Biomedical subjects

A Hara

Publications and source records attributed to A Hara.

At least 253 records · Page 14Linked to original sources

PubMed1996

Involvement of nitric oxide in spinally mediated capsaicin- and glutamate-induced behavioural responses in the mouse.

The intrathecal (i.t.) injection of capsaicin (0.1 nmol/mouse) through a lumbar puncture elicited scratching, biting and licking responses. Pretreatment with the nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester (L-NAME) (320 nmol), by i.t. injection, resulted in a significant inhibition of the behavioural response produced by i.t. capsaicin (0.1 nmol/mouse). Similar behavioural responses were induced by i.t. injections of NMDA (0.4 nmol), kainate (0.05 nmol) or AMPA (0.05 nmol), which were all inhibited by co-administration of L-NAME (20-80 nmol). L-Arginine (600 mg/kg, i.p.) but not D-arginine (600 mg/kg, i.p.) reversed the inhibitory effect of L-NAME on capsaicin-, NMDA-, kainate- and AMPA-induced behavioural response. Scratching, biting and licking responses induced by tachykinin receptor agonists, substance P, [Sar9,Met(O2)11]substance P, neurokinin A and neurokinin B were not affected by co-administration of L-NAME (40 and 80 nmol). These results suggest that spinal nitric oxide may play a significant role in mechanisms of the behavioural response to capsaicin, probably through the release of glutamate, but not tachykinins.

Animals↗

PubMed1996

Relationship between vitellogenin and its related egg yolk proteins in Sakhalin taimen (Hucho perryi).

Vitellogenin (Vg) and its three egg yolk protein products, lipovitellin (Lv), phosvitin (Pv) and beta'-component, were isolated from mature female Sakhalin taimen (Hucho perryi). Vg had an apparent molecular weight of 540 kDa and appeared as a major 240 kDa band in SDS-PAGE, which resolved into two major bands (165 and 125 kDa) after reduction. The estimated molecular weights of purified Lv, Pv, and beta'-component were 330, 23, and 30 kDa, respectively. Lv appeared as a main band of 150 kDa in SDS-PAGE which resolved into two smaller bands (92 and 29 kDa) after reduction. beta'-component appeared as a 34 kDa band before and as a 17kDa band after reduction. Except for Pv, the purified proteins all reacted with an antiserum to Vg. In SDS-PAGE, Pv appeared as a 23 kDa band and a second < 6.5 kDa diffuse band. An antiserum to Pv dephosphorylated by alkaline phosphatase (Ap) was prepared. In Western blots, the antiserum reacted with dephosphorylated Pv and Vg, but not with Lv and beta'-component. This is the first immunological proof that three egg yolk proteins (Lv, Pv, and beta'-component) are derived from Vg in fish.

Animals↗

PubMed1996

Clinoidal meningioma arising from falciforme process of optic canal: case report and review.

A 56-year-old female with an initial symptom of unilateral progressive visual disturbance presented with a clinoidal meningioma arising from a falciforme process of the optic canal. Magnetic resonance images with gadolinium-diethylenetriaminepenta-acetic acid demonstrated marked enhancement of a small mass, approximately 1 cm in diameter, roofing the right optic nerve at the intracranial opening of the optic canal. The clinoidal meningioma is rarely reported and the precise originating location of the neoplasm has not been clarified. This is the first case of clinoidal meningioma, which is confirmed by the operative observation as the origin of the neoplasm is a falciforme process of the optic canal. Magnetic resonance images with enhancement are useful for the diagnosis of such a small neoplastic lesion adjacent to bone structure and total removal without a deterioration of visual function is achieved by careful microsurgical procedure.

Animals↗

PubMed1996

Involvement of two basic residues (Lys-17 and Arg-39) of mouse lung carbonyl reductase in NADP(H)-binding and fatty acid activation: site-directed mutagenesis and kinetic analyses.

