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Biomedical subjects

A Watanabe

Publications and source records attributed to A Watanabe.

At least 415 records · Page 23Linked to original sources

First-trimester prenatal molecular diagnosis of infantile hypophosphatasia in a Japanese family.

We obtained a prenatal molecular diagnosis during the first trimester in a Japanese woman whose first child (the proband) had been a compound heterozygote for infantile hypophosphatasia. We examined chorionic villus DNA samples obtained at 10 weeks of gestation for the base substitutions detected in the proband DNA using polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) and PCR-allele-specific oligonucleotide (ASO) analysis. The genotype of the fetus was the same as that of the proband. The same mobility shift patterns of single strand conformation polymorphism (SSCP) bands were observed in the fetus and the proband. This molecular approach to prenatal diagnosis appears to be more accurate than the enzymatic method and also more accurate and more rapid than the conventional RFLP method.

Alleles↗

Dural caroticocavernous fistula with both ophthalmic arteries arising from middle meningeal arteries.

A 60-year-old woman presented with a dural caroticocavernous fistula (CCF) in whom both by ophthalmic arteries arose from the middle meningeal arteries instead of from the internal carotid arteries. Transarterial embolisation of the CCF was contraindicated because of the risk of blindness. Transvenous coil embolisation was therefore performed. Bilateral origin of the ophthalmic artery from the middle meningeal artery is extremely rare.

Brain Diseases↗

Prenatal diagnosis of the fetal RhD blood type using a single fetal nucleated erythrocyte from maternal blood.

OBJECTIVE: To develop a method that allows prenatal diagnosis of the fetal RhD blood type from maternal blood. METHODS: Maternal blood was obtained at 8-31 weeks' gestation, and nucleated erythrocytes were separated with Percoll using a discontinuous density gradient method, then collected individually by micromanipulation under microscopic observation. After whole genome amplification with primer extension pre-amplification, exon 7 of the RhD and RhCE as well as the ZFX/ZFY loci were further amplified by a nested polymerase chain reaction (PCR). RESULTS: Nucleated erythrocytes were detected in nine of ten maternal blood samples, and sex was determined in 13 of 21 nucleated erythrocytes. RhD genotype could be diagnosed in 12 of the 13 nucleated erythrocytes in which sex could be determined. The results of RhD blood type and sex in nucleated erythrocytes obtained from maternal blood were identical with those of newborns. Fetal RhD blood type could be determined in six of ten maternal blood samples. CONCLUSION: A new method for noninvasive prenatal diagnosis of the fetal RhD blood type using a single nucleated erythrocyte isolated from maternal blood was demonstrated. This diagnostic method offers extremely useful information for the management of Rh-negative pregnant women. Furthermore, this method of prenatal diagnosis can be applied to other genetic disorders and is expected to become the preferred method of noninvasive prenatal diagnosis of DNA.

Base Sequence↗

Therapeutic basis of glycyrrhizin on chronic hepatitis B.

Glycyrrhizin, a major component of a herb (licorice), has been intravenously used for the treatment of chronic hepatitis B in Japan and improves liver function with occasional complete recovery from hepatitis. This substance modifies the intracellular transport and suppresses sialylation of hepatitis B virus (HBV) surface antigen (HBsAg) in vitro. This study was designed to clarify the pharmacological basis for its effectiveness. The structure-bioactivity relationship of glycyrrhizin, glycyrrhetic acid 3-O-monoglucuronide and glycyrrhetic acid was determined, and glycyrrhetic acid was found to be the most active of them. The amounts of three substances bound to the liver were evaluated in guinea pigs after intravenous administration of glycyrrhizin. Glycyrrhizin and glycyrrhetic acid 3-O-monoglucuronide were detected at concentrations of 31.8-1.3 micrograms/g of liver, but glycyrrhetic acid was not detected. When glycyrrhizin attained these concentrations in the cellular fraction of the PLC/PRF/5 cell culture, it suppressed the secretion of HBsAg as reported previously. These results indicated that glycyrrhizin administered intravenously might bind to hepatocytes at the concentration at which glycyrrhizin could modify the expression of HBV-related antigens on the hepatocytes and suppress sialylation of HBsAg.

Animals↗

A postmortem study of glycine and its potential precursors in chronic schizophrenics.

