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Biomedical subjects

H Oda

Publications and source records attributed to H Oda.

At least 235 records · Page 13Linked to original sources

Oxidative stress response in iron-induced acute nephrotoxicity: enhanced expression of heat shock protein 90.

Iron overload with ferric nitrilotriacetate (Fe-NTA) induces acute renal proximal tubular necrosis, a consequence of oxidative tissue damage, that leads to a high incidence of renal adenocarcinoma in rodents. In the present study, we determined the proteins preferentially produced in response to the Fe-NTA-induced oxidative injury. A single intraperitoneal Fe-NTA treatment led to the enhanced production of a number of proteins with molecular masses of 85-95 kDa. These included heat shock protein 90 (HSP90) as determined by immunoprecipitation. The enhanced production of HSP90 was prominent in the renal tubular cells. Steady accumulation of HSP90 was observed in the subacute toxicity experiments with multiple injections of Fe-NTA, suggesting that the enhanced production of HSP90 is important in increasing resistance to subsequent injury caused by the Fe-NTA-induced oxidative stress.

Animals↗

Sequence analysis, chromosomal location, and developmental expression of the mouse preproendothelin-1 gene.

Recent studies have designated endothelins (ETs) as morphogenetic factors in embryonic development. In the present study, we cloned and characterized the mouse preproendothelin-1 (preproET-1) gene (Edn1) and examined its expression in reference to development. Edn1 comprises five exons, and the open reading frame encodes the 202-amino-acid preproET-1. The sequences and structural organization of Edn1 are highly homologous to those of other species, especially in the terminal 200-bp sequence of the 3'-noncoding region. Interspecific backcross mapping located Edn1 in the central region of chromosomal 13, where a mouse mutation, congenital hydrocephalus (ch), is also mapped. The highest expression of Edn1 mRNA is detected in the lung in adult mice, whereas Edn1 is predominantly expressed in the epithelium and mesenchyme of the pharyngeal arches and in the endothelium of the large arteries. Edn1 expression and ET-1 peptide levels in the lung progressively increased during the perinatal stage, whereas the expression of Edn3, a gene encoding ET-3, reciprocally decreases. These results suggest that Edn1 expression is developmentally regulated in different tissues and organs in mice in a spatial- and temporal-specific manner.

Amino Acid Sequence↗

Efficacy of marker wire for intracoronary stenting.

The Marker Wire was used for Palmaz-Schatz coronary stent implantation. The Marker Wire is useful in estimating lesion length and in determining the number of stents required, in addition to facilitating stent positioning.

Aged↗

Pertussis toxin-insensitive effects of mastoparan, a wasp venom peptide, in PC12 cells.

Recent studies have shown that mastoparan, an amphiphilic peptide derived from wasp venom, modifies the secretion of neurotransmitters and hormones from a variety of cell types. Mastoparan interacts with heterotrimeric guanine nucleotide-binding proteins (G proteins) such as Gi and G(o), which are ADP-ribosylated by pertussis toxin (PTX) and thereby uncoupled from receptors. Previously, some of the effects of mastoparan including secretion were reported to be modified selectively by PTX but not by cholera toxin (CTX). In the present study, we examined the influence of bacterial toxins on the effects of mastoparan in PC12 cells. Mastoparan stimulated [3H]noradrenaline (NA) release from prelabeled PC12 cells in the absence of CaCl2, although high K+ or ATP-stimulated the release in a Ca(2+)-dependent manner. Pretreatment with CTX, not PTX, for 24 h inhibited mastoparan-stimulated [3H]NA release. Mastoparan inhibited forskolin-stimulated cyclic AMP accumulation in a dose-dependent manner, although mastoparan had no effect by itself. Pretreatment with PTX completely abolished the inhibitory effect of carbachol via Gi on cyclic AMP accumulation and partially reduced the effect of mastoparan. However, the inhibitory effect of 20 microM mastoparan was not modified by pretreatment with PTX. Thus, we investigated the effect of mastoparan on CTX-catalyzed [32P]ADP-ribosylation of proteins in PC12 cells. A subunit of CTX (CTX-A) catalyzed [32P]ADP-ribosylation of many proteins in the cytosolic fraction of PC12 cells. One of these was a 20 kDa protein, named ADP-ribosylating factor (ARF). The addition of mastoparan to assay mixtures inhibited ADP-ribosylation of many proteins including ARF and CTX-A in the presence of the cytosolic fraction. In the absence of the cytosolic fraction, however, mastoparan slightly enhanced ADP-ribosylation of bovine serum albumin and auto-ADP-ribosylation by CTX-A. Mastoparan did not inhibit ADP-ribosylation of the alpha subunit of Gs in the membrane fraction. These findings suggest that 1) mastoparan interacts with PTX-insensitive and CTX-sensitive factor(s) to stimulate NA release, and 2) mastoparan interacts with ARF inhibiting its activity to enhance the ADP-ribosylation reaction by CTX. ARF may be an exocytosis-linked G protein.

