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Biomedical subjects

K Nakano

Publications and source records attributed to K Nakano.

At least 19 recordsLinked to original sources

Identification of NGF-response element in the rat neuropeptide Y gene and induction of the binding proteins.

Gene expression of the rat neuropeptide Y (NPY) increases by 100 times, as the PC12 cells differentiate into sympathetic neuron-like cells with NGF treatment and this increase is partly due to transcriptional activation of the NPY gene (Sabol and Higuchi, Mol. Endocrinol. 4, 384, 1990). To identify the NGF-response element, a transient expression assay was carried out by using the CAT reporter genes containing various lengths of the 5' upstream region of the NPY gene in the PC12 cells. The 48-base element (-80/-33 upstream of the Cap site) was identified as a NGF-response element (NGFRE). Gel shift assay indicated the existence of at least two DNA-binding proteins to NGFRE. The binding activity of the protein(s) (NDF1) to the upper region (-80/-63) was increased by 3-fold with NGF treatment for 24 h. These findings suggest that these nuclear proteins are involved in the enhanced transcription of the NPY gene by NGF.

Animals

Single retinal ganglion cells sending axon collaterals to the bilateral superior colliculi: a fluorescent retrograde double-labeling study in the Japanese monkey (Macaca fuscata).

Single retinal ganglion cells projecting bilaterally to the superior colliculi (SC) by way of axon collaterals were revealed in the Japanese monkey (Macaca fuscata). After injecting Fast blue into the SC on one side and Diamidino yellow into the SC on the opposite side, some retinal ganglion cells were double-labeled with both tracers. Most of them were large cells (more than 25 microns in diameter), and were localized in a narrow strip around the vertical meridian of the retina on each side. This retinal area roughly corresponds to the reported strip of nasotemporal overlap, where both crossed and uncrossed retinofugal projections arise.

Amidines

Non-dopaminergic neurons in the substantia nigra project to the reticular formation around the trigeminal motor nucleus in the rat.

The dorsolateral part of the substantia nigra (SN) of the rat was observed to send projection fibers to the reticular formation (RF) around the trigeminal motor nucleus (Vm), bilaterally with a clear-cut ipsilateral dominance, by the anterograde and retrograde tracing techniques with PHA-L and WGA-HRP. A combination of retrograde tracing and immunohistochemistry for tyrosine hydroxylase (TH) revealed that no SN neurons sending their axons to the RF around the Vm showed TH-like immunoreactivity.

Animals

Molecular cloning of dihydrolipoamide acetyltransferase of the rat pyruvate dehydrogenase complex: sequence comparison and evolutionary relationship to other dihydrolipoamide acyltransferases.

A complementary DNA (cDNA) clone of dihydrolipoamide acetyltransferase (E2) of the rat pyruvate dehydrogenase complex (PDC) was isolated from a lambda gt11 rat heart cDNA library. The amino acid sequence of a full mature protein of rat PDC-E2 was predicted by combination of the cDNA nucleotide sequence and the N-terminal amino acid sequence determined chemically. The amino acid sequence of rat PDC-E2 was well consistent with those of the E2 components of other alpha-ketoacid dehydrogenase complexes. These E2 components possess the sequence G-X-G-X-X-G, which is the consensus sequence for nucleotide binding sites of nucleotide binding proteins, in the E3 and/or E1 binding domains. The E2 components of the three alpha-ketoacid dehydrogenase complexes are suggested to be classified into three clusters separated during evolution.

Acetyltransferases

Nucleotide sequence of the genome of the bacteriophage alpha 3: interrelationship of the genome structure and the gene products with those of the phages, phi X174, G4 and phi K.

The complete nucleotide sequence of the genome of the circular single-stranded DNA (isometric) phage alpha 3 has been determined and compared with that of the related phages phi X174 and G4. The alpha 3 genome consists of 6087 nucleotides, which is 701 nucleotides longer than the nucleotide sequence of the phi X174 genome and 510 nucleotides more than that of the G4 genome. The results demonstrated that the three phage species have 11 homologous genes (A, A*, B, C, K, D, E, J, F, G and H), the order of which is fundamentally identical, suggesting that they have evolved from a common ancestor. The sequence of some genes and untranslated intergenic regions, however, differs significantly from phage to phage: for example, the degree of amino acid sequence homology of the gene product is averaged at 47.7% between alpha 3 and phi X174 and 46.9% between alpha 3 and G4, and alpha 3 has a remarkable longer intergenic region composed of 758 nucleotides between the genes H and A compared with the counterparts of phi X174 and G4. Meanwhile, in vivo experiments of genetic complementation showed that alpha 3 can use none of the gene products of phi X174 and G4, whereas the related phage phi K can rescue alpha 3 nonsense mutants of the genes B, C, D and J. These sequencing and in vivo rescue results indicated that alpha 3 is closely related to phi K, but distantly remote from phi X174 or G4, and supported an evolutional hypothesis which has been so far proposed that the isometric phages are classified into three main groups: the generic representatives are phi X174, G4 and alpha 3.

