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Biomedical subjects

S Ejiri

Publications and source records attributed to S Ejiri.

At least 37 records · Page 2Linked to original sources
PubMed2000

Region-specific bone mass changes in rat mandibular condyle following ovariectomy.

The effects of estrogen deficiency on the mandible have been indicated; however, there have been only a few reports on the mandibular condyle. To clarify the region-specific changes of condyles following estrogen deficiency, we histomorphometrically studied the anterior and posterior regions of condyles in sham-operated (Sham) and ovariectomized (Ovx) female Fischer rats, since it has been reported that rat occlusal force loads mainly onto the anterior part of the condyle. The anterior and posterior regions of condyles showed significantly different bone dynamics. The bone volume in the anterior region of the Sham group increased to 79.9% with aging, but that of the posterior region remained at ca. 60%. In the Ovx group, the bone volume of the posterior region significantly declined to 42.7% until 60 days post-ovariectomy, although no changes occurred in the anterior region. As compared with the Sham counterparts, both regions of the Ovx group showed high bone turnover and revealed significantly low bone volume at 60 days (p < 0.01). Regional differences between anterior and posterior regions were more emphasized in the Ovx than in the Sham group. The osteoclast number and amount of eroded surface in the Ovx were higher in the posterior than in the anterior region, with significance at 30 and/or 60 days (p < 0.05). The mineralized surface at 14 days and the bone formation rate at 60 days in the Ovx posterior region were significantly lower than those in the Ovx anterior region (p < 0.05). Consequently, a net bone loss occurred in the posterior region of the mandibular condyle in ovariectomized rats, owing to region specificity and estrogen deficiency.

Analysis of Variance↗
PubMed2000

Localization of alkaline phosphatase and osteopontin during matrix mineralization in the developing cartilage of coccygeal vertebrae.

We observed the manner in which alkaline phosphatase (ALPase) and osteopontin were localized in the cartilage and intramembranous bone of coccygeal vertebrae during matrix mineralization, shedding considerable light on the manner in which they develop. In the cartilage matrix of coccygeal vertebrae, we observed the localization of ALPase activity in the boundary of the proliferative and the hypertrophic zones. Granular nodules of mineralization were consistently found in the boundary of both zones, and increased in size when close to the hypertrophic zone. While osteopontin was rarely present in the early stages of mineralization, its localization along the margins of mineralized matrices in the hypertrophic zone was prominent. In contrast to cartilage, mineralized nodules in the intramembranous bone in the mid-portion of the vertebra displayed osteopontin-immunoreactivity, indicating its early synthesis and subsequent accumulation to early-stage mineralized nodules. When blood vessels, accompanied by osteoblastic and osteoclastic cell populations, invaded the cartilage, osteopontin was localized in the lower region of the hypertrophic zone, despite its maintaining the localization of ALPase and early-stage mineralization. Thus, our investigation demonstrated ALPase activity consistent with early-stage mineralization in the cartilage matrix. However, the fact that osteopontin-localization could not be pinpointed might account for its multifunctionality as concerns both the regulation of mineralization and the attachment of migrating osteogenic and osteoclastic cells to the mineralized matrix.

Acid Phosphatase↗
PubMed2000

Ultrastructural, cytochemical, and biophysical aspects of mechanisms of bone matrix calcification.

Primary calcification in embryonic ossification occurs as follows: crystallization within matrix vesicles, formation of calcified nodules, and finally the establishment of expansive calcified matrix. However, the participation of the matrix vesicles in other types of bone calcification, such as bone formation during bone remodeling in adults has not been examined sufficiently. We introduce our recent observations on the presence of matrix vesicles in aged bones. In addition, although it is well known that the extracellular fluid supersaturates the calcification crystal, hydroxyapatite, the specific mechanisms by which bone matrix calcify remain unclear. In order to further approach the mechanisms of bone matrix calcification, we also review ultrastructural and localizational alterations of the matrix organics according to the progression of calcification, and an evaluation of mineral micro-environment in the calcifying sites by energy-filter transmission electron microscopy.

Animals↗
PubMed1999

Changes of cancellous bone mass in rat mandibular condyle following ovariectomy.

