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M Niwa

Publications and source records attributed to M Niwa.

At least 199 records · Page 11Linked to original sources
PubMed1996

Involvement of an intercellular adhesion molecule 1-dependent pathway in the pathogenesis of secondary changes after spinal cord injury in rats.

The intercellular adhesion molecule 1 (ICAM-1) plays an important role in immune responses by promoting infiltration of neutrophils into tissues; however, its implication in the secondary destructive pathomechanism after the initial mechanical injury to the spinal cord has not been clarified yet. This study was conducted to examine the role of ICAM-1 in this process after spinal cord injury (SCI) in rats. The expression of ICAM-1 mRNA was investigated by the reverse transcription-PCR method and the effect of monoclonal antibody (mAb) to ICAM-1 on SCI was evaluated by measuring various parameters. ICAM-1 mRNA expression correlated with the severity of injury and reached its maximum level 6 h after SCI. Intravenous injection of ICAM-1 mAb (1 mg/kg) 30 min after SCI reduced motor disturbance and enhanced recovery. Moreover, it significantly suppressed myeloperoxidase activity by 43.0% and spinal cord edema by 1.1% in the injured spinal cord tissue. The posttraumatic drop in spinal cord blood flow was also improved. These results suggest that ICAM-1 is deeply involved in the secondary self-destructive process after mechanical injury of the spinal cord and should be an effective target for developing a pharmacological treatment for SCI.

Animals↗
PubMed1996

Low incidence of the stimulatory G protein alpha-subunit mutations in autonomously functioning thyroid adenomas in Japan.

Point mutations of the alpha-subunit of the stimulatory G protein (Gs alpha) (termed gsp) and the TSH receptor, leading to constitutive elevation of intracellular cAMP contents, have recently been described in autonomously functioning thyroid adenomas. Since we have recently found a clear difference in the incidence of such gain-of-function mutations of the TSH receptor between Japan and Europe, the present study was designed to evaluate the incidence of gsp mutations in a series of autonomously functioning thyroid adenomas in a Japanese population. Thirty-eight autonomously functioning thyroid adenomas were employed in the present study. Genomic DNA was extracted from archival formalin-fixed, paraffin-embedded tissue specimens. Exons 8 and 9 of the Gs alpha gene were examined for activating mutations at codons 201 and 227 by single-strand conformation polymorphism (SSCP) and nucleotide sequencing. Both exons 8 and 9 were successfully amplified in 28 of 38 samples by polymerase chain reaction. Of these only one sample for exon 9 displayed a distinct migration pattern in SSCP analysis, in which a point mutation of a G to C transversion was identified by nucleotide sequencing, resulting in substitution of His (CAC) for Gln (CAG) at codon 227. The wild-type sequence of exon 9 in 10 randomly selected samples with an identical migration pattern in SSCP analysis suggests that underestimation of the incidence of gsp mutations is unlikely in our work. In conclusion, the present data, together with our previous data on the TSH receptor, suggest that oncogenic mutations of the Gs alpha as well as the TSH receptor do not seem to play a major role in tumorigenesis of autonomously functioning thyroid adenomas in Japan.

Adenoma↗
PubMed1996

Expression and regulation of G protein-coupled receptor kinase 5 and beta-arrestin-1 in rat thyroid FRTL5 cells.

G protein-coupled receptor kinases (GRKs) and arrestins are implicated in homologous desensitization of G protein-coupled receptors. We have recently demonstrated that among six GRKs so far identified, GRK5 is the isoform predominantly expressed in the thyroid and appears to be mainly involved in homologous desensitization of thyrotropin receptor (TSHR) in FRTL5 cells. To further understand the molecular mechanisms of the TSHR desensitization, the expression and regulation of GRKs and arrestins together with those of the TSHR were examined in FRTL5 cells. Northern blot analysis of total RNA from FRTL5 cells with the available rat GRK cDNAs (GRK4, 5, and 6) as probes showed that only GRK5 mRNAs of approximately 3, 8, and 10 kilo bases (kb) in length were detectable. When probed with rat beta-arrestin-1 and beta-arrestin-2 cDNAs, beta-arrestin-1 mRNAs of approximately 7.5 and 2.5 kb long, but no (or possibly faint) approximately 2.4 kb beta-arrestin-2 mRNA, were observed, suggesting that in the thyroid, beta-arrestins appear to be predominantly of beta-arrestin-1 isoform. In studies on TSH-regulation of GRK5, beta-arrestin-1 and TSHR mRNAs, steady-state levels of GRK5 and TSHR mRNAs were 3- to 4-fold lower in the cells grown in the medium with TSH than in those without TSH, while betaarrestin-1 mRNA levels were unchanged. Downregulation of GRK5 and TSHR mRNAs by TSH was further confirmed by dose- and time-dependent experiments. Incubation with 1mM 8BrcAMP, a cAMP analog, for 24h fully reproduced this TSH inhibitory effect. A decrease in GRK5 protein by TSH was also confirmed with Western blot analysis. In summary, these data together with our previous data suggested that GRK5 and beta-arrestin-1 seem to be the isoforms predominantly expressed in the thyroid, and they appear to play a pivotal role in TSHR homologous desensitization. We also demonstrated TSH downregulation of GRKS, but not beta-arrestin-1, expression. Further studies will be necessary to elucidate how these phenomena are linked to thyroid pathophysiology.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1996

