Autocrine mechanism of epidermal growth factor in choriocarcinoma cell proliferation.

We examined four choriocarcinoma cell lines, NaUCC-1, NaUCC-3, NaUCC-4 and BeWo, for the presence of epidermal growth factor (EGF) by enzyme immunoassay and reverse transcription and polymerase chain reaction, and for EGF receptor (EGFR) by 125I-EGF binding assay. Specific EGF binding and EGF proteins were detected in these four choriocarcinoma cell lines. On the cell lines examined, NaUCC-4 had the greatest EGF binding capacity (18 x 10(5) sites/cell) and the highest amount of immunoreactive EGF (142 pg/ml).

Comparison of mutagenic activity of bile between Chilean and Japanese female patients having cholelithiasis.

The mutagenic activity of bile was compared between Chilean and Japanese female patients having cholelithiasis by the Ames assay using Salmonella typhimurium tester strain TA98 in the presence of S9 mix with blue rayon adsorption technique. A reason for conducting the present investigation is that Chile and Japan have the highest mortality rates for the gallbladder cancer (GBC) in the world. Of 24 bile samples collected in Chile, 20 (83.

Deletion of Src homology 3 domain results in constitutive activation of Tec protein-tyrosine kinase.

Tec protein-tyrosine kinase (PTK) is the prototype of a new subfamily of non-receptor type PTKs, and is abundantly expressed in hematopoietic tissues. We have revealed that Tec is inducibly tyrosine-phosphorylated and activated by stimulation with a wide range of cytokines. To get more insight into the signaling mechanism through Tec, we have generated a constitutively active form of Tec PTK.

Mutagenicity of 24-hour duplicate of Japanese diet.

In order to elucidate the genotoxicological characteristics of the Japanese diet, the mutagenicity of 24-h duplicate of the diet samples were investigated. The mutagenicity of blue rayon extract was examined in the Ames Salmonella/microsome assay. Thirty-two (91.

Tec protein tyrosine kinase is involved in the signaling mechanism of granulocyte colony-stimulating factor receptor.

Granulocyte colony-stimulating factor (G-CSF) is a critical cytokine to promote the growth, differentiation, and functional activation of myeloid cells. Despite its important roles, little is still understood how one cytokine can trigger such pleiotropic effects. By using mouse cell lines which can grow or differentiate in response to G-CSF, we investigated whether Tec protein tyrosine kinase is involved in either of the signaling pathways.

Bacterially expressed rat retinol-binding protein is functional for retinol and transthyretin bindings.

Retinol-binding protein (RBP) was expressed in Escherichia coli using the cDNA for rat RBP, and characterized. The expressed RBP was fused to maltose-binding protein (MBP) at the N-terminal end (MBP-RBP), and MBP was enzymatically removed from the MBP-RBP with proteinase factor Xa. The binding of retinol and transthyretin (TTR) to the recombinant RBP was monitored by means of gel filtration.

Human calcitonin has the same inhibitory effect on osteoclastic bone resorption by human giant cell tumor cells as salmon calcitonin.

Human calcitonin (hCT) has been reported to have a less hypocalcemizing effect on rats and to have a lower binding affinity for the receptor of mouse osteoclasts than salmon CT(sCT). In this study we comparatively examined the effect of hCT and sCT on osteoclastic bone-resorbing activity of unfractionated cells obtained from human giant cell tumor of bone and from rabbit and mouse long bones. We found that hCT had the same inhibitory effect as sCT on the bone-resorbing activity of human and rabbit osteoclastic cells, but a different one on that of mouse cells.

Mammalian mature osteoclasts as estrogen target cells.

The decrease in estrogen levels that follows the onset of menopause causes rapid bone loss, resulting in osteoporosis. However, the mechanism by which this occurs remains unclear, especially concerning the regulation of osteoclasts, i.e.

Cloning and characterization of mouse tec promoter.

Tec is a cytoplasmic protein-tyrosine kinase highly expressed in hematopoietic precursor cells. To investigate the mechanism regulating its expression, we cloned and characterized the mouse tec promoter. The promoter region does not contain the authentic TATA or CAAT box but has several consensus binding motifs for GATA and SP1.

Retinoic acid differentially up-regulates the gene expression of retinoic acid receptor alpha and gamma isoforms in embryo and adult rats.