Changes in cancellous bone of the rat mandibular condyle following estrogen deficiency were histomorphometrically examined with 120-day-old female Fischer rats. Sixty-four animals were either ovariectomized bilaterally (ovx) or subjected to sham surgery (sham), and eight from each group were killed at 7, 14, 30, and 60 days after surgery. Seven intact animals were killed on day 0. Before killing, tetracycline and calcein were administered to all animals. Following histological observation, bone histomorphometry of the mandibular condyle was done using a confocal laser scanning microscope and an image analyzer. The sampling site was divided into two regions for analysis: (1) a "subchondral region," formed by the region connected to cartilage; and (2) a "central region," formed by the region beneath the former. The changes in these two regions were analyzed separately. In the sham group's condyle, the bone volume of the subchondral and central regions increased with the passage of time, although the bone turnover became low. This bone gain could be due to the effects of growth and the mechanical stimulus by occlusal load. In the subchondral region of the ovx group's condyle, the bone volume decreased significantly at 7 days, but recovered to reach approximately the same value as the sham group from 14 days onward. In the central region of the ovx group's condyle, the bone volume was unchanged, but revealed a significantly lower value than that of the sham group at 60 days (p < 0.01). Thus, ovariectomy inhibited bone gain, which was observed in the sham group's condyle even though there was no bone loss. On the other hand, the trabecular separation in the ovx's condyle of both the subchondral and central regions increased considerably and small marrow cavities interconnected to form a large bone marrow. Therefore, the ovx rat mandibular condyles dynamically altered their structures under the effects of estrogen deficiency and occlusal loads. Consequently, estrogen deficiency induced transient subchondral bone loss and recovery, whereas, in the central region, it inhibited bone gain. This suggests that mechanical loading modulates the normal ovx-induced bone loss found in other parts of the skeleton.

Animals↗
PubMed1999

Expression of elongation factor 1 beta' in Escherichia coli and its interaction with elongation factor 1 alpha from silk gland.

Silk gland elongation factor 1 (EF-1) consists of four subunits: alpha, beta, beta', and gamma. EF-1 beta beta' gamma catalyzes the exchange of GDP for GTP on EF-1 alpha and stimulates the binding of EF-1 alpha-dependent aminoacyl-tRNA to ribosomes. The carboxy-terminal regions of the EF-1 beta subunits from various species are highly conserved. We examined the region of EF-1 beta' that binds to EF-1 alpha by in vitro binding assays, and examined the GDP/GTP exchange activity using deletion mutants of a GST-EF1 beta' fusion protein. We thereby suggested a pivotal amino acid region, residues 189-222, of EF-1 beta' for binding to EF-1 alpha.

Amino Acid Sequence↗
PubMed1998

A novel variant of translation elongation factor-1beta: isolation and characterization of the rice gene encoding EF-1beta2.

A rice gene encoding a novel isoform of translation elongation factor-1beta subunit (termed EF-1beta2) was isolated and characterized. The gene comprises of eight exons, and encodes a 226-amino-acid protein. Expression of EF-1beta2 mRNA is abundant in seeds and cultured cells, but is considerably low in the tissues of the rice seedling. Antiserum raised against an EF-1beta2 synthetic peptide detected a protein with a relative molecular mass of about 32 kDa, indicating the EF-1beta2 gene is actually expressed in rice tissues. EF-1beta2 showed a close similarity to the cognate subunits from plant (beta and beta').

Amino Acid Sequence↗
PubMed1998

Isolation and characterization of a rice cDNA encoding the gamma-subunit of translation elongation factor 1B (eEF1Bgamma).

We isolated a rice cDNA clone (refg) encoding the gamma-subunit of translation elongation factor 1B (eEF-1B gamma; the old designation was EF-1 gamma). The refg encodes an open reading frame of 419 amino acids which shows a similarity to the equivalent sequences from animals and yeast. Complex formation analysis, which showed the recombinant protein of refg (His-eEF1B gamma) and formed a complex with GST-eEF-1Bbeta, indicated that the refg encodes rice eEF1B gamma of the eEF1B alphabeta gamma complex. Expression analysis showed that refg mRNA is very abundant in suspension-cultured cells during the exponential phase of growth. A DNA blot analysis indicated that refg is located at a single locus in the rice genome.

Amino Acid Sequence↗
PubMed1998

The effect of aging and ovariectomy on mandibular condyle in rats.

