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Biomedical subjects

H Kurihara

Publications and source records attributed to H Kurihara.

At least 235 records · Page 13Linked to original sources

Impaired development of the thyroid and thymus in endothelin-1 knockout mice.

We have previously demonstrated that endothelin-1 (ET-1) is essential to the development of neural crest-derived craniofacial and cardiovascular structures. In this study we evaluated the intrauterine growth and development of the thyroid and thymus glands in Edn1-/- homozygous mice. Edn1-/- homozygous mice were smaller than their heterozygous or wild-type littermates (about 90% of normal body weight). The thyroid and thymus of Edn1-/- homozygous mice were smaller than those of normal mice and were not fused in the midline, resulting in two separate lobules. The thymus of Edn1-/- homozygous mice is not descended into the normal position. These results suggest that ET-1 is important for normal development of the glands in the neck region, as well as for systemic growth. The combination of abnormalities in Edn1-/- homozygous mice suggests that these mice might serve as an animal model for the human diseases DiGeorge syndrome and velocardiofacial syndrome.

Animals↗

Gene expression of endothelin isoforms and receptors in endothelin-1 knockout mice.

Gene expression of endothelin (ET) isoforms and receptors was examined in ET-1 knockout mice and compared with that of wild-type mice. ET-1 mRNA and peptide levels were in parallel with the number of Edn1 authentic alleles. ET-2 mRNA levels in intestine and ET-3 mRNA and/or peptide levels in lung, brain, and intestine were not different among Edn1+/+ wild-type, Edn1+/- heterozygous, and Edn1-/- homozygous mice. ET(A) and ETB receptor mRNA levels in several organs also were not different among Edn1+/+ wild-type, Edn1+/- heterozygous, and Edn1-/- homozygous mice. These results suggest that gene expression of ET-2, ET-3, and ET(A/B) receptors is not affected by levels of ET-1 production and is regulated independently. Neither compensation by redundant expression of other ET isoforms nor upregulation of ET receptors could explain the elevated blood pressure in Edn1+/- heterozygous mice.

Animals↗

Isolation and characterization of vascular endothelial cells derived from mice lacking endothelin-1.

Microvascular endothelial cells were isolated from endothelin (ET)-1 knockout mice. The ET-1-lacking endothelial cells represent normal shape and activity of acetyl-LDL uptake. The growth of ET-1-lacking endothelial cells was not different from that of control cells in the presence of serum. Compensatory production of other ET isoforms does not occur in these cells. Conversion of big ET-1 to ET-1 is not different between ET-1-lacking and control cells. These results suggest that ET-1 is not essential to endothelial morphology and growth, that the expression of ET isoforms is not redundant in endothelial cells, and that the machinery processing ET-1 is preserved despite of the absence of its substrate.

Animals↗

Heat shock protein 60 (GroEL) from Porphyromonas gingivalis: molecular cloning and sequence analysis of its gene and purification of the recombinant protein.

Porphyromonas gingivalis is associated with human periodontal disease. We cloned and sequenced the gene for heat shock protein 60 (GroEL, HSP60) from P. gingivalis FDC381. The identified clone carried a 2.6 kb DNA fragment which contained two open reading frames (ORFs) encoding a 9.6- and a 58.4-kDa protein. The translated amino acid sequence of these ORFs showed a high degree of homology with known sequences for GroES and GroEL from several bacterial species and humans. Escherichia coli carrying this clone expressed a 65-kDa protein which was recognized by anti-Mycobacterium leprae HSP60 monoclonal antibody. We purified the 65-kDa protein by DEAE-sepharose chromatography and hydroxyapatite chromatography. This protein was immunogenic and was recognized by sera from a number of patients with periodontal disease. This immunological reactivity and the existence of molecular mimicry between the P. gingivalis GroEL and other HSP homologs may indicate an important role for this molecule in periodontal lesion.

Amino Acid Sequence↗

Elevated blood pressure and craniofacial abnormalities in mice deficient in endothelin-1.

The endothelin-1 (ET-1) gene was disrupted in mouse embryonic stem cells by homologous recombination to generate mice deficient in ET-1. These ET-1-/- homozygous mice die of respiratory failure at birth and have morphological abnormalities of the pharyngeal-arch-derived craniofacial tissues and organs. ET-1+/- heterozygous mice, which produce lower levels of ET-1 than wild-type mice, develop elevated blood pressure. These results suggest that ET-1 is essential for normal mouse development and may also play a physiological role in cardiovascular homeostasis.

