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S Miyata

Publications and source records attributed to S Miyata.

At least 163 records · Page 9Linked to original sources

Expression of c-fos immunoreactivity in the hypothalamic magnocellular neurons during chronic osmotic stimulations.

The immunoreactivity of c-fos protein was transiently detected in magnocellular neurons of the supraoptic nucleus (SON) and paraventricular nucleus (PVN) of the rat hypothalamus after intraperitoneal injection of hypertonic NaCl solution. In contrast, c-fos-positive magnocellular neurons were persistently observed in the SON and PVN of the rats which were chronically stimulated by the drinking of hypertonic NaCl solution instead of water or by water deprivation. c-fos immunoreactivity was eliminated in the animals which were allowed to drink tap water for 24 h following the chronic osmotic stimulations. These results suggest that persistent expression of c-fos protein is closely associated with the neural plasticity of the hypothalamic magnocellular neuron.

Administration, Oral↗

Advanced Maillard reaction end products are associated with Alzheimer disease pathology.

During aging long-lived proteins accumulate specific post-translational modifications. One family of modifications, termed Maillard reaction products, are initiated by the condensation between amino groups of proteins and reducing sugars. Protein modification by the Maillard reaction is associated with crosslink formation, decreased protein solubility, and increased protease resistance. Here, we present evidence that the characteristic pathological structures associated with Alzheimer disease contain modifications typical of advanced Maillard reaction end products. Specifically, antibodies against two Maillard end products, pyrraline and pentosidine, immunocytochemically label neurofibrillary tangles and senile plaques in brain tissue from patients with Alzheimer disease. In contrast, little or no staining is observed in apparently healthy neurons of the same brain. The Maillard-reaction-related modifications described herein could account for the biochemical and insolubility properties of the lesions of Alzheimer disease through the formation of protein crosslinks.

Alzheimer Disease↗

Structural dynamics of neural plasticity in the supraoptic nucleus of the rat hypothalamus during dehydration and rehydration.

It has been known that magnocellular neuroendocrine cells (MNCs) of mammalian hypothalamus show structural plasticity in response to chronic osmotic stimulation. In this study, we investigated the relationships among plasma osmolarity and several structural changes such as alterations of soma size, juxtaposition, and synapses of the supraoptic nucleus (SON) in the rat hypothalamus during dehydration and rehydration. Male rats were osmotically stimulated by supplying with 2% NaCl solution instead of tap water for 10 days, and then they were rehydrated with tap water. Plasma osmolarity was gradually elevated with progress of salt loading and returned to control level on the seventh day of rehydration. Both the percentage of membrane contact (juxtaposition) and the soma size of MNCs were increased in response to the rise of plasma osmolarity, and decreased to control level on the seventh day of rehydration. The number of synapses including both single synapses and multiple synapses per 100 microns soma membrane was lower than control on the fifth day of dehydration, but it was not different from controls on the tenth day of dehydration, and on the seventh and fourteenth day of rehydration. The total number of synapses per 100 microns soma membrane, the synaptic density, was maintained relatively constant, although soma size was progressively changed during dehydration or rehydration. This synaptic reorganization seems to be mainly regulated by synaptic sprouting during dehydration and by degradation of synapses during rehydration.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Not only osmotic stress but also repeated restraint stress causes structural plasticity in the supraoptic nucleus of the rat hypothalamus.

Magnocellular neuroendocrine cells (MNCs) in the supraoptic nucleus (SON) of the hypothalamus have been known to undergo dramatic structural changes during chronic stimulation such as osmotic stress. In the present study, we examined whether this anatomical neural plasticity is associated with an another stress, such as restraint. Rats were chronically stimulated by either dehydration with 2% saline drinking instead of water or daily restraint with leg immobilization. The structural reorganizations of MNCs in the SON were analyzed morphometrically with use of light and electron microscopy. The results were compared to control animals that had free access to water and food. In restraint rats, the soma size of both oxytocin (OXT) and arginine vasopressin (AVP) neurons was enlarged, and the percent of soma-somatic/dendritic membrane contact (juxtaposition) was elevated significantly. The number of total synapses per 100 microns soma membrane was not changed, although soma profiles were enlarged. However, the number of multiple synapses (which contacted with more than one postsynaptic element) per 100 microns soma membrane was significantly increased. Similar structural changes were observed in dehydrated animals, and the degree of morphological changes was stronger than the restraint one. These findings indicate that NMCs undergo structural plasticity during not only osmotic stress but also restraint stress.