STATEMENT OF PROBLEM: It is important for dentists to understand the effect of systemic hormonal change on the osseous oral structures. PURPOSE: This study examined the effect of aging and ovariectomy on rat mandibular condyle. MATERIAL AND METHODS: Seventy-two 120-day-old female Fischer rats were killed at 7, 14, 30, and 60 days after bilateral ovariectomy or sham surgery. As the baseline control group, eight animals were killed on day 0 without surgeries. Changes in the bone mineral density and bone marrow area were detected through dual-energy x-ray absorptiometry and soft x-ray photography, respectively. RESULTS: No significant difference of bone mineral density was found between the bilateral ovariectomy and sham surgery groups with dual-energy x-ray absorptiometry, probably because the thickness of cortical bone obscured any possible changes in trabecular bone. Age-related osteosclerotic changes were found in the sham group with soft x-ray photography. In contrast, the bilateral ovariectomy group showed little change in bone marrow area in relation to time course; on the other hand, the value of their bone marrow area became significantly larger than that of the sham surgery group from 14 days after ovariectomy onward. CONCLUSIONS: It was inferred that estrogen deficiency caused the significantly large marrow area found in the rat mandibular condyle. Although much more research is necessary, this study allowed us to speculate that osteoporotic changes may occur in the mandibular condyle of postmenopausal women.

Absorptiometry, Photon↗
PubMed1998

Immunoreactive localization of transforming growth factor-beta type II receptor-positive cells in rat tibiae.

To identify the target cells of transforming growth factor-beta (TGF-beta) in normal bone tissue, we examined TGF-beta type II receptor expression using immunohistochemistry, reverse transcription-polymerase chain reaction (RT-PCR), and in situ hybridization in young rat tibiae. In the epiphyseal growth plate, the TGF-beta type II receptor cDNA was detected by RT-PCR and, alternatively, the TGF-beta type II receptor protein and mRNA expression were observed in the chondrocytes in the lower part of the proliferative cell layer, and in maturative and hypertrophic cell layers by immunohistochemistry and in situ hybridization. Of these, proliferative and maturative chondrocytes, in particular, revealed strong mRNA expression. In the cortical bone area, immunoreactivity for the TGF-beta type II receptor was detected in the fibroblastic cells near the osteoblasts on the endosteal surface of cortical bone. In conclusion, our findings suggest that target cells of TGF-beta in normal bone tissue could be considered mainly as extracellular matrix-producing chondrocytes and undifferentiated preosteoblasts, and TGF-beta may affect matrix production and differentiation of these cells.

Animals↗
PubMed1998

Isolation, characterization and mRNA expression of four cDNAs encoding translation elongation factor 1A from rice (Oryza sativa L.).

Four different cDNA clones encoding protein synthesis elongation factor 1A, eEF1A, were isolated from rice (Oryza sativa L.). The genes encoded by these cDNAs were designated rice elongation factor 1A genes refa1, refa2, refa3 and refa4. The genes encoded identical eEF-1A polypeptides and shared high amino acid identity with eEF1A of other eukaryotes. Southern blot analysis suggested that some of these refa genes may be organized in a cluster on the same chromosome within a short distance. PCR analysis of rice genomic DNA showed that refa1 and refa4, and refa3 and refa2 are in neighboring locations on the rice genome. The mRNAs of the four refa genes accumulated to nearly equal levels in a variety of tissues and at different stages of growth. Suspension-cultured cells were the most abundant in refa mRNAs. Dormant seeds contained a small amount of the four refa mRNAs. Transcript accumulation was highly induced after seed germination, and the same expression levels were maintained even in old leaf blades of mature plants.

Amino Acid Sequence↗
PubMed1997

Cell type- and positionally specific regulation of the aldolase P gene expression in rice seedlings.

We describe here different regulation of the AldP gene, a nuclear gene encoding chloroplast aldolase, in different tissues and growth ages of rice seedlings. Expression of the AldP gene is mesophyll cell-specific, and increases from the basal to the upper region in each leaf. The gene expression is repressed in the dark-grown leaf blade, but is induced by a short-term-exposure to light, to a level higher than that seen in the normal leaf blade. However, the light-inducibility differs among the tissues, and shows different patterns among leaf positions; i.e., the extent of light-induction is higher in the third leaf blade as compared with the earlier developed second leaf blade. Such positional differences in the regulation are also seen in the leaf sheath. These responses are not accompanied by changes of the cell type specificity in the expression.

