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Biomedical subjects

H Oda

Publications and source records attributed to H Oda.

At least 253 records · Page 14Linked to original sources

Echography of the inferior vena cava for estimating fluid removal from patients undergoing hemodialysis.

Echography was used to perform 118 consecutive measurements of the diameters of the inferior vena cava (IVC) in 28 chronic hemodialysis patients. There was a significant correlation between the percent changes in the IVC in the expiratory phase (dIVC-E) and the percent changes in body weight (dBW) (r = 0.620, p < 0.002, n = 118). The average IVC diameter in the expiratory phase before dialysis (IVC-E) was 15.3 +/- 4.6 mm and this value decreased gradually following ultrafiltration and reached an average diameter of 11.0 +/- 4.3 mm after dialysis. Three cases developed hypotension due to fluid removal. Their IVC-E value showed a rapid reduction below 11.0 mm in the early phase of hemodialysis, then maintained a plateau. The hypotension was attributed to hypovolemia due to excessive dehydration. Our results suggest that the IVC-E diameter in postdialysis would approach 11.0 +/- 4.3 mm, which could be interpreted as being close to the dry weight.

Adult↗

Immunofluorometric assay of prostaglandin D synthase in human tissue extracts and fluids.

A two-site sandwich-type assay for human prostaglandin D (PGD) synthase (beta-trace) was developed with two monoclonal antibodies and using time-resolved fluorometry as the detection technique. The assay is precise (CVs < 10%), accurate, and highly specific for PGD synthase and has a detection limit of 0.05 microgram/L. Using this assay, we measured PGD synthase concentrations in serum, urine, amniotic fluid, cerebrospinal fluid (CSF), seminal plasma, breast cyst fluid, breast discharge fluid, breast milk, and breast tumor extracts. The highest concentrations were found in CSF. We identified proteolytic degradation of PGD synthase in amniotic fluid. Fetal tissues contained various amounts of the enzyme, with the highest values being found in brain and heart. In placental extracts, PGD synthase content was greatest at 11-28 weeks of gestation-in accordance with the concentrations measured in amniotic fluids for this gestational period. We conclude that PGD synthase is ubiquitous and is present in many fluids and tissues of adults and fetuses. This first quantitative and sensitive assay of PGD synthase should facilitate expansion of knowledge on this enzyme and possibly will have applications for diagnosis and monitoring of human diseases.

Adult↗

Immunohistochemical localization of apolipoprotein E in renal amyloidosis.

To clarify the role of apolipoprotein E (apo E) in the formation of amyloid deposits, we examined specimens from 11 patients with renal amyloidosis who underwent renal biopsy by an immunohistochemical method using a monoclonal antibody (murine IgG1). Apo E was distributed in the amyloid deposits of all patients in a pattern similar to that obtained with Congo red staining. Strong positive staining for apo E was found on the amyloid deposits in the glomeruli. These results suggest that apo E is a common constituent of amyloid fibrils and that it may be a useful marker for immunohistochemical studies of systemic amyloidosis including renal amyloidosis.

Adult↗

Involvement of bacterial antigens in immunoglobulin A nephropathy.

To investigate the involvement of bacterial antigens in Immunoglobulin A (IgA) nephropathy, we measured IgA, IgG and IgM antibodies to gram-negative Escherichia coli (E.coli) and Haemophilus influenzae (H.influenzae) by ELISA in 24 patients (11 males and 13 females) with IgA nephropathy and 22 normal controls (11 males and 11 females). The titers of IgA and IgM antibodies for E.coli and H.influenzae were significantly higher in the IgA nephropathy group than in the controls. In addition, IgA and IgM antibody titers for E.coli and H.influenzae showed a significant positive correlation with serum IgA and IgM levels. These findings suggest that subclinical infection by these bacteria stimulates IgA production and that this may be a factor in the development and progression of IgA nephropathy.

Adolescent↗

Laminin-rich extracellular matrix maintains high level of hepatocyte nuclear factor 4 in rat hepatocyte culture.