Animals↗

A spore-lytic enzyme released from Bacillus cereus spores during germination.

The exudate of fully germinated spores of Bacillus cereus IFO 13597 in 0.25 M sodium phosphate buffer, pH 7.0, was found to contain a spore-lytic enzyme. This enzyme was found to cause loss of absorbance in coat-stripped spore suspensions and phase-darkening of the spores but had minimal activity on isolated peptidoglycan substrates. The enzyme was purified in an active form and identified as a 24 kDa protein which is either an amidase or a peptidase. The amino-terminal 19 residues had the following sequence: FSNQVIQRGASGEKVIELQ. The spore-lytic enzyme retained its activity in a medium of a relatively high ionic strength containing a non-ionic surfactant such as nonaethyleneglycol n-dodecyl ether. This activity was optimum at a salt concentration of about 30 mM in assay buffer at neutral pH. In contrast to the enzyme in a spore-bound form, the enzyme in solution was shown to be heat-sensitive and was readily inactivated by thiol reagents.

Amino Acid Sequence↗

Hypertrophic growth of cultured neonatal rat heart cells mediated by type 1 angiotensin II receptor.

Primary cultures of neonatal cardiac myocytes were used to determine the identity of second messengers that are involved in angiotensin II (ANG II) receptor-mediated effects on cardiac hypertrophy and the type of ANG II receptor that is involved in ANG II-induced cell growth. Treatment of myocytes with ANG II significantly increased the protein-to-DNA and the RNA-to-DNA ratios. ANG II accelerated rates of protein synthesis by 24.9%. Intracellular free calcium was transiently increased after ANG II exposure. The activity of protein kinase C in particulate fractions was transiently increased after exposure to ANG II but returned to control level. The activity of protein kinase C in the cytosol was significantly decreased at all times after exposure to ANG II. After ANG II treatment, the content of c-Fos mRNA was increased. The stimulatory effects of ANG II on these parameters were inhibited by the type 1 angiotensin II receptor (AT1) antagonist, losartan. These studies demonstrate that ANG II-induced hypertrophic growth is, at least in part, mediated through AT1 receptors.

Angiotensin II↗

[Bone union after lumbar postero-lateral fusion: comparative evaluation by x-ray and CT].

As yet there has been no report on a uniform and precise method for evaluating bone union after lumbar postero-lateral fusion (PLF). Here we report and compare the roentgenogram (X-P) and computerized tomography (CT) methods to evaluate the union and non-union of PLF. PLF without internal fixation was undertaken in 252 patients between 1978 and 1991. Of these, 125 patients underwent both X-P and CT scanning and were followed up for more than one year (mean 4 years and 2 months, range 1 to 14 years). In the cases of complete union, X-P demonstrated that the solid fusion mass showed a continuous trabecular pattern along the grafted segment, while CT demonstrated that the solid fusion mass appeared as a long bone with the mass incorporated into the transverse process and the superior facet. In such cases, there was no micromotion on stress roentgenograms. In the cases of incomplete union, the status of the non-union was classified into three types as; Type I showing atrophy and resorption of the grafted bone, Type II showing a lack in trabecular continuity of the fusion mass, and Type III showing a gap either cranially or caudally between the fusion mass and lumbar matrix (which includes the transverse process and superior facet). X-P could easily diagnose Type I and Type II, but could not clearly diagnose Type III. CT, however, could easily diagnose Type III. Therefore, the use of only X-P might lead to a misdiagnosis in cases of Type III. The application of CT was more uniform and precise method to evaluate the state of PLF in the lumbosacral spine.

Adolescent↗

Investigation of type IIC von Willebrand disease.

Type II von Willebrand disease (vWD) is characterized by qualitative abnormality of von Willebrand factor (vWF). It is characterized by the absence of the largest, or the largest and intermediate-size plasma vWF multimers. Type IIC vWD is a very rare variant subtype. We report the tenth case of type IIC vWD who is a 32-year-old Japanese with a history of lifelong mild bleeding problems. The multimeric analysis of vWF of the patient's plasma revealed that the multimers were composed of the increased smallest multimer and progressively decreased larger multimers, with each multimer replaced by a single band instead of a normal triplet structure. The family study highly suggested that the propositus is the first case of homozygote for type IIC vWD gene, although previous studies have suggested that type IIC vWD is due to double heterozygosity of two different mutant genes. The patient's von Willebrand factor antigen showed a slight response to 1-deamino-8-D-arginine vasopressin (DDAVP), whereas his ristocetin cofactor showed no response. The patient responded very well to an intermediate-purity factor VIII concentrate, and then underwent cholecystectomy without bleeding problems.

