[Pepsinogen I and II].
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Biomarkers, Tumor↗
Biomedical subjects
K Miki
Publications and source records attributed to K Miki.
Eight-year trend of new patients at Clinic for Disabled Patients, University Hospital, Faculty of Dentistry, Tokyo Medical and Dental University.
The trend of new patients was investigated at the Clinic for Disabled Patients, University Hospital, Faculty of Dentistry, Tokyo Medical and Dental University for eight years from 1989 to 1996. The purpose of this study was to explore the future role of this specialized clinic. The study revealed the following information: 1. A total of 1,388 new patients, corresponding to 173.5 new patients per year on average, visited the clinic. 2. The percentage of patients aged 30 or more increased. As a result, the mean patient age rose from 40.4 in 1989 to 47.6 in 1996. 3. Referrals accounted for more than 80% of new patients. Medical doctors referred more patients to our clinic than dentists did. 4. While the percentage of disabled patients tended to decrease, the percentage of medically compromised patients increased. 5. The predominant complaint was dental caries. Patients with cardiovascular diseases also complained of gingival bleeding and ill-fitting dentures. The growing percentage of medically compromised patients and the increasing mean age of patients over the eight-year period seems to reflect advances in medical science and the aging society. These tendencies could be more remarkable in the future. Therefore, much closer cooperation between dentists and medical doctors will be required.
Adolescent↗
Hepatocyte growth factor activator: a possible regulator of morphogenesis during fetal development of the rat gastrointestinal tract.
The role played by the hepatocyte growth factor activator (HGFA) during morphogenesis of the gastrointestinal tract was investigated in fetal rats between days 16 and 21 of gestation. By our recently established method using chelation and dissecting microscope, samples could be separated into epithelium and mesenchyme, essentially without cross-contamination. The expression of the gene for HGFA together with those for hepatocyte growth factor (HGF) and its receptor, c-met, was investigated in each tissue element by RT-PCR. In the fetal rat gastrointestinal tract, mRNA signals for the HGFA gene were observed only in epithelia expressing c-met mRNA. In contrast, expression of HGF mRNA was limited to the mesenchymal elements, indicating the presence of a local HGF system in the gastrointestinal tract; an inactive form of HGF (proHGF) is secreted from the mesenchyme and then cleaved into the active form by HGFA secreted by the target epithelia. During the period of morphogenesis and histodifferentiation in the gastrointestinal tract, enhanced expression of the genes for HGF and its receptor/c-met was evident, with elevated HGFA mRNA level observed throughout the gastrointestinal tract except in the forestomach, where mRNA expression was barely detectable. These results strongly suggest the possibility that morphogenesis of the gastrointestinal tract is regulated not only by a local increase in production of HGF, but also by enhanced proteolytic activation of proHGF. Thus, it is probable that locally synthesized HGFA plays a significant role as a regulator of the morphogenic action of HGF during gastrointestinal tract development.
Animals↗
Identification of tethering domains for protein kinase A type Ialpha regulatory subunits on sperm fibrous sheath protein FSC1.
The fibrous sheath is a unique cytoskeletal structure in the sperm flagellum believed to modulate sperm motility. FSC1 is the major structural protein of the fibrous sheath. The yeast two-hybrid system was used to identify other proteins that contribute to the structure of the fibrous sheath or participate in sperm motility. When FSC1 was used as the bait to screen a mouse testis cDNA library, two clones were isolated encoding the type Ialpha regulatory subunit (RIalpha) of cAMP-dependent protein kinase. Deletion analysis using the yeast two-hybrid system and in vitro binding assays with glutathione S-transferase-FSC1 fusion proteins identified two RIalpha tethering domains on FSC1. A domain located at residues 219-232 (termed domain A) corresponds to the reported tethering domain for a type II regulatory subunit (RII) of cAMP-dependent protein kinase, indicating that this binding domain has dual specificity to RI and RII. Another RIalpha tethering site (termed domain B) at residues 335-344 shows specific binding of RIalpha and had no significant sequence homology with known RII tethering domains. However, helical wheel projection analysis indicates that domain B is likely to form an amphipathic helix, the secondary structure of RII tethering domains of protein kinase A anchoring proteins. This was supported by the finding that site-directed mutagenesis to disrupt the amphipathic helix eliminated RIalpha binding. This is apparently the first report of an RIalpha-specific protein kinase A anchoring protein tethering domain.
Amino Acid Sequence↗
Effects of halothane, ketamine and nitrous oxide on dynorphin mRNA expression in dorsal horn neurons after peripheral tissue injury.
