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Biomedical subjects

N Ueda

Publications and source records attributed to N Ueda.

At least 235 records · Page 13Linked to original sources

Immunocytochemical localization of platelet-type arachidonate 12-lipoxygenase in mouse blood cells.

Arachidonate 12-lipoxygenase is an enzyme that oxygenates the 12 position of arachidonic acid to produce its 12-hydroperoxy derivative. We were interested in the tissue distribution and subcellular localization of the platelet-type 12-lipoxygenase, which is distinguished from the leukocyte type by several criteria. Antiserum was raised in rabbits against purified recombinant arachidonate 12-lipoxygenase of human platelets. When mouse bone marrow cells and lung were immunostained and observed by light and electron microscopy, the positively stained cells were platelets, megakaryocytes, and eosinophils. 12-Lipoxygenase was localized in the cytoplasm of platelets but was hardly detectable in the plasma membrane and intracellular organelles. The enzyme was found in the cytoplasm of immature megakaryocytes with kidney-shaped nuclei and a few profiles of demarcation membranes, as well as in the mature form with well-developed demarcation membranes. These results indicated the expression of 12-lipoxygenase at an early stage in the course of megakaryocytopoiesis.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗

Angiotensin II stimulates interleukin-6 release from cultured mouse mesangial cells.

Interleukin-6 (IL-6) is a multifunctional cytokine exerting a wide variety of biologic responses, including cell proliferation. Recently, IL-6 has been known to play a role in the pathogenesis of mesangial proliferative glomerulonephritis. IL-6 is now recognized as an autocrine growth factor for glomerular mesangial cells, and various inflammatory mediators have been shown to promote IL-6 release from mesangial cells. However, little is known about the noninflammatory stimuli of IL-6 release from mesangial cells. In this study, it was hypothesized that angiotensin II (AngII) is one of the noninflammatory mediators of IL-6 release in mesangial cells, and the effects of AngII on IL-6 release and mRNA expression in cultured mouse mesangial cells (CMMC) were investigated. It was demonstrated that AngII (10(-7) M or higher) caused IL-6 release and mRNA accumulation in CMMC. IL-6 release was detected at 4 h and reached a plateau at 8 h after the addition of AngII, whereas IL-6 mRNA expression peaked at 4 h. The effects of AngII on IL-6 release and gene expression were completely blocked by the AngII receptor type 1 (AT1 receptor) antagonist CV-11974. AngII and IL-6 were both shown to stimulate DNA synthesis in CMMC, and the blockade of IL-6 signaling with anti-IL-6 receptor antibody abolished the enhanced DNA synthesis induced by AngII. These results raise a possibility that the growth-promoting effect of AngII on mesangial cells is at least partially mediated by IL-6 released from mesangial cells.

Angiotensin II↗

Association of angiotensin I-converting enzyme gene polymorphism with susceptibility to antiproteinuric effect of angiotensin I-converting enzyme inhibitors in patients with proteinuria.

The antiproteinuric effect of angiotensin I-converting enzyme (ACE) inhibitors in patients with renal diseases of various origins has been well recognized. However, individual responses regarding the degree of decrease in urinary protein excretion appear to vary considerably. The mechanism underlying this variable response to ACE inhibitors has not been clarified yet. A possible role of ACE gene insertion/deletion (I/D) polymorphism in the responsiveness to antiproteinuric effect of ACE inhibitors is examined. Thirty-six patients with proteinuria (23 men and 13 woman; mean age, 47 +/- 13 yr) were studied. These patients were classified into two groups on the basis of the percent decrease in their urinary protein excretion: the effective group, those with a decrease in proteinuria (18 patients, -64 +/- 19%) and the noneffective group (18 patients, +13 +/- 40%). A 287-base pair (bp) I/D polymorphism in the ACE gene was examined by polymerase chain reaction. The allelic frequencies of the ACE gene were I/D = 0.53/0.47 in the effective group and I/D = 0.81/0.19 in the noneffective group. The difference in the allelic frequencies between the two groups was significant (chi 2 = 6.25, P = 0.0114 < 0.05). Furthermore, the difference in the responsiveness of proteinuria to ACE inhibition between genotype II versus genotype ID + DD was statistically significant (chi 2 = 4.05, P = 0.0442 < 0.05). There was no significant difference between the two groups with regard to initial urinary protein level, blood pressure, renal function, and daily sodium intake. The genetic susceptibility to the antihypertensive effect of ACE inhibitors was also studied, but no significant relation was observed. This study suggests the association of ACE gene I/D polymorphism with the antiproteinuric efficacy of ACE inhibitors in patients with proteinuria.