Adult↗

Cleavage arrest in Xenopus embryos injected with a human c-myc gene.

The c-myc protein has been implicated in cell proliferation and differentiation of various cell lines, but its function in early embryos of Xenopus is obscure. We introduced the c-myc gene into embryos using a pcDL-SR alpha plasmid with an efficient promoter of the gene. It was found that fertilized eggs injected with the c-myc gene were arrested at the 4-cell stage within 1 h of the injection, when the plasmid that carried c-myc gene but not the plasmid that carried defective c-myc gene was injected. These results also suggest that Xenopus embryos already have the ability to transcribe an exogenously injected c-myc gene at the 4-cell stage.

Animals↗

Nutritional status of antioxidant vitamins (A, E, and beta-carotene) in elderly Japanese.

The nutritional status with respect to vitamins A and E, and beta-carotene was examined in elderly Japanese subjects in two institutions at Osaka and Kyoto. Only the plasma vitamin E level has been determined in the majority of previous investigations. In this study, vitamin E levels were determined in red blood cells (RBCs), platelets (PLT), mononuclear cells (MN), polymorphonuclear cells (PMN), and buccal mucosal cells (BMC), using HPLC with electrochemical detection. Alpha-tocopherol levels in plasma and RBCs did not differ between elderly and young adults, while those in PLT, MN, and BMC were lower in the elderly. Thus, the vitamin E status of elderly Japanese individuals appears to be inadequate of the cellular levels. The daily vitamin E intake of the elderly subjects was below the recommended dietary allowance for the Japanese population. Plasma levels of retinol and beta-carotene were also assessed. The vitamin A status did not differ between elderly and young adults on the basis of the levels of retinol and retinol-binding protein (RBP). The daily intake of retinol (as retinol equivalent) by the elderly subjects was more than 2,000 IU. With respect to beta-carotene, there was a large sex difference (female > male), which was more prominent in the young adults and became smaller in the elderly. This sex difference was partly attributable to a difference in plasma total lipids. No clear age-related trend was noted.

Adult↗

The interaction of botrocetin with normal or variant von Willebrand factor (types IIA and IIB) and its inhibition by monoclonal antibodies that block receptor binding.

We have recently shown the existence of two distinct forms of botrocetin (one-chain and two-chain), and demonstrated that the two-chain species is approximately 30 times more active than the one-chain in promoting von Willebrand factor (vWF) binding to platelet glycoprotein (GP) Ib. The N-terminal sequence of two-chain botrocetin is highly homologous to sea-urchin Echinoidin and other Ca(2+)-dependent lectins (Fujimura et al., Biochemistry 1991; 30: 1957-64). Present data indicate that purified two-chain botrocetin binds to vWF from plasmas of patients with type IIA or IIB von Willebrand disease and its interaction is indistinguishable from that with vWF from normal individuals. However, an "activated complex" formed between botrocetin and IIB vWF expresses an enhanced biological activity for binding to GP Ib whereas the complex with IIA vWF has a decreased binding activity. Among several anti-vWF monoclonal antibodies (MoAbs) which inhibit ristocetin-induced platelet aggregation and/or vWF binding to GP Ib, only two MoAbs (NMC-4 and RFF-VIII RAG:1) abolished direct binding between purified botrocetin and vWF. This suggests that they recognize an epitope(s) on the vWF molecule in close proximity to the botrocetin binding site.

Antibodies, Monoclonal↗

In vivo production of a stable single-stranded cDNA in Saccharomyces cerevisiae by means of a bacterial retron.

Gram-negative bacteria such as Myxococcus xanthus, Stigmatella aurantiaca, and Escherichia coli contain retroelements called retrons. Retrons consist of the msr-msd region and the gene for reverse transcriptase (RT), which are essential for the production of the branched RNA-linked ms-DNA (multicopy single-stranded DNA). In this study, we attempted to produce msDNA in the yeast Saccharomyces cerevisiae. Retron Ec67 from E. coli, which is responsible for the production of msDNA-Ec67, was cloned under the GAL10 promoter in a 2-microns-based plasmid. msDNA thus produced was detected by extending the 3' end of the msDNA by avian myeloblastosis virus RT. This yielded a main product of 117 nucleotides. Treatment of this product with RNase A resulted in a DNA of 105 nucleotides. These results are in good agreement with the structure of msDNA-Ec67. The production of msDNA-Ec67 was further confirmed by Southern blot hybridization. The msDNA production was dependent upon the bacterial RT gene in the clone and was increased severalfold when the RT gene of retron Ec67 was placed in front of the msr-msd region. The potential of msDNA as a eukaryotic vector producing a stable single-stranded DNA as well as RNA is discussed.