Animals↗

Massive intestinal bleeding caused by an ileal arteriovenous malformation: report of a case.

We report herein the case of a 43-year-old man in whom gastrointestinal bleeding was found to be caused by a large arteriovenous malformation (AVM) in the ileum. Emergency angiography proved invaluable in disclosing the AVM as the bleeding source, after which the patient was successfully treated by surgery. Although the number of reports of AVM has recently been increasing, ileal AVMs are still relatively rare. Moreover, according to the Japanese literature, no other patient has had such marked and large feeding arteries and draining veins as those in the patient described in this report.

Adult↗

Endobronchial metastasis from a primary uterine osteosarcoma in a patient with multiple myeloma: report of a case.

We present herein the case of a 75-year-old woman with multiple myeloma who underwent a left lower lobectomy for endobronchial metastasis from an uterine osteosarcoma. She had initially been admitted to our hospital for chemotherapy more than 1 year earlier, soon after which a primary uterine osteosarcoma was discovered and a total abdominal hysterectomy and bilateral salpingo-oophorectomy performed. One year after the operation, the patient developed hemoptysis. A flexible bronchofiberscopy demonstrated a polypoid mass obstructing the left basal bronchus, and computed tomographic scans showed three pulmonary nodules. Surgery was performed to control the hemoptysis. At thoracotomy, two metastatic nodules were identified in the left lower lobe, and the endobronchial extension of the tumor was resected en bloc with the left lower lobe. The tumor was diagnosed as lung metastasis from the uterine osteosarcoma. Although further lung tumors have recently appeared, the patient has remained well for the 3 years since her last operation without any hemoptysis.

Aged↗

Factors affecting functional prognosis in survivors of primary central nervous system germinal tumors.

We evaluated 79 patients of whom 62 survived treatment for intracranial germinal tumors between 1964 and 1992. The survivors were assessed for their Karnofsky scores and intellectual and reproductive functions to verify factors associated with better prognosis. Thirty-one tumors were pineal, 17 were suprasellar, and ten were in the basal ganglia. These included 50 germinomas, five teratomas, and seven tumors of extra-embryonic origin producing alpha-fetoprotein or human chorionic gonadotropin. Although patients with germinoma or benign teratoma had better Kaplan-Meier survival rates than those with extra-embryonic origin tumors, the mean Karnofsky scores of the survivors did not differ significantly among the three histologic groups. On the other hand, although survival rates did not differ significantly among different tumor site groups, the mean Karnofsky score of the pineal group was higher than that of the suprasellar or basal ganglia group. Intellectual and reproductive functions were severely affected in the basal ganglia and suprasellar tumor groups, respectively. The Karnofsky score of survivors treated after 1977 was higher than for those treated before 1977. The score of patients who received less than 55 Gy irradiation was better than those who received more than 55 Gy. No patient who received less than 45 Gy developed a recurrence, suggesting that a lower dose may suffice for pure germinomas. Tumors of extra-embryonic origin, however, should be treated with combined chemotherapy to minimize subsequent irradiation. Factors affecting survivors' capabilities are discussed.

Adolescent↗

Stenotic bicuspid aortic valve associated with a ventricular septal defect in an adult presenting with congestive heart failure: a rare observation.

This report is concerned with an adult presenting with stenotic bicuspid aortic valve associated with a ventricular septal defect (VSD). The association between aortic regurgitation (AR) and VSD has often been described, but that between a stenosed aortic valve and VSD has been rarely observed, although bicuspid aortic valves and ventricular septal defects are probably the two most common congenital heart defects. The development of congestive heart failure in the presented case was considered to be due to an increase in the left to right shunt through the VSD. This was attributable to a progressive elevation in left ventricular pressure as a result of the development, with age, of stenosis of the bicuspid aortic valve.

Aged↗

pH profile of kinetic constants of RNase Rh from Rhizopus niveus and its mutant enzymes towards UpU, and possible mechanisms of RNase Rh.