Laminin-rich extracellular matrix, EHS-gel, has been demonstrated to keep a high level of liver-specific gene expression in cultured rat hepatocytes. To obtain information about the effect of EHS-gel on liver-specific functions, gene expression of liver-enriched transcription factors in rat hepatocytes was investigated. The apolipoprotein A-I and albumin mRNA levels were higher in hepatocytes cultured on EHS-gel than in those cultured on type I collagen (TIC). The levels of mRNA for HNF-4, C/EBP alpha and C/EBP beta were also higher on EHS-gel than on TIC. The level of HNF-4 mRNA in hepatocytes on EHS-gel was almost comparable to that in liver. The HNF-3 alpha mRNA level was lower on EHS-gel than on TIC. C/EBP beta mRNA was induced by dexamethasone in both EHS-gel and TIC. The induction of C/EBP alpha and HNF-4 by dexamethasone was observed only on TIC. These data suggest that EHS-gel leads hepatocytes to keep the phenotypic expression through high expression of liver-enriched transcription factors, such as HNF-4.

Animals↗

Interaction of the microtubule cytoskeleton with endocytic vesicles and cytoplasmic dynein in cultured rat hepatocytes.

In a recent study (Goltz, J.S., Wolkoff, A.W., Novikoff, P.M., Stockert, R.J., and Satir, P. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 7026-7030), we found that ligand- and receptor-containing endocytic vesicles bind to endogenous microtubules in vitro after 60 min of receptor-mediated endocytosis of asialo-orosomucoid. In the presence of ATP, ligand-containing endocytic vesicles are released from microtubules, while those containing receptor are not. We hypothesized that cytoplasmic dynein may associate with ligand-containing, but not receptor-containing, domains of endocytic vesicles and might be involved in the movement of ligand-containing vesicles along microtubules during sorting of ligand from receptor. Direct evidence in support of this hypothesis has been obtained in the present study. Binding of ligand-containing vesicles to microtubules correlates highly (p < 0.001) with binding of dynein, but not kinesin, under a variety of conditions. Binding of receptor-containing vesicles to microtubules is independent of both cytoplasmic dynein and kinesin binding. Tight association of cytoplasmic dynein with a population of ligand-containing vesicles is seen directly by immunoprecipitation. These results support the view that in receptor-mediated endocytosis, ligand-containing vesicles become bound to microtubules by cytoplasmic dynein. While receptor domains of endosomes remain attached to microtubules in an ATP-independent manner, ligand-containing domains might be moved away toward pericentrosomal lysosomes by this motor molecule.

Adenosine Triphosphate↗

Possible involvement of botulinum ADP-ribosyltransferase sensitive low molecular G-protein on 5-hydroxytryptamine (5-HT)-induced inositol phosphates formation in 5-HT2c cDNA transfected cells.

To clarify the involvement of botulinum ADP-ribosyltransferase sensitive low molecular G-proteins in 5-hydroxytryptamine (5-HT)-induced stimulation of phosphatidylinositol turnover, we examined the effects of 5-HT on inositol phosphates formation in COS 7 cells transfected with 5-HT2c receptor cDNA, but did not in non-transfected or vector-transfected cells. A typical 5-HT2c receptor antagonist mianserin (0.3-3 microM) inhibited the 5-HT-induced inositol phosphates formation. Treatment with botulinum toxin D preparation (20 micrograms/ml, 8 h) that contained botulinum C3 ADP-ribosyltransferase, blocked the 5-HT-induced inositol phosphate formation, although botulinum toxin A preparation that did not contain the enzyme did not have an influence. These results support our previous findings suggesting that low molecular weight G-proteins ADP-ribosylated by botulinum ADP-ribosyltransferase are involved in phospholipase C activity.

ADP Ribose Transferases↗

A mutational hot spot in the p53 gene is associated with hepatoblastomas.