Peripheral tissue injury is known to induce changes in gene expression in spinal neurons and result in a prolonged alteration of neuronal excitability. The purpose of this study was to examine the effect of halothane on the dynorphin mRNA expression in spinal dorsal horn neurons after peripheral tissue injury by formalin injection and compare the effect to that of ketamine and nitrous oxide. Male Sprague-Dawley rats were anesthetized with 1.3% halothane, ketamine, or 67% nitrous oxide. Fifteen minutes after induction of anesthesia, rats received an intraplantar injection of 150 microliter 5% formalin into the unilateral hindpaw. General anesthesia was maintained for 8 h, and the expression of preprodynorphin (PPD) and preproenkephalin (PPE) mRNAs in the spinal cord (L4-5) was examined by in situ hybridization. The degree of edema of the inflamed foot was not different among the three anesthesia groups and the control (no anesthesia) group. The number of neurons expressing PPD mRNA dramatically increased in the superficial dorsal horn ipsilateral to the formalin injection in the control group compared to the contralateral side. The number of neurons labeled for PPD mRNA in the halothane group was significantly less than the control group. However, the number of PPD mRNA-expressing neurons in both the ketamine and nitrous oxide groups was significantly less than the halothane group. The expression of PPE mRNA was not influenced by these anesthetics. These data indicate that the suppressive effect of halothane anesthesia on the induction of PPD mRNA in dorsal horn neurons was smaller than those of ketamine and nitrous oxide, suggesting an important supplemental way to control the alteration of gene expression in spinal neurons for clinical settings.
Anesthetics, General↗
Crystallization and preliminary X-ray diffraction study of lactonohydrolase from Fusarium oxysporum.
The lactonohydrolase from the fungus Fusarium oxysporum AKU 3702 was crystallized by the vapour-diffusion procedure in the presence of polyethylene glycol 4000 as a precipitant. The crystals belong to the monoclinic space group P21 with unit-cell parameters a = 156, b = 100, c = 94.1 A, beta = 91.7 degrees. Assuming that there are two lactonohydrolase-dimer molecules in the asymmetric unit, the crystal volume per unit molecular mass, Vm, is calculated to be 2.94 A3 Da-1.
Carboxylic Ester Hydrolases↗
Involvement of dendritic cell response to resistance of stomach carcinogenesis caused by N-methyl-N'-nitro-N-nitrosoguanidine in rats.
The involvement of immune response in the resistance of chemically induced stomach cancer was studied in a resistant rat strain (Buffalo) and a sensitive rat strain (ACI). Groups of 10 male Buffalo and ACI rats, 6 weeks of age, were given drinking water with or without N-methyl-N'-nitro-N-nitrosoguanidine (MNNG; 100 mg/l) for 14 days. Total RNA was isolated from the stomach pyloric mucosa from five rats, and cDNA was prepared with reverse transcriptase. Tissue sections of the stomach pyloric mucosa from five rats were stained with antibodies recognizing molecules expressed by various immune cells. Reverse transcription-PCR (RT-PCR), competitive RT-PCR, and Northern blot demonstrated that the expression of MHC class II group genes [MHC class II, MHC class II-associated invariant chain (Ii), CD4 and IgM (B cell marker)], MHC class I group genes (MHC class I and CD8), B7-1 (costimulator on dendritic cells), and CD28 (receptor to B7 on T cells) in the pyloric mucosa was elevated by MNNG in both rat strains but was elevated to a 4-7-fold greater extent in Buffalo rats than in ACI rats. These genes were scarcely expressed in control rats. Histochemical antibody staining after MNNG exposure showed a greater number of cells stained with monoclonal antibody to Ii, OX-62 (dendritic cell marker), and ED-1 (dendritic cell and macrophage common marker) in the interstitial tissue of the pyloric mucosa of Buffalo rats compared with ACI rats. Cell proliferation, as measured by 5-bromo-2-deoxyuridine (BrdUrd)-labeling indices, revealed the presence of BrdUrd-labeled cells only among epithelial cells in the proliferative zone; cells in the interstitial tissue were not labeled with BrdUrd. The results suggest the involvement of dendritic cell response in the resistance to the MNNG induction of stomach carcinogenesis in rats.
Animals↗
Interaction site for soluble cytochromes on the tetraheme cytochrome subunit bound to the bacterial photosynthetic reaction center mapped by site-directed mutagenesis.