In order to elucidate the mechanism of action of Rhizopus niveus RNase Rh, we investigated the pH profiles of the kinetic parameters of RNase RNAP Rh, a derivative of RNase Rh, and its mutant enzymes, i.e., RNase RNAP Rh H104F, RNase RNAP Rh E105Q, and RNase RNAP Rh D51N. Based on comparisons of their profiles we concluded that protonation of His104 is indispensable for the enzymatic activity and Glu105 accelerates the enzymatic activity, especially at acid pH centered at pH 3.5. Based on these data and the previous data on the chemical modification and enzymatic properties of other mutant enzymes, we propose the following as a possible mechanisms of RNase Rh action. (i) His109 participates in enzymatic action as a general base catalyst which removes the hydrogen of the 2'-OH of the ribose moiety. (ii) His46 participates in the reaction as a general acid catalyst which interacts with the 5'-oxygen atom of the scissile phosphodiester bond and becomes a proton donor to the departing nucleoside or nucleotide. (iii) His104 interacts with phosphate anion and its protonation is favorable for the enzymatic activity. (iv) Since the protonated form of Glu105 is more favorable for activity, we postulate two possible roles for Glu105: (a) its stabilizes the intermediate, and (b) it interacts with the oxygen atom of P = O and polarizes the phosphorus atom.

Dinucleoside Phosphates↗

Isolation and characterization of a 53 kDa major cell envelope protein antigen from Treponema denticola ATCC 35405.

A major cell envelope protein was purified from the cell envelope fraction of Treponema denticola ATCC 35405 by ion exchange chromatography after extraction with Zwittergent 3-14. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed a relative molecular mass of 53 kDa for this protein with a pI of 6.3-6.8. Amino acid analysis revealed that this protein contained high proportions of hydrophobic amino acids (40.4%), and no cysteine could be detected. The N-terminus of the protein was blocked to Edman degradation. Rabbit antiserum raised against the purified 53 kDa protein reacted with the outer envelope of the T. denticola cell surface as confirmed by immunoelectron microscopy. This rabbit antiserum reacted with 4 of the 11 strains of treponemes tested in this study. Sera from 9 to 18 periodontitis patients reacted strongly with this 53 kDa cell envelope protein of T. denticola as determined by immunoblotting analysis.

Amino Acids↗

Role of Ca2+ and protein kinase C in shear stress-induced actin depolymerization and endothelin 1 gene expression.

Vascular endothelial cells adapt to changes in blood flow by altering the cell architecture and by producing various substances. We have previously reported that low shear stress induces endothelin 1 (ET-1) expression in endothelial cells and that this induction is mediated by depolymerization of actin fiber. In the present study, we examined the role of Ca2+ and protein kinase C (PKC) in shear stress-induced actin depolymerization and subsequent ET-1 gene expression. Exposure of cultured porcine aortic endothelial cells to low shear stress (5 dyne/cm2) for 3 hours increased the ratio of G-actin to total actin from 54 +/- 0.8% to 80 +/- 1.0%. This shear stress-induced actin depolymerization was completely blocked by chelation of extracellular Ca2+ with EGTA and partially inhibited by intracellular Ca2+ chelation with the tetraacetoxymethyl ester of BAPTA (BAPTA/AM). Pretreatment with staurosporine, a PKC inhibitor, or desensitization of PKC by treatment with 12-O-tetradecanoylphorbol 13-acetate (TPA) for 24 hours also resulted in partial inhibition of shear stress-induced actin depolymerization. Although PKC activation by TPA mildly increased G-actin content, the effect of TPA and shear stress on actin depolymerization was not additive. Moreover, shear stress-induced ET-1 gene expression was inhibited by EGTA, BAPTA/AM, and staurosporine to a degree similar to the inhibition of actin depolymerization. In contrast, ET-1 gene expression induced by cytochalasin B, an actin-disrupting agent, was not affected by staurosporine.(ABSTRACT TRUNCATED AT 250 WORDS)

Actins↗

Recurrence of primary intracranial germinomas after complete response with radiotherapy: recurrence patterns and therapy.

