Biomedical subjects

M Nishi

Publications and source records attributed to M Nishi.

At least 55 records · Page 3Linked to original sources

PubMed2000

Increased susceptibility to fatigue of slow- and fast-twitch muscles from mice lacking the MG29 gene.

Mitsugumin 29 (MG29), a major protein component of the triad junction in skeletal muscle, has been identified to play roles in the formation of precise junctional membrane structures important for efficient signal conversion in excitation-contraction (E-C) coupling. We carried out several experiments to not only study the role of MG29 in normal muscle contraction but also to determine its role in muscle fatigue. We compared the in vitro contractile properties of three muscles types, extensor digitorum longus (EDL) (fast-twitch muscle), soleus (SOL) (slow-twitch muscle), and diaphragm (DPH) (mixed-fiber muscle), isolated from mice lacking the MG29 gene and wild-type mice prior to and after fatigue. Our results indicate that the mutant EDL and SOL muscles, but not DPH, are more susceptible to fatigue than the wild-type muscles. The mutant muscles not only fatigued to a greater extent but also recovered significantly less than the wild-type muscles. Following fatigue, the mutant EDL and SOL muscles produced lower twitch forces than the wild-type muscles; in addition, fatiguing produced a downward shift in the force-frequency relationship in the mutant mice compared with the wild-type controls. Our results indicate that fatiguing affects the E-C components of the mutant EDL and SOL muscles, and the effect of fatigue in these mutant muscles could be primarily due to an alteration in the intracellular Ca homeostasis.

Animals↗

PubMed2000

A region of the sulfonylurea receptor critical for a modulation of ATP-sensitive K(+) channels by G-protein betagamma-subunits.

To determine the interaction site(s) of ATP-sensitive K(+) (K(ATP)) channels for G-proteins, sulfonylurea receptor (SUR2A or SUR1) and pore-forming (Kir6.2) subunits were reconstituted in the mammalian cell line, COS-7. Intracellular application of the G-protein betagamma2-subunits (G(betagamma)(2)) caused a reduction of ATP-induced inhibition of Kir6.2/SUR channel activities by lessening the ATP sensitivity of the channels. G(betagamma)(2) bound in vitro to both intracellular (loop-NBD) and C-terminal segments of SUR2A, each containing a nucleotide-binding domain (NBD). Furthermore, a single amino acid substitution in the loop-NBD of SUR (Arg656Ala in SUR2A or Arg665Ala in SUR1) abolished the G(betagamma)(2)-dependent alteration of the channel activities. These findings provide evidence that G(betagamma) modulates K(ATP) channels through a direct interaction with the loop-NBD of SUR.

ATP-Binding Cassette Transporters↗

PubMed2000

Distribution of nociceptin/orphanin FQ precursor protein and receptor in brain and spinal cord: a study using in situ hybridization and X-gal histochemistry in receptor-deficient mice.

Nociceptin/orphanin FQ (N/OFQ) is an opioid-like heptadecapeptide agonist for the opioid receptor homolog, N/OFQ receptor. To explore the precise distribution of the peptide-receptor system, the authors examined the brain and spinal cord from receptor-deficient mice bearing the targeted mutation (morc(m1)), a lacZ insertional mutation in the N/OFQ receptor gene. Precursor protein N/OFQ (preproN/OFQ) mRNA was detected by using in situ hybridization, and the N/OFQ receptor was detected by using X-gal histochemistry. The N/OFQ receptor reflected by lacZ expression was observed at high levels in the dentate gyrus, lateral septum, subparafascicular thalamic nucleus, medial preoptic area, median preoptic nucleus, ventromedial preoptic nucleus, anterior hypothalamic area, paraventricular hypothalamic nucleus, ventromedial hypothalamic nucleus, auditory brainstem nuclei, pontine dorsal tegmentum, and nucleus of the solitary tract. In situ detection of the N/OFQ receptor mRNA by digoxigenin-labeled riboprobes coupled with tyramide signal amplification in normal and wild-type mice resulted in the regional distribution paralleling the lacZ expression in these regions. PreproN/OFQ mRNA was expressed at high levels in the subparafascicular thalamic nucleus, central gray, central tegmental field, auditory brainstem nuclei, caudal spinal trigeminal nucleus, and spinal dorsal horn. Furthermore, variable levels of expression of the peptide and receptor were seen in distinct sites of the brain and spinal cord. These data indicate a correspondence of the peptide and the receptor in local distribution at limbic, hypothalamic, and brainstem sites. Together with concurrent physiologic and behavioral studies in mutant mice, the results suggest functional roles for the N/OFQ system, including the central regulation of learning and memory, hearing ability, water balance, food intake, and blood pressure.

