Is a late graft biopsy at 3 years after transplantation effective for improving the long-term outcome of renal allografts using cyclosporine?
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Azathioprine↗
Biomedical subjects
K Uchida
Publications and source records attributed to K Uchida.
[Effects on hemodynamics and myocardial metabolism of nicardipine and nitroglycerin during aortic cross clamp in dogs with experimentally produced coronary stenosis].
We investigated the effects of hemodynamics and myocardial metabolism of nicardipine (NIC) and nitroglycerin (NTG) in experimentally coronary constricted dogs. Coronary constriction was produced by reducing 50% of the left anterior descending coronary blood flow. After stabilization of hemodynamics, we cross-clamped the descending aorta and administered nicardipine (10 micrograms.kg-1.min-1) or nitroglycerin (10 micrograms.kg-1.min-1). The drugs were infused for 15 minutes and were discontinued just before the clamp on the aorta was released. In NIC group, the cardiac output (CO) increased 32% and the heart rate (HR) decreased 17% significantly, compared with pre-clamping values. The myocardial blood flow (MBF) in ischemic area measured by hydrogen clearance method was maintained. The value of the myocardial oxygen extraction ratio (MO2EX) decreased significantly for 24.3% compared with the pre-clamping value of 41.4%. On the other hand, in NTG group, CO, HR, and MO2EX were unchanged, but MBF in ischemic area was well maintained. The results of these experimentally coronary constricted dogs suggest that both NIC and NTG maintain the cardiac function during the aortic cross-clamping, but regarding the improvement of the myocardial metabolism, NIC appears to be the better choice for blood pressure control.
Animals↗
[Role of endothelium-derived nitric oxide in mediating the natriuretic response to acute extracellular volume expansion].
To determine the role of endothelium-derived nitric oxide (EDNO) in mediating the natriuretic response to acute extracellular volume expansion (ECVE) with isotonic saline (3% of body weight per hour), the diuretic and natriuretic responses to ECVE were studied in anesthetized Sprague-Dawley rats during the intravenous infusion of an EDNO synthesis inhibitor, NW-nitro-L-arginine methyl ester (L-NAME). Intravenous infusion of L-NAME at the dose of 5 micrograms/kg/min significantly inhibited the diuresis and natriuresis in response to ECVE by 58% and 67%, without altering arterial pressure, effective renal plasma flow, glomerular filtration rate and basal excretory function. This inhibitory effect of L-NAME on the diuretic and natriuretic responses to ECVE was attenuated by the infusion of the EDNO synthesis precursor, L-arginine (1mg/kg/min), but not by D-arginine. In addition, pretreatment with 0.3 mg/kg of the angiotensin II receptor antagonist, L-158,809, normalized the diuretic and natriuretic responses to ECVE in L-NAME-treated rats, suggesting an angiotensin-II-dependency of the reduced renal excretory response to ECVE during EDNO synthesis inhibition. Neither L-arginine nor L-158,809 alone significantly altered the renal excretory response to ECVE compared with vehicle-treated control rats. These results suggest that EDNO might play an important role in the regulation of sodium and water excretion during ECVE, and indicate a possible interaction between EDNO and angiotensin II on the renal excretory function.
Angiotensin II↗
[Assessment of the distribution of renal cortical blood flow by contrast ultrasonography].