Extraction and characterization of the smooth-type lipopolysaccharide from Fusobacterium nucleatum JCM 8532 and its biological activities.

The lipopolysaccharide (LPS) of Fusobacterium nucleatum JCM 8532 was isolated by hot-phenol water extraction. Most of the LPS was extracted in the phenolic phase and shown to be the smooth-type, whereas the aqueous phase contained mainly rough-type LPS. The chemical composition of the LPS was similar to that reported in other studies, but D-quinovosamine, which may be a major component of O-antigenic polysaccharide, and 3-deoxy-D-manno-2-octulosonic acid (Kdoi) were detected for the first time by gas chromatography-mass spectrometry. The biological activities of smooth-type LPS, including limulus activity, lethal toxicity, pyrogenicity, and B lymphocyte mitogenicity, were comparable to those of entero-bacterial LPS. Smooth-type LPS inhibited the cell growth and DNA synthesis of adult and fetal human gingival fibroblasts in a dose-dependent manner, suggesting that LPS may play a role in the occurrence of human gingivitis.

Adult↗
PubMed1996

Localization of endothelin ETB receptors on the myenteric plexus of guinea-pig ileum and the receptor-mediated release of acetylcholine.

1. The type of endothelin (ET) receptor located on the myenteric neurones of guinea-pig ileum was determined by receptor autoradiography and function of the receptor was examined by release experiments of acetylcholine (ACh) from the longitudinal muscle myenteric plexus (LM-MP) preparations. 2. Specific [125I]-ET-1 binding sites were distributed in muscle layers, myenteric and submucous plexuses, and mucosa layers. High-grain densities were detected in both myenteric and submucous plexuses. 3. Binding in the myenteric plexus was abolished by incubation with either IRL 1620 (endothelin ETB receptor agonist) or BQ 788 (endothelin ETB receptor antagonist), but not with BQ 123 (endothelin ETA receptor antagonist). The [125I]-IRL 1620 binding sites were evident in the myenteric plexus. Thus, the endothelin receptor located on the myenteric neurones is of the ETB type. 4. ET-1 (10(-10)-3 x 10(-8) M) and ET-3 (10(-10)-3 x 10(-8) M) evoked 3H outflow from LM-MP preparations of ileum preloaded with [3H]-choline, in a concentration-dependent manner. There was no significant difference between maximum amounts of ET-1-evoked and ET-3-evoked 3H outflow. 5. ET-1 and ET-3 evoked outflow of 3H was BQ 788-sensitive, but BQ 123-insensitive. Both evoked outflows of 3H were Ca(2+)-dependent and tetrodotoxin-sensitive. 6. These results indicate that the endothelin ETB receptor is located on the enteric cholinergic neurones and that stimulation evokes the release of ACh.

Acetylcholine↗
PubMed1996

Comparative study of pharmacokinetics of two new fluoroquinolones, balofloxacin and grepafloxacin, in elderly subjects.