Animals↗

PubMed2000

Characterization of human junctophilin subtype genes.

Junctophilin (JP) subtypes, namely JP-1, 2, and 3, have been currently identified in excitable cells and constitute a novel family of junctional membrane complex proteins. Our studies have suggested that JPs take part in the formation of junctional membrane complexes by spanning the membrane of the intracellular Ca(2+) store and interacting with the cell-surface membrane. In this report we describe the primary structures, genomic organization, and tissue distribution of human JP subtypes. By cloning and analyzing human genomic DNA segments, the protein-coding sequence interrupted with four introns was defined in each JP gene. The deduced human JP subtypes shared characteristic structural features with their rabbit and mouse counterparts. Genomic mapping demonstrated that JP genes do not cluster on the human genome. RNA blot hybridization indicated that tissue-specific expression patterns of JP genes in human are essentially the same as those in mouse; skeletal muscle contained both JP-1 and JP-2 mRNAs, the heart predominantly expressed JP-2 mRNA, and the brain specifically contained JP-3 mRNA. In the light of this, we propose intramolecular domains of JP subtypes based on the structural and functional characteristics.

Amino Acid Sequence↗

PubMed2000

Direct evidence for the localized single-triplet excitations and the dispersive multitriplet excitations in SrCu2(BO3)2.

We performed inelastic neutron scattering on the 2D Shastry-Sutherland system SrCu2(11BO3)2 with an exact dimer ground state. Three energy levels at around 3, 5, and 9 meV were observed at 1.7 K. The lowest excitation at 3.0 meV is almost dispersionless with a bandwidth of 0.2 meV at most, showing a significant constraint on a single-triplet hopping owing to the orthogonality of the neighboring dimers. In contrast, the correlated two-triplet excitations at 5 meV exhibit a more dispersive behavior.

Journal Article↗

PubMed2000

Trophic interactions between brain-derived neurotrophic factor and s100beta on cultured serotonergic neurons.

Brain-derived neurotrophic factor (BDNF) and S100beta stimulate serotonergic neurons in fetal rat raphe primary cultures grown under serum-free conditions. BDNF (50 ng/ml) treatment for 3 h enhanced S100beta immunoreactivity in both raphe and hippocampal glial cells. Combined treatment with BDNF and S100beta for 3 days increased the soma area of 5-HT neurons, but not the neurite length. Our results suggest that BDNF and S100beta, which regulate different signal transduction cascades, interact to exert complimentary effects on neuronal maturation by acting sequentially, not concurrently.

Animals↗

PubMed2000

Congenital malformations and childhood cancer.

BACKGROUND: We investigated the epidemiology of congenital malformations and childhood cancer. PROCEDURE: By employing the cases of the Registry of Childhood Malignancies in Hokkaido Prefecture, Japan, from 1969 to 1996, the numbers of malignancies in cases (diagnosis at 0-14 years of age) with Down syndrome (DS), mental retardation (MR) excluding DS, luxatio coxae congenita (LCC), congenital heart disease (CHD) excluding DS, undescended testicle (UT), and cleft palate-lip (CPL) were calculated. Using the percentages of malignancies in the 2,349 cases without malformation, expected numbers of malignancies in the cases with the malformations were calculated. The observed numbers were statistically compared to expected ones. RESULTS: In the DS cases, leukemia developed with a significantly high frequency, but no UT cases developed leukemia. No brain tumor was preceded by DS. This could not be explained only by early death from coexisting diseases such as CHD, insofar as the CHD cases without DS developed brain tumors more frequently than expected. The ratio of acute lymphoblastic leukemia (ALL) to acute nonlymphoblastic leukemia (ANLL) was different among the malformations. The MR cases developed ANLL more frequently than expected, whereas the CPL cases developed ALL more frequently. The distribution of the age at diagnosis for Wilms' tumor was different according to the underlying malformation. CONCLUSIONS: Malformations might have some factors that inhibit or delay as well as promote the development of certain malignancies.