The importance of the distribution of intrarenal blood flow in the regulation of various renal functions, such as urine concentration and sodium excretion, has been well recognized. However, there have been no reliable methods to measure local flow in the kidney in vivo. The present study demonstrated the usefulness of contrast ultrasonography combined with injection of sonicated 5% albumin for assessment of the distribution of renal cortical blood flow in eleven mongrel dogs. The left kidney was displayed by tomographic echography, and microbubbles of sonicated albumin were injected into the abdominal aorta above the left renal artery. Video density time curves were generated and fitted to a time-intensity curve. Intrarenal infusion of acetylcholine (4.0 micrograms/kg/min) increased renal blood flow (RBF) from 2.5 +/- 0.3 to 4.6 +/- 1.0 ml/min/g kwt (p < 0.01), and norepinephrine (0.5 microgram/kg/min) decreased RBF from 2.5 +/- 0.3 to 1.2 +/- 0.5 ml/min/g kwt (p < 0.01). There were significant positive correlations between percent change in RBF and peak intensity and area under the curve, which were calculated with a time-intensity curve. Furthermore, the inner/outer renal cortex ratio of peak intensity significantly increased during acetylcholine infusion (0.72 +/- 0.11 vs 0.86 +/- 0.09; p < 0.01), whereas no significant change was observed during norepinephrine infusion. These results suggest that renal contrast ultrasonography may be useful for real-time assessment of the distribution of renal cortical blood flow in vivo.
Acetylcholine↗
Aberrant upregulation of a novel integrin alpha subunit gene at 3p21.3 in small cell lung cancer.
Our recent identification of homozygous deletions at 3p21.3 in lung cancer has provided further support for the presence of a tumor suppressor gene in this chromosomal region. As a part of our efforts for positional cloning of a tumor suppressor gene at 3p21.3, we have characterized a transcriptional unit within this region using genomic fragments with interspecies conservation. The identified gene was found to encode a novel integrin alpha subunit, termed alpha RLC, which is closely related to alpha 4 in structure but clearly different from alpha 4 in its expression pattern in the physiological and pathological setting of the lung. This finding and the exact localization of the gene suggest that it is a good candidate for a tumor suppressor gene in lung cancer, but our extensive search covering one third of the gene did not reveal any somatic mutations within the coding region. Interestingly, however, alpha RLC was abundantly expressed in fetal lung and lung cancers, particularly small cell lung cancers (SCLC). Its aberrant upregulation in the SCLC samples, both cell lines and primary tumors, which might have been caused by a yet unidentified mutations or by deletions of other gene, and its homology to alpha 4, which is thought to play a role in metastasis, suggest that altered alpha RLC expression may contribute to the acquisition of malignant phenotypes of this type of lung cancer.
Amino Acid Sequence↗
Isolation of the poly(ADP-ribose) polymerase-encoding cDNA from Xenopus laevis: phylogenetic conservation of the functional domains.
The complete nucleotide (nt) sequence of the Xenopus laevis poly(ADP-ribose) polymerase (PARP)-encoding cDNA was determined. The putative X. laevis PARP protein consists of 1008 amino acids (aa) with a molecular weight of 113 kDa. X. laevis PARP shares 74, 83, 73, 78 and 42% aa sequence homology with the human, bovine, mouse, chicken and Drosophila melanogaster PARPs, respectively. Comparison of the PARP aa sequences among these species showed conservation of two zinc-finger motifs in the DNA-binding domain, and an NAD-binding motif and a Rossmann fold in the catalytic domain. The first Leu of the putative leucine zipper of D. melanogaster PARP is substituted to Lys in X. laevis PARP. All the Glu residues in the leucine zipper are conserved in these six species.
Amino Acid Sequence↗
2-Oxo-histidine as a novel biological marker for oxidatively modified proteins.
We report a promising finding that oxidative modification of proteins by free radicals could be monitored by the formation of oxidized histidine that is detectable by reverse-phase HPLC with electrochemical detection (HPLC-ECD). When the N-protected histidine derivative (N-benzoylhistidine) was exposed to a free radical-generating system (copper/ascorbate), a number of products were detected by HPLC-ECD and the main product among them was found to be identical to N-benzoyl-2-oxo-histidine. The acid hydrolysis of N-benzoyl-2-oxo-histidine provided a single product (2-oxo-histidine) that was detected sensitively by HPLC-ECD. Thus 2-oxo-histidine was indeed generated as the main product in the oxidatively modified proteins by free radicals. Taken together, 2-oxo-histidine may be a useful biological marker for assessing protein modifications under oxidative stress.
Biomarkers↗
A radioimmunoassay for pancreatic elastase 1: use of alpha 1-antitrypsin-elastase 1 conjugate as both standard and tracer to eliminate the influence of anti-elastase 1 autoantibodies.