Nine germinoma patients are described who developed a recurrence after a complete response to radiation without adjuvant chemotherapy. Extraembryonic tumors producing alpha-fetoprotein and human chorionic gonadotropin were excluded from this study. Four patterns of recurrence are described with respect to mechanism and appropriate treatment. Type I germinoma recurrence, characterized by intracranial recurrence caused by an inadequate initial irradiation field was treated by total craniospinal irradiation. Type II recurrence, characterized by a benign teratoma caused by late growth of the teratoma component was treated by surgery alone. All patients with these patterns of recurrence are still alive. Type III local recurrence is characterized by human chorionic gonadotropin- or alpha-fetoprotein-producing tumors of extraembryonic origin. This pattern of recurrence should be treated by chemotherapy or radiosurgery, because all these patients died. Type IV germinoma recurrence consists of extraneural metastasis without evidence of intracranial recurrence. Two of these patients were treated with chemotherapy. In summary, four patients died after recurrence, whereas the remaining five patients survived. The classification of germinoma recurrence patterns should facilitate the selection of the most appropriate treatment. However, it has been difficult to identify the precise histopathology by biopsy or partial resection alone. Furthermore, chemotherapy is indicated in treating germinomas that have a ventriculoperitoneal shunt because of the risk of extraneural metastases.

Adolescent↗

Molecular basis of leukocyte adhesion molecules in early-onset periodontitis patients with decreased CD11/CD18 expression on leukocytes.

We analyzed the cell-cell adherence related to CD11/CD18 and CD18 mRNA in individuals with decreased CD11/CD18 expression on their neutrophil surface. Epstein Barr virus-transformed B cell lines were developed from one localized juvenile periodontitis (LJP) patient with decreased CD11/CD18 in the peripheral blood neutrophils and without systemic diseases; two siblings with generalized prepubertal periodontitis (GPP) caused by leukocyte adhesion deficiency (LAD); another LJP patient; one localized prepubertal periodontitis (LPP) patient; and two healthy subjects. Adhesion of leukocytes to each other was measured as cluster formation by aggregation assay. The length and the amount of CD18 mRNA expressed in the cell lines were analyzed by Northern blotting using the 32P-labeled CD18 cDNA. The coding region of the mRNA was analyzed by the reverse transcription-polymerase chain reaction method. Base-mismatches between CD18 mRNA and the 32P-labeled RNA probe synthesized from CD18 cDNA were analyzed by RNase protection assay. In the adherence assay, cells from the LJP patients with decreased CD11/CD18 formed more clusters of smaller size and fewer cells than those of the other subjects. The cells from GPP and LAD patients did not aggregate and did not form clusters either in the absence or presence of PMA. There were no differences in the length and the amount of mRNA between the LJP patients and the other subjects, while GPP-LAD patients expressed a small amount of long mRNA. The whole coding region (2,313 base pairs) of all subjects was amplified except for the GPP-LAD patients, and the 5'-region (1,119 base pairs) was amplified from all subjects.(ABSTRACT TRUNCATED AT 250 WORDS)

Aggressive Periodontitis↗

Cytokine-dependent synergistic regulation of interleukin-8 production from human gingival fibroblasts.

Human Gingival Fibroblasts (HGF) may have an important role in the orchestration of immuno-participant cells infiltrating the gingiva in response to continuously recurring bacterial infection. To examine the cytokine network regulating HGF-derived interleukin (IL)-8, a potent neutrophil chemotactic cytokine, we analyzed the effects of inflammatory cytokines alone and in combination on IL-8 production by HGF. IL-1 beta, tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), IL-6, and IL-8 were used as stimulants. HGF secreted IL-8 in a dose-dependent manner after stimulation with either IL-1 beta or TNF-alpha, but not with IFN-gamma or IL-6. Furthermore, IL-8 itself did not affect IL-8 mRNA accumulation in HGF in an autocrine manner. The combination of IL-1 beta and TNF-alpha synergistically enhanced the secretion of IL-8, whereas IFN-gamma suppressed IL-8 secretion by IL-1 beta- or TNF-alpha-stimulated HGF. These effects were also observed at each level of IL-8 mRNA expression in HGF. IL-8 secretion by cytokine-stimulated HGF was not influenced by the inhibition of PGE2 synthesis with indomethacin, indicating that endogenous PGE2 was not involved in IL-8 production by HGF. These results indicate that IL-8 production by HGF is synergistically stimulated by specific cytokines, IL-1 beta and TNF-alpha, and suggest that these stimulatory effects are down-regulated by IFN-gamma at the transcriptional level through PGE2-independent pathways. Thus, neutrophil-mediated processes in periodontal disease may be regulated in part by HGF in the cytokine network of immuno-participant cells.

Chemotaxis, Leukocyte↗