Comparative pharmacokinetics and tolerability were studied in healthy elderly volunteers for two new fluoroquinolones, balofloxacin (Q-35) and grepafloxacin (OPC-17116), the main excretion routes being the renal and hepatic routes, respectively. Both agents were well tolerated in elderly subjects. In comparison with previously reported data from healthy younger adults, the absorption of balofloxacin was slightly delayed and urinary excretion was delayed and diminished. As a significant linear correlation was observed between renal clearance of balofloxacin and creatinine clearance, the delayed and diminished urinary recovery was attributed to the reduced renal function of the elderly subjects enrolled in the study. The absorption of grepafloxacin was also delayed, and the maximum plasma drug concentration and area under the plasma drug concentration-time curve were increased in the elderly by 31 and 48%, respectively, over those in younger adults on the basis of dose normalized to body weight. The plasma terminal elimination half-life and urinary recovery remained unchanged. Decreases in distribution volume and total body clearance in the elderly were considered to be the primary factors contributing to these differences.

Aged↗
PubMed1996

Oxidative modification of a cephalosporin C acylase from Pseudomonas strain N176 and site-directed mutagenesis of the gene.

A cephalosporanic acid acylase from Pseudomonas strain N176 catalyzes hydrolysis of both glutarylcephalosporanic acid and cephalosporin C to 7-amino-cephalosporanic acid. Chemical modification of the enzyme with acidic hydrogen peroxide was performed to investigate residues which play important roles in enzymatic activity. The activity of the enzyme was reduced to 76% of the original by oxidation. From protein chemical analysis combined with site-directed point mutagenesis, modification of Met-164 was found to correspond to the reduction in activity. To study the effect of Met-164 on the enzymatic character, we prepared mutant acylases in which Met-164 was replaced with several other amino acids and obtained the following data: (i) there existed a trend of mutation to noncharged hydrophilic residues, resulting in an increase of activity against glutarylcephalosporanic acid; (ii) the mutation of Met-164 to Gly and Ala resulted in the lowering of both Km values and the optimal pHs against glutarylcephalosporanic acid; (iii) the mutation to Leu enhanced cephalosporin C acylase activity; and (iv) the mutation to Gln improved the k(cat) value for glutarylcephalosporanic acid. In particular, the mutation to Gln resulted in a high rate of conversion of glutarylcephalosporanic acid to 7-amino-cephalosporanic acid under conditions similar to those of a bioreactor system. These results may indicate that Met-164 is located in or near the cephalosporin compound binding pocket on the enzyme.

Amidohydrolases↗
PubMed1996

Comparison between adult and pediatric arteriovenous malformations treated by Gamma Knife radiosurgery.

More than 290 cases of cerebral arteriovenous malformation have been treated at our hospital by Gamma Knife radiosurgery since May 1991, of which 99 were followed with angiography for 1 year or more. The results from adult and pediatric age groups were compared. There were 70 adults and 23 children. Previous hemorrhage had occurred in 73.7% of the adults and 91.3% of the children. The mean volume of the nidus was similar in both groups: 4.2 cm3 in adults and 4.8 cm3 in children. In both groups approximately 70% of the cases fell into Spetzler and Martin grade III. The mean margin dose was 20.0 Gy in the adults and 20.5 Gy in the children. Complete nidus occlusion at 1 year occurred in 45% of the adults and 74% of the children. The rates at 2 years were 81 and 94%, respectively. Complications occurred only in adults and consisted of 2 rebleeds, 1 radiation edema and 1 radiation necrosis. Gamma Knife radiosurgery is considered to be safer and have a higher success rate in children than in adults.

Adolescent↗
PubMed1996

Two subtypes of endothelin receptors and endothelin peptides are expressed in differential cell types of the rat placenta: in vitro receptor autoradiographic and in situ hybridization studies.

We studied the localization of endothelin (ET) receptors and ET peptides in the rat placenta. In vitro receptor autoradiographic and in situ hybridization studies revealed the differential and cell-specific distribution of ET receptor subtypes, suggesting that each ET receptor plays a different role in the function of the placenta. The expression of the ETB receptor was concentrated to cytotrophoblasts and trophoblastic giant cells of the basal zone, in which fetal cells directly face maternal cells. The ETA receptor was confined to the decidual tissue and vascular wall. Both ET receptors coexisted in the labyrinth in an approximately 50:50 ratio. Prepro-ET-1 messenger RNA (mRNA) was detected in cytotrophoblasts and trophoblastic giant cells of the basal zone and endothelial cells of vessels, whereas ET-1-like immunoreactivity was present not only in trophoblasts and endothelial cells, but also in the decidual tissue and vascular wall. ET-3 mRNA was localized in migrating cells. We also found changes in the expression levels of ET receptors by means of a cold ligand saturation study. The number of specific [125I]ET-1-binding sites was increased in the basal zone and labyrinth with gestation, but not in the decidual tissue. The enhancement of ETA receptor, ETB receptor, and prepro-ET-1 mRNA levels was also supposed, based on data obtained by RT-PCR Southern hybridization. On the other hand, ET-3 mRNA levels were reduced with gestation. These findings support the idea that ETs, through interaction with ETA and ETB receptors, play an important role in the regulation of placental growth and fetoplacental circulation through autocrine and paracrine mechanisms.

