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H Okamura

Publications and source records attributed to H Okamura.

At least 361 records · Page 20Linked to original sources

NADPH-diaphorase and nitric oxide synthase in the canine superior cervical ganglion.

By means of NADPH-diaphorase (NADPH-d) histochemistry and nitric oxide synthase (NOS) immunohistochemistry, we demonstrate that considerable numbers of NADPH-d-positive neurons are distributed throughout the canine superior cervical ganglion (SCG). These neurons also show NOS immunoreactivity. This finding indicates that NADPH-d histochemistry, a simple and reliable technique, can be used as a reliable marker of NOS activity in the sympathetic innervation of canine head and neck. The present findings suggest that the participation of nitric oxide in the SCG differs greatly between species.

Amino Acid Oxidoreductases↗

Circadian change of VIP mRNA in the rat suprachiasmatic nucleus following p-chlorophenylalanine (PCPA) treatment in constant darkness.

Neuronal activity of the suprachiasmatic nucleus (SCN) is known to be regulated by two major extrinsic factors conveyed by three anatomically distinct pathways to the SCN: photic stimulus by the direct retinohypothalamic tract (RHT) and the indirect geniculohypothalamic tract (GHT), and information from the brainstem by ascending forebrain serotonergic (5-hydroxytryptamine: 5-HT) tract. It has been shown that VIP mRNA level in neurons of the SCN is altered by external light, but remains stable in constant darkness. In the present study, by using the in situ hybridization technique combined with computer-assisted image analysis, we examined VIP mRNA expression in the SCN of rats in which the two major factors were eliminated, i.e. photic stimulus by exposing animals in total darkness and 5-HT transmission by three-day successive administration of p-chlorophenyl-alanine methylester (an inhibitor of tryptophan hydroxylase, 200 mg/kg, daily). In saline-treated controls, VIP mRNA levels remained almost constant throughout the day. In contrast, in PCPA-treated rats, a significant rhythm of VIP mRNA was observed with a peak at CT 4 and a trough at CT 20. These observations suggest that the removal of photic and 5-HT influence induces VIP mRNA rhythm in the SCN, indicating that VIP mRNA is controlled not only by photic information but also by the circadian clock.

Analysis of Variance↗

Induction of VGF mRNA in neurons of the rat nucleus tractus solitarius and the dorsal motor nucleus of vagus in duodenal ulceration by cysteamine.

To investigate the possible role of the brainstem in cysteamine-induced peptic ulceration, we examined the expression of VGF mRNA, which is induced in PC12 cells following application of nerve growth factor [23], in the nucleus tractus solitarius (NTS)/dorsal motor nucleus of vagus (DMV) complex of the medulla oblongata by in situ hybridization histochemistry. In control saline-treated rats, weak VGF mRNA signals were only rarely detected in neurons of the NTS and none were observed in those in the DMV. After 12 h of cysteamine administration (450 mg/kg, s.c.), the time at which duodenal ulcer was detected in all cases, heavily labeled VGF mRNA-expressing neurons appeared in the NTS and DMV. By quantitative analysis on macroautoradiogram, the VGF mRNA signals of the NTS/DMV complex in cysteamine-treated rats were twice as much as those in saline-treated rats. In situ hybridization histochemistry combined with the use of the retrograde neuronal tracer cholera toxin-B subunit revealed that the induced VGF mRNA-expressing neurons of the DMV projected directly to the stomach. The present results suggest that ulceration accompanies the induction of VGF mRNA in neurons of vagal afferent and efferent areas of the brainstem.

Animals↗

Stimulation of cyclic adenosine 3',5'-monophosphate-dependent protein kinase with brain gangliosides.