Hepatoblastomas generally appear in children aged 2 or 3 years old and arise from apparently normal, non-cirrhotic liver. To elucidate any possible role of p53 mutations in their genesis, we amplified and sequenced exons 5 to 8 of the p53 gene in 10 cases of hepatoblastoma. Somatic mutations were detected in 9 cases, in eight of which a common point mutation at the first-base position of codon 157 was found, resulting in an amino-acid substitution of phenylalanine for valine. Two missense mutations in codon 244, and one each in codons 273 and 279, were also found, with 3 hepatoblastomas having double mis-sense mutations. Out of the total of 12 mutations, 11 were G-to-T transversions. One was a G-to-A transition and guanines were always present on the transcribed strand. Furthermore, p53 over-expression was immunohistochemically observed in 7 out of 9 cases with p53 gene mutations, although the staining pattern was focal and heterogeneous. The findings suggest that particular environmental mutagens may be involved in mutagenesis of the p53 gene in some cases of hepatoblastomas and that p53 mutations at a specific site may play an important role in the genesis of this disease.

Base Sequence↗

Mutations of the p53 gene and p53 protein overexpression are associated with sarcomatoid transformation in renal cell carcinomas.

Renal cell carcinomas sometimes show sarcomatoid transformation, thus comprising both sarcomatous and carcinomatous components. Such sarcomatoid renal cell carcinomas are highly malignant with pronounced proliferative activity. The present investigation was conducted to assess the mutational status of the p53 and H-ras genes independently in carcinomatous and sarcomatous portions of individual tumors, applying PCR, subcloning, and sequencing to 14 cases. Sarcomatoid portions showed an extremely high mutation rate for the 53 gene (11 of 14, 78.6%) with two mutational hot spots at codons 278 (8 of 14, 57.1%) and 244 (6 of 14, 42.9%). Five cases showed double mutations, four cases had mutations at codons 278 and 244, and one case had mutations at codons 278 and 248. In contrast, the carcinomatous portions demonstrated a low mutation rate for the p53 gene (2 of 14, 14.3%) and no double mutations were detected. Ten cases showed genetic heterogeneity in the p53 gene between the two tumor components. Furthermore, p53 overexpression was immunohistochemically observed only in those components with p53 mutations, mainly in the sarcomatoid portions. No H-ras mutations were observed. The findings strongly suggest that p53 mutations leading to overexpression of p53 protein are closely associated with sarcomatoid transformation in renal cell carcinomas.

Amino Acid Sequence↗

Serum 7 alpha-hydroxycholesterol as a new parameter of liver function in patients with chronic liver diseases.

To examine bile acid synthesis in chronic liver diseases, serum total 7 alpha-hydroxycholesterol level was measured by gas-liquid chromatography-mass spectrometry in patients with cirrhosis (n = 23), patients with chronic hepatitis (n = 21), and control subjects (n = 18). The serum 7 alpha-hydroxycholesterol levels were significantly lower in patients with cirrhosis than the controls (78 +/- 59 pmol/mL vs. 237 +/- 97 pmol/mL; mean +/- SD). However, in patients with chronic hepatitis, the level was fully retained (262 +/- 102 pmol/mL). Serum 7 alpha-hydroxycholesterol levels of 17 patients with cirrhosis classified as Child B and C ranged from 33 to 69 pmol/mL, and all were less than the normal range (between 104 and 466 pmol/mL), however, those levels of some patients classified as Child A were within the normal range. Serum 7 alpha-hydroxycholesterol levels significantly correlated with serum albumin, cholinesterase, total bile acid, direct bilirubin, alkaline phosphatase, indocyanine green (ICG) retention rate, hepaplastin test, and lecithin-cholesterol acyltransferase activities. We conclude that bile acid synthesis is well preserved in patients with chronic hepatitis and that it is decreased in most patients with cirrhosis. Serum 7 alpha-hydroxycholesterol may be a new parameter of liver function testing to assess hepatic bile acid synthesis in patients with chronic liver diseases.

Adult↗

A polymorphism at codon 160 of human O6-methylguanine-DNA methyltransferase gene in young patients with adult type cancers and functional assay.

