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Biomedical subjects

T Miyazawa

Publications and source records attributed to T Miyazawa.

At least 19 recordsLinked to original sources

Inhibition of mucosal lipid hyperoxidation by green tea extract in 1,2-dimethylhydrazine-induced rat colonic carcinogenesis.

Phosphatidylcholine hydroperoxide (PCOOH) measured using a chemiluminescence detector to examine colonic mucosal lipid hyperoxidation increased after injection of 1,2-dimethylhydrazine and green tea extract (GTE), which we previously showed inhibited carcinogenesis and oxidative DNA damage in the gastrointestinal tract. Therefore, the hyperoxidation of membrane phospholipids reflected well the degree of DNA damage and carcinogenic alteration, and may be a useful intermediate biomarker for initiation of carcinogenesis.

1,2-Dimethylhydrazine

Fluorescence polarization study on the dynamics and location of peroxidized fluorescent phospholipids in liposomes.

Motional properties of fluorescent substances produced by lipid peroxidation by a time-resolved fluorescence polarization technique were studied. When liposomes containing phosphatidylethanolamine (PE) and linoleic hydrocarbon chain were incubated at 37 degrees C, fluorophores absorbing maximally at 360 nm and emitting near 430 nm were produced. Their fluorescence anisotropy decay measured at 23 degrees C was fitted well with a sum of a fast relaxation and a time-independent residual term. With the increase of oxidation degree, the time constant of the relaxation term increased. This may be explained by alteration in the membrane structure or by modification of the fluorescent products themselves. Information on the location of the fluorescent products was obtained when their motional property was compared with those of various extrinsic probes that were incorporated at different positions of the lipid bilayer. It was found that the motional property of the fluorescent oxidation products is similar to that of 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene, a rod-shaped hydrophobic probe with a charged terminal. Other probes sensing the polar region or the hydrophobic region of the membrane were characterized by a lower order parameter. It is suggested that the fluorescent oxidation products have a polar moiety located at the membrane surface and attached to the amino group of PE while the tail part being buried in the hydrophobic region of the membrane. This picture is supported by fluorescence quenching experiments with the aqueous quencher Co2+. On the other hand, fluorophores produced by the reaction of malondialdehyde and PE suggested to have a chemical structure in which the angle between the absorption and emission dipole moments is very large. On the basis of these observations, the production pathway of fluorophores in oxidized membranes is discussed.

Fluorescence Polarization

Age-related change of phosphatidylcholine hydroperoxide and phosphatidylethanolamine hydroperoxide levels in normal human red blood cells.

The age-related occurrence of phosphatidylcholine hydroperoxide (PCOOH) and phosphatidylethanolamine hydroperoxide (PEOOH) in normal human red blood cells (RBC) was confirmed by using chemiluminescence detection-high performances liquid chromatography (CL-HPLC). The concentration (mean +/- S.D.) for the healthy young adult (22-27 of age, n = 20) was 93 +/- 17 pmol PCOOH and 121 +/- 20 pmol PEOOH/ml packed RBC, while for the aged adult (56-92 of age, n = 20) the peroxide content was significantly higher, 162 +/- 52 pmol PCOOH and 186 +/- 40 pmol PEOOH/ml packed RBC. These results indicate that oxidative stress occurs constantly on RBC, even in normal humans, and that the susceptivity significantly increases with age.

Adult

beta-carotene as a high-potency antioxidant to prevent the formation of phospholipid hydroperoxides in red blood cells of mice.

In order to investigate the antioxidant effect of beta-carotene in vivo, phospholipid hydroperoxides and beta-carotene isomers in red blood cells (RBC), plasma and tissue organelles were quantitatively measured after the oral administration of beta-carotene (94.8% all-trans-beta-carotene) to mice. Three groups of 24 mice each were fed for 1 week on a semisynthetic diet supplemented with either 0.6% or 3.0% beta-carotene/diet or maintained on a control (beta-carotene-unsupplemented) diet. The RBC phospholipid hydroperoxides showed a significant decrease followed by an increase of beta-carotene intakes; i.e., 201, 16 and 4 pmol of phosphatidylcholine hydroperoxide/ml packed RBC, and 108, 22 and 8 pmol of phosphatidylethanolamine hydroperoxide/ml packed RBC, in the mice given the control diet, 0.6% carotene diet and 3.0% carotene diet, respectively. The RBC beta-carotene increased from 14 to 43 pmol/ml packed RBC as followed by the increase of beta-carotene intakes. Such a potent antioxidant effect of beta-carotene as observed in RBC was not confirmed in the plasma, liver or lungs, although their beta-carotene contents increased. The beta-carotene ingestion increased the all-trans-beta-carotene and retinol contents in RBC, plasma, liver and lungs, but the alpha-tocopherol content decreased. In the beta-carotene-supplemented (6 g and 30 g/kg diet) mice, cis-beta-carotene content was relatively higher in the RBC (25-35% of total beta-carotene) than that in the plasma, liver and lungs. The present findings indicate that not only does beta-carotene act as a potent antioxidant in vivo but also its antioxidant effect is very specific in the RBC phospholipid bilayers rather than in the plasma and other tissue organelles.

