Search PubMed⌕ Search

Biomedical subjects

E Ogata

Publications and source records attributed to E Ogata.

At least 73 records · Page 4Linked to original sources
PubMed1996

Absence of mutations in parathyroid hormone (PTH)/PTH-related protein receptor complementary deoxyribonucleic acid in patients with pseudohypoparathyroidism type Ib.

To clarify the mechanism of resistance to PTH in patients with pseudohypoparathyroidism (PHP) type Ib, the complementary DNA (cDNA) for PTH/PTH-related protein (PTHrP) receptor was analyzed in skin fibroblasts from three patients with PHP Ib and compared with those from a patient with PHP Ia and a normal subject. We have divided the full coding region of PTH/PTHrP receptor cDNA into five parts and amplified the cDNA by reverse transcription-coupled PCR. There was no difference in the size of PCR products among these patients and the normal control. Single strand conformation polymorphism analysis of the PCR products also showed no aberrant bands in PHP Ib patients. Furthermore, no mutation in PTH/PTHrP receptor cDNA was found by direct sequencing of the PCR products from these patients. These results demonstrate that there is no mutation in PTH/PTHrP receptor cDNA from skin fibroblasts at least in the examined patients with PHP Ib. In addition, the expression of PTH/PTHrP receptor messenger ribonucleic acid was reduced in two patients but was increased in one patient with PHP Ib, suggesting that a reduction in PTH/PTHrP receptor expression cannot explain the resistance to PTH in all patients with PHP Ib. Elucidation of the pathogenesis of PHP Ib may require examination of tissue-specific abnormality in the PTH signal transduction system in the kidney.

Adolescent↗
PubMed1996

[Serum concentration of pyridinoline cross-linked carboxy-terminal telopeptide of type-I collagen (ICTP) and carboxyterminal propeptide of human type I procollagen (PICP) in the diagnosis of bone metastases].

Recently discovered bone metabolic markers are expected to play an additional role in the diagnosis of bone metastasis. We measured bone metabolic markers, serum pyridinoline cross-linked carboxy-terminal telopeptide of type I collagen (ICTP) and carboxyterminal propeptide of human type I procollagen (PICP) in 224 patients with breast cancer (106 with bone metastases), 61 patients with prostatic cancer (30 with bone metastases), 45 patients with lung cancer (17 with bone metastases) and 13 patients with miscellaneous cancers (7 with bone metastasis) and compared the values in the presence and absence of bone metastasis. ICTP and PICP increased significantly in patients with bone metastases. By the analysis of sensitivity and specificity, the cut-off levels were considered to be 5.0 ng/ml for ICTP and 140 ng/ml for PICP. In lung cancer (bone metastases are mostly of osteolytic), ICTP was excellent marker in detecting bone metastasis. In breast cancer (bone metastases are mostly of mixed type), ICTP was good in detecting bone metastases. In prostatic cancer (bone metastases are mostly of osteoblastic), ICTP and PICP were good markers in detecting high grade of bone metastases. Over all, ICTP was more sensitive in the diagnosis of bone metastases than PICP. However, both markers were not effective in detecting low grade bone metastases. ICTP and PICP should play a supportive role to imaging modalities in the diagnosis of bone metastases.

Aged↗
PubMed1995

Neoplastic transformation of normal rat embryo fibroblasts by a mutated p53 and an activated ras oncogene induces parathyroid hormone-related peptide gene expression and causes hypercalcemia in nude mice.