O6-methylguanine-DNA methyltransferase (MGMT) plays an important role in repair of alkylating agent-induced DNA damage. Among the alkylation products of DNA, O6-methylguanine is one of the most critical lesions leading to the induction of mutations. The enzyme MGMT transfers the methyl group from O6-methylguanine of DNA to its own cysteine residue. Although mutations of other DNA repair genes involved in nucleotide excision repair and mismatch repair have been proven to be related to human tumorigenesis, the question of whether MGMT gene mutation might play a role in human carcinogenesis has hitherto not been elucidated. If there is a population with decreased enzyme activity due to defective MGMT gene, the affected individuals should be at risk of developing cancer early in life because of an increased susceptibility to alkylating agents. To test this hypothesis, germ line mutations of the MGMT gene were investigated in 12 young patients with adult type cancers (mean, 16.7 years old, 8 hepatocellular carcinomas, 3 gastric cancers, 1 cholangiocellular carcinoma) and 28 elderly patients who died of non-cancer diseases as controls (mean, 66 years old). A point mutation at codon 160 in exon 5, GGA to AGA, converting glycine to arginine was found in three of the young patients (25%), while the same mutation was found in three out of 28 (10.7%) in the control group. The mutated codon was located 15 codons from a functional cysteine residue toward the carboxyl terminal. Investigation of enzyme function, even in cases of bi-allelic mutation, revealed comparable activities for both mutated and wild type MGMT. Thus, we conclude the mutation is a normal polymorphism of the MGMT gene, present in approximately 15% of the population, although this does not rule out a possible influence in other tissues.

Adolescent↗

Inhibitory effect of probucol on nephrotoxicity induced by ferric nitrilotriacetate (Fe-NTA) in rats.

Effects of dietary probucol on renal damage induced by a renal carcinogen, ferric nitrilotriacetate (Fe-NTA), in male Wistar rats were quantitatively investigated with a computer-mediated image analyzer. The kidneys of rats fed a 1% probucol diet were protected from necrosis and lipid peroxidation induced by a single i.p. treatment with Fe-NTA solution at 5 mg Fe/kg body wt and were significantly resistant to a higher dose. For the parameter of lipid peroxidation, Schiff's staining method was utilized to demonstrate the extent of formation of aldehydes, products of lipid peroxidation. Thus following injection of Fe-NTA solution at 10 mg Fe/kg body wt the average areas of tubular necrosis were 85.8% versus 56.9% and the positive areas for Schiff's staining were 15.3% versus 5.6% in the renal cortices of rats fed control of 1% probucol diets respectively. These results indicate that probucol provides protection against Fe-NTA-induced nephrotoxicity through its antioxidant properties. In addition to being a cholesterol-lowering drug, useful for the control of hypercholesterolemia, probucol may therefore be beneficial for prevention and treatment of various pathogenic processes including those leading to carcinogenesis.

Animals↗

Inhibitory effect of probucol on benzo[a]pyrene induced lung tumorigenesis.

The effects of probucol, a clinically used cholesterol lowering and antioxidant drug, on benzo[a]pyrene (B[a]P) induced pulmonary and forestomach tumorigenesis as well as induction of colonic aberrant crypt foci (ACF) in female A/J mice was investigated. Diet containing 1% probucol fed prior to, during and after 8 bi-weekly 1 mg/mouse oral intubations of B[a]P, reduced the number of pulmonary adenomas by 52% (P < 0.001) and the number of forestomach tumors by 31%. The 0.06% probucol diet also resulted in decreased tumor formation but the differences did not reach statistical significance. Both probucol diets significantly reduced the numbers of large ACF, putative preneoplastic lesions of the colon mucosa, but showed no effects on the total numbers of ACF. The results of this study suggest that probucol may also be useful as a chemopreventive agent, in addition to being a cholesterol lowering and anti-atherogenic drug with low toxicity.

Animals↗

Effects of protein kinase C and A activation on ATP-stimulated release of [3H]noradrenaline from PC12 cells.

