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S Shimoda

Publications and source records attributed to S Shimoda.

At least 73 records · Page 4Linked to original sources
PubMed1992

Localization of glycosylated matrix proteins in secretory porcine enamel and their possible functional roles in enamel mineralization.

The present study was undertaken to investigate glycosylation of porcine enamel proteins secreted in the secretory stage of amelogenesis and to gain insight into functional roles of glycosylated proteins in enamel mineralization. Enamel proteins, isolated from various zones of the secretory enamel, were separated by SDS-PAGE and then transferred on to a nitrocellulose membrane. The transblotted proteins were visualized with either antibodies against porcine amelogenins or various biotin-conjugated lectins. The lectins used were Con-A, GS-II, STA, WGA, s-WGA, GS-I, MPA, VVA, PNA, RCA-I, DBA, SJA, UEA-I, Lotus-A and LPA. The results of the immuno- and lectin blottings revealed that most of the lectins did not bind to porcine amelogenins, while a large number of non-amelogenins having various molecular masses were stained strongly with the conjugated WGA, Con A and MPA lectins. On the basis of the binding specificity with the lectins, porcine non-amelogenins were classified into two groups: WGA (and Con A)-binding moieties at 60-90 kDa (WGA-HMW); and MPA-binding moieties at 13-17 kDa (MPA-LMW). These two groups of non-amelogenins differed distinctly in terms of their localization and stability in the secretory tissue and their adsorption properties onto hydroxyapatite. The WGA-HMW were concentrated in the outer region adjacent to the ameloblasts and disappeared (due to degradation) in the underlying inner secretory enamel. In contrast, the MPA-LMW were found in all zones of the secretory enamel and their quantity remained relatively constant. Histochemical studies using FITC-conjugated WGA and MPA showed that the fluorescence-labelling of WGA was localized in the core region of prism rods, while the fluorescence-labelling of MPA was locally limited at the rim of prism rods or at the prism sheath. In separate adsorption studies, it was found that the WGA-HMW, as well as the intact amelogenins, displayed a high adsorption affinity on to apatite crystals, whereas the MPA-LMW showed only marginal adsorption on to apatitic surfaces. The overall results indicate that part of the heterogeneity found in porcine enamel proteins can be ascribed to variations of carbohydrate moieties attached to non-amelogenins.(ABSTRACT TRUNCATED AT 400 WORDS)

Adsorption↗
PubMed1992

Effects of inorganic iodide, epidermal growth factor and phorbol ester on hormone synthesis by porcine thyroid follicles cultured in suspension.

Porcine thyroid follicles cultured in suspension for 96 h synthesized and secreted thyroid hormones in the presence of thyrotropin (TSH). The secretion of newly synthesized hormones was assessed by determining the contents of thyroxine (T4) and triiodothyronine (T3) in the media and by paperchromatographic analysis of 125I-labelled hormones in the media where the follicles were cultured in the presence and absence of inhibitors of hormone synthesis. The hormone synthesis and secretion was modified by exogenously added NaI (0.1-100 microM). The maximal response was obtained at 1 microM. Thyroid peroxidase (TPO) activity in the cultured follicles with TSH for 96 h was dose-dependently inhibited by NaI. One hundred microM of NaI completely inhibited TSH-induced TPO activity. Moreover, both epidermal growth factor (EGF: 10(-9) and 10(-8) M) and phorbol 12-myristate 13-acetate (PMA: 10(-8) and 10(-7) M) inhibited de novo hormone synthesis. An induction of TPO activity by TSH was also inhibited by either agent. These data provide direct evidences that thyroid hormone synthesis is regulated by NaI as well as TSH at least in part via regulation of TPO activity and also that both EGF and PMA are inhibitory on thyroid hormone formation.

Animals↗
PubMed1992

Effects of epidermal growth factor, phorbol ester, and retinoic acid on hormone synthesis and morphology in porcine thyroid follicles cultured in collagen gel.