ADP-Ribosylation Factors↗

Experimental bilirubin pigmentation of rat dentine and its detection by a qualitative analytical method.

An animal model of bilirubinemia was used to determine whether bilirubin present in pigmented teeth can be extracted and qualitatively analysed. The bile ducts of 10 Long-Evans Agouti rats were ligated and bilirubin (14 mg/kg per day) was injected intraperitoneally for 4 days. When the animals were killed 2 weeks later, pigmented lower incisors were observed in three animals. These teeth were dried, powdered and bilirubin was extracted with chloroform/methanol/acetic acid, 30:10:0.5, v/v for 10 min under sonication. After centrifugation, the supernatant was collected and evaporated. The residue was dissolved in chloroform and its absorption spectrum measured before and after diazo reaction. This resulted in a shift of the absorption maximum from 450 to 540 nm and indicated the presence of bilirubin in pigmented teeth. No bilirubin was found in the lower incisors of untreated control rats. This technique may be useful in distinguishing bilirubin staning from other intrinsic discolorations of teeth.

Acetic Acid↗

Detection of ultraviolet photoproducts in mouse skin exposed to natural sunlight.

In the present study, we for the first time investigated the formation of ultraviolet (UV) photoproducts, cyclobutane pyrimidine dimers (CPDs), pyrimidine-pyrimidone (6-4) photoproducts (64PPs) and Dewar isomers, in vivo in shaved and depilated C3H/HeN mouse skin exposed to natural sunlight (NSL) at noon for 5 min to 1 h in mid-summer, using a highly sensitive immunohistochemical method. This method permits the quantitative analysis of UV-photoproducts in formalin-fixed, paraffinembedded sections with specific antibodies against CPDs, 64PPs and Dewar isomers. We demonstrated that the induction of CPDs in vivo in mouse skin by NSL was exposure time-dependent, but the accumulation of 64PPs or Dewar isomers was comparatively low in the skin sections from mice exposed to NSL in vivo. The results indicate that CPDs are the main photoproducts in vivo induced by sunlight and that their formation and repair may be important in connection with carcinogenesis in sun-exposed areas of human skin.

Animals↗

Mutational analysis of the p53 and K-ras genes and allelotype study of the Rb-1 gene for investigating the pathogenesis of combined hapatocellular-cholangiocellular carcinomas.

Because combined hepatocellular-cholangiocellular carcinoma is rare and its biological features and pathogenesis have not been well established, we investigated alterations of the p53, K-ras and Rb-1 genes, as well as expression patterns of carcinoembryonic antigen and keratin, in seven combined hepatocellular-cholangiocarcinomas out of 557 hepatocellular carcinomas autopsied at Tokyo University during 30 years. Mutations of the p53 gene were found in two cases, at codon 244 (GGC to TGC) in the cholangiocellular carcinoma component of case 1 (mixed type, showing an intimate intermingling of both elements) and at codon 234 (TAC to AAC) in both components of case 5 (combined type, consisting of contiguous but independent masses of both elements). Mutation of the K-ras gene (codon 12, GGT to GAT) was seen only in the cholangiocellular carcinoma component of clinically apparent double cancer, case 6. Allelic alteration of the Rb-1 gene was observed in two cases, deletion of both alleles in the hepatocellular carcinoma component of case 3 (combined type) and replication error of the same pattern in both components of case 4 (mixed type). Immunohistochemical analysis showed that the hepatocellular carcinoma components of five cases (cases 2, 3, 5, 6, 7) were immunoreactive for keratin, suggesting biliary epithelial transformation. In four of the five cases (cases 3 and 5 combined, case 7 mixed and case 6 double cancer), cholangiocellular carcinoma components were also positive for keratin. These results suggest that both components of combined hepatocellular-cholangiocarcinoma have the same genetic and phenotypic character and might have arisen from the same origin in some cases.

Aged↗

Retrospective survey of chronic Q fever in Japan by using PCR to detect Coxiella burnetii DNA in paraffin-embedded clinical samples.

