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Biomedical subjects

T Nagano

Publications and source records attributed to T Nagano.

At least 19 recordsLinked to original sources

Toxicity of 1-methyl-4-phenylpyridinium derivatives in Escherichia coli.

Several derivatives of 1-methyl-4-phenylpyridinium (MPP+), i.e., 1-methyl-4-(4'-nitrophenyl)pyridinium (1), 1-methyl-4-(4'-cyanophenyl)pyridinium (2), 1-methyl-4-(3'-nitrophenyl)pyridinium (3), 1-methyl-4-(4'-chlorophenyl)pyridinium (4), 1-methyl-4-(4'-acetamidophenyl)pyridinium (5), and 1-methyl-4-(4'-aminophenyl)pyridinium (6), were synthesized in order to compare their toxicity with that of paraquat (PQ2+) in Escherichia coli. Addition of compounds 1, 2, and 3 to aerobic E. coli cell suspensions caused extracellular ferricytochrome c reduction, which was inhibited by superoxide dismutase in the same manner as that in the case of PQ2+. The rate of the ferricytochrome c (cyt. c) reduction was in the order of PQ2+ greater than 1 greater than 2 greater than 3, which is the same as that of the redox potentials of these compounds. On the other hand, MPP+, 4, 5, and 6, which have more negative potentials, had no effect on the cyt. c reduction. Compound 1 inhibited the growth of E. coli under aerobic conditions, but not under anaerobic conditions. The results show that compound 1 can act as a mediator for production of superoxide (O2-.), which seriously injures E. coli cells. However, though compounds 2 and 3 catalyzed the production of O2-. in E. coli cells, their activity of O2-. production was much lower than that of compound 1 or PQ2+. Thus, compound 3 had no effect on growth or survival of E. coli at 1 mM, while compounds 2 and 4 had both bacteriostatic and bacteriocidal effects which were independent of dioxygen (O2). The results show that the toxic mechanism is different from that of compound 1. MPP+, 5, and 6 had no effect on growth of E. coli. This paper shows that compound 1 is a novel enhancer of intracellular superoxide production, though the mechanism of toxicity of compounds 2 and 4 is not clear yet. The results suggest that the redox potential is a crucial factor for manifestation of the activity.

1-Methyl-4-phenylpyridinium

Difference in superoxide toxicity between 4,7-dicyanobenzofurazan and paraquat.

The O2-. production by aerobically cultured Escherichia coli in the presence of benzofurazan (1), 4,7-dimethylbenzofurazan (2), 4,7-dibromobenzofurazan (3), 4-bromo-6-cyanobenzofurazan (4), and 4,7-dicyanobenzofurazan (5) was examined by using the cytochrome c reduction method in order to elucidate the mechanism of cytotoxicity of benzofurazans. Adding compound 5 to E. coli cell suspension caused cytochrome c reduction, which was completely inhibited by superoxide dismutase. The rate of cytochrome c reduction was in the order of 1 = 2 = 3 less than 4 less than 5, which correlates well with that of the reduction potentials of these benzofurazans. Adding glucose to the E. coli cell suspension-compound 5-cytochrome c system accelerated the rate of cytochrome c reduction. The formation of 4,7-dicyanobenzofurazan anion radical in the cell suspension-compound 5-glucose system in the absence of O2 was followed by ESR spectroscopy. The ESR signal of the anion radical disappeared when O2 was added. Compound 5 was shown to have an approximately 10-fold greater increasing effect on the flux of O2-. by E. coli than paraquat (PQ) by the cytochrome c reduction method. The results were confirmed by the electrochemical method with an oxygen electrode. However, compound 5 had a bacteriostatic, but not lethal, effect, while PQ had both effects. The effect of compound 5 and PQ on lethality of E. coli showed a dramatic difference when E. coli was exposed to these two compounds and washed prior to testing the effects of that exposure. This difference probably arose because compound 5 readily leaked from the cells during dilution and plating. Also, the reduced form of compound 5 exits from the cells more readily than the reduced form of PQ and then generates O2-. in the medium by autoxidation. This suggests the importance of the intracellular production of O2-., rather than the extracellular production of O2-., for lethal effect.