Amino Acid Sequence

Abnormally high activity of 3-hydroxyanthranilate 3,4-dioxygenase in brain of epilepsy-prone El mice.

Quinolinic acid (QUI), a structural analogue of neurotransmitters such as L-glutamate and L-aspartate, may act as an 'excitotoxin' when it is abundant in the brain. The compound has been causally related to various neurodegenerative disorders, including epilepsy. We tested the capacity of the brains of epilepsy-prone El mice to synthesize QUI. The activity of 3-hydroxyanthranilate 3,4-dioxygenase in the cerebral cortex of El mice was about 17 times that of ddY mice, the parent strain of El mice. The activity of this enzyme was undetectable in brains of BALB/cA mice and C3H/HeN mice. In El mice the sexes had comparable enzyme activity. The enzyme activity increased gradually as the animals aged. An injection of endotoxin caused a further increase in the enzyme activity. The enzyme activity in the spleen of El mice did not differ from that of ddY mice, and endotoxin did not affect the enzyme activity in the spleen. No strain-difference was observed in the activity of quinolinate phosphoribosyltransferase, a QUI-degrading enzyme, in the cerebral cortex. These results suggest that an increase in the synthesis of QUI in the brain is involved in the pathogenesis of epileptic seizures in El mice.

3-Hydroxyanthranilate 3,4-Dioxygenase

Descending projections from the subparafascicular thalamic nucleus to the lower brain stem in the rat.

In the course of our study on the neuronal connections of the subparafascicular nucleus (SPF) in the rat, descending projections from the SPF to the lower brain stem were examined by using the anterograde tracer PHA-L (Phaseolus vulgaris leukoagglutinin) and retrograde tracer WGA-HRP (horseradish peroxidase conjugated to wheat germ agglutinin). When PHA-L was injected into the magnocellular and/or parvicellular division of the SPF (SPFm and/or SPFp), presumed terminal labeling was seen, bilaterally with an ipsilateral dominance, in the mesencephalic and pontine central gray matter, peripheral shell regions of the inferior colliculus, cuneiform nucleus, and superior olivary complex (mainly in the superior paraolivary nucleus, and additionally in the nuclei of the trapezoid body). A few labeled axon terminals were also seen in the cochlear nuclei bilaterally with a contralateral dominance. In the second set of experiments, WGA-HRP was injected into the inferior colliculus, superior olivary complex, or cochlear nuclei. When WGA-HRP was injected into the peripheral shell regions of the inferior colliculus or the superior olivary complex, many labeled neuronal cell bodies were seen in the SPFm bilaterally with an ipsilateral dominance, and a moderate number of labeled neuronal cell bodies were observed in the SPFp (lateral SPF) bilaterally with an ipsilateral dominance. When WGA-HRP was injected into the cochlear nuclei, a moderate number of labeled neuronal cell bodies were observed in the SPFm and SPFp bilaterally with a contralateral dominance. The results indicate that the SPFm and SPFp (lateral SPF) of the rat send a considerable number of projection fibers to the lower brain stem. The target regions of these projection fibers include the auditory relay nuclei, such as the inferior colliculus, superior olivary complex, and cochlear nuclei.

Animals

Human gallbladder bile becomes lithogenic during short-term intravenous hyperalimentation.

Biliary sludge formation is reported to be one of the complications of intravenous hyperalimentation (IVH); however, the change in human biliary lithogenicity during IVH treatment has been little studied. To clarify the pathogenesis of IVH-induced biliary sludge, we determined biliary lipid composition, vesicular cholesterol concentration, and nucleation time using gallbladder bile samples collected from three groups: The IVH group comprised 9 patients who received IVH with fasting for a period of 2-8 days prior to surgery for gastrointestinal diseases. The control group comprised 10 patients operated after overnight fasting for gastrointestinal diseases. The cholesterol gallstone group comprised 14 patients surgically treated for cholesterol gallstone disease after overnight fasting. The nucleation time in the IVH group was significantly shorter (7.8 +/- 5.3 days, mean +/- SD) than that in the control group (17.3 +/- 5.5 days), while it did not reach the value in the cholesterol gallstone group (3.1 +/- 3.3 days). The cholesterol saturation index in the IVH group (1.01 +/- 0.27) was higher but not significantly different compared with the control group (0.80 +/- 0.21). The concentrations of biliary lipids and individual bile acid were similar in the IVH and control groups. The vesicular cholesterol concentration in the IVH group (3.0 +/- 2.1 mM) was significantly higher than that in the control group (0.9 +/- 1.0 mM). In conclusion, IVH with fasting causes a rapid cholesterol nucleation time and a higher vesicular cholesterol concentration, thereby inducing an initial stage of gallbladder sludge formation.