Base Sequence↗

Immunohistochemical detection of advanced glycosylation end products in diabetic tissues using monoclonal antibody to pyrraline.

Pyrraline is one of the major Maillard compounds resulting from the reaction of glucose with amino compounds at slightly acidic pH. For in vivo studies, monoclonal pyrraline antibodies were raised after immunization of Balb/c mice with keyhole limpet hemocyamin-caproyl pyrraline conjugate. Of 660 hybridoma clones from one donor, 260 produced an antibody to the free hapten, two of which named Pyr-A and Pyr-B also cross-reacted with L-lysyl pyrraline. Using Pyr-B antibody and an ELISA, a gradual increase in pyrraline immunoreactivity was observed in serum albumin incubated with glucose or 3-deoxyglucosone. Plasma pyrraline levels increased fourfold (P less than 0.001) in Sprague-Dawley rats upon induction of diabetes with streptozotocin and were twofold increased in randomly selected plasmas from diabetic humans. Highly specific pyrraline immunoreactivity was detected in sclerosed glomeruli from diabetic and old normal kidneys as well as in renal arteries with arteriolosclerosis and in perivascular and peritubular sclerosed extracellular matrix and basement membranes. The preferential localization of pyrraline immunoreactivity in the extracellular matrix strengthens the notion that the advanced glycosylation reaction may contribute to decreased turnover and thickening of the extracellular matrix in diabetes and aging.

Animals↗

Maillard reaction-mediated molecular damage to extracellular matrix and other tissue proteins in diabetes, aging, and uremia.

Recent progress in structure elucidation of products of the advanced Maillard reaction now allows probing specifically for the role of this reaction in the pathogenesis of age- and diabetes-related complications. Pyrraline is a glucose-derived advanced glycation end product against which polyclonal and monoclonal antibodies have been raised. Immunohistochemical localization studies revealed that pyrraline is found predominantly in the sclerosed extracellular matrix of glomerular and arteriolar renal tissues from both diabetic and aged nondiabetic individuals. Pentosidine and carboxymethyllysine are Maillard end products derived from both glucose and ascorbate. In addition, pentosidine can be formed from several other sugars under oxidative conditions, and in vitro studies suggest that a common intermediate involving a pentose is a necessary precursor molecule. The highest levels of these advanced Maillard products are generally found in the extracellular matrix, but these products are also present in lens proteins and in proteins with a fast turnover such as plasma proteins. Diabetes, and especially uremia, greatly catalyzes pentosidine formation. Both conditions are characterized by accelerated cataractogenesis, atherosclerosis, and neuropathy, suggesting that molecular damage by advanced Maillard reaction products may be a common mechanism in their development.

Aging↗

WS009 A and B, new endothelin receptor antagonists isolated from Streptomyces sp. no. 89009. I. Taxonomy, fermentation, isolation, physico-chemical properties and biological activities.

WS009 A and B novel endothelin receptor antagonists, have been isolated from the fermentation broth of Streptomyces sp. No. 89009. These antagonists were purified from the culture filtrate followed by Diaion SP-207, DEAE Toyopearl column chromatography and HPLC. WS009 A and B showed selective activity in an endothelin receptor binding assay with IC50 of 5.8 x 10(-6) M and 6.7 x 10(-7) M, respectively. On the basis of spectroscopic and chemical evidence, the structures of WS009 A and B have been established as 1 and 3, and are highly hydroxylated benz[a]anthraquinone chromophores.

Anthraquinones↗

WS009 A and B, new endothelin receptor antagonists isolated from Streptomyces sp. no. 89009. II. Biological characterization and pharmacological characterization of WS009 A and B.

WS009 A and B, produced by Streptomyces sp. No. 89009, were found to be competitive and specific antagonists against endothelin (ET)-1 receptors in in vitro studies and also active in in vivo studies. Furthermore, WS009 A and B were specific antagonists for vascular ET-1 receptors (ETA receptors) and significantly prevented the accumulation of intracellular inositol 1,4,5-triphosphate (IP3) in endothelin treated rat aorta tissues.

Animals↗