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Biomedical subjects

S Tajima

Publications and source records attributed to S Tajima.

At least 199 records · Page 11Linked to original sources
PubMed1996

Erythropoietin in mouse avascular yolk sacs is increased by retinoic acid.

Erythropoiesis begins first in the visceral yolk sac (VYS) of the embryo; however, the involvement of erythropoietin (EPO) in yolk-sac erythropoiesis has not been studied adequately. This study reports the expression of EPO in normal and hypoxic VYSs and alterations in yolk sac components induced by retinoic acid (RA) in mice. Gravid mice (plug day = day 0 of gestation) were given one oral dose of 60 mg/kg of RA in olive oil on days 6, 6.5, 7, 7.5, or 8 of gestation and were sacrificed 2.5, 3, or 3.5 days later. Control mice received olive oil without RA. None of the dams developed anemia, but more than 80% of the embryos of the dams that received RA on day 6, 6.5, or 7 of gestation had avascular yolk sacs (AVYs) and anemia. In these AVYs, the adenosine triphosphate (ATP) level was as low as 18-59% of that in the control VYSs. Reverse transcription-polymerase chain reaction and Southern analysis of products demonstrated that mRNA for EPO receptor (EPR) was expressed in both VYSs and AVYs on days 9-11 of gestation, and EPO mRNA was present in VYSs and AVYs on days 9 and 10 of gestation and in vehicle-exposed VYSs on day 11 of gestation. Furthermore, enzyme immunoassay of EPO indicated that AVYs contained more EPO protein than control VYSs. Light microscopy revealed that, in AVYs, in addition to the defective hemopoietic cells, the endodermal layer was exclusively altered: The presence of focal proliferated regions and the separation from the mesenchyme led to a single layer from which some immature cells seemed to be migrating. Immunolocalization of EPO showed its presence in all components of VYSs with a characteristic distribution pattern: In the endodermal layer, cells with positive EPO staining decreased as gestation advanced, and erythroid precursor cells showed positive staining. In AVYs, the proliferated endodermal cells had EPO in abundance; in the separated regions, the distinction between positive and negative EPO staining became clearer than that in the control VYSs, and the immature cells in the lumens also had EPO. EPR was seen on the cell surface of the corresponding cells that reacted to EPO. These findings suggest that VYSs not only produce EPO temporarily but also respond to the oxygen content in situ. EPO and EPR appear to be synthesized in the endodermal cells of the VYSs that are likely to respond to the circumstances induced by RA.

Adenosine Triphosphate↗
PubMed1996

Experience of a new surgical procedure for the treatment of unilateral obstructive lymphedema of the lower extremity: adipo-lymphatico venous transfer.

We report five cases of adipo-lymphatico venous transfer (ALVT) performed to treat unilateral obstructive lymphedema of the lower extremity. ALVT is a surgical procedure that utilizes the long saphenous vein and its surrounding lymphatic tissue from the unaffected limb for the treatment of lymphedema. Since ALVT does not necessitate anastomoses of lymphatic vessels, it can be performed regardless of the severity and duration of lymphedema, and stable long-term results can be obtained when the transferred lymphatic tissue shows viability.

Adipose Tissue↗
PubMed1996

Immunohistochemical studies of glycosaminoglycans in nanophthalmic sclera.

BACKGROUND: It has been observed that there may be some connection between scleral collagen changes, glycosaminoglycan (GAG) changes and nanophthalmic uveal effusion. In a previous paper we reported increased dermatan sulfate (DS) and chondroitin sulfate (CS) contents in nanophthalmos sclera using biochemical analysis. In the present work we used immunohistochemical methods to assess the GAG changes in nanophthalmos sclera. METHODS: Scleral specimens from three nanophthalmic patients and five controls were processed to immunostaining with peroxidase-antiperoxidase (PAP) methods. Monoclonal antibodies for chondroitin (Ch), chondroitin-4-sulfate (C4S), chondroitin-6-sulfate (C6S), DS and keratan sulfate (KS) were used. The specimens were observed under a light microscope. RESULTS: Nanophthalmos scleral samples exhibited strong KS staining compared with the control specimens. Both the nanophthalmos and the control samples were stained with anti-Ch, anti-C4S, anti-C6S and anti-DS antibodies to the same intensity. CONCLUSION: Our results suggest that an increased or abnormal distribution of KS may contribute to abnormalities in collagen fibers.

Adolescent↗
PubMed1996

Inhibition of collagen expression by azelastine hydrochloride in cultured skin fibroblasts from normal individuals and scleroderma patients.