Adolescent↗

PubMed2000

Effects of near-infrared low-level laser irradiation on microcirculation.

BACKGROUND AND OBJECTIVE: Recently, there has been an increase in the clinical application of low-level laser irradiation (LLLI) in various fields. The present study was conducted to explore the effects of LLLI on microcirculation. STUDY DESIGN/MATERIAL AND METHODS: We investigated the effects of LLLI on rat mesenteric microcirculation in vivo, and on cytosolic calcium concentration ([Ca2+]i) in rat vascular smooth muscle cells (VSMCs) in vitro. RESULTS: LLLI caused potent dilation in the laser-irradiated arteriole, which led to marked increases in the arteriolar blood flow. The changes were partly attenuated in the initial phase by the superfusion of 15 microM L-NAME, but they were not affected by local denervation. Furthermore, LLLI caused a power-dependent decrease in [Ca2+]i in VSMCs. CONCLUSION: The circulatory changes observed seemed to be mediated largely by LLLI-induced reduction of [Ca2+]i in VSMCs, in addition to the involvement of NO in the initial phase.

Animals↗

PubMed2000

A polymorphic marker in the leptin gene associated with Japanese morbid obesity.

The prevaleance of morbid obesity (body mass index of 35.0 or greater) is low in Japan (0.2-0.3%), and little systematic investigation of its cause in this population has been carried out. Leptin plays a central role in regulation of body weight; mice deficient in leptin develop marked obesity. We sought mutations in the leptin gene in 53 morbidly obese Japanese (maximum body mass index 35-60) including 46 with type 2 diabetes. Direct DNA sequencing was performed following polymerase chain reaction amplification. Apart from a silent mutation at codon 25 (CAA/CAG, glutamine) detected in eight subjects, no mutations were detected. We found a significantly higher prevalence of the variant leptin 25CAG allele among the 53 obese subjects (0.085) studied than in 132 nonobese control subjects (0.011, P<0.001). In Japanese populations mutations in the protein coding sequence of the leptin gene are unlikely to be a major cause of morbid obesity. However, the leptin 25CAG allele may be linked to morbid obesity in this population. Specifically, genetic variation located near the leptin gene may be involved in pathogenesis. The leptin polymorphism 25CAG appears to be a new genetic marker for obesity susceptibility, at least in Japanese.

Body Mass Index↗

PubMed2000

Molecular cloning of cDNA encoding a drosophila ryanodine receptor and functional studies of the carboxyl-terminal calcium release channel.

Ryanodine is a plant alkaloid that was originally used as an insecticide. To study the function and regulation of the ryanodine receptor (RyR) from insect cells, we have cloned the entire cDNA sequence of RyR from the fruit fly Drosophila melanogaster. The primary sequence of the Drosophila RyR contains 5134 amino acids, which shares approximately 45% identity with RyRs from mammalian cells, with a large cytoplasmic domain at the amino-terminal end and a small transmembrane domain at the carboxyl-terminal end. To characterize the Ca(2+) release channel activity of the cloned Drosophila RyR, we expressed both full-length and a deletion mutant of Drosophila RyR lacking amino acids 277-3650 (Drosophila RyR-C) in Chinese hamster ovary cells. For subcellular localization of the expressed Drosophila RyR and Drosophila RyR-C proteins, green fluorescent protein (GFP)-Drosophila RyR and GFP-Drosophila RyR-C fusion constructs were generated. Confocal microscopic imaging identified GFP-Drosophila RyR and GFP-Drosophila RyR-C on the endoplasmic reticulum membranes of transfected cells. Upon reconstitution into the lipid bilayer membrane, Drosophila RyR-C formed a large conductance cation-selective channel, which was sensitive to modulation by ryanodine. Opening of the Drosophila RyR-C channel required the presence of microM concentration of Ca(2+) in the cytosolic solution, but the channel was insensitive to inhibition by Ca(2+) at concentrations as high as 20 mM. Our data are consistent with our previous observation with the mammalian RyR that the conduction pore of the calcium release channel resides within the carboxyl-terminal end of the protein and further demonstrate that structural and functional features are essentially shared by mammalian and insect RyRs.

Amino Acid Sequence↗

PubMed2000

In vitro and in vivo immunocytochemistry for the distribution of mineralocorticoid receptor with the use of specific antibody.