Parathyroid hormone-related peptide (PTHRP) is a 141-amino acid protein identified in various carcinomas associated with humoral hypercalcemia of malignancy (HHM). Although the causal role of PTHRP in HHM syndrome has been established, the molecular and cellular mechanism by which PTHRP gene is overexpressed in certain malignancies remains unknown. We have demonstrated in the present study that PTHRP secretion was markedly induced concomitantly with the formation of transformed foci after normal rat embryo fibroblasts (REFs) were co-transfected with an activated ras (ras) and a mutated form of p53 (p53-mt) genes. In either ras- or p53-mt-transfected (nontransformed) cells, only modest or barely detectable secretion of PTHRP was observed, respectively. Northern blot analysis revealed that PTHRP mRNA was markedly induced in fully transformed cells 11 days after transfection with both ras and p53-mt genes. Inhibition of RNA synthesis with actinomycin D resulted in almost complete disappearance of PTHRP mRNA at 2-3 h, suggesting a transcriptional mechanism. Transient transfection experiments revealed that PTHRP promoter activity was induced in ras + p53-mt transfectants. REFs transformed by ras and p53-mt genes and thereby induced to secrete PTHRP in vitro produced aggressively growing tumors associated with HHM syndrome when injected into nude mice. These results suggest that activation of PTHRP gene is closely related to malignant transformation of normal mammalian cells and that ras and p53 may be important regulators of PTHRP gene transcription. The transfection-focus formation system of REFs should provide an excellent model to study the molecular and cellular mechanism underlying concomitant overexpression of PTHRP gene with carcinogenesis.

Animals↗
PubMed1995

Binding of vitamin D to low-density-lipoprotein (LDL) and LDL receptor-mediated pathway into cells.

The present study was undertaken to identify serum components other than vitamin D-binding proteins that bind to 1,25(OH)2 D3, and its analog. The binding rate of 1,25(OH)2 D3, 22-oxa-1,25(OH)2D3 (OCT) or 25(OH) D3 to total lipoprotein(TLP) represented 16.7%, 4.65%, and 3.11% of total counts added, respectively. Polyacrylamide gel electrophoresis of the TLP revealed that 1,25(OH)2 D3 and OCT were associated with LDL. The binding studies of OCT-bound LDL to the fibroblasts showed specific pathway to the cells mediated by LDL-receptor. These findings may have important implications in understanding the mechanisms of the diverse biological actions of 1,25(OH)2 D3 and in designing a novel delivery system for vitamin D analogs.

Apolipoprotein B-100↗
PubMed1995

Ligand-dependent G protein coupling function of amyloid transmembrane precursor.

Amyloid precursor protein (APP), a transmembrane precursor of beta-amyloid, possesses a function whereby it associates with G(o) through its cytoplasmic His657-Lys676. Here we demonstrate that APP has a receptor function. In phospholipid vesicles consisting of baculovirally made APP695 and brain trimeric G(o), 22C11, a monoclonal antibody against the extracellular domain of APP, increased GTP gamma S binding and the turnover number of GTPase of G(o) without affecting its intrinsic GTPase activity. This effect of 22C11 was specific among various antibodies and was observed neither in G(o) vesicles nor in APP695/Gi2 vesicles. In APP695/G(o) vesicles, synthetic APP66-81, the epitope of 22C11, competitively antagonized the action of 22C11. Monoclonal antibody against APP657-676, the G(o) binding domain of APP695, specifically blocked 22C11-dependent activation of G(o). Therefore, APP has a potential receptor function whereby it specifically activates G(o) in a ligand-dependent and ligand-specific manner.

Amyloid beta-Protein Precursor↗
PubMed1995

In vivo evidence for progressive activation of parathyroid hormone-related peptide gene transcription with tumor growth and stimulation of osteoblastic bone formation at an early stage of humoral hypercalcemia of cancer.

The present study was undertaken to clarify in vivo the temporal profile of parathyroid hormone-related peptide (PTHRP) gene expression as well as bone histomorphometric features as a function of tumor growth, using an athymic rat model associated with humoral hypercalcemia of malignancy (HHM). Tumor-bearing animals exhibited hypercalcemia, hypophosphatemia, and increased circulating levels of PTHRP, and died within 3 weeks. Steady-state PTHRP mRNA levels and the transcription rate of PTHRP gene in the tumors were markedly increased with tumor growth. RNAse mapping analysis revealed that both upstream and downstream promoters of the human PTHRP gene were utilized in the tumors and became progressively activated with time. Bone histomorphometric analysis showed that osteoclastic bone resorption was progressively increased throughout the course, whereas osteoblastic bone formation was stimulated more than 2-fold at a very early stage (day 6 after tumor implantation) and then markedly suppressed thereafter on day 12 and day 18 compared with age-matched control animals. These results provide in vivo evidence that PTHRP gene transcription is progressively activated with tumor growth and that activation of osteoblasts does occur at a very early phase of HHM syndrome in contrast to the marked suppression of bone formation at later stages.