We have measured the concentrations of glycine and its potential precursors, serine and threonine, in 20 areas of the postmortem brains of chronic schizophrenics and controls using high-performance liquid chromatography by pre-column derivatization with dimethyl-amino-azobenzene sulphonyl chloride. The regional distribution pattern of glycine in the postmortem brains with and without the disease was more similar to that of serine (r = 0.874, P < 0.0001) than to that of threonine (r = 0.476, P < 0.01). A multiple regression analysis with regressor variables including diagnosis, age at death and interval between death and freezing revealed that there is a significant difference between schizophrenics and controls in the contents of these amino acids in a number of brain areas. The level of glycine in the orbitofrontal cortex of schizophrenics was found to be significantly increased in schizophrenics, with a tendency to an increase in that of serine. The increase in glycine was also significantly high in the off-drug group of schizophrenics who had not taken antipsychotics more than 40 days before death. Prominent decreases in both glycine and serine were observed in the somesthetic cortex of the on-drug schizophrenics. Serine was found to be significantly decreased in the putamen of the off-drug schizophrenics. A marked decrease in threonine was also observed in the supramarginal cortex and posterior portion of the lateral occipitotemporal cortex of the off-drug group of schizophrenics and in the putamen of all schizophrenics. The highly similar distribution pattern of glycine and serine in the postmortem brains supports the close coupling of synthesis and metabolism between these chemicals in human brains. The increased content of glycine in the orbitofrontal cortex, the reduced level of serine in the putamen and the decrease in threonine in the cerebral cortices, which were prominent in the off-drug schizophrenics, may be involved in the pathophysiology of schizophrenia.

Adult↗

Methamphetamine modifies the photic entraining responses in the rodent suprachiasmatic nucleus via serotonin release.

We examined whether methamphetamine modifies the photic entraining responses in the rat suprachiasmatic nucleus. Optic nerve stimulation increased vasoactive intestinal polypeptide release from rat suprachiasmatic nucleus slices, and methamphetamine inhibited this increase in a concentration-dependent manner. Optic nerve stimulation has been reported to evoke field potentials in rat suprachiasmatic nucleus slices. Methamphetamine attenuated this field potential, and maximal inhibition (75.5%) was achieved at a concentration of 100 microM. Systemic administration of methamphetamine (1-5 mg/kg) inhibited light (300 lux, 1h)-induced Fos expression in the suprachiasmatic nucleus; methamphetamine at a dose of 5 mg/kg, i.p. caused 40% inhibition of light-induced Fos expression. We examined whether the inhibitory effect of methamphetamine on photic entraining responses mediates serotonin release from the suprachiasmatic nucleus. High-performance liquid chromatographic analysis revealed that methamphetamine application increased serotonin release from rat suprachiasmatic nucleus slices in a concentration-dependent manner, but did not affect noradrenaline release. In addition, reduction of serotonin content attenuated the effect of methamphetamine on field potential induced by optic nerve stimulation in vitro and also light-induced phase advances of wheel running activity rhythm in vivo. The present results support the idea that methamphetamine produces an inhibitory effect on photic entrainment in the suprachiasmatic nucleus via serotonin release.

Animals↗

Contralateral lung injury associated with single-lung ischemia-reperfusion injury.

BACKGROUND: There have been very few studies on the effect of single-lung ischemia-reperfusion on the function of the contralateral lung. This study was designed to clarify the effect. METHODS: Fifteen mongrel dogs were divided into two groups. In group 1 (n = 7), the left lung was subjected to ischemia without ventilation for 90 minutes, and then reperfused. In group II (n = 8), the lung was not subjected to ischemia, and was ventilated during the 90-minute ischemia of group I. Arterial blood gas, hemodynamics, extravascular lung water, and airway pressure were measured. Pulmonary biopsy was performed to evaluate adenine nucleotide levels. The protein concentration and phosphorous concentration of phospholipids in bronchoalveolar lavage fluid were measured. RESULTS: Group I, with perfusion and ventilation of the right lung alone, was significantly inferior to group II with respect to arterial blood gas, right pulmonary compliance, extravascular lung water of the right lung, and the protein concentration in the bronchoalveolar lavage fluid of the right lung after the 90-minute period. CONCLUSIONS: These results indicate that 90 minutes of warm ischemia and reperfusion of the left lung caused deterioration of not only the left but also contralateral right pulmonary function.