The holoenzyme of cAMP-dependent protein kinase (cAMP-kinase) partially purified from the particulate fraction of rat brain was stimulated by gangliosides. Among various gangliosides tested, GM1 was most potent, giving Ka value of 19.5 microM. The maximal activation of the kinase was obtained with 100 microM GM1 using kemptide as substrate. Gangliosides inhibited the kinase activity of the catalytic subunit of cAMP-kinase. Of various substrates tested, the ganglioside-stimulated cAMP-kinase could phosphorylate microtubule-associated protein 2, synapsin I and myelin basic protein, but not histone H1 and casein. The molecular mechanisms of the stimulatory effect of gangliosides were investigated. The kinase activated with GM1 was inhibited by the addition of PKItide, a specific inhibitor for cAMP-kinase. However, GM1 did not dissociate the holoenzyme into the catalytic and regulatory subunits and did not interfere with the binding ability of cAMP to the holoenzyme. These results suggest that the gangliosides can directly activate cAMP-kinase in a different manner from cAMP.

Animals↗

Sympathetic preganglionic neurons contain nitric oxide synthase and project to the superior cervical ganglion: combined application of retrograde neuronal tracer and NADPH-diaphorase histochemistry.

Nitric Oxide (NO), which was initially identified as an endothelium-derived relaxing factor, has recently been demonstrated to be a neuronal messenger in central and peripheral nervous systems. In the present study, we examined the possibility of NO producing neurons in teh intermediolateral (IML) cell collum of the thoracic spinal cord (Th) project to the superior cervical ganglion (SCG). First, we observed the NADPH-diaphorase-positive/nitric oxide synthase (NOS)-immunoreactive neurons of the IML and the dorsal part of the central canal at the level of Th1-Th3, and numerous fiber-stainings in the superior cervical ganglion. Second, after injecting WGA-HRP (wheat germ agglutinin-horse radish peroxidase complex), a retrograde neuronal tracer, into the SCG, and developing WGA-immunohistochemistry and the NADPH-diaphorase histochemistry in the same sections, we detected double-labeled neurons in the IML. These findings provide evidence that sympathetic preganglionic NO producing neurons directly innervate to the SCG.

Amino Acid Oxidoreductases↗

Neurotransmitters for the canine inferior pharyngeal constrictor muscle.

The inferior pharyngeal constrictor muscle plays an important role at the pharyngeal phase of deglutition and is anatomically composed of the thyropharyngeal muscle and cricopharyngeal muscle. In this study we investigated the distribution pattern of neuropeptidergic and catecholaminergic nerve fibers in the thyropharyngeal muscle and cricopharyngeal muscle of seven puppies by immunohistochemistry. Some of the calcitonin gene-related peptide-, substance P-, vasoactive intestinal polypeptide-, and tyrosine hydroxylase-immunoreactive nerve fibers were found to lie parallel to the muscle fibers in both the thyropharyngeal muscle and cricopharyngeal muscle. Nerve fibers with immunoreactivity to all substances examined were found to be associated with blood vessels in both the thyropharyngeal muscle and cricopharyngeal muscle, and the number of calcitonin gene-related peptide, neuropeptide Y, and tyrosine hydroxylase nerve fibers was higher than the number of substance P, vasoactive intestinal polypeptide, and galanin nerve fibers. Motor end plate-like structures with calcitonin gene-related peptide immunoreactivity were found in both the thyropharyngeal muscle and cricopharyngeal muscle. These structures in the cricopharyngeal muscle were clearly less than those in the thyropharyngeal muscle. Some clusters of neurons were detected only in the cricopharyngeal muscle of all dogs examined. Substance P-, vasoactive intestinal polypeptide-, galanin-, and neuropeptide Y-immunoreactive neurons were found in this ganglion, and the vasoactive intestinal polypeptide-immunoreactive neurons were the most abundant. Abundant calcitonin gene-related peptide- and vasoactive intestinal polypeptide-immunoreactive nerve fibers, and some substance P- and galanin-immunoreactive nerve fibers were distributed in the ganglion.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

A 6-kb upstream region of the human transthyretin gene can direct developmental, tissue-specific, and quantitatively normal expression in transgenic mouse.