To examine the distribution of mineralocorticoid receptor (MR) and the interactions with glucocorticoid receptor (GR) in the brain, we raised a polyclonal antibody against the transcriptional modulation domain of rat MR using the GST-fusion system. Immunoblotting analysis revealed that this antibody recognized a band with the molecular mass of MR in MR-transfected COS-1 cells and in a homogenate of rat hippocampus, and showed no cross-reactivity with GR. In vitro immunocytochemistry of both primary cultured hippocampal neurons and MR-transfected cells revealed immunoreactivity detected by this antibody in both the cytoplasm and nucleus in the absence of aldosterone (ALD), a specific agonist of MR. After 1 h of treatment with 10(-7) M ALD, the MR-immunoreactivity was accumulated in the nuclear region. In the case of GR-transfected cells, our anti-MR antibody either detected no immunopositive cells in the presence or absence of GR agonist. In our in vivo study, MR-immunoreactivity was observed in the rat hippocampus, where cell nuclei showed immunopositive reactions. These results suggest that our antibody against rat MR shows high specificity for the receptor both in liganded and unliganded forms, with no cross-reactivity to GR, and will be useful for cell biological and neuroanatomical investigations of MR.

Aldosterone↗

PubMed2000

Role of nociceptin systems in learning and memory.

This article summarizes our recent finding that the nociceptin system is involved in the regulation of learning and memory. The nociceptin-knockout mice show greater learning ability in the water maze task, an enhanced latent learning in the water finding task, better memory in the passive avoidance task, and further, larger long-term potentiation in the hippocampal CA1 region than wild-type mice. Nociceptin itself induces an impairment of passive avoidance task in wild-type mice, which is reversed by naloxone benzoylhydrazone (NalBzoH). Thus, the nociceptin system seems to play negative roles in learning and memory, and NalBzoH may act as a potent antagonist for the nociceptin receptor.

Animals↗

PubMed2000

Junctophilins: a novel family of junctional membrane complex proteins.

Junctional complexes between the plasma membrane (PM) and endoplasmic/sarcoplasmic reticulum (ER/ SR) are a common feature of all excitable cell types and mediate cross-talk between cell surface and intracellular ion channels. We have identified the junctophilins (JPs), a novel conserved family of proteins that are components of the junctional complexes. JPs are composed of a carboxy-terminal hydrophobic segment spanning the ER/SR membrane and a remaining cytoplasmic domain that shows specific affinity for the PM. In mouse, there are at least three JP subtypes: JP-1, -2, and -3. JP-2 is abundantly expressed in the heart, and mutant mice lacking JP-2 exhibited embryonic lethality. Cardiac myocytes from the mutant mice showed deficiency of the junctional membrane complexes and abnormal Ca2+ transients. Our results suggest that JPs are important components of junctional membrane complexes.

Amino Acid Motifs↗

PubMed2000

Early normal menstrual cycle pattern and the development of uterine leiomyomas.

It is unclear if women who develop uterine leiomyomas have had menstrual regularity or irregularity in their reproductive life. This case-control study examines the recalled menstrual cycle patterns throughout the reproductive life among women requiring hysterectomy for myomas. One hundred twenty-two women with myomas and 244 age-matched healthy controls without myomas were enrolled in Japan. The incidence of normal menstrual cycle pattern in their teens among patients with myoma was significantly higher than that among healthy controls (p < 0.01). The subset results for parous women were the same as those for all subjects. The size of the uterus in myoma patients with teenage menstrual regularity was not larger than in those with teenage menstrual irregularity. Women who developed myomas later in life tended to have early normal menstrual cycle pattern. Early menstrual regularity may enhance leiomyoma growth in early reproductive life.

Adult↗

PubMed2000

Constituents of a fern, Diplazium subsinuatum. III. Four new hopane-triterpene lactone glycosides.