Adenine Nucleotides↗

Expression and localization of basic fibroblast growth factor (bFGF) in the repair process of rat liver injury.

BACKGROUND/AIMS: To clarify the expression and localization of basic fibroblast growth factor in the repair process of liver injury, acute liver injury was induced by administration of carbon tetrachloride, D-glactosamine hydrochloride, or dimethylnitrosamine to rats. METHODS: We measured basic fibroblast growth factor protein in the liver tissue by radioimmunoassay, evaluated the expression of basic fibroblast growth factor mRNA by the reverse transcriptase polymerase chain reaction, and identified basic fibroblast growth factor-positive cells by immunostaining. RESULTS: In the carbon tetrachloride injured liver, the basic fibroblast growth factor protein contents began to increase 2 days after administration when liver injury was most marked, and reached a peak after 4 days, decreasing thereafter. In the carbon tetrachloride-injured liver, basic fibroblast growth factor mRNA expression was observed from 12 h after administration, prior to an increase in the protein content. In the D-galactosamine hydrochloride-injured liver, basic fibroblast growth factor protein also increased. On the other hand, in the dimethylnitrosamine-injured liver, the basic fibroblast growth factor protein content decreased 2 days after administration when liver injury was marked, but increased after 7 days. In the regenerating liver after partial hepatectomy, the basic fibroblast growth factor protein content did not increase. Among cell fractions, the Ito cell fraction obtained from the carbon tetrachloride-injured liver after 4 days showed expression of basic fibroblast growth factor mRNA. In cells cultured for 24 h, this fraction was immunopositive for basic fibroblast growth factor. Ito cells in the liver tissue markedly increased in the carbon tetrachloride-injured liver and increased after 7 days in the dimethylnitrosamine-injured liver. CONCLUSIONS: This study confirmed basic fibroblast growth factor production in the liver tissue in the repair process of liver injury. Our results suggest that basic fibroblast growth factor is primarily produced in Ito cells, acts on sinusoidal wall cells including Ito cells by the autocrine and paracrine mechanisms, and promotes extracellular matrix production and vascularization, involving the repair process of liver injury.

Animals↗

Acute arterial thrombosis due to platelet aggregation in a patient receiving granulocyte colony-stimulating factor.

We describe a 44-year-old man with non-Hodgkin's lymphoma receiving granulocyte colony-stimulating factor (G-CSF) who developed an acute arterial thrombosis. The removed thrombus contained large amounts of platelet aggregation. A rapid increase of platelets and increased adenosine diphosphate (ADP)- and collagen-induced platelet aggregation were observed at the time of the thrombotic event. A challenge test of G-CSF showed an increase in the platelet count and an augmentation of ADP- and collagen-induced platelet aggregation. In the use of GCSF. patients who produce a rapid increase in platelet levels could be at greater risk for thrombotic events and need to be followed-up carefully.

Adult↗

Scarpa's adipofascial flap for repair of wide scalp defects.

Scarpa's fascia is a prominent superficial fascial system of the body. It consists of a single membrane between the superficial fatty layer and deep fatty layer, and lies widely in the lower abdominal wall. We describe a case with a wide scalp defect resulting from a resection of a dermatofibrosarcoma, and reconstruction of the defect with Scarpa's adipofascial flap (i.e., a combined paraumbilical perforator-based adipofascial flap-groin adipofascial flap). The primary advantage of Scarpa's adipofascial flap for scalp defects is that (1) the donor site is most acceptable for a free flap with a minimal donor scar and minimal dysfunction; (2) even in cases in which large flaps are used, donor defects can be closed directly without skin grafting; (3) in the obese patient, this flap is preferable because of cosmetic improvement of the abdominal wall; (4) the donor area has so many perforators that an extended adipofascial flap can be obtained with a combination of these perforators; and (5) the flap may be nourished with one of several arteries, such as the superficial or deep inferior epigastric artery, or the superficial or deep circumflex iliac artery. The disadvantages of this flap are that the territory with a single artery may be smaller than a skin flap with the same artery and oversurfacing of the graft results in a poor cosmetic appearance. Scarpa's adipofascial flap is indicated when the defects are in an exposed area, especially in children, young patients, and females, and when this procedure is combined with a skin-expanding method in the secondary repair.

