↗

Inhibition of in vitro T cell activation by corneal endothelial cells.

Cells and tissues of the anterior uvea and aqueous humor express activities which inhibit immune responses. These activities include soluble factors such as TGF-beta and uncharacterized cell surface interactions. Relatively little is known regarding the immunologic activities of corneal endothelium, despite its potentially important role in contributing to the immune privilege of the anterior chamber and the high success rate of corneal transplantation. In this report, in vitro studies of cultured rat corneal endothelial (CE) cells were done using S-antigen-specific LEW rat T cell lines, or S-antigen-specific T cell hybridomas, to examine the immunologic capabilities of CE cells. Monolayers of LEW rat CE cells were unable to present antigen or a mitogen, Con A, to T cell lines or hybridomas as assessed by the lack of a proliferative response or IL-2 secretion. Furthermore, the CE cells exerted a potent inhibitory effect when added to in vitro proliferation assays of T cell lines stimulated with antigen or Con A. When T cells were preactivated on conventional antigen presenting cells and then transferred to wells containing CE cells, their proliferation was not inhibited. Although CE cells inhibited activation of T cell lines and hybridomas, they did not inhibit the growth of T cell hybridomas or CTLL cells, nor did the CE cells adversely affect the viability of resting T cells cultured on CE monolayers. The inhibitory effect was reversible as preincubation of T cells on CE cells for up to 6 days followed by washes restored T cell responsiveness when assayed on splenocytes. The inability to stimulate proliferative responses was not affected by preincubation of the CE cells with lymphokines which increase MHC antigen expression. The inhibition observed in these assays was not MHC-restricted as CE cells from both LEW and BN rats were equally inhibitory. CE cells from rabbits and cats were also potent inhibitors of T cell activation, suggesting that the mechanism is evolutionarily conserved. The mechanism of inhibition of CE cells is unknown at this time.

Amino Acid Sequence↗

Purification and NH2-terminal amino acid sequences of human and rat kidney fatty acid omega-hydroxylases.

A cytochrome P-450 (P-450), designated P-450HK omega, has been isolated and purified from human kidney microsomes to a specific content of 13 nmoles of P-450/mg of protein. P-450HK omega showed an apparent molecular weight of 52,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Absolute spectra of the oxidized form indicated that this P-450 was largely in the low-spin state and partly in the high-spin state. It catalyzed the omega- and (omega-1)-hydroxylation of fatty acids such as laurate, myristate, and palmitate, with no activity toward prostaglandin A1, benzphetamine, 7-ethoxycoumarin, or 7-ethoxyresorufin. The first 35 NH2-terminal amino acid sequence of P-450HK omega had about 70% homology with those of rabbit kidney fatty acid omega-hydroxylases of the P-450 IVA gene subfamily, P-450ka-1, P-450ka-2, and P-450kd, except for four undetermined residues. Moreover, Western blot and immuno-inhibition studies showed that P-450HK omega reacted with an antibody against the rabbit kidney fatty acid omega-hydroxylase. The results suggest that P-450HK omega is a member of the same P-450 gene family (IVA subfamily) as the rabbit enzymes. In addition, the terminal sequence of P-450HK omega also showed 54% homology with that of P-450k-2, a fatty acid omega-hydroxylase from rat kidney microsomes. To our knowledge, this is the first time that a P-450 specific for fatty acid omega-hydroxylase activity has been isolated to homogeneity from human tissues.

Adult↗

Inhibitory effect of curcumin on fatty acid desaturation in Mortierella alpina 1S-4 and rat liver microsomes.

An extract of rhizomes of Curcuma longta L. (turmeric) inhibited the desaturation of dihomo-gamma-linolenic acid (DGLA) in the arachidonic acid (AA) producing fungus Mortierella alpina 1S-4. The factor responsible for this phenomenon was isolated and identified as curcumin (diferuloyl methane). Mycelial DGLA levels increased about two-fold (22.3 mg/g dry weight) with a concomitant decrease in AA levels when the fungus was cultivated with curcumin. The 50% inhibitory concentration against delta 5 desaturase was 27.2 microM. Curcumin also inhibited rat liver microsomal delta 5 and delta 6 desaturases.

8,11,14-Eicosatrienoic Acid↗

A simple method to assess osteoclast-mediated bone resorption using unfractionated bone cells.

To determine osteoclastic bone resorption we established a simple assay system in which unfractionated cells obtained from femora of 13-day-old mice were cultured on a dentine slice and the number of osteoclasts and their induced pit area on the slices were measured. When the bone cells (1 x 10(5) cells/dentine slice) were cultured in the presence of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] or human parathyroid hormone (hPTH) for 4 days, at which time newly-formed osteoclasts were not detected, the pit area was dose-dependently increased, being a 4.3- or 4.1-fold respective increase over the control at a 10(-8) M concentration of hormones. Chick calcitonin (cCT) inhibited the osteoclastic bone resorption induced by either of these hormones. cCT alone also suppressed the bone resorption by the cells (3 x 10(5) cells/dentine slice). These findings indicate that 1,25(OH)2D3 or hPTH may mainly activate pre-existing osteoclasts, resulting in increased bone resorption, and that cCT may suppress this osteoclastic activity. When 1,25(OH)2D3 or hPTH was added to the cells pre-cultured in factor-free medium for 6 days, at which time pre-existing osteoclasts had almost degenerated, new osteoclasts were formed, resulting in an increase in pit formation. Thus this system is a useful method which could more sensitively evaluate the effects of hormones or factors on osteoclast formation and activation than other previous systems.