Epidermal growth factor (EGF), phorbol esters (PEs), and retinoic acid (RA) inhibit differentiated functions of thyrocytes. In the present study the inhibitory effects of these growth-promoting factors on hormone synthesis were studied in thyroid follicles cultured in type-I collagen gel, and morphologic alteration by these factors was examined by light and electron microscopy (EM). Porcine open thyroid follicles obtained by treatment with 0.1% collagenase were embedded in collagen gel and cultured in Ham's F12 medium supplemented with 6H (insulin, hydrocortisone, somatostatin, transferrin, glycyl-his-lys, and thyrotropin) + 0.5% fetal bovine serum (FBS). After 1 week these open follicles developed to closed follicles, and the medium was changed to one containing 6H + 0.5% FBS + 0.1 microM sodium iodide (NaI). Some media were supplemented with either EGF, phorbol 12-myristate 13-acetate (PMA), or all-trans RA. The closed follicles retained ability for hormone synthesis for 2 weeks after the medium change in the presence of 6H + FBS + NaI. The amounts of T4 and T3 secreted into the culture medium from day 9 to day 12 after the medium change were 60% and 45% of those from day 0 to day 4, respectively. EGF reduced production of T4 and T3 by 61% and 69%, respectively; PMA, by 87% and 99%; and RA, by 55% and 44%. In the medium supplemented with 6H + 0.5% FBS, the follicles exhibited intact polarity. Apical surfaces with microvilli were oriented to the follicular lumen and tight junctions were on the apical side of cell-to-cell contacts. Desmosomes were found on both the apical and basal halves of the cell contacts.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Labile or surface pools of magnesium, sodium, and potassium in developing porcine enamel mineral.

The present study was undertaken to assess the labile or surface pools of Mg, Na, and K ions in porcine enamel tissues at various developmental stages. The enamel samples, corresponding to the outer and the inner secretory, the early maturing, and the mature hard enamel, were dissected from the labial sides of permanent incisors of 6- to 8-month-old piglets. Each enamel sample was extracted successively with solutions of de-ionized water and 50 mmol/L Tris-4 mol/L guanidine buffer (for removal of organic matrix proteins, mainly amelogenins). The labile (free or organically bound) pools of Mg, Na, and K were assessed by the total amounts of these ionic species extracted by the water and Tris-guanidine buffer. The surface (adsorbed onto enamel mineral) pool of Mg was assessed directly by determination of the adsorption of Mg onto enamel mineral at various developmental stages. The results showed that: (i) 30-40% of the Mg in the secretory and early maturation enamel was in the surface pool (adsorbed onto the enamel mineral); (ii) 25 to 40% of the total sodium in the enamel samples was in labile forms; and (iii) most (around 70-80%) of the total potassium was readily extracted in water and appeared to originate from the enamel fluid; only marginal portions remained in the solids. The present adsorption studies also indicated that the maximum uptake of magnesium in the early maturation enamel was due mostly to an increase of the occupancy by Mg ions of adsorption sites on the crystal surfaces, which become accessible with a massive removal of enamel matrix proteins.(ABSTRACT TRUNCATED AT 250 WORDS)

Adsorption↗
PubMed1992

Effect of a phorbol ester on immunoreactive endothelin-1 release from cultured porcine aortic endothelial cells.

This study was designed to examine how protein kinase C (PKC) regulates the release of endothelin-1 (ET-1) from cultured porcine aortic endothelial cells. We measured the release of immunoreactive (IR)-ET-1 from cells cultured for up to 72 h in the presence or absence of a phorbol ester TPA. The release of IR-ET-1 from control cells (no TPA) increased according to time for up to 72 h. In the presence of TPA, the release of IR-ET-1 from the cells was higher than the control level for up to 8 h, but was lower thereafter and reached a plateau after 48 h. TPA dose-dependently stimulated IR-ET-1 release during incubation for 4 h, but suppressed it after incubation for 72 h. Stimulation of PKC by diacylglycerol mimicked the early (4 h) action of TPA. On the other hand, pretreatment of cells with TPA to downregulate PKC significantly suppressed basal and thrombin- or FCS-stimulated IR-ET-1 release. These findings suggest that the activation of PKC is related to the stimulation of ET-1 release and that down-regulation of PKC leads to the suppression of ET-1 release from cultured endothelial cells.

Animals↗
PubMed1992

Neuroendocrine protein 7B2 in Prader-Willi syndrome.

7B2 is a neuroendocrine polypeptide of unknown function, the gene for which is sited near or within the chromosomal region deleted in Prader-Willi syndrome and Angelman's syndrome. Plasma immunoreactive 7B2 levels were measured in 26 individuals with Prader-Willi syndrome, and appropriate controls. Plasma 7B2 levels were within normal limits compared to the control groups, in adults with Prader-Willi syndrome. 7B2 levels in children with Prader-Willi syndrome were higher, this age-dependent variation having been previously reported in normal children.

Adolescent↗
PubMed1992

[Acute autonomic neuropathy and the other related diseases].