Benzoxazoles

Changes in the distribution of filipin-sterol complexes in the boar sperm head plasma membrane during epididymal maturation and in the uterus.

The distribution of filipin-sterol complexes (FSCs) and intramembranous particles (IMPs) in the plasma membrane of the late spermatid of the boar and of the sperm obtained from the epididymides, ejaculates, and uterus 2 hours after mating was examined by a freeze-fracture replica technique. In the late spermatid, the FSC density was found to be very low. A majority of the FSCs in the acrosomal plasma membrane (APM) appeared as protuberances on the E face in the epididymal, ejaculate, and uterine sperm. The density of the FSCs in the principal segment (PS) of the APM was 291 +/- 44 FSC/microns2 (mean +/- standard deviation, S.D.), 322 +/- 41 FSC/microns2 and 355 +/- 31 FSC/microns2 in the caput, corpus, and cauda epididymidis, respectively. In comparison with the cauda epididymal sperm, the FSC density gradually decreased in the PS of the ejaculated (277 +/- 39 FSC/microns2) and uterine sperm (243 +/- 50 FSC/microns2). The reduction was especially remarkable in the equatorial segment (ES), where the density of FSCs in ejaculated and uterine sperm decreased to about half and less than half of that in the cauda epididymal sperm, respectively. Large (13 nm) and small (8 nm) IMPs were distributed evenly and densely in the P face of the APM in the late spermatid, epididymal, and ejaculated sperm. In the uterine sperm, IMP-free areas were observed in the P face of the plasma membrane, a feature thought to represent one of the capacitation changes of the boar sperm.

Animals

Acute myelomonocytic leukemia with marrow eosinophilia showing 5q- and 16q22 mosaicism.

We report an 82-year-old Japanese female with acute myelomonocytic leukemia with dysplastic marrow eosinophilia (FAB M4Eo) exhibiting a partial deletion of the long arm of chromosome 5, del(5)(q13q31) and a derivative chromosome 16 with a breakpoint at band 16q22, in addition to 5q-. The patient had a history of a preleukemic phase for several months with marked dysplasia in trilineage marrow cells, in addition to leukemic features compatible with M4Eo. These findings strongly suggested that the leukemic cells in this case were derived from a preleukemic clone with a del(5q).

Aged

Thiobacillus novellus cytochrome c oxidase contains one heme alpha molecule and one copper atom per catalytic unit.

The minimal structural unit of cytochrome c oxidase purified from Thiobacillus novellus was composed of one molecule each of two subunits with molecular masses of 32 and 23 kDa, respectively, and the unit had one molecule of heme a and one atom of copper. In the presence of n-octyl-beta-D-thioglucoside, the oxidase existed as the monomeric form of the unit, while it occurred as the dimeric form of the unit in the presence of Tween 20. The monomeric form showed an active cytochrome c oxidizing activity and reduced molecular oxygen to water with ferrocytochrome c. Namely, it has been shown that the bacterial cytochrome c oxidase with one heme a molecule and one copper atom per molecule can catalyze oxidation of ferrocytochrome c with concomitant reduction of molecular oxygen to water.

Bacillus

Analysis of antigenic determinants shared by two different allergens recognized by human T cells: house dust mite (Dermatophagoides pteronyssinus) and chironomid midge (Chironomus yoshimatsui).

To analyse the cross-reactivity of T-cell-mediated immunity between Dermatophagoides pteronyssinus (Dp) and Chironomus yoshimatsui (Cy), the most common allergens in Japan, we established antigen-specific human T-cell lines and clones. Some but not all of the Cy-induced T-cell lines showed a significant proliferative response not only to Cy, but also to Dp. No T-cell line responded to other unrelated antigens. When we stimulated the Dp-induced T-cell clones with Cy, 3 of the 40 clones (7.5%) showed a significant proliferation, and 2 of the 3 clones produced interleukin-4 and interferon-gamma, indicating their helper function. Cross-reactivity was diminished significantly after the absorption of Dp antigen in an anti-Cy affinity column. The cross-reactive epitopes were thought to be expressed on the Dp molecule of 45-53 kD. The presence of helper T cells reactive to both allergens suggests a possibility that this cross-reactivity might be involved in part in the high incidence of allergy to the 2 major allergens.