We have recently reported the occurrence of anti-elastase 1 autoantibodies in human sera (Asada et al., Biochim Biophys Acta, 1991;1080:34-39). The usual radioimmunoassay of elastase 1 in autoantibody-positive sera gave abnormally high levels of elastase 1 and low recoveries of elastase 1 added to the sera. Now we have established a new radioimmunoassay system for determining pancreatic elastase 1 in serum, in which alpha 1-antitrypsin elastase 1 conjugate, the major conjugation form of elastase 1 in serum, is used as standard and alpha 1-antitrypsin-[125I]elastase 1 as tracer, because we found that the conjugate could bind to the elastase 1 specific antiserum but not to the autoantibodies. Thus the new assay system completely eliminate the unfavorable effects of the autoantibodies. The elastase 1 levels determined by the new assay method exhibited a good correlation with those obtained by a commercial assay kit in the autoantibody-negative sera, while no correlation was observed in the autoantibody-positive sera.
Acute Disease↗
Immunochemical detection of 4-hydroxynonenal protein adducts in oxidized hepatocytes.
We report here the development of an immunochemical procedure that uses an antibody specific to the 4-hydroxynonenal (HNE) moiety for the detection of HNE-protein adducts. The HNE-specific antibody was prepared by immunizing rabbits with a HNE-keyhole limpet hemocyanin conjugate and purifying the rabbit serum on an affinity gel prepared by covalent attachment of a HNE-conjugated heptapeptide. When various preparations of glyceraldehyde-3-phosphate dehydrogenase containing 0-7.0 equivalent of HNE-histidine residues per subunit were obtained by incubating samples of glyceraldehyde-3-phosphate dehydrogenase with increased amounts of HNE and subjected to immunoblotting with the HNE-specific antibody, the intensities of the blots were directly proportional to the number of HNE-histidine adducts as measured directly by amino acid analysis. Binding of the HNE-conjugated glyceraldehyde-3-phosphate dehydrogenase to the HNE-specific antibody could be completely inhibited by HNE-N-acetylhistidine, HNE-N-acetyllysine, or HNE-glutathione, suggesting that the antigenic determinant recognized by the antibody is the HNE moiety, not the HNE-amino acid conjugates, such as HNE-histidine, HNE-lysine, and HNE-cysteine. The utility of the HNE-specific antibody was demonstrated by its ability to react selectively with a number of HNE-protein adducts in immunoblot analyses of crude homogenates of rat liver hepatocytes that had been exposed to HNE or oxidative stresses with tert-butylhydroperoxide or metal-ion-catalyzed oxidation systems.
Aldehydes↗
Quantitation of HTLV-I provirus among seropositive blood donors: relation with antibody profile using synthetic peptides.
To study the pathophysiological state of HTLV-I carriers and the quantitative aspect of the risk of HTLV-I infection, we measured the absolute amounts of HTLV-I provirus in 39 seropositive blood donors. The amount of provirus varied from less than one to more than 10(4) molecules per 10(5) peripheral-blood mononuclear cells (PBMC). The average amount of HTLV-I provirus among seropositive blood donors was estimated as 3.7 x 10(3) molecules per 10(5) PBMC. The titer of the antibody against only the synthetic peptide (aa 100 to 130) of the gag p19 region of the HTLV-I provirus showed a weak but significant correlation with the amount of the provirus.
Adult↗
In vitro degradation of very low density lipoprotein from diabetic patients by lipoprotein lipase.
Fatty acid release by incubation with lipoprotein lipase (LPL) in vitro from very low density lipoproteins (VLDL) obtained from diabetic patients was low compared with that from healthy subjects, though the compositions were similar in both VLDL. Percentages of the large size VLDL decreased and those of the small size VLDL increased after the incubation with LPL. At the same time, on polyacrylamide gel disk electrophoresis, the smaller catabolic products from these VLDL appeared at a similar position to that of low density lipoproteins (LDL) and at the running front where high density lipoproteins (HDL) had migrated. The amount of the small size VLDL and the LDL-like lipoproteins produced from diabetic VLDL were less than those from normal VLDL and inversely correlated with the percent decrease of the large original size VLDL. This fact suggests that VLDL from diabetic patients are poor substrates for LPL compared with normal VLDL.