To ascertain whether a 6-kb upstream region of the human transthyretin (TTR) gene contains the cis-element(s) required for proper specificity and level of expression, transgenic mice carrying the human mutant TTR gene containing either 6-kb (6.0-hMet30) or 0.6-kb (0.6-hMet30) of the upstream region were produced and studied. The 6.0-hMet30 gene was expressed in the yolk sac, liver, and choroid plexus, where the mouse endogenous TTR gene is also expressed. In contrast, expression of the 0.6-hMet30 gene was restricted to the yolk sac and liver. The expression levels of the 6.0-hMet30 gene in the liver and serum were similar to those of the mouse TTR gene, and about 10-fold those of the 0.6-hMet30 gene. Before birth, the developmental profiles of the expression of both transgenes in each tissue were similar to those of the mouse TTR gene. However, the expression levels of the 6.0-hMet30 gene in the liver and serum increased after birth to reach adult levels at an age of 4 weeks, while expression of the 0.6-hMet30 gene remained at a low level after birth. These results suggest that the 6-kb upstream sequence contains the cis-elements required for developmental, tissue-specific, and quantitatively normal expression.

Animals↗

A survey of Japanese physicians' attitudes and practice in caring for terminally ill cancer patients.

To clarify the psychiatric liaison issues in cancer care, questionnaires were distributed to physicians at 31 teaching hospitals in Japan, including cancer centers and psychiatrists at 197 teaching hospitals. Data from 329 physicians and 156 psychiatrists showed that the majority of the physicians felt troubled by the psychiatric problems of terminally ill patients. However, actual psychiatric referrals were infrequent. An important factor that interferes with appropriate psychiatric referrals for cancer patients is that most physicians do not usually inform patients of a cancer diagnosis. This, it seems that close communication between physicians and psychiatrists is essential in caring for terminally ill cancer patients in the context of Japanese culture, when the psychiatric consultations are offered.

Adult↗

Growth inhibition of Ureaplasma urealyticum by the proton pump inhibitor lansoprazole: direct attribution to inhibition by lansoprazole of urease activity and urea-induced ATP synthesis in U. urealyticum.

The proton pump inhibitors (PPIs) omeprazole and lansoprazole and the acid-activated analog of lansoprazole AG-2000, which potently inhibit the urease of Helicobacter pylori (K. Nagata, H. Satoh, T. Iwahi, T. Shimoyama, and T. Tamura, Antimicrob. Agents Chemother. 37:769-774, 1993), also inhibited the urease activities of cell-free extracts as well as intact cells of Ureaplasma urealyticum. The 50% inhibitory concentrations were between 1 and 25 microM. These compounds also inhibited the ATP synthesis induced by urea in ureaplasma cells. The 50% inhibitory concentrations for ATP synthesis were close to those for urease activity, but they were lower than those of urease inhibitors, such as acetohydroxamic acid, hydroxyurea, and thiourea. In addition, one of the metabolites of lansoprazole found in human urine, M-VI, also inhibited ureaplasmal urease activity and the ATP synthesis induced by urea at almost the same concentrations as those of lansoprazole. The inhibition of PPIs against ureaplasma urease was very similar to those against H. pylori urease, suggesting that the inhibitory mechanism against these ureases was due to the blockage of the SH residues on the cysteine of the enzyme. Omeprazole, lansoprazole, AG-2000, and M-VI inhibited the growth of U. urealyticum. Since ureaplasma urease is thought to be involved in the pathogenicity of this organism in the urogenital tract, PPIs and their analogs may be useful as chemotherapeutic agents against diseases caused by U. urealyticum.

2-Pyridinylmethylsulfinylbenzimidazoles↗

Inhibitory action of lansoprazole and its analogs against Helicobacter pylori: inhibition of growth is not related to inhibition of urease.