Animals

Stapes fixation associated with symphalangia.

We describe a 10-year-old boy and his 9-year-old sister, both of whom had bilateral hearing loss associated with ankylosis of the proximal interphalangeal joints (symphalangia). They had no other abnormal findings except hearing loss and ankylosis of the joints of some fingers and toes on systemic examination. The cause of conductive hearing loss in both cases was bony fusion between the stapes and the bone of the oval window niche. There were no other anatomical abnormalities in the middle or the external ear. The patients' hearing improved markedly after stapes surgery. Histopathologic examination of the stapes revealed an abnormal zone of ossification near the anterior annular ligament and calcification in the annular ligament, which seemed to be the cause of the stapes fixation.

Child

Regulatory properties of the integrated long terminal repeat of the feline immunodeficiency virus.

To examine the regulatory properties of feline immunodeficiency virus (FIV) long terminal repeat (LTR) integrated into host chromatin, Crandell feline kidney cells were stably transfected with the FIV LTR that directs the bacterial chloramphenicol acetyltransferase (CAT) gene. Using these cells, we examined the effects of treatment with several chemical agents, infection with feline viruses, or transfection with effector plasmids expressing FIV gene products on FIV LTR-directed gene expression. Among them, treatment with the phorbol ester (a strong activator of protein kinase C), forskolin (an inducer of cyclic-AMP), 5-azacytidine (a DNA methylation antagonist), or infection with feline herpesvirus type 1 (FHV-1), resulted in induction of CAT activity in the cells. These results suggest that the integrated FIV LTR is stimulated by cellular transcriptional factors induced by phorbol ester, forskolin and FHV-1, and is also inactivated by DNA methylation. Furthermore, this permanent cell line can be used as a screening system of activators of the FIV LTR.

Animals

Neurotoxicity of gadolinium contrast agents for magnetic resonance imaging in rats with osmotically disrupted blood-brain barrier.

The neurotoxicity of intravenously injected Gadolinium (Gd) complexes to rats with disrupted blood-brain barrier (BBB) was evaluated. After disruption of the BBB by infusion of mannitol solution, one of several contrast agents tested was injected intravenously at a dose of 1 or 3 mmol Gd/kg, and neurological symptoms were graded. The concentrations of Gd in brain and plasma were also measured. Injection of Gd-DTPA at a dose of 3 mmol Gd/kg did not change behavior. On the other hand, Gd-DTPA-BMA, Gd-DO3A-butrol, and Gd-DO3A-HP each induced behavioral impairments, and some animals died within 1 h after injection. Gd-DO3A-HP showed lethal effect even at a dose of 1 mmol/kg. The concentration of Gd in the brain of the animals injected with Gd-DO3A-HP at 3 mmol Gd/kg was essentially the same as that of animals injected with Gd-DTPA at the same dose. The neurotoxicity of the contrast agents tested was graded as follows: Gd-DTPA < or = Gd-DTPA-BMA = Gd-DO3A-butrol < Gd-DO3A-HP.

Animals

Pontine glioma with osteoblastic skeletal metastases in a child.

BACKGROUND: The development of systemic metastases from primary intracranial gliomas is rare. We report here a rare case of pontine glioma with osteoblastic skeletal metastases. CASE: This 12-year-old boy presented with a 4-month history of hoarseness, dysphagia, and a progressively ataxic gait. Cranial computed tomography (CT) and magnetic resonance imaging (MRI) revealed a brain stem tumor that was diagnosed as a low grade glioma by stereotactic biopsy. Twelve months later following chemotherapy and radiotherapy, neurologic examination and neuroradiologic studies disclosed a recurrence of the pontine glioma. Skeletal roentgenograms revealed widespread osteoblastic metastases in the skull, vertebral bodies, pelvis, and long bones. A specimen from the iliac bone demonstrated cells that were immunoreactive glial fibrillary acidic protein (GFAP). DISCUSSION: The mechanism of how glioma cells determine their biologic behavior at bony metastatic sites is not known. Infratentorial gliomas, which occur frequently in young patients and demonstrate active bony metabolism, may stimulate osteoblastic cells, and induce osteoblastic changes.