Adipose Tissue↗

Differentiation induction therapy with all-trans retinoic acid (ATRA) in a patient with secondary 11q23 leukemia.

PURPOSE: Frequently recurrent leukemia is encountered in childhood and is usually resistant to multidrug therapy. We evaluated the efficacy of differentiation induction therapy in a child with leukemia. PATIENT AND METHODS: We administered all-trans retinoic acid (ATRA) alone, and then a combination of low-dose 1-beta-D-arabinofuranosylcytosine (Ara-C), to a 7-year-old boy who had leukemia secondary to etoposide with chromosomal abnormality 11q23. RESULTS: The patient exhibited a good response to ATRA and had a complete remission after combination therapy of ATRA and low-dose Ara-C. CONCLUSION: Differentiation induction therapy with ATRA was effective in treating a patient with recurrence of leukemia without t(15;17) translocation, and which is resistant to conventional cytotoxic treatment.

Antineoplastic Combined Chemotherapy Protocols↗

Interleukin-1 beta and tumor necrosis factor-alpha stimulate synergistically the expression of monocyte chemoattractant protein-1 in fibroblastic cells derived from human periodontal ligament.

The present study demonstrates that interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) induce and synergistically stimulate monocyte chemoattractant protein-1 (MCP-1) expression in fibroblasts from human periodontal ligament. IL-1 beta and TNF-alpha induced in a dose-dependent manner the expression of the MCP-1 gene in the fibroblasts from the human periodontal ligament. However, such an inducing effect was not observed with IL-6 and interferon-gamma. The peak expression of the MCP-1 gene by IL-1 beta or TNF-alpha was observed at 3 h after initiation of their treatment. Furthermore, IL-1 beta in combination with TNF-alpha synergistically stimulated the MCP-1 gene expression in the cells. We also observed significant chemotactic activity for human monocytes in conditioned medium of fibroblasts from the human periodontal ligament treated with both cytokines. The stimulated chemotactic activity induced by these cytokines depended on both dose and treatment time. The chemotactic activity in conditioned medium of IL-1 beta-treated fibroblasts from the human periodontal ligament was neutralized by antiserum specific for MCP-1 protein. The MCP-1 gene product in conditioned medium of IL-1 beta-treated fibroblasts from the human periodontal ligament was shown to have a molecular mass of 11,000 Da by immunoprecipitation with the specific antiserum, and IL-1 beta also stimulated synergistically MCP-1 protein expression in combination with TNF-alpha. These results suggest that IL-1 beta and TNF-alpha may contribute to the infiltration of monocytes into inflammatory sites of periodontal ligament tissues via the MCP-1 gene product produced by fibroblasts from the human periodontal ligament.

Chemokine CCL2↗

Improvement of bone disease with increased dose of glucocerebrosidase in a Gaucher disease patient who had a bone lesion presenting during low-dose enzyme replacement therapy.

In recent years, enzyme replacement therapy has been shown to be useful for the treatment of Gaucher disease. A 10 year old Japanese boy with Gaucher disease underwent splenectomy at the age of 5 years and received enzyme replacement therapy from the age of 6 years. He had avascular necrosis of the bilateral femoral heads, which was not seen at the beginning of the therapy, without deterioration of hematological variables during maintenance therapy. The enzyme dosage was increased from 20 to 120 IU/kg per month resulting in an improvement of the clinical symptoms and bone lesion. In enzyme replacement therapy, dose increase is considered to be essential for improvement in bone disease; however, it is important to watch for the development of bone lesion.

Child↗

CD14-mediated signal pathway of Porphyromonas gingivalis lipopolysaccharide in human gingival fibroblasts.