Mouse lung carbonyl reductase, a member of the short-chain dehydrogenase/reductase (SDR) family, shows a strong coenzyme preference for NADP(H) over NAD(H), and is uniquely activated by fatty acids. Previous chemical modification and X-ray crystallography studies show that interactions responsible for the coenzyme specificity include salt linkages between the 2'-phosphate of NADPH and side-chains of Lys-17 and Arg-39 of the enzyme. Although Arg-39 is highly conserved in NADP(H)-dependent enzymes of the SDR family, Lys-17 is substituted with Arg in about half of the NADP(H)-dependent enzymes. The present study shows that mutations of Lys-17 to His (K17H) or Ser (K17S) and of Arg-39 to Ala (R39A) bring about decreases (from 5 to 90-fold) of the affinities for NADP(H), but minor changes in the affinity for NAD+. The binding energy arising from the mutations on the binding of the 2'-phosphate of NADP+ was decreased by 38-66% from the value of 4.8 kcal/mol calculated for the wild-type enzyme. In contrast, the mutation of Lys-17 to Arg (K17R) had little effect on the kinetic or thermodynamic properties. The activation by fatty acids was completely attenuated by the mutations of K17H and K17S, but not by K17R or R39A. These results indicate that the 2'-phosphate group of NADP(H) is recognized by both Lys-17 and Arg-39, of which Lys-17 is a component of the binding site for the activator, probably interacting with the negatively charged carboxylate group of fatty acids, and also suggest that the existence of a positively charged residue (either Lys or Arg) at position 17 is required for both NADP(H) specificity of the SDR family enzymes and fatty acid activation of the pulmonary carbonyl reductase.

Alcohol Oxidoreductases↗

PubMed1996

Rapid method of preparation of lysoglycosphingolipids and their confirmation by delayed extraction matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

We have recently developed a rapid method for the preparation of lysoglycosphingolipids on a small scale, in high yield. This procedure of microwave-mediated saponification of about 1 mg of glycosphingolipid with 0.5 ml of 0.1 M NaOH in methanol for two minutes can be easily repeated if larger amounts of lyso-compounds are needed. We have also found that the new methodology of delayed extraction matrix-assisted laser desorption ionization time-of-flight mass spectrometry in the reflector mode is extremely effective for the confirmation of different lysoglycosphingolipids together with their long chain base components. The combined method of preparation and confirmation of lysoglycosphingolipids is also important for the identification of long chain bases of various sphingolipids, because the usual analytical method of long-chain bases of sphingolipids depends on acidic methanolysis, which results in the formation of by-products such as O-methylsphingosines and threo-sphingosines.

Animals↗

PubMed1996

Propofol attenuates hydrogen peroxide-induced mechanical and metabolic derangements in the isolated rat heart.

BACKGROUND: Oxygen-derived free radicals are involved in tissue damage during myocardial ischemia and reperfusion. Recent in vitro studies have demonstrated that a beneficial effect of propofol lies on its free radical scavenging properties. The current study, therefore, examined whether propofol is effective against the mechanical and metabolic damage induced by exogenously administered hydrogen peroxide in the isolated rat heart. METHODS: Rat hearts were perfused aerobically with Krebs-Henseleit bicarbonate buffer at a constant flow rate according to Langendorff's technique, while being paced electrically. Hearts were studied in control Krebs-Henseleit bicarbonate buffer, with Intralipid vehicle, with 25 microM or 50 microM propofol for 40 min, and with 50 microM propofol for 30 min followed by Intralipid for 10 min. A similar set of hearts was treated with hydrogen peroxide for 4 min, either in the absence of or beginning 10 min after Intralipid or propofol infusion. Left ventricular pressure was recorded as an index of mechanical function. The tissue concentrations of adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, and creatine phosphate were measured as indices of energy metabolism. The tissue concentration of malondialdehyde was measured to evaluate lipid peroxidation. RESULTS: Hydrogen peroxide (600 microM) significantly increased the left ventricular end-diastolic pressure, decreased the left ventricular developed pressure (i.e., it produced mechanical dysfunction), and decreased tissue concentrations of adenosine triphosphate and creatine phosphate (i.e., metabolic damage). Hydrogen peroxide also increased the tissue concentration of malondialdehyde. These mechanical and metabolic alterations induced by hydrogen peroxide were significantly attenuated by propofol (25 microM or 50 microM), while the increase in malondialdehyde was completely suppressed by propofol. CONCLUSIONS: The current study demonstrates that in the isolated heart, propofol attenuates both mechanical and metabolic changes induced by exogenously applied hydrogen peroxide. The beneficial action of propofol is probably correlated with reduction of the hydrogen peroxide-induced lipid peroxidation.