From a whole plant of a fern, Diplazium subsinuatum, three new hopane-triterpene lactone glycosides, diplaziosides V-VII (1-3), were isolated, together with a new monoacetyl derivative (4) of diplazioside VII (3). Compounds 1-3 were defined as the respective 3-O-[beta-D-glucopyranosyl-(1-->2)]-beta-D-glucopyranosides of 3beta,24-dihydroxyhopan-28,22-olide (1), of 3beta,17,24-trihydroxyhopan-28,22-olide (2), and of (22R)-3beta,24,30-trihydroxyhopan-28,22-olide (3), and 4 as the 6"-O-acetate of 3, respectively, on the basis of spectral evidence. 1-3 are new in their glycoside structures but also in their triterpene structures. Furthermore, in compounds 1-4, the coupling between the 24-hydroxy proton and one of the 24-methylene protons showed a very large J-value (11.4 Hz); based on this 1H-NMR evidence, etc., preferred conformations of the 24-hydroxymethylene groups in 1-4 are also inferred. In a similar manner, 1H-NMR coupling patterns of the 24-hydroxymethylene in the monoglucoside (2a) and aglycone (2b), derived from 2, are also reported and discussed here.

Glycosides↗

PubMed2000

Factors influencing the visual acuity of primary school pupils.

To investigate the factors influencing the visual acuity of primary school pupils, an epidemiological study of 480 pupils in the 6th grade (11-12 years of age) was conducted in 8 primary schools in Sapporo City, Japan. Questionnaires were used to inquire into their current and past visual acuity and related factors. Dividing the subjects into those whose visual acuity of both eyes was 0.7 or more and those whose visual acuity of at least one eye was less than 0.7, odds ratios of various factors were calculated. Lifestyle and dietary factors showed no significant odds ratios. Parental myopia and age of parents at the birth of subjects showed significant odds ratios. Visual acuity of the pupils whose parents were myopic or older than 30 years when they were born tended to have worse visual acuity as they got older. Till the ages 11 or 12, hereditary factors seem much more contributory to visual acuity than environmental ones.

Chi-Square Distribution↗

PubMed2000

Relationship between suicide and holidays.

Employing the vital statistics from January 1, 1979 to December 31, 1994 in Hokkaido Prefecture, Japan, we investigated the relationship between suicide (ICD 9 code, El15) and days of the week. In one day 1.97 males and 0.98 females committed suicide on average. On Saturdays the number of suicides per day was the smallest. On Mondays it was the highest. When the days were classified into (1) a holiday, (2) the day before a holiday, (3) the day after a holiday (4) the day both before and after a holiday and (5) others, the rate of suicide was the lowest in the days before a holiday. In the days after a holiday, however, the rate was the highest. The same tendency was also noted when the subjects were classified into several subgroups from the viewpoint of ages, seasons or calendar years. The time relation of holidays seemed to have something to do with the intention to commit suicide.

Chi-Square Distribution↗

PubMed2000

Power Doppler sonography: evaluation of hepatocellular carcinoma after treatment with transarterial embolization or percutaneous ethanol injection therapy.

OBJECTIVE: The aim of this study was to compare tumor detectability by assessing the vascularity on power and color Doppler sonography and CT after transarterial embolization or percutaneous ethanol injection therapy or both in hepatocellular carcinoma. SUBJECTS AND METHODS: Forty-seven nodules of hepatocellular carcinoma (size, 28 +/- 7 mm [mean +/- standard deviation]; range, 20-40 mm) in 38 patients were treated with transarterial embolization (n = 6), percutaneous ethanol injection therapy (n = 23), and transarterial embolization plus percutaneous ethanol injection therapy (n = 9). Power Doppler sonography, color Doppler sonography, and CT were performed before and 2 weeks, 3 months, and 6 months after the treatments. The existence of hepatocellular carcinoma was confirmed by positive findings for color signals on both Doppler sonography techniques and for tumor stains on CT. All the tumors were determined to be malignant by microscopic examination of biopsy specimens. RESULTS: Before the treatments, power Doppler sonography (100%) and CT (100%) were significantly more effective than color Doppler sonography (61.7%) (p < 0.001, for both). Six months after the treatments, the sensitivity of power Doppler sonography (87.5%) was significantly better than that of color Doppler sonography (12.5%) but was not significant in comparison with CT (66.6%). However, power Doppler sonography detected color signals in two of three tumors in which iodized oil was accumulated and no tumor stain appeared on CT, and the two lesions detected with power Doppler sonography were carcinomas. CONCLUSION: Power Doppler sonography can be considered the most sensitive technique in assessing the viability of hepatocellular carcinoma treated with transarterial embolization or percutaneous ethanol injection therapy or both.

Adult↗