Adult↗

[A case of cystic fibrosis in a Japanese student].

A 16-year-old boy was admitted to our hospital because of coughing, sputum, and exertional dyspnea. Seven months after birth cystic fibrosis had been diagnosed. The chest roentgenogram on admission showed diffuse reticulonodular shadows and overinflation. Pulmonary function tests revealed obstructive and restrictive impairment. Erythromycin and Lomefloxacin were administered by mouth, and aminoglycosides were administered by inhalation. His symptoms were alleviated, and he is now an outpatient. In Japan, cystic fibrosis is rare, and this patient is extremely rare because he has grown up to be a 16-year-old. In this case, low-dose and long-term erythromycin administration was very effective.

Administration, Inhalation↗

Characterization of arachidonate 12-lipoxygenase found in the liver of mongrel dog and its immunohistochemical localization in neutrophils.

The cytosol fraction of non-parenchymal cells isolated from the liver of adult mongrel dogs converted arachidonic acid to 12S-hydroxy-5,8,10,14-eicosatetraenoic acid. The arachidonate 12-lipoxygenase enzyme reacted with linoleic and alpha- and gamma-linolenic acids as well as arachidonic acid. The enzyme was immunoprecipitable with an antibody against the leukocyte 12-lipoxygenase, but not with an antibody against the platelet 12-lipoxygenase. By immunohistochemical observation with anti-leukocyte 12-lipoxygenase antiserum, hepatocytes, Kupffer cells and sinusoidal endothelial cells were not immunostained, but a large number of neutrophils located in sinusoidal cavities were positively stained. The 12-lipoxygenase activity thus detected may be attributed predominantly to the neutrophils appearing in sinusoidal cavities rather than the non-parenchymal cells in the liver of mongrel dog upon infection. In agreement with this finding there was essentially no neutrophil accumulation in the liver of Beagle dog which had no bacterial and parasitic infection.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗

Well-differentiated hepatocellular carcinoma: clinicopathological features and results of hepatic resection.

OBJECTIVES: To clarify clinicopathological features and surgical results in patients with well-differentiated hepatocellular carcinoma (wd-HCC) corresponding to Edmondson's cell grading I/I-II. METHODS: The clinicopathological characteristics, diagnosis, and results of surgery were studied in 14 wd-HCC patients who underwent limited hepatic resection. RESULTS: All patients had liver cirrhosis and solitary tumors. The tumor sizes ranged from 10 to 25 mm. Serum alpha-fetoprotein levels were normal in 13 patients. The detection rates of sonography, computed tomography (CT), angiography, CT during arterial portography, 1-wk lipiodol CT, and 3-wk lipiodol CT were 100%, 43%, 36%, 69%, 57%, and 0%, respectively. High echoic pattern on sonography and disappearance of lipiodol 3 wk after injection were characteristic findings. There was no operative death. Although seven patients developed intrahepatic tumor recurrence, early treatment including ethanol injection therapy and arterial embolization/infusion therapy controlled the recurrences in five patients. Eleven patients are surviving 25-83 mo after surgery (mean, 44 mo); the remaining three died of pneumonia or hepatic insufficiency 44-62 mo after surgery. Eight patients received successful direct interruption surgery for concurrent esophagocardia varices. Pathological features of wd-HCCs included lack of tumor invasiveness or capsule, nuclear crowding, microacinar formation, fat deposition, presence of portal tracts, and surrounding adenomatous hyperplasia. CONCLUSION: Wd-HCCs present atypical findings in various imagings and histology because of the earlier developmental stage of HCCs. Hepatic resection for wd-HCCs provides excellent patient survival.

Carcinoma, Hepatocellular↗

[Normal-sized ovary carcinoma syndrome: histopathological analysis of 14 cases].