We used PCR to detect Coxiella burnetii DNA in paraffin-embedded tissues obtained from patients with chronic endocarditis in which the etiological agent had been unknown. On the basis of the published nucleotide sequence of the C. burnetii htpB gene, primers were chosen to produce an amplified fragment of 285 bp. A total of 60 samples from 56 patients were tested for the presence of C. burnetii DNA. Five samples from four patients were found to be positive. All of the amplified DNA fragments possessed a TthHB8I restriction site, as predicted from the published sequence of C. burnetii. In one of the four positive patients, rickettsia-like particles were found in sections of tissue stained by Gimenez's method. This is the first report of chronic Q fever in Japan.

Base Sequence↗

Nicardipine hydrochloride suppresses DNA synthesis in human mesangial cells stimulated with recombinant human (rh) platelet-derived growth factor AA, rh interleukin-1 alpha, or rh tumor necrosis factor-alpha.

In order to define the role of cytokines in mesangial cell pathophysiology, we measured the mitogenic activity of recombinant human platelet-derived growth factor AA (rhPDGF-AA), rh interleukin-1 alpha (rhIL-1 alpha) and rh tumor necrosis factor-alpha (rhTNF-alpha) in cultured human mesangial cells, and investigated the effect of the calcium channel blocker nicardipine hydrochloride on their cell mitogenic activity. DNA synthesis in mesangial cells, stimulated by rhPDGF-AA, rhIL-1 alpha or rhTNF-alpha, was measured using [3H]TdR up take and similar investigations, with nicardipine hydrochloride added to the above, were conducted. The results showed DNA synthesis in cultured human mesangial cells were stimulated by rhPDGF-AA, rhIL-1 alpha and rhTNF-alpha, and this effect was reduced by the addition of nicardipine hydrochloride. Since rhPDGF-AA, rhIL-1 alpha and rhTNF-alpha are released by the inflammatory cells which infiltrate glomeruli, these cytokines may be involved in the mechanisms of mesangial cell proliferation observed in immune-mediated mesangial proliferative glomerulonephritis. Nicardipine hydrochloride reduced DNA synthesis in cultured human mesangial cells in response to these cytokines, suggesting possible application in medical therapy for immune-mediated mesangial proliferative glomerulonephritis.

Calcium Channel Blockers↗

Plasma thrombomodulin: usefulness as a blood access failure marker in hemodialysis patients.

In 144 patients on hemodialysis (76 males and 68 females, median age 55.7 +/- 14.1 years, mean period on dialysis 44.1 +/- 33.3 months), thrombomodulin was determined by enzyme immunoassay prior to initiation of hemodialysis. The results showed that the mean thrombomodulin value of hemodialysis patients was 13.59 +/- 3.63 ng/ml which was significantly higher than the control value (3.20 +/- 0.90 ng/ml). The thrombomodulin values were significantly higher in patients with blood access failure (15.27 +/- 4.45 ng/ml) than in those without (13.11 +/- 3.31 ng/ml), and the rate of blood access failures was also significantly higher in those with thrombomodulin values of 15.0 ng/ml or higher than in those with values < 15.0 ng/ml. It was evident that there is a higher risk of blood access failure in patients with severe systemic vascular endothelial injury, and thrombomodulin is a useful marker of such an injury.

Adult↗

Platelet-derived growth factor, interleukin (IL)-1 beta, IL-6R and tumor necrosis factor-alpha in IgA nephropathy. An immunohistochemical study.

We clarified the presence of platelet-derived growth factor (PDGF), interleukin (IL)-1 beta, IL-6, IL-6 receptor (IL-6R) and tumor necrosis factor-alpha (TNF-alpha) in renal biopsy specimens from 62 patients with IgA nephropathy, and discuss their relationship with mesangial cell proliferation, degree of histological damage and various clinical factors. Mesangial proliferation was determined histologically by PAS staining and the positive rate of proliferating cell nuclear antigen (PCNA). Renal biopsy specimens were stained using an enzyme-antibody method to determine the presence of cytokines and the receptor. PCNA-positive cells in the glomeruli significantly increased in patients positive for PDGF, IL-6, IL-6R and TNF-alpha. The degree of histological damage increased with the positive rates of PDGF, IL-6 or IL-6R and the number of PCNA-positive cells in the glomeruli. In the PDGF-A-positive patients, total urinary protein (TUP), urinary beta2-microglobulin (u-beta2-m) and systolic and diastolic blood pressure were significantly higher, and creatinine clearance (Ccr) was significantly lower than in the PDGF-A-negative patients. In the PDGF-B-positive patients, TUP, serum creatinine (s-Cr) and urinary and serum beta2-m increased significantly and Ccr decreased significantly. Il-1beta was not related to any clinical factors. In the IL-6-positive patients, TUP was significantly higher than in the IL-6-negative patients. In the IL-6R-positive patients, TUP, s-Cr, urinary beta2-m and systolic blood pressure were significantly higher than in the IL-6R-negative patients. In conclusion, PDGF, IL-6 and IL-6R may be closely related to mesangial cell proliferation, histological changes and deterioration of various clinical factors in patients with IgA nephropathy.