The diverse biological effects of retinoic acid (RA) are exerted by its nuclear receptor-mediated gene expression. One of the two nuclear retinoic acid receptor subfamilies is composed of three subtypes of the all-transretinoic acid receptor (RAR-alpha, RAR-beta, and RAR-gamma). Furthermore, several isoforms are generated from each of three RARs by differential promoter usage and/or alternative splicing.

High expression of the tec gene product in murine testicular germ cells and erythroblasts.

Tec is a novel non-receptor-type protein tyrosine kinase that was originally identified from a murine liver cDNA library. While the function of Tec remains unknown, it was shown recently that two Tec-related kinases are involved directly in the growth and differentiation of bone marrow stem cells. As the localization of Tec protein has not been reported yet, immunohistochemical and immunochemical studies of various murine organs were conducted in the present study to clarify which cells express this kinase protein.

Tec protein-tyrosine kinase is an effector molecule of Lyn protein-tyrosine kinase.

The Tec family is a recently emerging subfamily among nonreceptor type protein-tyrosine kinases (PTKs) consisting of Tec, Txk, Btk, Bmx, and Itk/Tsk/Emt. They have a long amino-terminal unique region containing a pleckstrin homology domain and a Tec-homology domain. We could previously show that, through the Tec-homology domain, Tec is bound to Lyn kinase both in vitro and in vivo.

Vitamin K2 inhibits osteoclastic bone resorption by inducing osteoclast apoptosis.

In contrast to vitamin K1(VK1), vitamin K2(VK2) inhibited osteoclastic bone resorption by unfractionated bone cells and isolated osteoclasts. To investigate the mechanism of inhibition of osteoclastic bone resorption by VK2, we examined the effect of this vitamin on osteoclast apoptosis using a DNA-binding fluorescent dye, Hoechst 33258. In unfractionated bone cells and isolated osteoclasts on dentin slices, we first demonstrated that VK2 induced osteoclast apoptosis, but VK1 did not.

Physical mapping of the Tec and Gabrb1 loci reveals that the Wsh mutation on mouse chromosome 5 is associated with an inversion.

In the mouse, mutations in the c-Kit proto-oncogene, a member of the receptor tyrosine kinase (RTK) gene family, have pleiotropic effects on hematopoiesis, pigmentation and fertility (dominant spotting, W). However, in the Wsh allele the defect is confined to abnormal pigmentation caused by the disruption of 5' regulatory sequences of Kit leaving an intact structural gene. In this report, the previously published physical map around the Pdgfra-Kit-Flk1 RTK loci is extended by mapping the loci encoding the GABAA (gamma-aminobutyric acid) receptor subunit beta 1, Gabrb1 and a cytoplasmic kinase (Tec) 3 Mb proximal to Kit.

Tandem immunoaffinity chromatography for plasma 1 alpha,25-dihydroxyvitamin D3 utilizing two antibodies having different specificities: a novel and powerful pretreatment tool for 1 alpha,25-dihydroxyvitamin D3 radioreceptor assays.

We report here a novel and powerful pretreatment method for radioreceptor assays (RRAs) for human plasma 1 alpha,25-dihydroxyvitamin D3 (1,25(OH)2D3) based on "Tandem" immunoaffinity chromatography (Tandem IAC). Two antibodies having different specificities were each immobilized on agarose gel with cyanogen bromide to produce immunosorbents which were stable and repeatedly usable. An ethyl ether extract of plasma was applied to the first affinity column, from which 1,25(OH)2D3 could be preferentially eluted and separated from 1 alpha-deoxy type metabolites.

Interleukin 3 and erythropoietin induce association of Vav with Tec kinase through Tec homology domain.

Although hematopoietic cytokine receptors lack tyrosine kinase domains, the binding of their ligands to the receptors induce rapid tyrosine phosphorylation of various cellular target proteins. The specific tyrosine kinases which phosphorylate these substrates, however, have not been identified, other than that JAK kinases which phosphorylate STAT proteins and the receptors. We found that the c-vav proto-oncogene product, Vav, is rapidly and transiently tyrosine-phosphorylated in response to erythropoietin and IL3 stimulations and that Tec kinase is also transiently activated by these cytokines.

Retinoic acid directly stimulates osteoclastic bone resorption and gene expression of cathepsin K/OC-2.