Animals↗
PubMed1996

Neuroprotective effect of TTC-909, an isocarbacyclin methyl ester incorporated in lipid microspheres, on hippocampal delayed neuronal death of stroke-prone spontaneously hypertensive rats.

TTC-909 is a newly developed isocarbacyclin methyl ester (TEI-9090) incorporated in lipid microspheres. The neuroprotective effect of TTC-909 was histologically examined in the pyramidal cell layer of the hippocampus CA1 subfield 7 days after transient forebrain ischemia using stroke-prone spontaneously hypertensive rats. TTC-909, given intravenously 10 min after the transient forebrain ischemia, dose-dependently protected against ischemia-related delayed neuronal death. The blood pressure remained unchanged following TTC-909 administration. This finding suggests that TTC-909 has a neuroprotective action on ischemic delayed neuronal death in the hippocampus.

Animals↗
PubMed1996

Coagulation and fibrinolysis study after local thrombolysis of a cerebral artery with urokinase.

Coagulation and fibrinolysis factors were studied in six patients after local thrombolysis with urokinase (720,000 IU). Transient abnormalities, such as prolonged prothrombin time, decreased plasminogen and alpha 2-antiplasmin activities, decreased fibrinogen, and increased fibrin degradation products were seen on the day after thrombolysis, but tended to return to the normal range on the 4th day except for one patient who suffered from disseminated intravascular coagulation. Antithrombin III activity did not change so much. Therefore, the dosage of urokinase should be as low as possible to prevent fluctuations in the coagulation and fibrinolysis system.

Aged↗
PubMed1996

[Cariostatic mechanisms of fluoride and its effects on human beings].

Fluorine is ubiquitous in man's environment, rocks, soil, water, air, etc, and mainly is in the form of inorganic fluoride (compounds of fluorine) and utilized extensively for industrial and medical purposes. It is also known that fluoride has both beneficial and toxic effects in man. One of the most beneficial effects of fluorine on human health is the application of fluoride solution for the prevention or reduction the incidence of dental caries. The major cariostatic mechanisms is believed to be that fluoride reduces dental caries through influencing the morphology of teeth by reducing the solubility of the enamel and promoting remineralization, and through its effect on oral cariogenic bacteria. Recently, the possible beneficial effects of fluoride on osteoporosis have been studied. Chronic effects of fluoride, described by many epidemiological studies, are only dental fluorosis and skeletal fluorosis. Most cases of acute poisoning in human subjects have been associated with the suicidal, accidental or experimental (by researcher himself) ingestion of a fluoride solution.

Dental Caries↗
PubMed1996

[Radiosurgery of cavernous sinus meningiomas with gamma-knife].

The treatment results of cavernous sinus meningioma with gamma-radiosurgery are reported. There were 18 cases of cavernous sinus meningioma, including 2 males and 16 females, whose age ranged from 39 to 63 with an average of 51.0 years. As prior treatments, operative tumor resection or biopsy had been carried out in 14 cases, and the pathology was verified. The other 4 cases were diagnosed clinically with radiological studies. The mean tumor diameter was 28.3mm (17.7-35.0) during the radiosurgery. The maximum dose ranged from 22 to 36Gy (mean 28.0Gy), with the marginal tumor dose ranging from 11 to 18Gy (mean 13.9Gy). Irradiation to the near-by optic nerves was less than 10Gy. Follow-up period ranged from 12 to 50 months with a mean of 25.5 months. MRI showed a minor tumor shrinkage in 9 (50.0%) and no obvious change in 8 (44.4%), and tumor progression in 1 (5.6%), which required a 2nd radiosurgery. Neurologically facial pain and facial dysesthesia were well improved (7/13). However the ophthalmoparesis was usually unchanged and only 1 out of 11 (9.1%) improved after radiosurgery. Deterioration of neurological signs was rare. Symptomatic edema presenting neurological signs was not seen. In conclusion, radiosurgery with a gamma-knife is one of the useful alternatives to operative intervention in the treatment of cavernous sinus meningiomas, not only for tumor control, but also for relief from the symptoms.