Adolescent↗

Cadherin-mediated cell adhesion and cell motility in Drosophila trachea regulated by the transcription factor Escargot.

Coordination of cell motility and adhesion is essential for concerted movement of tissues during animal morphogenesis. The Drosophila tracheal network is formed by branching, migration and fusion of tubular ectodermal epithelia. Tracheal tip cells, located at the end of each branch that is going to fuse, extend filopodia to search for targets and later change their cell shape to a seamless ring to allow passage of lumen. The cell adhesion molecule DE-cadherin accumulates at the site of contact to form a ring that marks the site of lumen entry and is essential for the fusion. DE-cadherin expression in tip cells of a subset of branches is dependent on escargot, a zinc finger gene expressed in all tip cells. Such escargot mutant tip cells failed to adhere to each other and continued to search for alternative targets by extending long filopodia. We present evidence indicating escargot positively regulates transcription of the DE-cadherin gene, shotgun. Overexpression of DE-cadherin rescued the defect in one of the fusion points in escargot mutants, demonstrating an essential role of DE-cadherin in target recognition and identifying escargot as a key regulator of cell adhesion and motility in tracheal morphogenesis.

Animals↗

Three-dimensional organization of rat hepatocyte cytoskeleton: relation to the asialoglycoprotein endocytosis pathway.

Analysis by confocal microscopy has revealed features of the microtubule network of rat hepatocytes in culture, establishing the three-dimensional disposition of the microtubule-based cytoskeleton, its relation to the actin-based cytoskeleton and to ligand-containing endosomes during receptor-mediated endocytosis and the alterations in its structure and disposition by the microtubule pertubant, Taxol. By co-localization studies, we have been able to demonstrate that the microtubules have a significant role in receptor-mediated endocytosis of asialoglycoproteins in this cell. Asialoorosomucoid-containing endosomes attach to widely spaced arrays of microtubules running under the baso-lateral surface of the hepatocytes 5-15 minutes after the initiation of endocytosis and then travel along microtubule paths to become concentrated with microtubules near the centrosome and at bile canaliculi after 30-60 minutes of receptor-mediated endocytosis. Receptor-mediated endocytosis is affected, but not abolished by Taxol, which inhibits the rate of asialoorosomucoid degradation at the same concentrations as those that disrupt microtubule and cytoplasmic dynein distribution, and that prevent the concentration of endosomes centrally. The results support suggestions that asialoorosomucoid-containing endosomes are captured by microtubules just below the actin layer at the cell periphery and these are actively transported centrally along microtubules, possibly by cytoplasmic dynein, so that the concentration of endosomes near the centrosome, and the subsequent efficient lysosomal degradation of ligand, are consequences of the confluence of microtubules in this region.

Actin Cytoskeleton↗

[Monoclonal antibody against an urinary protein secreted from the renal tubules--immunohistochemical analysis of renal cell carcinoma].

BACKGROUND: Monoclonal antibodies (mABs) against urinary proteins originated from renal tubules were obtained in order to find a possible tumor marker for renal cell carcinoma. METHODS: Urine samples were collected from healthy adult males. After removal of Thamm-Horsfall protein (THP) by sodium chloride precipitation, supernatant urine was ultrafiltered, thus, THP-free urinary proteins were obtained. Then, proteins of plasma origin were removed by circulating the urinary proteins through an affinity column which contained immobilized rabbit anti-human whole serum immunoglobulins. BALB/c mice were immunized with the THP- and plasma protein-free urinary proteins. The spleen cells were fused with mouse myeloma cells. Hybridomas were screened by indirect immunofluorescence of fresh frozen kidney tissue sections using culture supernatants. RESULTS: Five hybridomas producing mAbs which reacted with renal tubular segments were established. One mAb reacted strongly with distal renal tubules. Using this mAb, formalin fixed and paraffin embedded sections of 39 renal cell carcinomas were stained by immunoperoxidase method. Twenty-six of the 39 cancer tissues (66.7%) were stained positive. According to the histological classification, positive staining rate of clear cell subtype, and mixed subtype was 75% (6/8), 80% (8/10) and 66.7% (6/9), respectively. According to the tumor grade, positive staining rate of G1, G2, G3 was 66.7% (6/9), 71.4% (15/21) and 55.5% (5/9), respectively. CONCLUSION: These findings indicate that most of renal cell carcinomas were originated from common precursor cells for both proximal and distal tubules, because normal proximal tubular cells were not stained by the mAb.

Adult↗