Fourteen cases of "normal-sized ovary carcinoma syndrome" (diffuse metastatic malignant disease of the abdominal cavity of the female, with normal-sized ovaries, with no origin assigned definitively by intraoperative or preoperative evaluation, Feuer et al., 1989) were analyzed histologically, histochemically, immunohistochemically and ultrastructurally. Through these studies, 14 cases were divided into 11 primary peritoneal tumors (4 diffuse malignant mesotheliomas and 7 serous surface papillary carcinomas) and 3 metastatic peritoneal tumors (2 ovarian tumors and 1 appendicular tumor). To distinguish serous surface papillary carcinomas from epithelioid diffuse malignant mesotheliomas, examinations such as hyaluronidase digestion test, electron microscopy, and immunohistochemical studies using antibodies for Ber-EP4 and Vimentin were found to be useful. In order to achieve an accurate prognosis of normal-sized ovary carcinoma syndrome, it seems necessary for us to accumulate more information on this syndrome.

Carcinoma, Papillary↗

[Calcitonin gene-related peptide and human atrial natriuretic hormone levels in response to cardiac operation under high dose fentanyl anesthesia].

To elucidate a role of calcitonin gene-related peptide (CGRP) in anesthesia and surgery with cardiopulmonary bypass (CPB), we measured CGRP which is reported to be a marker for fluid overload, simultaneously with HANP (human atrial natriuretic hormone) in 12 patients during high dose fentanyl anesthesia (50-70 microgram. kg-1). Plasma concentration of CGRP increased to 3 times of the value during preanesthetic phase at 30 min after initiation of CPB. A 3-fold increase compared with control in CGRP occurred 30 min after initiation of CPB. A 3-fold increase in HANP also occurred just before termination of CPB. But, there was no correlation between plasma levels of CGRP and HANP. The changes in CGRP did not relate with those of pulmonary capillary wedge pressure. The results of the present study suggest that the mechanism for the increase is unclear, and CGRP could be influenced during cardiac or coronary artery surgery using CPB.

Adult↗

[Clinical study of bronchial asthma in adult, intractable asthmatics after introduction of guideline therapy].

Introduction of Guideline for asthma treatment proposed by the committee of Japanese allergology have a tremendous impact on patients with bronchial asthma. Intractable asthmatics who have had to take some oral steroid to overcome disease severity, may have also some merit by this treatment, so that some of them might be no longer considered as intractable asthmatics. To clarify this, multicenter study was conducted. In this study, a case who have had more than 5 mg of prednisolone and/or 800 mu g of beclomethasone dipropionate throughout the year, was diagnosed as intractable asthmatics. In 845 case, 14.7%, 123 cases were diagnosed as intractable. These cases were significantly to be non-atopic and adult onset. Also, they have a significant tendency to be deteriorated by infection and careless drug administrations. Using the multiquantification method to examine the most powerful factor on intractable asthmatics, type of asthmatics was the most important and the past history of severe attack was the second. When intractable asthmatics diagnosed mainly by their BDP usage (BDP-intractable) were compared with intractable diagnosed by oral PSL (PSL-intractables), BDP-intractables were significantly atopic compared to PSL-intractables.

Asthma↗

cDNA cloning of a cytosolic protein tyrosine phosphatase (RKPTP) from rat kidney.

A rat cDNA encoding a non-receptor type phosphotyrosine phosphatase (PTPase; EC 3.1.3.48) was identified. The 1608 bp cDNA contains a single open reading frame that predicts a 382 amino acid protein with M(r) 44,438. The predicted protein has no apparent signal or transmembrane sequences, suggesting that it is a cytosolic protein. The C-terminal region has a PTPase catalytic domain that has 40-50% nucleic acid homology to other known PTPases. The N-terminal region has little amino acid sequence homology to any other known sequences. The recombinant protein of the cloned cDNA expressed in Escherichia coli was shown to possess PTPase activity using myelin basic protein, tyrosine phosphorylated by p43v-abl tyrosine kinase, as a substrate.

Amino Acid Sequence↗

Column-switching liquid chromatographic method for the simultaneous determination of iothalamic acid and creatinine in biological fluids.