Bone Neoplasms

Feline immunodeficiency virus can infect a human cell line (MOLT-4) but establishes a state of latency in the cells.

Infectivity of feline immunodeficiency virus (FIV) in feline and human lymphoblastoid cell lines was examined using homogeneous populations of FIV derived from infectious molecular clones of strains TMZ and Petaluma, and two recombinant chimeric clones carrying gag, pol, vif and ORF-A from the heterologous virus. FIV from the clones with the env region of the Petaluma strain was shown to infect and establish provirus in a human lymphoid cell line (MOLT-4), although the FIV-infected cells did not produce any infectious viruses. By treatment of the infected MOLT-4 cells with a phorbol ester, infectious virus was rescued. To examine which stage of the life-cycle of FIV is blocked in these cells, we analysed transcription of FIV-14 in the cells by RT-PCR. FIV-specific RNA expression could not be detected. These results strongly suggest that latency of the virus in MOLT-4 cells is due to a failure in transcription.

Antigens, Viral

Genetic diversity of Argentine isolates of feline immunodeficiency virus.

We report the nucleotide sequence and genetic diversity of part of the envelope (env) gene of four strains of feline immunodeficiency virus (FIV) isolated from Argentine domestic cats. The DNA encoding the V3 to V5 regions of the env gene of the FIV isolates were amplified by PCR, cloned and sequenced. Phylogenetic analysis revealed that the Argentine isolates did not cluster into a single group; one isolate clustered with subtype B FIV isolated in the USA and Japan, whereas the others formed a new cluster of FIV which might represent a prototype sequence for subtype E.

Amino Acid Sequence

Eustachian tube function and middle ear barotrauma associated with extremes in atmospheric pressure.

Eustachian tube (ET) function was studied by means of sonotubometry and tubotympano-aerodynamography (TTAG) prior to and following exposure to hypobaric or hyperbaric conditions. Forty normal adults were subjected to hypobaric pressure. Fifty adults who underwent hyperbaric oxygen (HBO) therapy also were studied. Following hypobaric exposure, 14 of 80 ears (17.5%) exhibited middle ear barotrauma. Following hyperbaric exposure, 34 of 100 ears (34%) exhibited middle ear barotrauma. Dysfunction of the ET, characterized by altered active and passive opening capacity, was more prevalent following exposure to extremes in atmospheric pressure compared to baseline. The ET function, which was impaired after the first HBO treatment, improved gradually over the next 2 hours. Overall, however, ET function was worse after the seventh treatment. The patients who developed barotrauma exhibited worse ET function prior to hypobaric or hyperbaric exposure. Thus, abnormal ET function can be used to predict middle ear barotrauma prior to exposure to hypobaric or hyperbaric atmospheric pressure.

Adult

[Synthesis and antibacterial properties of quinoxaline 1,4-dioxide derivatives].

Novel quinoxaline 1,4-dioxide derivatives were synthesized from benzofuroxans and the enolic form of 1,3-diketones or 3-oxoalkanoic esters or 3-oxoalkanamides or butanedioic esters catalyzed by silica gel or molecular sieves and their antibacterial activities were evaluated. As the results of antibacterial screening tests in vitro, quinoxaline 1,4-dioxides revealed strong activities against Bacteroides fragilis.

Bacteria

Persistence of high virus neutralizing antibody titers in cats experimentally infected with feline immunodeficiency virus.

The development of virus neutralizing (VN) antibody is one of the most effective host defense mechanisms against virus infection. In the present study, we developed a new VN assay against feline immunodeficiency virus (FIV) using a feline T-lymphoblastoid cell line, MYA-1 cells, based on inhibition of viral reverse transcriptase production. This assay is applicable to strains of FIV which can not infect CRFK cells. By using the assay, we examined long-term responses of VN antibody in cats experimentally infected with FIV. VN antibody titers increased progressively during first 30 weeks post inoculation and remained at high titers thereafter for 7 years of observation periods.

Animals

Phospholipid hydroperoxides and lipid peroxidation in liver and plasma of ODS rats supplemented with alpha-tocopherol and ascorbic acid.