Animals↗

PubMed1996

Chemopreventive effect of a xanthine oxidase inhibitor, 1'-acetoxychavicol acetate, on rat oral carcinogenesis.

The effect of a xanthine oxidase inhibitor, 1'-acetoxychavicol acetate (ACA), on 4-nitroquinoline 1-oxide (4-NQO)-induced oral carcinogenesis was investigated in male F344 rats. All rats except those in the ACA-alone and untreated groups were given 4-NQO (20 ppm) In the drinking water for 8 weeks to induce oral cancer. Starting 1 week before the 4-NQO exposure, animals were fed diet containing 100 ppm or 500 ppm ACA for 10 weeks, followed by the basal diet without ACA for 22 weeks. Other groups were fed the diet containing ACA at 100 ppm or 500 ppm for 22 weeks, starting 1 week after the cessation of 4-NQO exposure. The remaining groups consisted of rats given 500 ppm ACA alone or untreated rats. At the termination of the experiment (32 weeks), the incidences of tongue neoplasms and preneoplastic lesions, polyamine levels in the tongue tissue, and cell proliferation activity estimated in terms of 5-bromodeoxyuridine (BrdU)-labeling index and by morphometric analysis of silver-stained nucleolar organizer regions' protein (AgNORs) were compared among the groups. Feeding of ACA at the two doses during initiation or postinitiation significantly decreased the development of tongue carcinoma (93-100% reduction, P < 0.001) and preneoplasia (43-50% reduction for hyperplasia and 34-48% reduction for dysplasia, P < 0.05). There were no such lesions in rats fed ACA alone or those in the untreated control group. The number of AgNORs per cell nucleus was significantly decreased by feeding of ACA at a high dose (500 ppm) (29% inhibition, P < 0.05). The BrdU-labeling index was also reduced by dietary administration of ACA (23-32% inhibition, P < 0.01). In addition, ACA feeding reduced tongue polyamine levels (35-40% inhibition, P < 0.05). These results indicate that ACA inhibited rat oral carcinogenesis, and such inhibition might be related to suppression of cell proliferation in the oral mucosa by the xanthine oxidase inhibitor.

4-Nitroquinoline-1-oxide↗

PubMed1996

Suppressing effects of 6-(2,5-dichlorophenyl)-2,4-diamino-1,3,5-triazine and related synthetic compounds on azoxymethane-induced aberrant crypt foci in rat colon.

The modifying effects of dietary administration of 6-(2,5-dichlorophenyl)-2,4-diamino-1,3,5-triazine and 5 related compounds on the occurrence of azoxymethane (AOM)-induced colonic aberrant crypt foci (ACF) were investigated in rats. Male F344 rats were given s.c. injections of AOM (15 mg/kg body weight) once a week for 3 weeks to induce ACF. They also received the diet containing 200 ppm test compound for 5 weeks, starting one week before the first dosing of AOM. At the termination of experiment, all of the compounds had caused a significant reduction in ACF frequency, which might be associated with suppression of the expression of proliferation biomarkers. The apoptotic index in the colonic mucosal epithelium of rats killed at 6 h after the first AOM exposure revealed no blocking activity of the compounds.

Animals↗

PubMed1996

Early alterations of apoptosis and cell proliferation in azoxymethane-initiated rat colonic epithelium.