Aged

Effects of a new aldose reductase inhibitor, (2S, 4S)-6-fluoro-2',5'-dioxospiro[chroman-4,4'-imidazolidine]-2-ca rboxamid e (SNK-860), on the slowing of motor nerve conduction velocity and metabolic abnormalities in the peripheral nerve in acute streptozotocin-induced diabetic rats.

The effects of a new aldose reductase inhibitor (ARI), (2S,4S)-6-fluoro-2',5'-dioxospiro[chroman-4,4'-imidazolidine]-2-ca rboxamide (SNK-860), on the slowing of motor nerve conduction velocity (MNCV) and metabolic abnormalities in sciatic nerve were investigated in acute streptozotocin (STZ)-induced diabetic rats. MNCV in the diabetic rats was significantly slower 2 weeks after STZ injection. In the following 2 weeks, treatment with SNK-860 improved MNCV in a dose-dependent manner. The efficacy of 1 mg/kg SNK-860 was equipotent to that of 20 mg/kg sorbinil. Four weeks after STZ injection, increases in sorbitol levels, decreases in myo-inositol levels, and reductions in Na+, K(+)-adenosine triphosphatase (ATPase) activity were observed in sciatic nerves of diabetic rats. Administration of SNK-860 for 14 days beginning 2 weeks after the induction of diabetes inhibited these metabolic abnormalities in a dose-dependent manner. SNK-860 restored all of these parameters to normal levels at a dose of 2 mg/kg. In addition, close correlations were observed between MNCV and sorbitol levels (r = -.95) and between MNCV and myo-inositol levels (r = .93) in the sciatic nerve; a close correlation was also observed between sorbitol and myo-inositol levels in the sciatic nerve (r = -.86). Therefore, it is suggested that the effect of SNK-860 on the slowing of MNCV results from normalizing the above-mentioned metabolic abnormalities in the sciatic nerve of diabetics. Thus, SNK-860 may be useful in the treatment of diabetic neuropathy.

Aldehyde Reductase

Cranial fasciitis of childhood: a case report.

Cranial fasciitis of childhood is very rare, only 17 cases having been reported in the literature. We report an additional case of this rare disease. The patient was a 5-year-old boy who complained of left exophthalmos and double vision. Computed tomography (CT) and magnetic resonance imaging (MRI) revealed a large epidural mass in the left frontal region that had invaded into the underlying anterior skull base. The tumor showed homogeneous, low density with nonhomogeneous contrast enhancement on the CT scans, and low intensity on the T1-weighted and high intensity on the T2-weighted MRI images. A whitish-pink, elastic, hard tumor was revealed in the epidural space in the left anterior cranial fossa, which was totally excised with curettage of the affected anterior skull base. The origin of the tumor was suspected to be the fibrous connective tissue of the sphenofrontal suture. The histological diagnosis was that of cranial fasciitis. There was no evidence of recurrence 1 year postoperatively.

Child, Preschool

An autoradiographic study of cortical projections from motor thalamic nuclei in the macaque monkey.

The special areal and laminar distributions of cortical afferent connections from various thalamic nuclei in the monkey (Macaca fuscata) were studied by using the anterograde axonal transport technique of autoradiography. The following findings were obtained. The superficial thalamocortical (T-C) projections, terminating in the (superficial half of) cortical layer I, arise mainly from the nucleus ventralis anterior, pars principalis (VApc) and nucleus ventralis lateralis, pars oralis (VLo), and possibly from the nucleus ventralis lateralis, pars medialis (VLm) and nucleus ventralis anterior, pars magnocellularis (VAmc). The VApc gives rise to the superficial T-C and deep T-C projections onto the postarcuate premotor area around the arcuate genu and spur, and onto the dorsomedial part of the caudal premotor area as well as the supplementary motor area (SMA). The VApc also gives rise to only deep T-C projections onto the remaining premotor area and onto the rostral bank of the arcuate sulcus as well as the ventral bank of the cingulate sulcus at the level of the premotor area. The VLo gives rise to the superficial T-C projections onto the ventrolateral part of the motor area (mainly to the forelimb motor area) and onto the dorsomedial part to the mesial cortex at the rostral level of the motor area. The VAmc gives rise to the superficial T-C projections onto the banks of the arcuate genu and adjacent region of area 8. Area X, the nucleus ventralis posterolateralis, pars oralis (VPLo), nucleus ventralis posterolateralis, pars caudalis (VPLc), nucleus ventralis posteromedialis (VPM) and possibly the nucleus ventralis lateralis, pars caudalis (VLc) send only deep T-C projections. The dorsal and medial parts of the VLc project onto the premotor area, the rostral part of the motor area and the SMA, and also the ventral bank of the cingulate sulcus. Area X projects onto the premotor area, the SMA, and the caudal part of area 8. The thalamic relay nuclei projecting onto the frontal association cortex were found to be the VAmc, medial VLc and area X.