Forty-five mutant male ODS rats, unable to synthesize ascorbic acid, were fed nine diets containing 5, 50 or 250 mg of vitamin E/kg diet and 150, 300 or 900 mg of vitamin C/kg diet for 21 days. The concentrations of vitamins C and E increased in liver and plasma in relation to the level of these vitamins in the diet. Vitamin C dietary supplementation increased the plasma vitamin E content at low levels of vitamin E intake, supporting the concept of an in vivo synergism between both antioxidant vitamins. Vitamin C, at the dietary levels studied, did not affect the lipid peroxidation. Vitamin E decreased liver and plasma endogenous levels of thiobarbituric acid-reactive substances and liver sensitivity to non-enzymatic lipid peroxidation. This was confirmed by a highly specific assay of lipid hydroperoxides using high performance liquid chromatography with chemiluminescence detection. The hepatic concentration of both phosphatidylcholine and phosphatidylethanolamine hydroperoxides decreased as the vitamin E content of the diet increased. The results show for the first time the capacity of vitamin E to protest against peroxidation of major phospholipids in vivo under basal unstressed conditions.

Animals

High phosphatidylcholine hydroperoxide level in plasma of guinea pigs with low and excess supplementation of ascorbic acid.

Graded amounts (0, 50, 500 and 5,000 mg/liter) of ascorbic acid (AsA) were given in drinking water to guinea pigs for 21 days to prepare AsA-deficient, low-AsA, moderate-AsA and excess-AsA animals, and the plasma phospholipid hydroperoxide level and lipid concentration were quantitatively determined to investigate the antioxidant effect of AsA in vivo. Phosphatidylcholine hydroperoxide (PCOOH) was a predominant phospholipid hydroperoxide present in the plasma, and the PCOOH concentration was significantly higher in AsA-deficient, low-AsA and excess-AsA animals (80.4 nM, 54.8 nM and 42.2 nM, respectively) as compared with that in moderate-AsA animals (27.2 nM). Hyperlipidemic plasma characterized as high cholesterol and high triacylglycerol concentrations was confirmed in AsA-deficient animals. Molar ratios of plasma AsA and alpha--tocopherol against 10(4) moles of phospholipids were significantly lower in AsA-deficient and low-AsA animals (0.6-2.1 and 5.5-8.5, respectively) than in moderate-AsA and excess-AsA animals (14.2-18.0 and 11.2-11.9, respectively). In plasma, a high correlation coefficient (r = 0.979) was observed between PCOOH and AsA for which there was optimum AsA level to keep the low PCOOH and such correlation was stronger than that (r = 0.558) observed with alpha-tocopherol. The results indicated that AsA has an important function to control the phospholipid hydroperoxide level in plasma and that moderate supplementation of AsA is required to reveal its optimal antioxidant effect in vivo. The present study also showed that AsA-deficiency especially invites an increase in plasma PCOOH together with a hyperlipidemic state which are risk factors in developing atherogenesis.

Animals

[Tuberculous aneurysm of the descending aorta--successful surgical treatment].

A 62-year-old woman visited her primary physician because of high fever and vomiting. From a chest roentgenogram miliary tuberculosis was diagnosed. Anti-tuberculosis drugs were prescribed. Mycobacterium tuberculosis was cultured from samples of sputum. Although she took drugs for 3 months, her symptoms did not resolve and back pain developed. A chest roentgenogram showed an egg-sized mass in the posterior part of the mediastinum. Examination of a chest CT scan showed that the mass was an aortic aneurysm. A graft replacement was done, and histologic examination of the resected specimen showed that it was a tuberculous aneurysm of the descending aorta. Her symptoms disappeared, and she recovered fully. Tuberculous aneurysms are rare: they account for only 0.3% of all aneurysms. This is only the 9th reported case of tuberculous aneurysm successfully treated with surgery in Japan. We must realize that aneurysms are life-threatening complication of miliary tuberculosis.

Aortic Aneurysm, Thoracic

Intracarotid blood pressure changes during contrast medium injection.

PURPOSE: To investigate changes in blood pressure during intracarotid injection of contrast material. METHODS: Two catheters were inserted into the ipsilateral common carotid artery of dogs. The proximal catheter was used for injection of contrast material and the other was positioned distally to monitor blood pressure. RESULTS: Distal intracarotid pressure rose significantly during injection of contrast material at all rates (5, 7, and 10 mL/s) and at all doses (0.2, 0.5, 0.7, and 1.0 mL/kg). In addition, these blood pressure elevations were shown to be correlated with injection rates and doses. Even when the pressure-monitoring coaxial catheter was advanced into smaller arteries, no decline in the impact of injection was observed. CONCLUSION: Intracarotid injection of contrast material causes a temporal elevation of cerebral blood pressure in dogs.

Animals