Lipopolysaccharide (LPS) induces expression of inflammatory cytokines in monocytes/macrophages via CD14, one of the LPS receptors, which is expressed predominantly in these cells. It has been demonstrated that Porphyromonas gingivalis LPS (P-LPS) also is able to induce inflammatory cytokines in human gingival fibroblasts. Therefore, it is important to determine whether CD14 is expressed in gingival fibroblasts and to define the P-LPS-mediated signal-transducing mechanism in the cells. In this study, we observed unexpectedly by immunohistochemical, Western blotting (immunoblotting), and Northern (RNA) blotting assays that CD14 is expressed at high density in human gingival fibroblasts. P-LPS-induced expression of the monocyte chemoattractant protein 1 (MCP-1) gene in the cells was inhibited markedly by treatment with anti-human CD14 antibody and was completely inhibited by herbimycin A, a potent inhibitor of tyrosine kinase. The inhibitor also dramatically inhibited monocyte chemotactic activity of and MCP-1 production by the cells. Furthermore, P-LPS-induced expression of the MCP-1 gene in the cells also was blocked by inhibitors of two transcription factors, i.e., curcumin, an inhibitor of AP-1, and pyrolidine dithiocarbamate, an inhibitor of NF-kappaB. Both inhibitors inhibited monocyte chemotactic activity in the culture supernatant of P-LPS-treated cells. Gel shift mobility assay showed stimulation of the AP-1 and NF-kappaB contents in P-LPS-treated cells. This study is the first to demonstrate the expression of CD14 in human gingival fibroblasts and to show that the signal-transducing pathway of P-LPS in the cells is mediated by CD14.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

Ca2+ sensitizer Org-30029 reverses acidosis- and BDM-induced contractile depression in canine myocardium.

Effects of the Ca2+ sensitizer N-hydroxy-5,6-dimethoxy-benzo[b]thiophene-2-carboximidamide hydrochloride (Org-30029) on the myocardial contractile depression induced by acidosis and 2,3-butanedione monoxime (BDM) were investigated in aequorin-loaded canine ventricular myocardium. The peak Ca2+ transient-peak force relation during administration of Org-30029 (10(-4) to 10(-3) M) was shifted to the left and upward compared with the relation for elevation of the extracellular Ca2+ concentration ([Ca2+]o) (2.5-12.5 mM). Acidosis (pH 6.6) depressed the force with a small increase in the peak Ca2+ transient. BDM (3 mM) depressed the force with no change in the peak and duration of the Ca2+ transient, indicating that BDM may inhibit selectively the cross-bridge interaction. During acidosis or in the presence of BDM, elevation of [Ca2+]o increased the peak Ca2+ transient to the same extent as that in the control, but the force was inhibited. In contrast, Org-30029 increased the force to a level equivalent to the control with a slight change in the peak Ca2+ transient. In addition, during acidosis, Org-30029 (10(-3) M) increased the force in association with a slight decrease in the peak Ca2+ transient. Thus Org-30029 can reverse the myocardial contractile depression induced by a decrease in the Ca2+ sensitivity of myofilaments, as occurs in pathophysiological situations such as acidosis in cardiac ischemia. Org-30029 may exert the Ca(2+)-sensitizing effect by an increase in the affinity of troponin C for Ca2+ and by a direct action on the cross-bridge interaction.

Acidosis↗

Inhibitory effect of CV4151, a thromboxane A2 synthetase inhibitor, on ventricular arrhythmias induced by coronary artery occlusion in rats.

The purpose of this study was to determine whether thromboxane A2 (TXA2) is involved in the development of ventricular arrhythmias produced by coronary artery occlusion. Ventricular arrhythmias were induced by coronary artery occlusion in 66 male Sprague-Dawley rats. Rats were separated into 4 groups, and saline (n = 19) or CV4151 (a TXA2 synthetase inhibitor)(10 mg/kg, n = 14; 30 mg/kg, n = 15; or 100 mg/kg, n = 18) was injected intravenously 5 min before coronary artery occlusion. The antiarrhythmic effect of CV4151 was assessed in terms of the number of ventricular premature complexes (VPCs), the combined duration of ventricular tachycardia (VT) and ventricular fibrillation (Vf), the incidence of Vf, and the mortality rate within 30 min after occlusion. The total number of VPCs was as follows; control: 1789 +/- 330 beats: 10 mg/kg group: 1289 +/- 302 beats: 30 mg/kg group: 1008 +/- 229 beats: 100 mg/kg group: 986 +/- 275 beats, with no significant differences between groups. The incidence of Vf was significantly reduced in the 30 mg/kg and 100 mg/kg groups, as was the combined duration of VT and Vf and the mortality rate. Our results indicate that the TXA2 synthetase inhibitor CV4151 reduces the incidence of lethal arrhythmias induced by coronary artery occlusion in rats.

Animals↗