Fructose-Bisphosphate Aldolase↗
PubMed1996

Haploinsufficiency of parathyroid hormone-related peptide (PTHrP) results in abnormal postnatal bone development.

Although apparently phenotypically normal at birth, mice heterozygous for inactivation of the gene encoding parathyroid hormone-related peptide (PTHrP) develop haplotype insufficiency by 3 months of age. In addition to histologic and morphologic abnormalities similar to those seen in homozygous mutants, heterozygous animals demonstrated alterations in trabecular bone and bone marrow. These included metaphyseal bone spicules which were diminished in volume, irregularly distributed, and less well developed than those seen in age-matched controls as well as bone marrow, which contained an inordinate number of adipocytes. A substantial reduction in PTHrP mRNA was detected in heterozygous tissue, while circulating parathyroid hormone (PTH) and calcium concentrations were normal. Thus, while a physiologic concentration of PTH was capable of maintaining calcium homeostasis, it was incapable of compensating for PTHrP haploinsufficiency in developing bone. In normal animals, both PTHrP and the PTH/PTHrP receptor were expressed predominantly in chondrocytes situated throughout the proliferative zone of the tibial growth plate. In the metaphysis, the PTH/PTHrP receptor was identified on osteoblasts and preosteoblastic cells situated in the bone marrow, while PTHrP was expressed only by osteoblasts. These observations indicate that postnatal bone development involves susceptible pathways that display exquisite sensitivity to critical levels of PTHrP and imply that the skeletal effects of PTH are influenced by locally produced PTHrP. Moreover, identification of both the ligand and its N-terminal receptor in metaphyseal osteoblasts and their progenitors suggests an autocrine/paracrine role for the protein in osteoblast differentiation and/or function. Impairment in this function as a consequence of PTHrP haploinsufficiency may critically influence the course of bone formation, resulting in altered trabecular architecture and perhaps low bone mass and increased bone fragility.

Age Factors↗
PubMed1996

Genomic structure of the rice aldolase isozyme C-1 gene and its regulation through a Ca 2+ -mediated protein kinase-phosphatase pathway.

Complementary and genomic DNA clones coding for aldolase C-1, the fourth-type isozyme of aldolase in rice Oryza sativa L., have been characterized. The organization of the gene is quite similar to those encoding rice aldolase C-a and a maize cytoplasmic-type aldolase, in that introns are located in the same position. Amino acid sequences are highly conserved among cytoplasmic aldolases in plants. Expression of the gene in rice callus is activated by a protein phosphatase inhibitor okadaic acid, and is inhibited in the presence of thapsigargin, a reagent which increases calcium influx into the cytoplasm. The inhibition is rescued by the simultaneous addition of protein kinase inhibitor H-7. Thus, it is suggested that expression of the aldolase C-1 gene is regulated through a signal transduction pathway involving a Ca 2+ -mediated protein kinase-protein phosphatase system.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1996

17 beta-estradiol increases calcium content in fetal mouse parietal bones cultured in serum-free medium only at physiological concentrations.

Using a bone organ culture system that shows mineralization in vitro, we investigated whether 17 beta-estradiol dose-dependently increases calcium content in cultured calvarial bones in serum-free medium. Fetal mouse parietal bones (3 x 3 mm) were cultured in phenol red-free BGJ medium containing phosphate (3-4 mmol/L), calcium (1-1.25 mmol/L), insulin (6 micrograms/ML), and transferrin (6 micrograms/mL) for 4-5 days. Under these culture conditions, the calcium content of the cultured bones (at dissection 34.0 +/- 4.6 micrograms/bone [mean +/- SD], n = 50) increased by 15-20 micrograms during 4-5 days of culture. 17 beta-Estradiol increased the calcium content significantly at 10(-12) to 10(-11) mol/L, but not at lower (10(113) mol/L) or higher (10(-10) to 10(-9) mol/L) concentrations. 17 alpha-Estradiol had no effect. The stimulatory effect of 17 beta-estradiol was completely inhibited by the antiestrogen agent ICI-182,780. The anabolic effect of 17 beta-estradiol was elicited not only in bones from females but also in those from males. 17 beta-Estradiol had no significant effect on 45Ca release from prelabeled parietal bones. Furthermore, light- and electron-microscopic examinations revealed that bone mineralization proceeded through formation of matrix vesicles, without any metastatic or dystrophic calcification. These in vitro findings suggest that 17 beta-estradiol elicits small, but reproducible, direct effects on calcium content in the parietal bones not only in female but also in male fetal mice at physiological-free E2 concentrations (10(-12)-10(-11) mol/L), which is attainable in serum of normal human subjects. In contrast to in vivo studies, pharmacological doses of 17 beta-estradiol had no anabolic effect on parietal bones. The mechanism of such a biphasic effect of estrogens remains to be elucidated.