The crystallographic structure of the Blastochloris (formerly called Rhodopseudomonas) viridis tetraheme cytochrome subunit bound to the photosynthetic reaction center (RC) suggests that all four hemes are located close enough to the surface of the protein to accept electrons from soluble cytochrome c2. To identify experimentally the site of this reaction we prepared site-directed mutants of Rubrivivax gelatinosus RCs with surface charge substitutions in the bound cytochrome subunit and studied the kinetics of their reduction by soluble cytochromes (mitochondrial horse cytochrome c, Blc. viridis cytochrome c2, and Rvi. gelatinosus cytochrome c8). In comparison with the wild-type, the mutants E79K (glutamate-79 substituted by lysine), E93K (glutamate-93 substituted by lysine), and E85K (glutamate-85 substituted by lysine) located near the solvent-exposed edge of low-potential heme 1, the fourth heme from the special pair of bacteriochlorophyll, exhibited decreased second-order rate constants for the reaction between the tetraheme subunit and the soluble cytochromes. Double charge substitutions in this region: E79K/E85K (glutamate-79 and -85 both replaced by lysine) and E93K/E85K (glutamate-93 and -85 both replaced by lysine) appeared to show an additive inhibitory effect. Mutations in other charged regions did not alter the kinetics of electron transfer between bound and soluble cytochromes. In light of the available structural information on Blc. viridis RC, these results indicate that the cluster of acidic residues immediately surrounding the distal heme 1 of the RC-bound tetraheme subunit forms an electrostatically favorable binding site for soluble cytochromes. Thus, all four hemes in the subunit seem to be directly involved in the electron transfer toward the photo-oxidized special pair of bacteriochlorophyll. On the basis of these findings, a model is proposed for the hypothetical cytochrome c2-RC transient complex for Blc. viridis.
Amino Acid Substitution↗
Presence of polysialic acid and HNK-1 carbohydrate on brain glycoproteins from beta-1,4-galactosyltransferase-knockout mice.
Polysialic acid and HNK-1 carbohydrate are expressed on Gal beta 1-->4GlcNAc outer chains of N-linked sugar chain of neural cell recognition molecules at certain developmental stages and involved in neural tissue formation. Targeted inactivation of the mouse beta-1,4-galactosyltransferase (beta-1,4-GalT) gene resulted in short life of the mice which supposedly do not have such carbohydrate antigens but have no defects in neural tissue formation. Analysis of the mutant mouse brain glycoproteins revealed that polysialic acid and HNK-1 carbohydrate are normally expressed in an age-dependent manner. In support of this, protein bands reacted with Ricinus communis agglutinin-I, which interacts with oligosaccharides terminated with the Gal beta 1-->4GlcNAc group, and beta-1,4-GalT activity toward GlcNAc beta-S-pNP were detected in the mutant mouse brain, indicating that brain contains another functional beta-1,4-GalT important for the expression of the carbohydrate antigens.
Animals↗
Alternative splicing generates isoforms of human neuron-derived orphan receptor-1 (NOR-1) mRNA.
Neuron-derived orphan receptor-1 (NOR-1) is a novel member of the Nur77/NGFI-B subfamily within the nuclear receptor superfamily. Recently, several proteins closely related to NOR-1 have been described. To elucidate the relationships between NOR-1 and these closely related proteins, we analyzed the human NOR-1 gene and its transcripts by molecular cloning. We identified two variant NOR-1 transcripts in human skeletal muscles. One variant has a different 5'-untranslated region, and the other lacks C-terminal amino acid sequences corresponding to the putative ligand binding domain. These variant sequences share the common exon-intron boundary with NOR-1 mRNA, suggesting that they were generated from a single gene by alternative splicing, with the divergent points conserved between the rat and human. We also examined the promoter activities of the 5'-flanking regions of the two NOR-1 transcripts (NOR-1alpha and NOR-1beta mRNAs) by luciferase gene transfection. We demonstrated that the 5'-flanking region of the previously described NOR-1 gene, which has characteristics of a promoter found in housekeeping genes, showed potent promoter activity although the promoter for NOR-1beta mRNAs could not be determined.
Alternative Splicing↗
Risk factors for hyperamylasemia after hepatectomy using the Pringle maneuver: randomized analysis of surgical parameters.