A column-switching liquid chromatographic method for simultaneous determination of iothalamate and creatinine in human serum and urine was developed. Iothalamate and creatinine were separated on a weakly acidic ion-exchange column (C1) by ion-exclusion chromatography and iothalamate excluded from the column was purified by gel chromatography on a hydrophilic gel column (C2) and then by ion-exchange chromatography on a weakly basic ion-exchange column (C3). Creatinine that was eluted from C1 after iothalamate was transferred to a hydrophilic gel column (C4) and then to a strongly acidic ion-exchange column (C5). The mobile phase for C1-C4 was a pH 3.8 propionate buffer (propionic acid-NaOH = 0.35 + 0.035 mol/kg in water) and a pH 5.6 propionate buffer (propionic acid-NaOH = 0.04 + 0.035 mol/kg in water) was used for C5. Diluted serum and urine samples could be injected directly on to C1, as the matrix of C1 is hydrophilic and C1 is backflushed after the transfer of the creatinine fraction from C1 to C4. Iothalamate and creatinine in the eluates were determined by measuring their ultraviolet absorption at 245 and 234 nm, respectively. The precision (R.S.D.) of the chromatographic method was 1.6% (n = 7) and 0.36% (n = 6) for diluted serum and urine with iothalamate concentrations of 1.0 and 10.0 mumol/l, respectively, and 0.85% (n = 7) and 0.55% (n = 7) for diluted serum and urine with creatinine concentrations of 5.77 and 272 mumol/l, respectively.

Chromatography, Gel↗

G protein beta gamma subunits. Simplified purification and properties of novel isoforms.

The beta and gamma subunits of heterotrimeric guanine nucleotide-binding regulatory proteins (G proteins) form tightly associated complexes. To examine functional differences among the large number of possible combinations of unique beta and gamma subunits, we have synthesized and characterized beta gamma complexes containing gamma 5 and gamma 7, two widely distributed gamma subunits. When either gamma 5 or gamma 7 is expressed concurrently with beta 1 or beta 2 subunits in a baculovirus/Sf9 cell system, all four subunit complexes support pertussis toxin-catalyzed ADP-ribosylation of rGi alpha 1 (where "r" indicates recombinant), indicating formation of functional complexes. Each of the complexes was purified by subunit exchange chromatography, using the G203A mutant of rGi alpha 1 as the immobilized ligand. The purified preparations were compared with other recombinant beta gamma subunits, including beta 1 gamma 1 and beta 1 gamma 2, for their ability to modulate type I and II adenylyl cyclase activities; stimulate phosphoinositide-specific phospholipase C beta; support pertussis toxin-catalyzed ADP-ribosylation of rGi alpha 1 and Go alpha; and inhibit steady-state GTP hydrolysis catalyzed by Gs alpha, Go alpha, and myristoylated rGi alpha 2. The results emphasize the unique properties of beta 1 gamma 1. The properties of the complexes containing gamma 5 or gamma 7 were similar to each other and to those of beta 1 gamma 2.

Adenylate Cyclase Toxin↗

Difference in sero-diagnostic values among KL-6-associated mucins classified as cluster 9.

KL-6 classified as Cluster 9 (MUC-I) is a circulating high-molecular-weight mucin-like molecule. Serum level of KL-6 was measured by a sandwich assay using KL-6 antibody as not only a catcher but also as a tracer. We established 2 additional monoclonal antibodies (MAbs), LISA 101 and EH-123, reacting with KL-6 epitopes different from the epitope recognized by KL-6 antibody. The KL-6-associated mucins detected by the sandwich assay using LISA 101 or EH-123 antibody as a catcher and KL-6 antibody as a tracer were designated as LISA 1-6 and CAM 123-6 respectively. The diagnostic values as the serum markers of KL-6, LISA 1-6 and CAM 123-6 were evaluated measuring their levels in the same serum from healthy individuals and from patients with pulmonary, pancreatic and breast adenocarcinomas. KL-6 was increased abnormally at high rates of more than 50% in pancreatic cancer and in benign lung diseases, LISA 1-6 only in pancreatic cancer, and CAM 123-6 only in pulmonary adenocarcinoma. In benign lung diseases, however, LISA 1-6 and CAM 123-6 were increased abnormally at the rates of only 5.3% and 0% respectively. These observations clearly indicate that LISA 1-6 and CAM 123-6 constitute a part of KL-6, but that they are superior to KL-6 as tumor markers for pancreatic cancer and for pulmonary adenocarcinoma respectively, because of their much lower false-positive rates.

Adenocarcinoma↗