Analysis of Variance↗
PubMed1995

Humoral hypercalcemia of malignancy: some enigmas on the clinical features.

Humoral hypercalcemia of malignancy (HHM) is a common paraneoplastic syndrome mediated by tumor-derived parathyroid hormone-related peptide (PTHRP), which bears structural and functional similarities to PTH. Thus the clinical features of HHM are very similar to those of primary hyperparathyroidism (1 degree HPT), a prototype of humoral hypercalcemia caused by PTH. On the other hand, HHM syndrome differs from 1 degree HPT in several aspects, including serum 1,25(OH)2D levels, acid-base balance, and bone remodeling process, the reason of which remains largely unknown. We approached these questions using a unique animal model of HHM, nude rats implanted with PTHRP-overproducing human carcinomas. In this review we will summarize the results and discuss the implications in understanding the disease mechanism.

Animals↗
PubMed1995

[A drug-induced transient Fanconi syndrome associated with pure red cell aplasia: pathophysiology of the electrolyte disorders].

A 55-year-old female with a long history of pure red cell aplasia temporarily manifested Fanconi syndrome with hypophosphatemia, hypouricemia, glycosuria, acidic amino aciduria, but not metabolic acidosis nor hypokalemia. These abnormalities completely resolved in 3-4 weeks after withdrawal of several drugs, suggesting that the cause of her Fanconi syndrome could be attributed to a drug or a combination of multiple drugs, though lymphocyte stimulating tests revealed negative results for all possible drugs. Postmortem specimen of her kidneys showed mild mesangial proliferation without significant changes in tubular and interstitial regions. Massive ferrous precipitations were found in the zona glomerulosa of the adrenal glands. The pathophysiology of Fanconi syndrome shown in the patient was likely to have been a drug-induced transient and mild dysfunction of the Na+/K+ pump of the renal proximal tubules, which might also explain the selective amino aciduria. The absence of hypokalemia corroborates well with both a lack of bicarbonaturia and hemochromatosis-induced adrenal insufficiency. Patients with a renal dysfunction associated with electrolyte derangements and without proteinuria or azotemia should be under vigilant observation when using many drugs.

Fanconi Syndrome↗
PubMed1994

Evidence for the uptake of a vitamin D analogue (OCT) by a human carcinoma and its effect of suppressing the transcription of parathyroid hormone-related peptide gene in vivo.

The present study was undertaken to clarify the pharmacokinetics of 22-oxa-1,25-dihydroxyvitamin D3 (22-oxa-1,25-(OH)2D3, OCT), a vitamin D3 analogue with little calcemic activity, and its effect on the transcription of parathyroid hormone-related peptide (PTHRP) gene in nude mice bearing a human carcinoma (FA-6) associated with humoral hypercalcemia. FA-6 tumor expressed vitamin D receptor (VDR) mRNA, and its nuclear extract contained a specific and saturable 1,25-(OH)2D3 binding activity. Although [3H]OCT administered intravenously into FA-6 tumor-bearing nude mice was cleared from the circulation more rapidly than [3H]1,25-(OH)2D3, the uptake of [3H]OCT into the tumor tissue, relative to the radioactivity in the circulation, was greater than that of [3H]1,25-(OH)2D3. Intravenous or oral administration of OCT reduced the steady-state levels of PTHRP mRNA in FA-6 tumor, and nuclear run-off assays demonstrated that the effect of OCT on PTHRP gene expression occurred at a transcriptional level. RNase mapping analysis revealed that both upstream and downstream promoters of the human PTHRP gene were down-regulated by OCT. Finally, OCT exerted a preventive as well as therapeutic effect on cancer-associated hypercalcemia with a marked prolongation of the survival time in tumor-bearing animals. These results suggest that OCT is effectively taken up by a VDR-positive human carcinoma in vivo and has a therapeutic potential for cancer-associated hypercalcemia through suppression of PTHRP gene transcription.