Vitamin A metabolites such as all-trans-retinoic acid (all-trans-RA) affect several steps of metabolic processes in vertebrates. In the last few years, several studies have shown the effect of RA on bone formation and metabolism. However, mechanisms of its action still remain unclear, especially with respect to the regulation of bone cells.

Of the GATA-binding proteins, only GATA-4 selectively regulates the human interleukin-5 gene promoter in interleukin-5-producing cells which express multiple GATA-binding proteins.

Interleukin-5 (IL-5) is produced by T lymphocytes and known to support B-cell growth and eosinophilic differentiation of the progenitor cells. Using ATL-16T cells which express IL-5 mRNA, we have identified a region within the human IL-5 gene promoter that regulates IL-5 gene transcription. This cis-acting sequence contains the core binding motif, (A/T)GATA(A/G), for GATA-binding family proteins and thus suggests the involvement of this family members.

Hepatocyte growth factor is involved in formation of osteoclast-like cells mediated by clonal stromal cells (MC3T3-G2/PA6).

Osteoclast formation from hemopoietic precursors has been shown to require the support of stromal cells in bone tissue. In this study, we demonstrated that hepatocyte growth factor (HGF) is one of the stromal cell-derived molecules responsible for osteoclast-like cell formation. For our experiments, we used a coculture system for osteoclastic cell formation and activation in which hemopoietic blast cells are cocultured with calvaria-derived stromal MC3T3-G2/PA6 (PA6) cells on dentine slices in the presence of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3].

Expression of p210bcr/abl by metallothionein promoter induced T-cell leukemia in transgenic mice.

The p210bcr/abl chimeric protein is considered to be implicated in the pathogenesis of Philadelphia chromosome-positive human leukemias. To investigate its biologic function in vivo, we generated transgenic mice expressing p210bcr/abl driven by the metallothionein enhancer/promoter. Two of six founder mice and the transgenic progeny developed leukemias several months after birth.

Association and activation of Btk and Tec tyrosine kinases by gp130, a signal transducer of the interleukin-6 family of cytokines.

Interleukin-6 (IL-6), leukemia inhibitory factor, oncostatin M, IL-11, and ciliary neurotrophic factor constitute the IL-6 family of cytokines and play important roles in hematopoiesis, immune response, and nervous system. The receptors for the IL-6 family of cytokines share gp130 through which signals are generated, although the cytoplasmic region of gp130 does not contain any catalytic domain. In this study we show that in addition to Jak family tyrosine kinase, the stimulation of gp130 by IL-6 plus soluble IL-6 receptor alpha induced the activation of Btk and Tec tyrosine kinases, whereas IL-3 and granulocyte colony-stimulating factor activated Tec but not Btk in a pro-B cell line.

Tec protein-tyrosine kinase is involved in interleukin-3 signaling pathway.

Among cytoplasmic protein-tyrosine kinases (PTKs) Tec now forms a novel subfamily with recently identified Tec-related PTKs (Btk and Itk/Tsk). Tec is known to be abundantly expressed in myeloid cells, and multiple forms of Tec protein can be generated via the mechanism of alternative splicing. In this report, we have investigated 5'-terminal diversity of the tec messages to demonstrate a predominant form of the Tec protein in mouse hematopoietic cell lines.

Increased tyrosine-phosphorylation of 55KDa proteins in beta-actin/Tec transgenic mice.

Protein-tyrosine kinases are considered to play important roles in cell proliferation and differentiation. Tec is a cytoplasmic protein-tyrosine kinase expressed in liver and hematopoietic tissues. To better understand Tec function in vivo, we generated transgenic mice expressing tec driven by the cytomegarovirus enhancer and beta-actin promoter.

Geographical variations in mutagenicity of blue rayon extracts of Japanese human bile.

The mutagenicity of human bile was investigated in the Ames test after blue rayon treatment. In the present study, 52 and 59 bile samples were collected from the high and the low risk population for gallbladder cancer (GBC), respectively. The bile mutagenicity was detected only when the blue rayon extracts of bile were assayed with Salmonella typhimurium TA98 in the presence of S9 mix.

In vivo isomerization of retinoic acids. Rapid isomer exchange and gene expression.

The in vivo isomerization of all-trans- and 9-cis-retinoic acids (RAs) was evaluated by high performance liquid chromatography after oral administration to rats. All-trans (2 ng/ml)- and 13-cis (1.8 ng/ml)-RAs, but not 9-cis-RA, were detected in the serum of normal rats.