In 1969, Young et al. described a patient with an acquired, pure pan-dysautonomia of unknown etiology, involving both the sympathetic and the parasympathetic nervous systems without other neurological manifestations. Since then, a number of similar cases have been described. More recently, reports have appeared of predominantly cholinergic dysautonomia, of dysautonomia associated with infection, and of dysautonomia with other impairment of nervous function. As an etiology, immunological disorder has been suggested but the precise mechanism has not been clarified. By accumulating more cases, a more accurate etiology and more satisfactory therapy will be developed.

Acute Disease↗
PubMed1991

Elevation of plasma endothelin concentrations during endotoxin shock in dogs.

The effect of endotoxin on the release of endothelin, a novel potent vasoconstrictor peptide, was examined in anesthetized dogs and in cultured endothelial cells. Administration of 2.63 mg lipopolysaccharide, E. coli 0111:B4/kg body weight caused shock in the animals and produced a long-lasting increase in the plasma immunoreactive endothelin-1 level that remained higher than the basal level (1.83 pg/ml as mean level) from 30 to 120 min after the injection, with a peak at 90 min (8.15 pg/ml as mean level). In vitro immunoreactive endothelin-1 in a culture medium, in which calf pulmonary artery endothelial cells were incubated in the presence of 10% fetal bovine serum, increased dose dependently with the concentration of added lipopolysaccharide between 0.01 and 10 micrograms/ml. These data indicate that plasma endothelin increases during endotoxin shock and that stimulation by endotoxin, per se, in the presence of serum participates at least partially in the mechanism for its release.

Adrenocorticotropic Hormone↗
PubMed1991

Effect of glucose and insulin on immunoreactive endothelin-1 release from cultured porcine aortic endothelial cells.

We measured the release of immunoreactive endothelin-1 (IR-ET-1) by cultured porcine aortic endothelial cells under normoglycemic (5.5 mmol/L) and hyperglycemic (27.5 and 55 mmol/L) conditions. Compared with cells incubated in the presence of a normal glucose concentration, cells incubated in 27.5 mmol/L glucose medium released 52% less IR-ET-1, and those incubated in 55 mmol/L glucose medium released 54% less IR-ET-1. The observed effects of elevated glucose on IR-ET-1 release were both sugar-specific and not due to increased osmolarity. Fetal calf serum (FCS)-stimulated IR-ET-1 release in the presence of elevated glucose was also less than that in the presence of a normal glucose concentration. In addition, the effects of two hormones, insulin and insulin-like growth factor 1 (IGF-1), on IR-ET-1 release were examined. Both insulin and IGF-1 dose-dependently stimulated IR-ET-1 release. Twenty micrograms/mL insulin and 10(-8) mol/L IGF-1 increased IR-ET-1 release by 38% and by 44%, respectively. These results indicate that hyperglycemic condition results in reduction of IR-ET-1 release from cultured porcine aortic endothelial cells and that insulin and IGF-1 stimulate its release. The possible relevance of these observations to physiological regulation of ET-1 release in vivo and pathological processes in diabetes remains to be established.

Animals↗
PubMed1991

A 3' splice site mutation in the thyroglobulin gene responsible for congenital goiter with hypothyroidism.

A case of congenital goiter with defective thyroglobulin synthesis has been studied in molecular terms. The patient is the fifth of a kindred of six, three of which have a goiter. The parents are first cousins. Segregation of thyroglobulin alleles in the family was studied by Southern blotting with a probe revealing a diallelic restriction fragment length polymorphism (RFLP). The results demonstrated that the three affected siblings were homozygous for the RFLP. Northern blotting analysis of the goiter RNA with a thyroglobulin probe suggested that thyroglobulin mRNA size was slightly reduced. Polymerase chain reaction amplification of the 8.5-kb thyroglobulin mRNA as overlapping cDNA fragments demonstrated that a 200-bp segment was missing from the 5' region of the goiter mRNA. Subcloning and sequencing of the cDNA fragments, and of the patient genomic DNA amplified from this region, revealed that exon 4 is missing from the major thyroglobulin transcript in the goiter, and that this aberrant splicing is due to a C to G transversion at position minus 3 in the acceptor splice site of intron 3. The presence in exon 4 of a putative donor tyrosine residue (Tyrosine nr 130) involved in thyroid hormone formation provides a coherent explanation to the hypothyroid status of the patient.

Adult↗
PubMed1991

Changes in acid-phosphate content in enamel mineral during porcine amelogenesis.