Alterations of apoptosis and cell proliferation in the colonic epithelium of rats after exposure to azoxymethane (AOM) were estimated by means of the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) method, measurement of 5-bromo-2'-deoxyuridine (BrdU) incorporation, immunohistochemical staining for proliferating cell nuclear antigen (PCNA), and counting of mitotic cells. F344 male rats were given a single s.c. injection of AOM (15 mg/kg body weight) at 6 week of age, and killed 4 h, 8 h, 3 days, and 7 days after the AOM treatment. At 4 h after the treatment, many damaged cells were already observed in the colonic epithelium, and they were positive by TUNEL staining. At 8 h, the number of TUNEL-positive cells was largest. The reduction of DNA synthesis in the colonic epithelium, confirmed by BrdU incorporation, was not distinct in comparison with the mitotic inhibition. There was no remarkable change in PCNA labeling index, except that strong expression of PCNA was detected in many damaged cells. On the 3rd day, the appearance of cell death became infrequent and an increase of cell proliferation occurred. On the 7th day, the expression of TUNEL and the cell proliferation biomarkers were at almost normal levels. These findings suggest that AOM induces apoptosis, which is associated with synchronous inhibition of mitosis. The data also indicate that PCNA immunostaining does not reflect the true proliferation state in the early phase after AOM exposure, probably due to the occurrence of cell cycle arrest or DNA repair.

Animals↗

PubMed1996

[A new approach to the understanding of the mechanism of ischemia/reperfusion damage in the heart and the effects of anti-ischemic drugs].

The classical understanding of the mechanism of anti-anginal or anti-ischemic drugs is an increase in blood supply to the heart and/or a decrease in oxygen consumption of the heart, maintaining energy balance in the heart between supply and demand and hence maintaining the tissue levels of high-energy phosphates. This scheme is reasonable. During reperfusion following ischemia, however, there is more serious damage to the heart, although the tissue levels of high-energy phosphates increase. This is probably because toxic substances are generated in the heart during ischemia/reperfusion. We propose that both lysophosphatidylcholine and palmitoyl-L-carnitine that accumulate in the myocardium during ischemia/reperfusion are candidates for the toxic substances that accelerate ischemia/reperfusion damage to the heart. Therefore, drugs that have anti-lysophosphatidylcholine and/or anti-palmitoyl-L-carnitine effects are promising for the treatment of ischemic heart diseases. We found that K-7259, a novel derivative of dilazep having a minimal effect on the normal heart, is a drug that attenuates the deleterious effects of both lysophosphatidylcholine and palmitoyl-L-carnitine on the heart, and therefore attenuates the ischemia/reperfusion damage.

Adrenergic beta-Antagonists↗

PubMed1996

Human T-cell lymphotropic virus type-I associated myelopathy complicated by optic neuritis.

A 48-year-old Japanese woman from Kyushu was admitted to the Nippon Medical School First Hospital with complaints of numbness in both legs, gait disturbance, and urinary problems. On examination, her lower extremities were spastic with increased reflexes and positive Babinski sign. Sensation was absent below the T4 spinal level. The cerebrospinal fluid contained HTLV-I antibodies and she was diagnosed with HTLV-I associated myelopathy. Her symptoms were resolved with prednisolone, but six months later a visual disturbance of the left eye edema developed. Orbital CT scans showed that left optic nerve was edematous. T2 enhanced MRI revealed a high intensity lesion of the left optic nerve. ERG was normal and no VEP was detected in response to flashing stimulation to the left eye. The HTLV-I antigen titer of CBF was very high. Her optic neuritis improved following oral administration of prednisolone and retrobulbar injections of dexamethasone. This is the first case report of HTLV-I associated myelopathy complicated by optic neuritis.

Anti-Inflammatory Agents↗

PubMed1996

Distribution of dystrophin and dystrophin-associated protein 43DAG (beta-dystroglycan) in the central nervous system of normal controls and patients with Duchenne muscular dystrophy.

In skeletal muscles of patients with Duchenne muscular dystrophy (DMD), the absence of dystrophin was thought to lead to the large reduction in all of the dystrophin-associated proteins (DAPs). Of the seven types of DAPs identified in skeletal muscle, only the 43-kDa glycoprotein (beta-dystroglycan) has recently been found in the monkey brain. To clarify the distribution and characterization of dystrophin and beta-dystroglycan in the brain of humans, we carried out immunostaining and immunoblotting studies on tissues from three DMD patients with intellectual disturbances (ages 17,22, and 26 years) and in five controls (age range, 42-74 years). An antidystrophin antibody revealed dystrophin to be localized in neuronal cells and in the vascular wall in control brains, but it was absent from these tissues in DMD patients. In contrast, beta-dystroglycan was distributed throughout neuronal cells and in the vascular wall of control brains, and was well preserved in the brain of patients with DMD.