Analysis of Variance↗
PubMed1996

Purification and properties of tyrosinase isozymes from the gill of Lentinus edodes fruiting body.

Six tyrosinase isozymes were purified from the browned gill of the fruiting body of Lentinus edodes by ammonium sulfate fractionation, DEAE-Sephacel and Q-Sepharose column chromatography, and partially denaturing SDS-PAGE. At the step of Q-Sepharose column chromatography, two active fractions (A and B) were obtained. Each fraction was separated to three further fractions, A1, A2, and A3, and B1, B2, and B3, respectively, by partially denaturing SDS-PAGE. All these isozymes consisted of two types of polypeptides: alpha polypeptide (A alpha or B alpha) and either beta (A beta or B beta) or gamma polypeptide (A gamma or B gamma). The alpha polypeptide contained the consensus amino acid sequence of the active site of known tyrosinases, which is considered to act as a catalytic subunit. From the results of peptide mapping and the amino acid composition, A alpha and B alpha polypeptides were considered to be different proteins. The kinetic properties of the purified tyrosinase isozymes differed greatly according to whether they contained beta or gamma polypeptide, indicating these polypeptides to be a possible regulatory subunit.

Amino Acid Sequence↗
PubMed1996

Effect of age on physiologic and mechanically stressed rat alveolar bone: a cytologic and histochemical study.

The effect of age on physiologic and mechanically stressed rat alveolar bone was assessed with special focus on bone formation and resorption in maxillary molar sections of 1-, 4-, 9-, 16-, and 24-month-old male Fischer rats. Tetracycline- and calcein-labeled undecalcified sections were used for observation by confocal laser scanning microscopy and contact microradiography. The localization of osteoclasts and the morphological change of osteoblasts were observed on decalcified sections. Mechanical stress was applied by an orthodontic elastic inserted between the first and second maxillary molars for 7 days. Results showed that bone-formative activity of osteoblasts and bone-resorptive activity of osteoclasts declined with age around the maxillary molar section of physiologic rat alveolar bone. However, results of mechanically stressed rat alveolar bone revealed no evidence of numerical difference in number, size, and activity of osteoclasts and osteoblasts among the ages, suggesting that mechanical stress provokes a higher activity of osteoblasts and osteoclasts similarly in both adult and younger rat alveolar bone.

Age Factors↗
PubMed1995

The promoter from the rice nuclear gene encoding chloroplast aldolase confers mesophyll-specific and light-regulated expression in transgenic tobacco.

The rice genome contains at least four separate loci that encode aldolase isozymes. Among these, the aldolase P (AldP) gene, a nuclear gene coding for chloroplast aldolase, is expressed predominantly in the leaf blade mesophyll cells in rice. To dissect promoter elements that regulate such tissue- or cell type-specific expression, we constructed various AldP promoter-beta-glucuronidase (GUS) fusion genes and transferred them into Nicotiana tabacum (tobacco) plants. Analysis of GUS activities in the transgenic tobacco revealed the presence of at least two elements within 2.0 kb AldP promoter region. One is located within the segment from position -2.0 kb to -1.2 kb and acts as a negative element. The other is a positive element located between -1.2 kb and -0.31 kb that confers developmentally regulated, mesophyll cell-specific expression. In addition, the 1.2 kb rice promoter segment flanking the transcription start site contains an element(s) that serves as target for light induction in tobacco. The results suggest that the AldP gene promoter of rice, a monocot promoter, can function in an essentially physiological manner in the dicot tobacco plant.

Blotting, Northern↗