The present study was undertaken to investigate changes in the acid-phosphate content of porcine enamel mineral during its development and to assess separately the HPO4(2-) pools in labile and stable forms. Enamel samples at the secretory and maturing stages of amelogenesis were obtained from the permanent incisors of five- to six-month-old slaughtered piglets. Human enamel from erupted, extracted teeth, synthetic hydroxyapatite, and carbonatoapatite containing acid phosphate were included as references. The acid-phosphate content of each sample was determined chemically through its pyrolytic conversion to pyrophosphate. The assessment of HPO4(2-) in labile forms was made by analysis of samples preequilibrated with solutions containing 3 mmol/L phosphate at pH11 (to de-protonate the HPO4(2-) species on crystal surfaces). The analytical results of porcine enamel samples showed that: (a) the outermost secretory (youngest) enamel contained the highest HPO4(2-), corresponding to about 16% of the total phosphate; (b) the acid-phosphate content decreased gradually to 10% in the inner (older) secretory and to 6% in the maturing tissue; (c) a substantial part of the HPO4(2-) in developing enamel tissue (50-60% of the HPO4(2-) for the secretory enamel) was in labile forms; and (d) the pool of the labile HPO4(2-) decreased with the growth of enamel mineral. In parallel studies with mature human enamel, it was ascertained that the total acid phosphate was only about 3% of the total phosphate, much lower than in developing porcine enamel, and that the labile pool of HPO4(2-) was also small, corresponding to about 15% of the total acid phosphate determined.(ABSTRACT TRUNCATED AT 250 WORDS)

Amelogenesis↗
PubMed1991

[Study of parameters to evaluate the grade of malignancy of colorectal cancer with submucosal invasion].

Histological sections were prepared from 69 cases of colorectal cancer and the following 3 parameters were determined: A) the area of malignant change (mm2), B) the area of malignancy in the submucosal layer (mm2), and C) the proportion of B to A (%). The relationship between these parameters and biological malignant findings (lymph node metastasis and lymphatic and/or venous invasion) was investigated. There was no relationship between A and biological malignant findings. Lesions with B values of less than 7.5 mm2 were not associated with biological malignant findings but those with values more than 30 mm2 always were. A significant difference in B values was noted between patients with and without these findings (22.0 +/- 13.2 mm2 vs. 6.5 +/- 7.0 mm2, p less than 0.01). C values of more than 45% were always associated with biological malignant findings but values less than 20% never were. A significant difference in C values was observed between patients with and without these findings (57.6 +/- 22.1% vs. 14.1 +/- 11.3%, p less than 0.01). These results suggested that the area of malignancy in the submucosal layer and its percentage to the entire area of malignant change on the histological sections were useful parameters to evaluate the grade of malignancy of invasive colorectal carcinoma.

Colorectal Neoplasms↗
PubMed1990

[Altered alpha atrial natriuretic peptide (ANP) concentrations in plasma, atria and hypothalamus in experimentally induced hyper- and hypothyroid rats].

In order to assess a possible involvement of thyroid hormone in atrial natriuretic peptide (ANP), experimentally induced hyper- and hypothyroid rats were employed, and the immunoreactive rat ANP (IR-ANP) concentrations in plasma, atria and brain regions including the hypothalamus were measured by a specific radioimmunoassay. Plasma IR-ANP concentration in hypothyroid rats was 14.5 +/- 2.9 (mean +/- SD) fmol/ml, significantly lower than that in control rats (p less than 0.05 vs control of 24.9 +/- 9.7 fmol/ml). Plasma IR-ANP concentration in hyperthyroid rats was 66.4 +/- 9.7 fmol/min, significantly higher than that in the controls (p less than 0.01). Atrial IR-ANP concentration in hyperthyroid rats was significantly lower than that in the controls (79.9 +/- 11.1 nmol/g vs 133.5 +/- 21.2 nmol/g (control), p less than 0.05), though no significant change was observed in atrial IR-ANP concentration in hypothyroid rats. While hypothalamic ANP concentration in hypothyroid rats was significantly lower than that in the controls (17.5 +/- 3.5 pmol/g vs 31.9 +/- 1.9 pmol/g (control), p less than 0.05), there was no significant change of that in the hyperthyroid rats. On reverse phase high performance liquid chromatography, the major peak in plasma and hypothalamus extract was thought to be identical to synthetic alpha-rat ANP (1-28). These results may suggest that in the hyperthyroid state an excessive amount of ANP is released from atria into the blood, and that in the central nervous system thyroid hormone involve ANP metabolism being different from the atrium.

Animals↗
PubMed1990

Immunoreactive endothelin concentrations in maternal and fetal blood.