Adolescent↗

PubMed1996

The effect of methylprednisolone on acoustic trauma.

The effect of methylprednisolone (mPSL) upon acoustic trauma was studied using albino guinea pigs which were exposed to 2 kHz pure tone of 110, 115 or 120 dB SPL for 10 min. After the exposure to intense sound, mPSL of 6, 12 or 40 mg/kg was intraperitoneally given daily for 7 days and the threshold of the compound action potential (CAP) was examined on the 8th day. Compared with CAP of the control animals given physiological saline solution, no significant difference was observed in the CAP threshold shift between the mPSL group and the controls following exposures to the sound of 115 and 120 dB SPL, respectively. However, when the animals were exposed to the sound of 110 dB SPL, the CAP threshold shift was significantly smaller in the mPSL animals than in the controls. The present results indicate that mPSL possesses a therapeutic effect in cases of mild acoustic overstimulation.

Acoustic Stimulation↗

PubMed1996

Protective effects of dilazep and its novel derivative, K-7259, on mechanical and metabolic derangements induced by hydrogen peroxide in the isolated perfused rat heart.

The effect of dilazep, a potentiator of the adenosine-mediated effects, on the hydrogen peroxide (H2O2)-induced mechanical and metabolic derangements was studied in the isolated rat heart, and compared with that of K-7259, a dilazep derivative having less potentiating action on the adenosine-mediated effects. The heart was perfused aerobically by Langendorff's technique at a constant flow and driven electrically. H2O2 (600 microM) decreased the left ventricular developed pressure and increased the left ventricular end diastolic pressure in the heart (i.e, mechanical dysfunction), decreased the tissue ATP level and increased the tissue AMP level (i.e., metabolic change) and increased the tissue level of malondialdehyde (i.e., lipid peroxidation). These mechanical and metabolic alterations induced by H2O2 were attenuated by dilazep (1 microM), and the effect of dilazep was not modified by 8-(p-sulfophenyl)-theophylline (20 microM), a nonselective adenosine receptor antagonist. K-7259 (1 microM) also attenuated the H2O2-induced mechanical and metabolic derangements. Nevertheless, neither dilazep nor K-7259 modified the tissue malondialdehyde level, which was increased by H2O2, and the mechanical function and energy metabolism of the normal (H2O2-untreated) heart. These results suggest that both dilazep and K-7259 attenuate mechanical and metabolic derangements induced by H2O2. The protective action of dilazep and K-7259 on the H2O2-induced derangements is not due to potentiation of adenosine-mediated effects, reduction of lipid peroxidation or preservation of energy.

Animals↗

PubMed1996

Effects of 6-(3-dimethylaminopropionyl)forskolin hydrochloride on energy metabolism in dog hearts in situ.

The effect of NKH477 (6-(3-dimethylaminopropionyl)forskolin hydrochloride, CAS 138605-00-2) [formula: see text], a newly developed forskolin derivative, on mechanical function, carbohydrate metabolism and energy metabolism was examined in the dog heart, and compared with that of dobutamine. Intravenous injection of NKH477 (10 or 30 micrograms/kg) or dobutamine (1 or 3 micrograms/kg) increased the maximum rate of rise of left ventricular pressure (LVdP/dtmax), producing a positive inotropic effect. The duration of the effect of NKH477 was longer than that of dobutamine. In addition, NKH477 (10 or 30 micrograms/kg) increased heart rate (positive chronotropic effect). Nevertheless, neither NKH477 (10 or 30 micrograms/kg) nor dobutamine (1 or 3 micrograms/kg) modified the tissue levels of adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, creatine phosphate and lactate. Glycolytic flux (as expressed by the ratio of ([glucose-6-phosphate] + [fructose-6-phosphate]) / [fructose-1,6-diphosphate]) and redox state (as expressed by the ratio of [lactate] / [pyruvate]) in the myocardial cells were not influenced by NKH477 (10 or 30 micrograms/kg) or dobutamine (1 or 3 micrograms/kg). These results suggest that NKH477 produces both positive inotropic and positive chronotropic effects, while it does not interfere with the myocardial energy and carbohydrate metabolism.