Animals↗

A simple method for the release of asparagine-linked oligosaccharides from a glycoprotein purified by SDS-polyacrylamide gel electrophoresis.

A simple method for the release of oligosaccharides from glycoproteins separated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) has been developed. Asialo-alpha 1-acid glycoprotein, which was tritiated at the nonreducing terminal D-galactopyranosyl residue by reduction with sodium borotritide after incubation with D-galactose oxidase, was used as a model compound. After electrophoretic separation of the glycoprotein, oligosaccharides were released by the use of a gas-phase hydrazinolysis apparatus. In the first method, the gel was stained with Coomassie Blue and the glycoprotein together with the gel was directly subjected to gas-phase hydrazinolysis after removal of water in a P2O5 desiccator. The recovery of released oligosaccharides was 25.9 +/- 2.4%, based on the amount of the glycoprotein loaded on the gel within the range of 3.5-28.5 micrograms. In the second method, the glycoprotein was electroblotted onto an Immobilon transfer membrane and was visualized by staining with Coomassie Blue. A small piece of the membrane with the corresponding band was cut out, dried in a desiccator and subjected to gas-phase hydrazinolysis. In this case, the recovery of released oligosaccharides was 15.2 +/- 1.0%. These procedures, particularly the first one, should be widely applicable for the isolation of oligosaccharides from glycoproteins separated by SDS-PAGE.

Animals↗

Structure-activity relationships in human interleukin-1 alpha: identification of key residues for expression of biological activities.

To identify the sites important for the different biological activities of human interleukin-1 alpha (hIL-1 alpha), 56 single-amino acid-substituted mutants of hIL-1 alpha were produced in Escherichia coli using site-directed mutagenesis, and were examined for their biological activities such as mouse lymphocyte activating factor activity (LAF activity), cytostatic activity against human melanoma cells A-375 (A375 activity) and prostaglandin E2 (PGE2) inducing activity in human osteosarcoma cells MG-63 (PEI activity). Two amino acid residues, Asp26 and Asp151, were found to be important for these activities. The replacement of Asp26 by Val caused a decrease in LAF and PEI activities by one or two orders of magnitude and a slight decrease in A375 activity. The Tyr or Phe substitution for Asp151 caused decreases in LAF and A375 activities by one or two orders of magnitude and complete loss of PEI activity. The change from Asp151 to Lys or Arg resulted in marked decrease in LAF activity and complete loss of A375 and PEI activities. Since Asp26 and Asp151 are close to each other in the three-dimensional structure, the region involving these amino acids seems to be important for the biological activities of hIL-1 alpha.

Amino Acid Sequence↗

Production of Dihomo-gamma-Linolenic Acid by a Delta5-Desaturase-Defective Mutant of Mortierella alpina 1S-4.

A mutant, which has low Delta5-desaturase activity, of an arachidonic acid-producing fungus, Mortierella alpina 1S-4, was shown to be a novel potent producer of dihomo-gamma-linolenic acid (DHGA). On submerged culture under optimal conditions for 6 days at 28 degrees C in a 10-liter fermentor, the mutant produced 3.2 g of DHGA per liter of culture broth (123 mg/g of dry mycelia), which accounted for 23.4% of the total mycelial fatty acids. Mycelial arachidonic acid amounted to only 19 mg/g of dry mycelia (0.5 g/liter of culture broth), which accounted for 3.7% of the total mycelial fatty acids. The other major mycelial fatty acids were palmitic acid (11.0%), stearic acid (12.8%), oleic acid (22.7%), linoleic acid (8.9%), gamma-linolenic acid (6.5%), and lignoceric acid (7.8%). More than 97 mol% of the DHGA produced was found in the triglyceride fraction irrespective of the growth temperature employed (12 to 28 degrees C).

Journal Article↗

[The use of a metallic stent in 32 patients with benign prostatic hypertrophy. Preliminary results and 3 months of follow-up].

A prospective noncontrolled study of the safety and potential efficacy of the metallic stent was performed on 32 patients with benign prostatic hypertrophy. Mean age was 76.6 years (range, 56-98 years), and mean prostatic volume was 24.2 cm3. The patients were selected on the basis of a quantitative symptom score (QSS), uroflowmetry measurements, and residual urine volume (RU). Nineteen patients had urinary retention and remaining 13 patients had moderate symptoms and signs of prostatism. Placing the stent was successfully done in 31 patients (97%). It took 15 minutes to place the stent using transabdominal and/or endorectal sonography. After 3 months, 27 patients (87%) showed improved QSS. In patients with dysuria, maximum flow rate (MFR) and RU before treatment were 6.9 +/- 1.7 ml/sec and 112.3 +/- 61.8 ml, respectively. After treatment, they improved to 12.3 +/- 2.7 ml/sec and 12.7 +/- 6.7 ml, respectively. On the other hand, all patients who had urinary retention were able to urinate just after treatment, and MFR and RU were 12.9 +/- 3.6 ml/sec and 24.4 +/- 43.3 ml, respectively. Evaluation on the basis of improvement in MFR and reduction in RU showed that the stent was effective in 71% of total patients (22 out of 31 patients), 94% of the patients with urinary retention (17 out of 18 patients). The overall clinical efficacy of this stent was 68% (21 patients). There were no major complications such as urge incontinence and urinary tract infection during follow-up. Although proximal migration of the stent was observed in 6 patients, the stent could be taken out and replaced in 4 patients. From the above results, we conclude that the metallic stent is useful for the treatment of prostatism and urinary retention.

Aged↗