OBJECTIVES: To determine whether the increased portal venous pressure caused by use of the Pringle maneuver contributes to inducing posthepatectomy hyperamylasemia and, subsequently, to evaluate risk factors for its development. DESIGN: Randomized study. SETTING: University hospital. PATIENTS: Forty patients who were going to undergo hepatectomy were assigned prospectively to either a superior mesenteric artery clamp (n=20) or a nonclamp (n=20) group by the random-block method. INTERVENTIONS: The Pringle maneuver was used during hepatectomy, and in the superior mesenteric artery clamp group the superior mesenteric arteries were clamped simultaneously. MAIN OUTCOME MEASURES: Amylase activity, isozyme, and creatinine levels in the blood and urine samples were measured before and after surgery, and the amylase creatinine clearance ratio was estimated. RESULTS: The serum amylase activity levels of the superior mesenteric artery clamp and nonclamp groups did not differ significantly during the 7 postoperative days. The serum amylase activity levels exceeded 250 U/L in 14 patients (group 1) and remained below this level in 26 (group 2). The salivary-type isozyme levels of group 1 increased significantly compared with those of group 2, and the levels of group 2 remained normal. The total amount of amylase excreted in the urine samples of group 1 patients also increased significantly, with the salivary-type isozyme predominating. All the mean amylase creatinine clearance ratios before and after surgery remained normal. The mode chi2 of the logistic model including the indocyanine green retention rate at 15 minutes and the ratio of the resected liver weight to the whole liver volume showed a significantly increased risk (P=.01). CONCLUSION: It is not the increased portal venous pressure caused by use of the Pringle maneuver but the liver function and the extent of liver resection that are considered risk factors for inducing posthepatectomy salivary-type hyperamylasemia.
Aged↗
Antroduodenal motor function and gastro-oesophageal reflux in neurologically impaired children, adolescents and young adults.
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Adolescent↗
Mechanism of lower oxidizability of eicosapentaenoate than linoleate in aqueous micelles. II. Effect of antioxidants.
We have reported that the peroxyl radicals derived from methyl eicosapentaenoate (20:5n-3) are more polar than those from methyl linoleate (18:2n-6) since the former peroxyl radicals have at least two molecules of oxygen in a molecule while the latter peroxyl radical has one. This lowers the oxidizability for 20:5n-3 in aqueous Triton X-100 micelles by enhancing the termination reaction rate for peroxyl radicals and by reducing the rate of propagation since there may be more polar peroxyl radicals derived from 20:5n-3 at the surface than within the micelle core. In this study, we measured the effect of three antioxidants, di-tert-butyl-4-methylphenol (BHT), 2,2,5,7,8-pentamethyl-6-chromanol (PMC) and 2-carboxy-2,5,7,8-tetramethyl-6-chromanol (Trolox), on the oxidation of lipids in aqueous micelle. Antioxidants give a clear induction period during oxidation of 18:2n-6 initiated with a water-soluble radical initiator, and its induction length decreases in the order of BHT > PMC > Trolox. This is consistent with the proposed location of three antioxidants: being in the core of micelle, at the surface, or in aqueous phase, respectively. However, BHT does not inhibit the oxidation of 20:5n-3 efficiently, and its rate of oxidation is slower than that observed in the oxidation of 18:2n-6, supporting the idea that polar peroxyl radicals derived from 20:5n-3 are preferentially located at the surface of the micelle. Similar results were obtained when oxidation was initiated with a lipid-soluble radical initiator except antioxidants had lesser effect on the oxidation rate of 20:5n-3.
Aerobiosis↗
The sugar permeability test reflects disease activity in children and adolescents with inflammatory bowel disease.
OBJECTIVES: To investigate the relationship of intestinal permeability in children and adolescents with inflammatory bowel disease (IBD) to disease activity, disease extent, and response to therapy. STUDY DESIGN: Patients with new and established diagnoses of IBD (12 Crohn's disease [CD] and 18 ulcerative colitis [UC]) were studied. Intestinal permeability was evaluated by measuring with high-performance liquid chromatography 5-hour urinary excretion ratio of lactulose/L-rhamnose (L/Rh). RESULTS: In 8 of 9 patients with active CD, the L/Rh ratio was higher than the reference range (0.006 to 0.074, n = 36). In inactive CD (n = 3) the L/Rh ratio was within the reference range. In 6 of 7 patients with active extensive UC, the L/Rh ratio was elevated. In inactive extensive UC (n = 6) the normal permeability ratio was shown. In both active CD and active extensive UC, the frequency of elevated intestinal permeability was significantly greater than values in both inactive forms. The permeability ratio was normal in 4 of 5 patients with active left-sided colitis. In 5 of 7 patients (3 CD, 4 UC), repeat permeability values entered the reference range after acute phase therapy. Two patients with persistently elevated intestinal permeability (1 CD, 1 UC) had a disease flare-up within 6 months. CONCLUSIONS: Intestinal permeability is a marker of disease activity in CD and extensive UC. Serial permeability test may be useful in monitoring disease activity.
Adolescent↗
Liver allograft functional reserve estimated by total asialoglycoprotein receptor amount using Tc-GSA liver scintigraphy.