Diabetes Mellitus, Type 2↗
Antitumor and antimicrobial activities of enterococcal preparation orally administered to mice.
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Administration, Oral↗
[Aldosterone response to various stimuli in hyperthyroidism: in vivo and in vitro studies].
Responses of plasma aldosterone (PA) to alpha-ACTH-(1-24) (250 micrograms, im) injection and graded angiotensin II (AII) infusions (2, 4 and 8ng/kg/min for 30 min at each dose) on a constant sodium intake (170mEq daily) were assessed in 17 patients with Basedow's disease and 13 age-matched normal subjects. Aldosterone production in response to ACTH, AII and potassium in adrenal zona glomerulosa cells from L-thyroxine-induced hyperthyroid rats (H-rats) were also examined. Basal levels of plasma renin activity (PRA) and urinary aldosterone excretion were significantly higher (p < 0.01 and p < 0.05, respectively) in the patients with Basedow's disease than in the normal subjects, whereas basal PA level was similar in the two groups. The ACTH injection induced similar increases in plasma cortisol, plasma 18-hydroxycorticosterone (18-OHB) and PA in the two groups. The graded AII infusions also produced increases in plasma 18-OHB and PA in the two groups. Responses of these two corticosteroids to AII were, however, significantly lower (p < 0.05) in the patients with Basedow's disease than in the normal subjects. In the experimental animal study, basal PRA levels and the adrenal glomerulosa cell count/adrenal were significantly higher (p < 0.05) in the H-rats than in the control rats, whereas basal PA levels were similar in the two groups. Aldosterone production in response to AII, ACTH, and potassium increased in a dose-dependent manner in the two groups. Responses of aldosterone production to AII were, however, significantly lower (p < 0.05) in the H-rats than in the control rats. These results suggest that the impaired responsiveness of adrenal zona glomerulosa cells to AII, as well as an increased metabolic clearance rate of aldosterone, may be involved in the abnormal aldosterone metabolism in hyperthyroidism.
18-Hydroxycorticosterone↗
High-field magnetoabsorption study of heterointerface effects in CdTe/Cd0.6Mn0.4Te multiple quantum wells.
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Journal Article↗
Isolation of cDNAs encoding the catalytic domain of poly(ADP-ribose) polymerase from Xenopus laevis and cherry salmon using heterologous oligonucleotide consensus sequences.
We have isolated and sequenced cDNAs encoding the catalytic domain of poly(ADP-ribose) polymerase (PARP) from Xenopus laevis and Oncorhyncus masou (cherry salmon). The cDNAs were amplified by polymerase chain reaction using heterologous oligonucleotides corresponding to the conserved sequences of mammalian cDNAs as primers. The deduced amino acid sequences of Xenopus laevis and cherry salmon cDNA showed 84.4% and 75.6% similarities to that of human PARP, respectively. In both species, mRNA for PARP was identified as a single band of 4 kb, and PARP mRNA was abundant in ovary and brain. Thus, mixed oligonucleotide-primed amplification is a useful method in the cloning of cDNAs from different species, and the catalytic domain of PARP is conserved structurally among phylogenetically different species, suggesting an importance of poly(ADP-ribosyl)ation.
Amino Acid Sequence↗
MHC-linked diabetogenic gene of the NOD mouse: molecular mapping of the 3' boundary of the diabetogenic region.