Adenine Nucleotides↗

PubMed1996

Dilazep and its derivative, K-7259, attenuate mechanical derangement induced by palmitoyl-L-carnitine in the isolated, perfused rat heart.

The effect of dilazep, a potentiator of the adenosine-mediated effects, on the palmitoyl-L-carnitine (PALCAR)-induced mechanical derangement was studied in the isolated rat heart and compared with that of K-7259, a dilazep derivative having less potentiating action on the adenosine-mediated effects. The heart was perfused aerobically by the Langendorff's technique at a constant flow and driven electrically. PALCAR (5 microM) decreased the left ventricular developed pressure and increased the left ventricular end diastolic pressure in the heart (i.e., mechanical dysfunction). These mechanical alterations induced by PALCAR were attenuated by dilazep (1 microM) and K-7259 (1 microM). In contrast, adenosine (10 or 100 microM) did not attenuate the PALCAR-induced mechanical derangement. On the other hand, neither dilazep nor K-7259 modified the mechanical function of the normal (PALCAR-untreated) heart. These results suggest that dilazep and K-7259 attenuate the PALCAR-induced mechanical derangement and that the protective action of both drugs is not due to potentiation of adenosine-mediated effects.

Adenosine↗

PubMed1996

In vivo implantation of human osteosarcoma cells in nude mice induces bones with human-derived osteoblasts and mouse-derived osteocytes.

Two human osteosarcoma cell lines, Hu09 and OST, were suspended in Matrigel (Becton Dickinson Labware, Bedford, Massachusetts) and implanted subcutaneously in the backs of nude mice. To study phenotypic changes of tumor cells and host cells, expression of mRNA for osteopontin (OPN), osteocalcin (OC), and osteonectin (ON) was analyzed by in situ hybridization. Bone tissue was formed in the tumors derived from Hu09 cells. OPN mRNA was transcribed predominantly in osteocyte-like cells within the bone, whereas OC mRNA was transcribed in osteoblast-like cells that surrounded the bone. ON mRNA was detected in both types of cells. The similarity of the expression pattern of OPN, OC, and ON during osteogenesis of Hu09 cells to that of normal skeletal development suggests that the bone formed in Hu09-implanted mice is the same as normal bone tissue. By DNA-DNA in situ hybridization using a human-specific Alu probe and a mouse-specific m-L1 probe, osteoblast-like cells in Hu09 tumorous bone were, however, of human origin, whereas osteocyte-like cells were of mouse origin. In the tumors derived from OST cells, no osteogenesis was observed during the experimental period, and the expression of OPN, OC, and ON was not detected in tumor cells. An endochondral bone formation was not evident when these cells were simply implanted into muscle tissue. An endochondral bone was, however, reactively induced in the host mUscle tissue either when 1 alpha-hydroxyvitamin D3 and all-transretinoic acid were administered to OST-implanted mice or when Hu09 cells were pretreated with dexamethasone before implantation. Hu09 implantation seems to be a useful tool not only for the study of the differentiation of osteosarcoma cells but also for the investigation of the mechanism of bone formation. This system, using Hu09 and OST, may provide us with a new tool for the isolation of the unidentified factors that induce or inhibit osteogenesis in vivo.

Animals↗

PubMed1995

DNA fragmentation in granular cells of human cerebellum following global ischemia.

Vulnerability of human cerebellum in two autopsy cases following global brain ischemia was examined histologically by using a specific in situ nick-end labeling method for DNA breaks. In both cases, DNA fragmentation was observed in approximately one-third of the granular cells in cerebellar cortex, whereas Purkinje cells were still alive and no DNA fragmentation was recognized in the nuclei. The present study suggests that some granular cells of cerebellar cortex are more vulnerable to transient ischemia than Purkinje cells and death of granular cells is induced by an apoptotic DNA fragmentation following global brain ischemia.

Adult↗