The effects of azelastine hydrochloride on cell proliferation and collagen synthesis in cultured human skin fibroblasts were studied. Azelastine inhibited cell proliferation during proliferating cell phases. Azelastine was found to inhibit collagen synthesis without altering cell proliferation during quiescent phases. It did not alter the ratio of type I to III collagen synthesis. Northern blot analysis of collagen chain mRNAs revealed that the levels of alpha1 (I), alpha1 (III) and alpha1 (VI) mRNAs were reduced by azelastine treatment, whereas the level of alpha2 (VI), alpha3 (VI) mRNAs were not significantly changed. These results suggest that azelastine modulates collagen synthesis at a pretranslational level. Azelastine inhibited collagen synthesis in fibroblasts from scleroderma patients to the same extent as in normal skin fibroblasts. This drug may be useful in the treatment of fibrotic diseases.

Case-Control Studies↗
PubMed1996

Differential in vitro responses of elastin expression to basic fibroblast growth factor and transforming growth factor beta 1 in upper, middle and lower dermal fibroblasts.

Elastin mRNA levels were measured in cultured skin fibroblasts derived from upper, middle and lower dermal layers. The elastin mRNA levels were highest in the fibroblasts from the upper dermal layer and lowest in the lower dermal fibroblasts. Modulation of elastin expression by basic fibroblast growth factor and transforming growth factor beta 1 in the dermal fibroblasts was also studied. Basic fibroblasts growth factor downregulated elasin expression in the upper dermal fibroblasts but did not significantly change elastin expression in the middle and lower dermal fibroblasts. Upregulation of elastin expression by transforming growth factor beta 1 was greater in the upper dermal fibroblasts than in the middle and lower dermal fibroblasts. Platelet-derived growth factor induced no significant changes in the three types of dermal fibroblasts. The results suggest that the differential responses of elastin expression to potent modulators may be at least partially responsible for the abnormal elastin metabolism specifically observed in the upper dermal layer.

Adult↗
PubMed1996

Immunolocalizations of human gelatinase (type IV collagenase, MMP-9) and TIMP (tissue inhibitor of metalloproteinases) in normal epidermis and some epidermal tumors.

The matrix metalloproteinases (MMPs) MMP-2 and MMP-9 (gelatinases) have been suggested as serving an important role in cleaving the basement membrane structure. Tissue inhibitors of metalloproteinases TIMPs (particularly TIMP-1) are known to inhibit MMPs. Based on this background, we raised monoclonal antibodies against human gelatinase (MMP-9) and human recombinant TIMP (TIMP-1), and immunostained these two components in skin from patients with squamous cell carcinoma (SCC), Bowen's disease (BD) and keratoacanthoma (KA). MMP-9 showed positive staining mainly in the granular layer of normal epidermis. In some cases of SCC and BD, MMP-9 showed positive staining in the dysplastic lesions even in the basal layer. TIMP showed a thorough positivity in normal epidermis. Unstained regions with this antibody were observed in SCC and BD. These results suggest that an altered staining pattern for MMP-9 and TIMP may be closely related to the malignant transformation of SCC and BD.

Animals↗
PubMed1996

Role of glycoprotein 130 and c-Kit signaling in proliferation and differentiation of human hematopoietic progenitor cells.

Glycoprotein (gp) 130, a receptor component for interleukin 6 (IL-6), can associate with a soluble IL-6 receptor (sIL-6R)-IL-6 complex. To examine the role of gp130 signaling in human hematopoietic progenitor-cell proliferation and differentiation, we studied the effects of the sIL-6R-IL-6 complex in combination with other cytokines on human CD34+ cells in clonal and suspension cultures. The sIL-6R-IL-6 complex, but not sIL-6R or IL-6 alone, in the presence of stem-cell factor (SCF) produced dramatic increases in the populations of various cell lineages, including erythroid cells and various hematopoietic progenitors, in suspension culture. Significant numbers of colonies of (particularly) multilineage and blast cells were generated in methylcellulose culture supplemented with a combination of sIL-6R-IL-6 complex and SCF. Addition of anti-gp130 monoclonal antibodies (MAbs) and anti-IL-6R MAbs to the above-mentioned cultures dose-dependently inhibited the generation of cells of various lineages and of progenitor cells in suspension culture and completely blocked multilineage colony production in methylcellulose culture; an anti-erythropoietin antibody did not cause inhibition. These findings demonstrate that both proliferation and differentiation of hematopoietic progenitor cells can be induced through gp130 and c-Kit signaling, indicating that progenitor cells are responsive to the sIL-6R-IL-6 complex, even though they do not express IL-6R. Together with previous studies showing that detectable levels of sIL-6R, IL-6, and SCF are present in human serum, these results suggest that gp130 signaling may play an important role in human hematopoiesis in vivo.