Adult↗
PubMed1996

[Gamma knife treatment of AVM of the basal ganglia and thalamus].

Arteriovenous malformatios (AVMs) in the basal ganglia (BG) and thalamus (Thal) are difficult to treat by microsurgery or intravascular embolization alone, and the role of stereotactic gamma radiosurgery (gamma knife) of these AVMs is discussed. We have treated 324 cases of AVM with gamma knife since May 1991, and in 71 of these cases (19%) the AVM was in the BG or Thal. The results of gamma radiosurgery on AVMs of the BG and Thal were compared with the results of treating AVMs at other intracranial locations by gamma radiosurgery. The nidi were small (mean diameter: 16.4 mm), and they were treated with a mean maximum dose of 36.4 Gy and marginal dose of 19.9 Gy. The results were evaluated angiographically in 39 (55%) of the 71 cases, with a mean follow-up period of 23 months. The complete obliteration rate of AVMs in the BG and Thal 1 and 2 years after treatment was 54.3% and 92.0%, respectively, and the rate at the other locations was 42.9% and 76.0%, respectively. Adverse effects of this treatment in the AVM cases overall were rebleeding from the nidus in 5 cases (1.5%) and radiation necrosis in 4 cases (1.2%). In conclusion, AVMs of the BG and Thal were effectively and safely treated with the gamma knife, and stereotactic radiosurgery is a definitive alternative treatment for deep seated AVMs.

Adolescent↗
PubMed1996

[A sensitive fluorescent assay for the detection and quantification of mitochondrial 3243 mutation].

A mutation at nucleotide 3243 in the mitochondrial DNA (mtDNA) specifying tRNALeu(UUR) has been shown to be related with diabetes mellitus. Since mtDNA shows heteroplasmy and its composition would change among various tissues, it is important to quantitate the percentage of mutant mtDNA precisely. Here we report a rapid and sensitive method to determine the mutant mtDNA. A fragment of mtDNA containing nucleotide 3243 was amplified by PCR using a rhodamine-labeled primer and the product was digested with ApaI. The PCR product from normal gene was resistant to ApaI digestion, while a mutation from A to G generated an ApaI site in the mutant mtDNA and its PCR product was cleaved into two fragments. The digested product was separated by acrylamide gel electrophoresis and each product was quantitated by a fluorescence analyzer. PCR analysis using standard mixtures of normal and mutant mtDNAs indicated that this method can be used to detect as little as 1% mutant mtDNA.

DNA Primers↗
PubMed1996

[The results of gamma knife radiosurgery for malignant skull base tumors].

The results of gamma knife radiosurgery for malignant skull base tumors were analyzed using repeated magnetic resonance imagings and neurological examinations. Nineteen malignant skull base tumors were treated and followed up for 22.3 months (5-40 months) using MR imagings. The mean age was 54.4 years old (ranging from 16-85). Ten were male and 9 were female. Prior to the radiosurgery, removal of the tumors in 17 cases, conventional radiation therapy in 7, and chemotherapy in 4 etc. were performed. The pathological diagnoses were chordoma in 6 cases, metastatic tumors in 5, epipharyngeal carcinoma in 2, adenoid cystic carcinoma in 2, and others in 4. The locations of tumors were the clivus in 8, the parasellar region in 5, the epipharynx in 2, the paranasal sinus in 2, C-P angle in 1, and intraorbital region in 1 (14 were intracranial and 5 were extracranial). The mean diameter of the tumor was 33.5mm. The mean maximum dose was 26.8Gy and the mean marginal dose was 12.9Gy during treatment. Repeated MR imagings revealed decrease of tumor size in 12 cases, no change in 1, and increase of tumor size in 5 (unknown in 1). Follow-up neurological examinations showed improvement in 3 cases, no change in 9, and deterioration in 7. There were 11 deaths during a mean follow-up period of 17.8 months (5-32 months) and another 8 cases are alive for a mean follow-up of 30.5 months (20-40 months) after the radiosurgery. Although the tumor size was large at the time of treatment, the results of gamma knife radiosurgery were promising. Considering the quality of life of patients with malignant skull base tumors, it is emphasized that gamma knife treatment is the method of choice compared with radical removal of the tumors.

Adolescent↗