The influence of activation of protein kinase C (PKC) and cyclic AMP on noradrenaline (NA) release in the neurosecretory rat pheochromocytoma PC12 cell line was investigated. External ATP induced [3H]NA release from prelabeled PC12 cells, in the presence of extracellular CaCl2. The potency order of ATP analogs was adenosine 5'-O-(gamma-thiotriphosphate) > or = ATP > 2-methylthio ATP > 2',3'-O-(4-benzoyl)benzoyl ATP. alpha,beta-Methylene ATP, beta gamma-methylene ATP, and 8-bromo ATP were inactive. Neither ADP, GTP, nor ITP was active. The addition of phorbol 12-myristate 13-acetate (PMA) or agents elevating the cyclic AMP content, such as vasoactive intestinal peptide (VIP) or an adenosine analog, also stimulated [3H]NA release. Not only high K(+)- but also ATP-stimulated [3H]NA release was enhanced by co-addition with PMA or agents elevating the cyclic AMP content. PMA and VIP had no effect on the cytosolic free Ca2+ concentration ([Ca2+]i) or on the ATP-stimulated [Ca2+]i rise, although both stimulatory effects on [3H]NA release were dependent on extracellular CaCl2. The addition of PMA stimulated [3H]NA release dose-dependently, and enhanced 300 microM (maximal dose) ATP-stimulated [3H]NA release without changing the affinity for ATP. The effect of PMA was inhibited by PKC inhibitors such as calphostin C and in PKC-depleted cells, and potentiated by elevation of cyclic AMP. These data suggest that the process of ATP-stimulated NA release, not ATP-stimulated Ca2+ influx, is regulated by the dual, PKC- and cyclic AMP-dependent mechanisms, positively and independently. Treatment with pertussis toxin had no effect on the ATP-stimulated [Ca2+]i rise or [3H]NA release.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗

Quantitative detection of ultraviolet light-induced photoproducts in mouse skin by immunohistochemistry.

UVB-induced cyclobutane pyrimidine dimers (CPDs) and pyrimidine-pyrimidone (6-4)photoproducts [(6-4)photoproducts] in mouse skin DNA were quantitatively measured using an immunohistochemical approach with a computer-aided color image analyzer. The skins of the C3H/HeN mice were irradiated with ultraviolet B (UV-B, 280-320 nm), and processed to give conventional formalin-fixed, paraffin-embedded histologic sections. Routine immunohistochemistry clearly demonstrated a dose-dependent induction of both photoproducts. CPDs were detectable at doses > or = 125 J/m2, while for (6-4)photoproducts, the minimal dose at which they were detectable was 250 J/m2 in the present study. A time course study showed that the repair of (6-4)photoproducts was more rapid than that of CPDs, and that epidermal cells had a higher capacity for their removal than dermal cells. About half of the (6-4)photoproducts were excised within the first 24 h after the irradiation, and the process was essentially complete by 72 h. In contrast, there was no apparent removal (less than 10%) of CPDs in the first 24 h and they only completely disappeared from the epidermal cells at 120 h after irradiation. The effect of DNA dilution due to increased turnover of epidermal cells after UV-B irradiation was evaluated by quantitative immunohistochemical measurement of the time course of bromodeoxy-uridine (BrdUrd) incorporated into nuclei at 2 days post irradiation when the proliferation reaches a peak. The removal of photoproducts was more marked than the decrease in BrdUrd staining. Our results suggest that mouse skin cells can repair both (6-4)photoproducts and CPDs, but with considerably lower efficiency, especially in the latter case, then human or monkey skin cells.

Animals↗

Glomerulocystic kidney disease in a young adult.

We report an 18-year old woman who had glomerulocystic kidney disease (GCKD) without a family history of renal disease or hypertension and no known congenital abnormalities. Her renal function was normal. Renal biopsy showed cystic dilatation of the Bowman's spaces and atrophy of the glomerular tufts. Electron microscopy revealed specific changes in the basement membranes of noncystic glomeruli, suggesting a congenital origin for her renal pathology. This relatively rare case contrasts with the usual presentation of GCKD in neonates or children.

Adolescent↗