To localize and characterize the MHC-linked diabetogenic gene of the NOD mouse, we studied the class III region of the MHC in the NOD mouse and related strains. Hsp70, Bat5, Tnfa and Tnfb loci were studied by microsatellite polymorphism analysis and/or restriction mapping. The CTS mouse had the same allele as the NOD mouse at the Hsp70 locus, but different alleles at the Bat5, Tnfa and Tnfb loci from those of the NOD mouse. Our previous studies indicated that in the CTS mouse, class II MHC was the same as that of the NOD mouse, but that class I MHC was different at both K and D loci, and that CTS MHC was diabetogenic in the presence of NOD background genes. These data map major genetic susceptibility to type 1 diabetes to the segment flanked by class I K and class III Bat5 loci. Moreover, since the diabetogenic effect of the CTS MHC is weaker than that of the NOD MHC, these data suggest the presence of a second MHC-linked gene or gene complexes that modulate susceptibility to type 1 diabetes outside this segment.
Animals↗
Cloning of cDNA encoding Drosophila poly(ADP-ribose) polymerase: leucine zipper in the auto-modification domain.
We have isolated cDNA clones for a Drosophila poly(ADP-ribose) polymerase (PARP; EC 2.4.2.30) by screening a lambda gt11 cDNA library with a Drosophila partial cDNA fragment. The Drosophila PARP probe was obtained by the polymerase chain reaction with heterologous primers deduced from conserved amino acids in the mammalian, chicken, amphibian, and fish sequences. The Drosophila PARP mRNA is 3.2 kb in length and is expressed in the early stages of development. The PARP protein of 994 amino acids contains two zinc-finger motifs and an NAD-binding motif, which are conserved among different species. Interestingly, the heptad leucine repeat in an alpha-helix was found in Drosophila PARP. Alignments of the auto-modification domains of various species showed the repeated hydrophobic amino acids on the same face of the helix that make the coiled-coil configuration in the mammalian and chicken sequences. The presence of a leucine-zipper motif in the auto-modification domain suggests that this motif might be responsible for protein-protein interaction between PARP and physiological acceptors. PARP may have novel functions, possibly involving its homo- and/or heterodimerization with other nuclear leucine-zipper proteins and its regulation by ADP-ribosylation.
Amino Acid Sequence↗
Covalent attachment of 4-hydroxynonenal to glyceraldehyde-3-phosphate dehydrogenase. A possible involvement of intra- and intermolecular cross-linking reaction.
Cytotoxic action of membrane lipid peroxidation product 4-hydroxynonenal (HNE) is due mainly to its facile reactivity with proteins (Esterbauer, H., Schaur, R. J., and Zollner, H. (1991) Free Radical Biol. Med. 11, 77-80). In the present study, the detailed mechanism of HNE modification of a key enzyme in intermediary metabolism, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), is studied mainly focusing on the formation of HNE-amino acid adducts in the enzyme. When GAPDH (1 mg/ml) was treated with 0-2 mM HNE in sodium phosphate buffer (pH 7.2) for 2 h at 37 degrees C, the enzyme was inactivated by HNE in a concentration-dependent manner. The loss of enzyme activity was associated with the loss of free sulfhydryl groups. Following its reduction with NaBH4, amino acid analysis of the HNE-modified enzyme demonstrated that histidine and lysine residues were also modified. At concentrations lower than 0.5 mM, HNE reacts preferentially with cysteine and lysine residues. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the HNE-modified enzyme suggested the formation of intra- and intermolecular cross-links of the enzyme subunit. The HNE-dependent loss of amino acid residues was accompanied by the generation of protein-linked carbonyl derivatives as assessed by reduction with NaB[3H]H4 and reaction with 2,4-dinitrophenylhydrazine. Thus, the conjugation of all the amino acids appears to involve Michael addition type reactions in which the carbonyl function of HNE would be preserved. The modified histidine residues were quantitatively recovered as the HNE-histidine adduct. However, only 28% of the missing lysine could be accounted for as the HNE-lysine derivative, and only 15.6% of the modified cysteine could be accounted for as the HNE-cysteine thioether derivative. It is proposed that the carbonyl groups of the HNE-derived Michael addition products may undergo secondary reactions with the amino acid groups of lysine residues to yield inter- and intrasubunit cross-links.
Aldehydes↗