The proton pump inhibitors omeprazole and lansoprazole and its acid-activated derivative AG-2000, which are potent and specific inhibitors of urease of Helicobacter pylori (K. Nagata, H. Satoh, T. Iwahi, T. Shimoyama, and T. Tamura, Antimicrob. Agents Chemother. 37:769-774, 1993), inhibited the growth of H. pylori. The growth was inhibited not only in urease-positive clinical isolates but also in their urease-negative derivatives which had no urease polypeptides. AG-1789, a derivative of lansoprazole with no inhibitory activity against H. pylori urease, also inhibited the growth of both strains even more strongly than the urease inhibitors lansoprazole and AG-2000. Furthermore, the antibacterial activity of omeprazole and lansoprazole was not affected by glutathione or dithiothreitol, which completely abolished the inhibitory activity of lansoprazole against H. pylori urease. These results indicated that the inhibitory action of these compounds against the growth of H. pylori was independent from the inhibitory action against urease.

2-Pyridinylmethylsulfinylbenzimidazoles↗

A novel costimulatory factor for gamma interferon induction found in the livers of mice causes endotoxic shock.

Administration of monoclonal anti-CD3 antibody to mice treated with Propionibacterium acnes induced secretion of a high level of gamma interferon (IFN-gamma) into the circulation system, while it induced no significant release in untreated mice. In order to analyze this high-level induction of IFN-gamma in these bacterium-treated mice, we investigated the factors that might be involved. An activity that induces IFN-gamma in T cells was observed in the liver extracts of mice treated with P. acnes and subsequently challenged with lipopolysaccharide. Here, we purified an IFN-gamma-inducing factor from the liver extract to homogeneity and characterized it. Its molecular mass was 18 to 19 kDa, and its pI was 4.9. The amino acid sequence of the NH2-terminal portion was determined and shown to have no similarities to any protein in the EMBL, GenBank, and PIR data bases. The same molecule was also demonstrated in the serum factor that was previously reported to have an IFN-gamma-inducing activity and to have an apparent molecular mass of 75 kDa. Moreover, the activity of this serum factor was recovered in the fraction containing the 18- to 19-kDa protein under reducing conditions and was shown to have the same NH2-terminal amino acid sequence as that of the factor from the liver extract. In addition to the ability to induce IFN-gamma, this protein augmented T-cell proliferation and NK activity in the spleen cells. Thus, several of its biological activities were apparently similar to those of interleukin-12. These results indicated that this novel protein, which exhibited marked costimulatory activity on IFN-gamma production in vitro, was elevated vivo in response to P. acnes treatment. This factor, probably released from the producing cells by lipopolysaccharide stimuli, may be involved in the high-level induction of IFN-gamma in the P. acnes-treated mice.

Amino Acid Sequence↗

Prolidase from Xanthomonas maltophilia: purification and characterization of the enzyme.

Prolidase (iminodipeptidase, EC 3.4.13.9) was purified from an extract of Xanthomonas maltophilia, by ammonium sulfate fractionation and sequential chromatographies on DEAE-Toyopearl, Toyopearl HW65C, FPLC-Hiload Superdex 200 pg, and FPLC-Hitrap Q columns, which an activity recovery of 2.3%. The enzyme was the most active at pH 7.5 with Leu-Pro as substrate. It was stable between pH 6.0 and 8.5 for 60 min at 37 degrees C and retained half of activity after 60 min at 37 degrees C. The isoelectric point of the enzyme was 3.7. Its molecular weight was estimated to be 100,000 by gel filtration on FPLC-Hiload Superdex 200 and 51,000 by SDS-PAGE, suggesting that it is a dimer. It hydrolyzed dipeptides only if proline is located at the carboxyl terminal position. The enzyme was inhibited by PCMB and o-phenanthroline, and was activated by Mn2+.

Chromatography, Ion Exchange↗

Immunohistochemical localization of epidermal growth factor and its effect on granulosa cell proliferation in rat ovary.