Antigens, CD34↗
PubMed1996

Ascorbic acid reduces tropoelastin levels in culture medium of chick skin fibroblasts.

Ascorbic acid reduced tropoelastin levels in the culture medium of embryonic chick skin fibroblasts to one-third or one-fourth of their original value in a dose- and exposure time-dependent manner under conditions in which collagen synthesis was stimulated 1.8-fold. The steady-state level of elastin mRNA remained essentially constant, whereas the collagen mRNA content increased in proportion to the increase in collagen synthesis with ascorbic acid treatment. These results suggest that the reduction of tropoelastin levels is not controlled by the elastin mRNA level but, rather, is related to the posttranslational modification of the tropoelastin molecule.

Actins↗
PubMed1996

Melanogenesis, biosynthetic phenotype of fibronectin and collagen, and migrating activity in cloned B16 mouse melanoma cells.

Three clones, melanotic (M3), amelanotic (A4 and A7) cells were isolated from B16 F10 mouse melanoma cell line. These clones exhibited different biosynthetic activities of fibronectin and collagen: A4 clone showed relatively active synthesis of both collagen and fibronectin, and A7 clone exhibited most active fibronectin synthesis, whereas no significant synthesis of these molecules was observed in M3 clone. No significant difference in growth rate was observed in these three clones. Migrating activities measured by basement membrane matrix-coated dishes were greater in A7 clone than in A4 and M3 clones. Messenger RNA levels of collagen and fibronectin paralleled collagen and fibronectin synthesis in these clones whereas tyrosinase mRNA level was unaltered between melanotic (M3) and amelanotic (A4 and A7) cells. These results indicate that B16 melanoma cells have heterogeneous cell populations consisting of different biosynthetic and metastatic properties. These clones may provide good tools for studying the relationship between the phenotypes of melanogenesis biosynthesis of extracellular macromolecules and migrating activity of melanoma cells.

Animals↗
PubMed1996

Fibrous-long spacing fiber formation by collagen and non-collagenous acidic components from calf skin.

Fibrous-long spacing (FLS) fibers were reconstituted in the supernatant fraction after disaggregation of calf insoluble collagen fibers. Macromolecular components of the fibers were biochemically analyzed. Amino acid analysis demonstrated that the major component of FLS was collagen consisting of types I and III collagen in the relative percentage of 92 and 8% respectively. Hyaluronic acid and dermatan sulfate were the major components of glycosaminoglycans present in the ratio of 1:2. Polyacrylamide gel electrophoresis (PAGE) revealed several minor non-collagenous proteins which were not derived from serum components. The results suggest that non-collagenous components including glycosaminoglycans, are involved in the formation of FLS.

Acids↗
PubMed1996

Analysis of connective tissue macromolecular components in Ishibashi rat skin: role of collagen and elastin in cutaneous aging.

The Ishibashi (IS) rat, established from cross-breeding between Wistar and wild rats, has a unique skin appearance, with wrinklings and furrows indicative of skin aging appearing at the age of 12 weeks. To understand the underlying mechanism of the formation of wrinkles, macromolecular components of connective tissue, collagen and elastin, in the young (5-6-week-old) and the aged (23-30-week-old) IS rat skins were biochemically analyzed. Hydroxyproline and isodesmosine contents in the aged IS rats were reduced 22% (P < 0.05) and 37% (P < 0.05) compared to the young rats, whereas no significant differences in the contents of both macromolecules in control Sprague-Dawley (SD) rats were seen. The relative content of type III collagen was unaltered between the young and aged skins of both IS and SD rats. A relative decrease in the intact elastin molecule (65 kDa) and a relative increase in the elastin fragments with lower molecular weights were observed in the aged IS rat skin by immunoblotting method. These results indicate that the reduction in collagen and elastin contents and increased degradation of elastin molecules in the aged IS rat skin could be related to the formation of wrinkles. Thus, the IS rat may provide a useful model for the study of skin aging.

Aging↗
PubMed1996

Non-disulfided pro alpha 1(IV) chain in B16 melanoma cell culture.