Animals↗
PubMed1994

Bidirectional regulation of c-fos promoter by an oncogenic gip2 mutant of G alpha i2. A novel implication of retinoblastoma gene product.

G alpha i2 is a tissue-specific proto-oncogene product, whose activated mutant gip2 induces transformation through less defined downstream pathways. We found that c-fos promoter is a target of gip2 in multiple kinds of cells. Serum response element was shown to be the positive enhancer element that mediates gip2-induced c-fos expression. We further demonstrated that gip2 stimulates the negative silencer activity of the retinoblastoma (Rb) control element (RCE) and inhibits the c-fos promoter activity through RCE located in the c-fos promoter region. The effect of gip2 on RCE was shown to be mediated by the Rb gene product (pRb). Furthermore, gip2 augmented underphosphorylated active form of pRb by promoting pRb expression and by affecting the phosphorylation state of pRb. gip2 therefore propagates both positive and negative signals to the c-fos promoter through two different elements, and pRb mediates the negative signal of gip2. We conclude that gip2 has bifunctional roles in transformation which pRb critically regulates. Given that Rat-1 cells, which gip2 can transform, lack the sensitivity to the gip2/pRb-mediated negative pathway, this study provides a novel insight into oncogenesis by gip2 and its tissue specificity.

Animals↗
PubMed1994

A redox factor protein, ref1, is involved in negative gene regulation by extracellular calcium.

A rise in extracellular calcium (Ca2+e) suppresses not only secretion of parathyroid hormone (PTH) but also expression of the PTH gene to ensure constant plasma Ca2+ level. A nuclear protein(s) in a wide variety of cells bound to the specific DNA elements (negative Ca2+ responsive elements, nCaREs) in the human PTH gene in sequence-specific and Ca2+e concentration-dependent manners. Our Southwestern cloning revealed that a redox factor protein (ref1), which was known to activate several transcription factors via alterations of their redox state, belonged to an nCaRE binding protein. The level of ref1 mRNA as well as of its protein was elevated by an increase in Ca2+e concentration. In gel shift assay, anti-ref1 antibody eliminated formation of the nCaRE-protein complex. We also found that there was another protein(s) interacting with nCaREs and ref1. Further, experiments with an antisense-ref cDNA expression vector introduced into cultured cells suggested that DNA (nCaRE)-ref1 interaction led to Ca2+e-mediated transcriptional suppression. Thus, it is concluded that ref1 possesses transcription repressor activity in addition to its function as a transcriptional auxiliary protein.

Animals↗
PubMed1994

Inhibition of inwardly rectifying K+ current by external Ca2+ ions in freshly isolated rabbit osteoclasts.

1. Regulation of membrane potential by extracellular Ca2+ concentration ([Ca2+]o) was examined in freshly isolated rabbit osteoclasts. 2. The resting membrane potential of osteoclasts was close to the K+ equilibrium potential in 1 mM Ca2+ medium. An elevation of [Ca2+]o caused membrane depolarization, accompanied by a decrease in the membrane conductance. 3. The inwardly rectifying K+ current observed under voltage clamp was dose-dependently inhibited by an elevation of [Ca2+]o, which explained the membrane depolarization caused by high [Ca2+]o. 4. Other divalent cations also inhibited the inwardly rectifying K+ current with the following order of potency: Ca2+ < Ni2+ < or = Co2+ < Cd2+. 5. In the presence of intracellular GTP gamma S the inwardly rectifying K+ current was irreversibly inhibited by [Ca2+]o, whereas the inhibition of the inwardly rectifying K+ current was greatly attenuated by intracellular application of GDP beta S. 6. Pertussis toxin (PTX) treatment did not abolish the inhibition of the inwardly rectifying K+ current caused by [Ca2+]o. 7. These results suggest that inwardly rectifying K+ channels in osteoclasts were regulated by a PTX-insensitive G-protein, which was coupled to the putative Ca2+ receptor or sensor on the cell membrane.