Allergens

Effects of norepinephrine on the oxidative pentose phosphate pathway in the rat heart.

To examine whether stimulation of alpha-adrenergic receptors may affect the oxidative pentose phosphate pathway (PPP) in the rat heart, norepinephrine (NE) and the alpha-adrenergic agonist norfenephrine were used. NE was administered as a continuous intravenous infusion in awake rats for 3 days. It stimulated the activity of cardiac glucose-6-phosphate dehydrogenase (G-6-PD), the first and regulating enzyme of the oxidative PPP, in a dose-dependent manner. With the highest dose (0.2 mg.kg-1.hr-1), there was also a time-dependent enhancement. The increase observed after 48 hours was attenuated partially by the beta-receptor blocker metoprolol and the alpha-receptor blocker prazosin. It was entirely abolished when both drugs were administered. Carvedilol, a beta-adrenergic blocker and vasodilator with alpha 1-blocking activity (0.5 mg.kg-1.hr-1), prevented the NE-induced increase in cardiac G-6-PD activity, in functional parameters (heart rate, left ventricular systolic pressure, and left ventricular dP/dtmax), and in the heart weight/body weight ratio. The alpha-adrenergic stimulator norfenephrine increased myocardial G-6-PD activity; prazosin prevented this stimulation. NE and norfenephrine also elevated the available pool of cardiac 5-phosphoribosyl-1-pyrophosphate. G-6-PD activity was enhanced in cardiac myocytes freshly isolated from the left ventricle of rats that had received NE infusion for 3 days (12.3 +/- 1.4 units/g protein) compared with control rats (1.5 +/- 0.4 units/g protein). The activity of 6-phosphogluconate dehydrogenase, one of the enzymes in the oxidative PPP, was elevated only moderately from 12.7 +/- 0.7 to 19.1 +/- 1.4 units/g protein. Combined alpha- and beta-receptor blockade with carvedilol attenuated these effects.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenergic beta-Antagonists

Bacteriostatic effect of 4,7-dicyanobenzofurazan due to inactivation of 2,3-dihydroxyisovalerate dehydratase.

As part of our research on benzofurazans (BZs), we have reported the bacterioses of BZs in Escherichia coli, which may be due to O2-. produced within E. coli in the presence of dioxygen (O2). Incubation of E. coli with 4,7-dicyanobenzofurazan (1) lowered the 2,3-dihydroxyisovalerate dehydratase activity detectable in extracts from these cells. Addition of branched chain amino acids such as valine and leucine protected E. coli from growth inhibition by compound 1, though it could not protect E. coli from the damage by paraquat (PQ). Addition of Fe(III)-tris[N-(2-pyridylmethyl)-2-aminoethyl]amine (Fe-TPAA), a novel superoxide dismutase mimic, protected the dehydratase in a dose-dependent manner, which confirms that inactivation of the dehydratase is largely due to production of O2-.. The possibility was discussed that the bacteriostatic effect of compound 1 is due to the inactivation of 2,3-dihydroxyisovalerate dehydratase.

Bacteria

New method of detecting nitric oxide production.

Nitric oxide (NO) reacts with H2O2 to produce a rise in potent luminol-dependent chemiluminescence under neutral pH conditions. We applied this reaction to the continuous detection of NO released from the rat isolated perfused kidney with simultaneous pressure changes. Acetylcholine increased NO release dose-dependently with the pressure reduction. An NO synthesis inhibitor, NG-monomethyl-L-arginine, abolished the release. This inhibition was reversed by the addition of L-arginine.

Animals

Application of hemoglobin analysis by CO-oximeter to medico-legal practice with special reference to diagnosis of asphyxia.