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Adolescent↗
Calcitonin gene-related peptide increase in the rat spinal dorsal horn and dorsal column nucleus following peripheral nerve injury: up-regulation in a subpopulation of primary afferent sensory neurons.
Calcitonin gene-related peptide in sensory primary afferent neurons has an excitatory effect on postsynaptic neurons and potentiates the effect of substance P in the rat spinal dorsal horn. It has been established that calcitonin gene-related peptide expression in dorsal root ganglion neurons is depressed, and the effect of calcitonin gene-related peptide on dorsal horn neurons is attenuated, following peripheral nerve injury. We report here that a subpopulation of injured dorsal root ganglion neurons show increased expression of calcitonin gene-related peptide. Using in situ hybridization and the retrograde tracer, FluoroGold, we detected an increased number of medium- to large-sized rat dorsal root ganglion neurons projecting to the gracile nucleus that expressed alpha-calcitonin gene-related peptide messenger RNA following spinal nerve transection. Immunohistochemistry revealed a significant increase in calcitonin gene-related peptide immunoreactivity in the gracile nucleus and in laminae III-IV of the spinal dorsal horn. These results indicate that a subpopulation of dorsal root ganglion neurons express alpha-calcitonin gene-related peptide messenger RNA in response to peripheral nerve injury, and transport this peptide to the gracile nucleus and to laminae III-IV of the spinal dorsal horn. The increase of the excitatory neuropeptide, calcitonin gene-related peptide, in sites of primary afferent termination may affect the excitability of postsynaptic neurons, and have a role in neuronal plasticity following peripheral nerve injury.
Animals↗
Induction of precocious pepsinogen synthesis by glucocorticoids in fetal rat gastric epithelium in organ culture: importance of mesenchyme for epithelial differentiation.
Glucocorticoids significantly affect both proliferation and differentiation of gastric epithelial cells in vivo. Here we examined the mechanism of action of glucocorticoids on the cells in vitro, with special reference to the epithelial-mesenchymal interaction. When 16.5-day fetal rat gastric explants were maintained in organ culture, the epithelial cells began to invaginate into mesenchyme on days 3 to 4, and formed glandular structures on days 5 to 6 in culture. Immunohistochemical analysis with specific antibodies revealed that pepsinogen-synthesizing cells first appeared on day 2, and they increased in number with epithelial morphogenesis to about 20%-30% of total epithelial cells on days 4 to 6, and that these cells were localized at the base of glandular structures in control media. When the explants were treated with hydrocortisone (1 microgram/ml), epithelial morphogenesis was mostly suppressed, but epithelial cytodifferentiation was significantly stimulated, indicating that epithelial morphogenesis is not necessary for their cytodifferentiation. In glucocorticoid-treated explants, pepsinogen-synthesizing cells first appeared on day 1, and more than 90% of the cells were positively stained with the antibodies from days 3 to 5 in culture. Biochemical analysis showed that much higher acid protease activity could be detected in glucocorticoid-treated explants than in controls from days 2 to 6 in culture, and analysis by zymography indicated that the synthesis of pepsinogen 1 but not cathepsin E was stimulated by the hormone. Northern blotting analysis showed that the level of pepsinogen 1 mRNA was greatly increased by glucocorticoids. Examination of the effect of the hormone on the epithelial proliferation showed that hydrocortisone (1 microgram/ml) significantly inhibited the epithelial growth from days 1 to 3 in culture. To investigate the role of epithelial-mesenchymal interaction in the glucocorticoid-induced differentiation of the gastric epithelial cells, effects of the hormone on the proliferation and differentiation of the cells in the absence of mesenchyme were examined, using a recently established primary culture system. The epithelial cells synthesized cathepsin E but not pepsinogen in cell culture, irrespective of glucocorticoid treatment, and the level of acid protease activity was not affected by the hormone, indicating that mesenchyme is necessary for the hormone to induce pepsinogen gene expression in the epithelial cells. In the cell culture system, glucocorticoids did not inhibit but significantly stimulated epithelial proliferation. This suggests that the hormone indirectly inhibited epithelial proliferation in organ culture, probably via mesenchyme. The mechanism of action of glucocorticoids on the epithelial-mesenchymal interaction in the fetal glandular stomach is discussed.
Animals↗
Reconstruction of an extensive scalp defect using the split latissimus dorsi flap in combination with the serratus anterior musculo-osseous flap.
A combination of split latissimus dorsi musculocutaneous flap with the serratus anterior musculo-osseous flap was used to cover a very large defect of the scalp. This technique offers a new alternative for single-stage reconstruction of extensive scalp defects.
Bone Transplantation↗