Immunoreactive-endothelin (ir-ET) concentrations were determined in peripheral maternal blood and in umbilical cord blood just after delivery. The concentrations in both the umbilical artery (2.83 +/- 1.36 pmol/l plasma, Mean +/- SD) and vein (3.37 +/- 1.53 pmol/l) were significantly higher than those found in maternal venous blood (1.43 +/- 1.02 pmol/l). On the other hand, ir-ET levels in maternal blood were not significantly different when compared with those found in non-pregnant women (1.50 +/- 0.83 pmol/l). No significant difference of ir-ET levels between the umbilical artery and vein was observed. A highly significant correlation (r = 0.60, p less than 0.01) of ir-ET levels between the umbilical artery and vein was observed. Also, a significant correlation (r = 0.48, p less than 0.01) between umbilical vein and maternal vein ir-ET levels with a weaker correlation (r = 0.36, p less than 0.05) between umbilical artery and maternal vein ir-ET levels was demonstrated. The present study indicates that ir-ET may be actively secreted in fetal circulation and the plasma levels in maternal and fetal circulation may have a possible relation.

Adult↗
PubMed1990

Inhibition by protein kinase-C inhibitor and cycloheximide of phorbol ester- and epidermal growth factor-induced arachidonic acid metabolism in cultured porcine thyroid cells.

In an attempt to elucidate possible mechanism(s) for stimulated arachidonic acid metabolism by phorbol 12-myristate 13-acetate (PMA) and epidermal growth factor (EGF) in porcine thyroid cells, we examined the effects of protein kinase inhibitors, isoquinolinesulfonamide derivatives (H-7 and HA-1004), and cycloheximide. The production of PGE2 stimulated by either PMA or EGF was strongly inhibited by H-7, with an ID50 value of approximately 20 to 25 mumol/L in each case, as well as by cycloheximide, with an ID50 value of less than 0.5 micrograms/mL in each case. In contrast, 100 mumol/L of HA-1004 showed less inhibition of PGE2 production provocated by either PMA or EGF. On the other hand, PGE2 production in basal or stimulated condition by exogenously added arachidonic acid, was inhibited to an even lesser extent by both H-7 and cycloheximide. The EGF- and PMA-stimulated release of 3H-arachidonic acid from the cells was also strongly inhibited by H-7 and cycloheximide. These results suggest an induction of synthesis of some proteins responsible for the release of arachidonic acid, which might be attributed to protein kinase-C activation in arachidonic acid metabolism stimulated by PMA or EGF. Moreover, PGE2 production was potently induced by PMA and slightly by EGF in the cyclooxygenase-inactivated cells by acetyl salicylate pretreatment, which also suggests that both agents might induce the synthesis of cyclooxygenase in cultured porcine thyroid cells, although we did not measure its activity.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1990

Presence of high affinity receptor for interleukin-1 (IL-1) on cultured porcine thyroid cells.

Using 125I-interleukin-1 beta (125I-IL-1 beta) as a ligand, a specific receptor of high affinity dissociation constant (1.1 +/- 0.2 x 10(-10) M) with binding sites (350 +/- 40/cell) for interleukin-1 beta (IL-1 beta) has been demonstrated on cultured porcine thyroid cells. IL-1 alpha almost equally cross-reacted with the receptor (Kd = 1.2 +/- 0.3 x 10(-10) M and 350 +/- 50 binding sites/cell). TSH, IL-2 and other peptide hormones did not inhibit the binding of 125I-IL-1 beta to thyroid cells. Crosslinking study revealed a major band (approximately 95 kD) with a corrected molecular mass of approximately 78 kD. Moreover, both IL-1 beta and IL-1 alpha stimulated prostaglandin E2 production of cultured porcine thyroid cells, although the potency of IL-1 alpha was slightly greater than that of IL-1 beta. These results suggest that IL-1 may be involved in the regulation of thyroid cell function.

Animals↗
PubMed1990

Increased endothelin concentration in CSF from patients with subarachnoid hemorrhage.

A considerable quantity of endothelin-like immunoreactivity was demonstrated as present in the CSF of patients with subarachnoid hemorrhage. The endothelin levels in the CSF raised from 0.4 +/- 0.2 (Mean +/- SD) pmol/L at Day 0-1 to 2.2 +/- 0.6 pmol/L at Day 6 and the levels decreased gradually. The result suggest that endothelin may contribute the generation of vasospasm often observed in subarachnoid hemorrhage.

Endothelins↗