Hemoglobin analysis by CO-oximeter is more easily and rapidly available method than other spectrophotometric or GC methods. In the present study, the possibility of diagnosis of asphyxia by CO-oximeter was examined experimentally using rabbits. When measured value of oxyhemoglobin (O2Hb) was lower than about 20% and that of deoxyhemoglobin (HHb) was, on the other hand, higher than about 80%, being accompanied by low measured values of both methemoglobin (MetHb) and CO-hemoglobin (COHb) (< 5%), asphyxia should be firstly considered as the cause of death. To the corpses found at the scene of fire, three chief factors causing death such as burns, suffocation due to oxygen deficiency and intoxication by poisonous gases may have occurred simultaneously, and through measuring the values of O2Hb, COHb, MetHb and HHb, the main lethal factor could be selected out among these three factors above and the cause of death could be examined much more closely. Because of easy and rapid analysis, it could be considered that CO-oximeter was useful for forensic practices and police activities.

Animals

Ultrastructural analysis of platelet-like particles from a human megakaryocytic leukemia cell line (CMK 11-5).

Ultrastructural observations were performed to further characterize the human megakaryocytic leukemia cell line, CMK, and its subclone, CMK11-5. We found that particles derived from CMK11-5 cells had cytoplasmic projections and no nucleus, and that some particles contained alpha granules. Incubation with ADP induced fibrinogen receptors on the surface of these particles. Furthermore, the particles had glycoprotein Ib antigen on their surfaces, and attached nonreversibly to rabbit aortic subendothelium, showing associated morphological changes similar to those observed in normal platelets. CMK and CMK11-5 are the first megakaryocytic cell lines that have been found to release particles that have some of the same functions as normal platelets. In particular, CMK11-5 seems to be a useful model for studying megakaryocyte function.

Blood Platelets

The immunocytochemical localization of tumour necrosis factor and leukotriene in the rat liver after treatment with lipopolysaccharide.

After administration of bacterial lipopolysaccharide, there is an increase in the number of leucocytes which adhere to the endothelial cell surface of the hepatic vessels and pass through the endothelial layer by comparison with controls. There is also marked endothelial cell damage including intracytoplasmic oedema, increased numbers of autophagic vacuoles and dilatation of the intercellular junction in LPS-treated samples. The presence of immunocytochemical products of leukotriene (LTR) and tumour necrosis factor (TNF) was examined using in both LPS-treated and control samples. Immunoreactions of LTR which were seen in specific granules of neutrophils and monocytes attached to the endothelial cell surface may indicate the onset of endothelial cell damage. Positive immunoreactions of TNF on the endothelial cell surface, seen only in LPS-treated samples, indicate that TNF may enhance the passage of blood cells through the endothelia and also increase the endocytotic activity of the liver parenchymal cells, as revealed by the present marker experiment using horseradish peroxidase. Positive reactions of TNF in lysosomes of the endothelial cells suggest that they are able to produce TNF and transport it to the cell surface.

Animals

[A case report of replacement of the extracardiac conduit for tetralogy of Fallot: reconstruction with autogenous tissue around the conduit].

Replacement of the valved extracardiac conduit with autogenous tissue was performed on a 13-year-old female. The procedure was based on that reported by Danielson in 1987. She underwent the first operation for tetralogy of Fallot with pulmonary atresia using a valved extracardiac conduit (#16 mm Björk-Shiley valve) 8 years ago. Recently, she complained exertional dyspnea and chest pain. Reoperation was scheduled because of bleeding tendency, somatic growth of the patient, and severe distal anastomotic stenosis of the conduit by right ventriculogram. At operation, the valved conduit was removed under cardiopulmonary bypass, and autogenous connective tissue around the conduit was preserved as the posterior wall of the new conduit. An onlay patch (pericardial monocusp patch) was sutured to form the roof of the new tract. Postoperative catheterization showed no residual stenosis or pressure gradient between the right ventricle and the pulmonary artery. This technique is useful as the replacement of extracardiac conduit because of its simplicity, no necessity of postoperative anti-coagulant therapy, and possibility to make a generous-sized new outflow tract.

Adolescent

[Surgical treatment under extracorporeal circulation for renal cell carcinoma with tumor thrombus in inferior vena cava].