We have previously reported that two species of pro alpha 1(IV) collagen chains, disulfided (500 kDa) and non-disulfided (180 kDa), were produced by B16 melanoma cells (J Biochem (1994) 116; 1039-1043). The mechanism by which the non-disulfided pro alpha 1(IV) chain is produced was studied. No significant difference in prolyl hydroxylation in both polypeptides was observed. When the culture was treated with alpha, alpha 'dipyridyl', a potent inhibitor for hydroxylation of collagen, the secretion of the disulfided alpha 1(IV) chain was inhibited, but non-disulfided alpha 1(IV) chain secretion was not affected. Short pulse and pulse-chase experiments demonstrated that both chains appeared at the same time in the culture medium and the relative amounts of both chains in the medium were unaltered with increasing chase periods. These results indicate that the non-disulfided alpha 1(IV) chain is fully hydroxylated and that it's secretion is independent of it's hydroxylation level. A marked susceptibility of the 180 kDa alpha 1(IV) chain to pepsin at 4 degrees C suggests that the non-disulfided chain may be present in a denatured form.

2,2'-Dipyridyl↗
PubMed1996

Ascorbic acid preferentially enhances type I and III collagen gene transcription in human skin fibroblasts.

Ascorbic acid is a potent stimulator for type I and III collagen expression in human skin fibroblasts; stimulation of type I and III collagen synthesis and their mRNA levels by ascorbic acid has been reported previously. Nuclear run-on experiments demonstrated that ascorbic acid enhanced the transcription of type I and III collagen genes 4- and 3.4-fold respectively, whereas transcription of type IV collagen was slightly stimulated (1.7-fold). The results suggest that ascorbic acid preferentially enhanced type I and III collagen transcription.

Ascorbic Acid↗
PubMed1996

Role of endothelin-1 in hyperpigmentation in seborrhoeic keratosis.

Seborrhoeic keratosis (SK) is a benign epidermal tumour with a varying degree of pigmentation. We have recently demonstrated that endothelin-1 (ET-1) is a strong keratinocyte-derived mitogen and melanogen for human melanocytes in UVB-induced melanosis. To clarify the role of ET-1 in hyperpigmentation in SK, we have used immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR) to see whether the production of ET-1 is accentuated in SK. Immunohistochemical analysis in SK (n = 7; acanthotic and deeply pigmented types) revealed marked immunostaining with anti-ET-1 in almost all basaloid and basal cells as compared with definite staining confined to basal cells in the perilesional normal controls. In parallel, RT-PCR of ET-1 mRNA demonstrated accentuated expression of ET-1 transcript in SK (n = 4) in comparison with that in the perilesional normal controls, accompanied by a similarly accentuated expression of tyrosinase mRNA. These findings suggest that ET-1 plays a part in the hyperpigmentation seen in SK.

Adult↗
PubMed1996

A compound heterozygote for familial hypercholesterolaemia with a homozygous mother.

Homozygous familial hypercholesterolaemia (FH) is a rare disorder in which the patients develop severe hypercholesterolaemia and premature coronary atherosclerosis from childhood. Here we report a unique family with clustering of homozygous FH. The proband was a 25-year-old man, who showed marked hypercholesterolaemia, multiple xanthomas and severe coronary atherosclerosis. His mother also showed the typical characteristics of homozygous FH. Sequencing analysis of the low-density lipoprotein receptor gene revealed that he was a compound heterozygote, carrying two different point mutations. One was a novel mutation, FH Wakayama (Cys-->Ser at 317), derived from his mother, and the other was a recurrent mutation, FH Niigata (T-->C at 1845 + 2, 5' splice signal in intron 12), derived from his father. The proband we report seems to be a very rare case of an FH homozygote born from a homozygous mother.

Adult↗
PubMed1996

Isolation and expression of a Xenopus laevis DNA methyltransferase cDNA.

A Xenopus DNA methyltransferase cDNA was isolated from a Xenopus oocyte cDNA library by screening with the mouse DNA methyltransferase cDNA as a probe. The elucidated nucleotide sequence gave a 4,470 nucleotide open reading frame, and the predicted protein was composed of 1,490 amino acid residues, showing high homology to animal DNA methyltransferases, especially in the catalytic domain in the carboxyl-terminal region. The cysteine-rich region and the Lys-Gly repeat which were first found in the mouse sequence were conserved in Xenopus. However, 200 amino acid residues at the amino-terminus of Xenopus DNA methyltransferase were quite different from those of mouse and human, but showed 70% homology with those of chicken. The cloned Xenopus DNA methyltransferase cDNA expressed in COS1 cells showed a significant DNA methyltransferase activity. The size of the translation product of Xenopus DNA methyltransferase cDNA expressed in COS1 cells was identical with that of the endogenous DNA methyltransferase in Xenopus A6 cells and also with the size of newly synthesized DNA methyltransferase in Xenopus oocytes. However, a slightly larger immunoreactive band of about 205 kDa, and a small immunoreactive band of about 100 kDa, which were poorly labeled by short incubation with radiolabeled amino acids, were the main bands in stage I-III and stage IV-VI oocytes, respectively.

Amino Acid Sequence↗