Afferent Pathways

Determination of monosaccharides and sugar alcohols in tissues from diabetic rats by high-performance liquid chromatography with pulsed amperometric detection.

A sensitive and simple high-performance liquid chromatographic method has been developed to determine the concentration of monosaccharides and sugar alcohols in animal tissues. Five neutral monosaccharides (D-glucose, D-galactose, D-mannose, D-fructose, and D-ribose) and three neutral sugar alcohols (myo-inositol, glycerol, and D-sorbitol) predominate in the renal cortices and sciatic nerves of rats. These monosaccharides and sugar alcohols were extracted with distilled water, purified by deproteinization with ethanol, a Sep-Pak C18 cartridge, and columns of Dowex 50W-X8 and Amberlite CG-400, then separated on Ca2+ and Pb2+ cation-exchange columns, eluted with deionized distilled water at 80 degrees C, and detected using integrated pulsed amperometry. About 10 pmol of each sugar was detectable with a signal-to-noise ratio of 10:1. D-Glucose, D-fructose, D-sorbitol, and D-mannose were higher in both the renal and sciatic tissues of diabetic rats than in those of normal animals. D-Ribose and glycerol were higher in the renal cortex of diabetic animals.

Animals

Heavy ion-induced chromosome breakage studied by premature chromosome condensation (PCC) in Syrian hamster embryo cells.

We have studied induction and repair of chromosome damage induced by high linear energy transfer (LET) heavy ions in G1/G0 interphase Syrian golden hamster embryo (SHE) cells as revealed by the premature chromosome condensation (PCC) technique. The number of chromosome breaks in condensed chromosomes induced by high LET heavy ions was higher than those induced by 137Cs gamma-rays. Compared with 137Cs gamma rays, the relative biological effectiveness (RBE) for PCC breaks was 1.5 for 35 keV/microns 4He ions, 1.9 for 77keV/microns 4He ions, and 2.5 for 530keV/microns 14N ions. Although 95% of the PCC breaks induced by gamma-rays rejoined during 8 h post-irradiation incubation, only 35-45% of fragments induced by high LET radiations rejoined in the same time. These results suggest that there is a difference, spatial or qualitative, in the initial chromosome damage produced by high LET radiations and low LET radiations.

Animals

Effect of multiple irradiation with low doses of gamma-rays on morphological transformation and growth ability of human embryo cells in vitro.

We have measured expression of transformed phenotypes in human embryo (HE) cells repeatedly irradiated with a dose of 7.5 cGy per week throughout the life span of these cells in vitro. Irradiation was repeated until the cells had accumulated 195 cGy at which time the cells had reached the equivalent of their 26th passage and samples of cells at several passages were assayed for cell survival by colony formation, for mutation at hypoxanthine guanine phosphoribosyl transferase (HGPRT) locus and for transformation by focus formation. The lifespan (mean population doublings) of multiple irradiated cultures with a total dose of 97.5 cGy was slightly, but significantly, prolonged over that of controls. For example, if cells had accumulated 195 cGy, the maximum number of cell division of HE cells in vitro extended to 130-160% of non-irradiated control. Although transformed foci were not observed with cells until cells had accumulated 97.5 cGy, it increased with increasing accumulated dose. No cells, however, showed unlimited life span in vitro and also expressed tumorigenicity.

Cell Survival

Spinous process-splitting laminoplasty using hydroxyapatite spinous process spacer.

Between February 1986 and August 1989, 45 patients with cervical myelopathy were treated by spinous process-splitting laminoplasty. Hydroxyapatite intraspinous spacers were used to maintain the enlargement of the cervical spinal canal. The shape of this spacer is trapezoidal. After sagittal splitting of the spinous process, the spacer was inserted between the two halves and affixed with the wire. Histologic study showed there was good fusion between the spacer and bone. In all cases, good enlargement of the cervical spinal canal was achieved. Spacer displacement, wire breakage, and postoperative infection were not seen. There was no postoperative neurologic deterioration. Computed tomography showed that the width of the cervical spinal canal was maintained.

Animals