Three cases of renal cell carcinoma with tumor thrombus extending into the inferior vena cava are reported. Radical nephrectomy and thrombectomy were performed under extracorporeal circulation in all the cases. The level of tumor thrombus was preoperatively determined by computed tomography, magnetic resonance imaging or venacavography. The tumor thrombus extended into the right atrium in one, and above the hepatic vein in two cases. One patient whose thrombus reached the right atrium died of multiple metastasis of renal cell carcinoma 5 months after operation. Another patient with lung metastasis was given interferon-alpha and is alive 5 months after operation. The other patient is clinically free of disease and in good health 7 years after operation. We believe that extracorporeal circulation allows an opportunity to resect the tumor thrombus in a controlled situation, and makes the operation safer.

Aged

Light-induced retinal damage in mice. Hydrogen peroxide production and superoxide dismutase activity in retina.

The oxidative mechanism in retinal damage due to exposure to intense light was investigated histochemically and biochemically. SMA mice (albino mice) of 2 to 3 months of age were exposed to intense light (1000-1400 lux). In the cyclic light-reared group (without dark adaptation), the outer and inner segments of the photoreceptor cells were damaged after 3 days of exposure, and severe outer nuclear layer damage was observed after 5 to 7 days of exposure. Hydrogen peroxide (H2O2) production in the outer nuclear layer increased with the progress of retinal damage. In the dark-reared group (dark adaptation of 16-18 hours), outer and inner segment damage was noted after 4 hours of light exposure, and severe outer nuclear layer damage was noted after 12 hours of light exposure. H2O2 production increased in the outer nuclear layer with retinal damage. Superoxide dismutase (SOD) activity did not change before the occurrence of retinal damage, and decreased by 25% after 3 days of exposure to light in the cyclic light-reared group. The decrease in total SOD activity corresponded to that of manganese-SOD (Mn-SOD). In the dark-reared group, SOD activity did not change, even after 1 day of exposure. There appears to be some relationship between retinal light damage and H2O2 production in the outer nuclear layer. Superoxide dismutase activity failed to provide protection against retinal oxidative damage due to intense light exposure.

Animals

Diagnosis of paraaortic and pelvic lymph node metastasis of gynecologic malignant tumors by ultrasound-guided percutaneous fine-needle aspiration biopsy.

Transabdominal ultrasonography was done in patients with gynecologic malignancies to detect paraaortic and pelvic lymphadenopathy. Of the 78 patients who underwent ultrasonic examination of pelvic lymph nodes, pelvic lymphadenopathy was detected in 24, of whom ultrasound-guided percutaneous fine-needle aspiration biopsies (FNAB) were done in 16. Of these 16 patients, the results of 13 biopsies were positive. Of the 41 patients who underwent ultrasonographic examination of the paraaortic lymph nodes, 27 had paraaortic lymphadenopathy. Ultrasound-guided percutaneous FNAB were done in 26 of these 27 patients, with positive findings in 21 of the 26. It was concluded that ultrasound-guided percutaneous FNAB is useful in detecting metastatic lymph nodes in gynecologic malignancies.

Abdominal Neoplasms

Cytochromes c of Nitrobacter winogradskyi and Thiobacillus novellus: structure, function and evolution.

The amino acid sequences of Thiobacillus novellus and Nitrobacter winogradskyi cytochromes c have been compared with those of cytochromes c from several other organisms. The two bacterial cytochromes resemble eukaryotic cytochromes c; 49 amino-acid residues are identical between T. novellus and horse cytochromes c, and 50 residues identical between N. winogradskyi and horse cytochromes c. However, their reactivity with cow cytochrome c oxidase is about 80% lower than the reactivity of eukaryotic cytochromes c with the cow mitochondrial oxidase, while they react with yeast cytochrome c peroxidase as rapidly as eukaryotic cytochromes c. The numbers of identical amino-acid residues between T. novellus and animal cytochromes c are 45-53 and those between N. winogradskyi and animal cytochromes c 47-53, while those between the two bacterial cytochromes and yeast and protozoan cytochromes c are around 40. Thus, N. winogradskyi and T. novellus cytochromes c are more similar to animal cytochromes c than to yeast and protozoan cytochromes c on the basis of the amino-acid sequence.

Amino Acid Sequence