The localization of epidermal growth factor (EGF) in the ovary and its effect on proliferation of granulosa cells were investigated in gonadotrophin-primed immature female rats. Immunoreactions with anti-rat EGF monoclonal antibody were observed sparsely in the granulosa layer and antrum of follicles, but not in the theca layer or stromal tissue. The EGF-positive cells were round or oval shaped and often larger than granulosa cells. The localization and morphological appearances of these cells in the follicles were in good agreement with those of macrophages. Although EGF alone did not promote granulosa cell growth in vitro, the labelling index with [3H]thymidine of granulosa cells cultured with 0.1 ng/ml EGF and 0.1 ng/ml basic fibroblast growth factor was significantly greater than that without the growth factors (18.4% vs. 15.8%, P < 0.01). These results suggest that macrophages in follicles may modulate follicular development through a paracrine mechanism by secreting EGF and other growth factors.

Animals↗

Effects of macrophage colony-stimulating factor on folliculogenesis in gonadotrophin-primed immature rats.

The effect of macrophage colony-stimulating factor (M-CSF) on folliculogenesis and ovulation was studied. Folliculogenesis and ovulation were induced in immature female rats with a s.c. injection of equine chorionic gonadotrophin (eCG), followed 48 h later by human chorionic gonadotrophin (hCG). The ovulation rate was measured after the following treatments. (1) Graded doses of human M-CSF (1-300 x 10(3) iu per rat) were administered i.p. daily for 3 consecutive days. (2) M-CSF (100 x 10(3) iu) was administered i.p. at designated times between 96 h before and 10 h after hCG injection. (3) Rabbit anti-human M-CSF polyclonal antibody (5 micrograms) was administered into the left ovarian bursa at designated times between 49 h before and 10 h after hCG injection. In addition, the effect of M-CSF on ovarian macrophages was investigated using immunohistochemistry with mouse anti-rat macrophage monoclonal antibody, TRPM-3. The treatment with M-CSF (> 30 x 10(3) iu per rat) significantly increased the ovulation compared with controls in a dose-dependent manner. This stimulatory effect of M-CSF was observed when it was administered between 96 h and 49 h before hCG injection. The ovarian intrabursal administration of anti-M-CSF antibody significantly inhibited the number of ovulated ova from the treated ovaries compared with either those from control rats or from the contralateral untreated ovaries between 24 h before and 3 h after hCG injection. The immunohistochemistry revealed that M-CSF increased the number of ovarian macrophages in growing follicles.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Adenylyl cyclase activity in human decidua and myometrium during pregnancy and labour.

Adenylyl cyclase activity was studied in human decidua and myometrium in early pregnancy and at term before and after the onset of labour. Decidual basal, prostaglandin-, catecholamine- and forskolin-stimulated adenylyl cyclase activities at term before the onset of labour were significantly lower than those in early pregnancy. After the onset of labour at term, decidual basal, prostaglandin-, catecholamine-, NaF- and forskolin-stimulated adenylyl cyclase activities significantly increased compared with those at term before the onset of labour. Myometrial prostaglandin- and catecholamine-stimulated activities did not alter during pregnancy, except for basal and forskolin-stimulated activity. Myometrial basal, prostaglandin-, catecholamine-, NaF- and forskolin-stimulated activities at term showed no change after the onset of labour. At term, before the onset of labour, myometrial basal, prostaglandin, catecholamine-, NaF- and forskolin-stimulated activities were the same as those in the decidua. However, after the onset of labour at term, decidual basal and the stimulated activities were significantly higher than those in the myometrium. These results suggest that decidual prostaglandin- and catecholamine-stimulated adenylyl cyclase may play an important role in the initiation or maintenance of human labour or in both processes.

Adenylyl Cyclases↗

Retrieval of a migrated detachable coil--case report.

A 74-year-old female presented with intraventricular and subarachnoid hemorrhage due to probable rupture of a basilar artery (BA) aneurysm or a superior cerebellar artery (SCA) aneurysm. She was treated by endovascular therapy using detachable coils through the BA. The BA aneurysm was completely occluded, but part of a coil migrated into the BA from the SCA aneurysm during the procedure. The migrated coil was retrieved using a snare type endovascular retrieving device. The snare loop required concentric closure around the coil by simultaneous pulling of the corewire and a slight forward movement of the tip of the snare catheter. The SCA aneurysm was successfully occluded 1 week later using a shorter coil.

Aged↗