Animals↗
PubMed1994

An intrinsic guanine nucleotide exchange inhibitor in Gi2 alpha. Significance of G-protein self-suppression which antagonizes receptor signal.

The alpha subunit of Gi2 (Gi2 alpha) is a member of the heterotrimeric G protein family, which transduces receptor signals as a proto-oncogene product. We have found a novel self-suppressive region in Gi2 alpha near its C terminus. A polypeptide consisting of residues 338-352 of Gi2 alpha (Gi2 alpha-339-352) antagonizes receptor- and receptor peptide-stimulated Gi2 alpha activation, without affecting basal activity. Antagonism by Gi2 alpha-338-352 is attributable to an interaction with activated Gi2 alpha, which is not competitive with receptor polypeptides. Combined with the reports suggesting the presence of self-suppressive domains in a juxta-C-terminal portion of Gi2 alpha and G(o) alpha, this study supports the hypothesis that Gi2 alpha-338-352 constitutes an intrinsic guanine nucleotide exchange inhibitor, which in turn antagonizes receptor stimulation, suggesting that G proteins are activated by receptors through relaxation of a self-suppressive conformation.

Amino Acid Sequence↗
PubMed1994

Effects of activin A on ionic channels in human FSH-secreting tumour cells.

1. Effects of activin A on ionic channels were examined in human FSH-secreting tumour cells using electrophysiological techniques. 2. Under voltage clamp with the conventional whole-cell clamp technique, the voltage-gated Na+ channel, the T- and L-type Ca2+ channels, the delayed K+ channel and the A-channel were observed. 3. With the nystatin-perforated whole-cell clamp technique, the same voltage-gated channels were recorded. Activin A (10(-7) M) increased the amplitude of the L-type Ca2+ current, whereas it decreased the amplitude of the delayed K+ current. 4. Under current clamp with the perforated whole-cell clamp technique, more than 80% of the cells exhibited spontaneous action potentials. Application of 10(-7) M activin A depolarized the membrane with a conductance increase and augmented action potential frequency. The reversal potential of the activin A-induced current was -20 to 0 mV. The activin A-induced current was abolished in a Na(+)-free extracellular solution, indicating that the membrane depolarization caused by activin A was due to the conductance increase to Na+ ions through non-selective cation channels.

Activins↗
PubMed1994

Bimodal effect of transforming growth factor-beta on insulin secretion in MIN6 cells.

The effects of transforming growth factor-beta (TGF-beta) on insulin secretion were investigated using a glucose-responsive clonal cell line, MIN6. One hundred pM TGF-beta stimulated insulin release during 0.5-24 h of incubation in the presence of 5.5 mM glucose, but not after 48 h; 1 nM TGF-beta also stimulated insulin release up to 2 h of exposure, but the effect was not seen after 6 h of exposure. When cells were incubated with 25 mM glucose for 24 h, 100 pM TGF-beta significantly inhibited glucose-stimulated insulin release, whereas insulin release was not altered at 0 or 2.8 mM glucose. On the contrary, forskolin- (10 microM) and tolbutamide- (40 microM) induced insulin release were not affected by TGF-beta. TGF-beta affected neither the cell growth nor the cellular insulin content. An addition of 1 microM nitrendipine abolished TGF-beta-induced insulin secretion at 5.5 mM glucose. The presence study shows that TGF-beta exerts a bimodal effect on glucose-induced insulin secretion from MIN6 cells, depending on dose, time of exposure and concentrations of coexisting glucose. These effects might be mediated by the Ca(2+)-dependent mechanism.

Animals↗