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Biomedical subjects

K Arai

Publications and source records attributed to K Arai.

At least 397 records · Page 22Linked to original sources
PubMed1995

Epidemiological analysis of strains of methicillin-resistant Staphylococcus aureus (MRSA) infection in the nursery; prognosis of MRSA carrier infants.

Forty-five neonates who carried methicillin-resistant Staphylococcus aureus (MRSA) were studied. Retrospective molecular analysis using pulsed-field gel electrophoresis showed three separate MRSA epidemics in the nursery. Strains of MRSA isolated from the neonates were also isolated from the hospital environment and health care providers. Clinical manifestations included skin pustules (eight patients), conjunctivitis (four patients), or other minor infections (two patients). No neonate developed systemic infection. The prevalence of MRSA decreased with age. At one year, three (14.3%) of 21 infants that had carried MRSA at six days remained carriers and only two (1.1%) of 180 infants in a control 'S. aureus-negative at six days' group carried MRSA.

Age Distribution↗
PubMed1995

Synergistic action of interleukin-10 (IL-10) with IL-3, IL-4 and stem cell factor on colony formation from murine mast cells in culture.

We examined the effects of interleukin-10 (IL-10) on colony formation from two different types of murine mast cells, bone marrow-derived mast cells (BMMC) and serosal mast cells (SMC), using a methylcellulose culture method. IL-10 alone did not induce colony formation from either phenotype. However, BMMC and SMC produced colonies in the presence of IL-10 in combinations with IL-3, IL-4 or stem cell factor (SCF), but they responded in different manners. IL-10 enhanced the IL-3-dependent colony formation from BMMC and induced the colony formation from BMMC synergistically with IL-4 or SCF dose-dependently, although IL-4 and SCF were not active alone. The most significant synergism was observed between IL-10 and IL-4. The addition of IL-10 to the cultures of BMMC in the presence of two or three factors enhanced the colony formation induced by IL-3 plus IL-4, and inhibited the colony formation induced by IL-3 plus SCF or IL-3, IL-4 plus SCF. In the colony formation from SMC, IL-10 synergized with IL-3 but not with IL-4 or SCF. IL-10 in combination with two or three factors enhanced the colony formation from SMC induced by IL-3 plus IL-4, but did not affect the colony formation induced by other combinations among IL-3, IL-4 and SCF. These findings indicate that IL-10 plays an important role in the proliferation of murine mast cells.

Animals↗
PubMed1995

Mice deficient for the IL-3/GM-CSF/IL-5 beta c receptor exhibit lung pathology and impaired immune response, while beta IL3 receptor-deficient mice are normal.

The receptors for IL-3, GM-CSF, and IL-5 share a common beta subunit (beta c), and mice have an additional IL-3 beta subunit (beta IL3). We have independently generated mice carrying null mutations of each molecule. beta c mutant bone marrow showed no response to GM-CSF or IL-5, whereas IL-3 stimulation of beta c or beta IL3 mutant bone marrow was normal. beta c mutant mice showed lung pathology consisting of lymphocytic infiltration and areas resembling alveolar proteinosis, and also exhibited low basal numbers of eosinophils. Infection of beta c mutant mice by Nippostrongylus brasiliensis resulted in the absence of blood and lung eosinophilia. Animals repopulated with beta c mutant bone marrow cells showed slower leukocyte recovery and reduced eosinophil numbers. These data define the role of beta c in vivo, and show a phenotype that is likely to be the cumulative effect of loss of GM-CSF and IL-5 stimulation.

Animals↗
PubMed1995

Differential expression of granulocyte-macrophage colony-stimulating factor and IL-3 receptor subunits on human CD34+ cells and leukemic cell lines.

Cytokines transduce their signals through specific receptors. Receptors for granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-3, and IL-5 share the common signal transducing subunit (beta c), whereas the alpha subunits function as specific ligand binding components. In this study we prepared specific mouse monoclonal antibodies against human GM-CSF receptor-alpha subunit (hGMR alpha) by immunizing mice with Ba/F3 cells transfected with hGMR alpha complementary DNA. Using these anti-hGMR alpha antibodies in combination with antibodies against IL-3 receptor-alpha (IL-3R alpha), beta c subunits, and c-kit, we examined expression patterns and modulation of these receptor subunits on several human hematopoietic cells, including CD34+ cells and leukemic cell lines. GMR alpha and IL-3R alpha were expressed on GM-CSF- and IL-3-responsive cell lines, such as TF-1 and UT-7, whereas the expression levels were much lower on UT-7E, a GM-CSF- and IL-3-unresponsive subline of UT-7. The GMR alpha subunit was expressed only on mature granulocytes and monocytes, and IL-3R alpha was expressed on monocytes but not on mature granulocytes, and none of these subunits were expressed on lymphocytes. For CD34+ cells, GMR alpha was expressed more abundantly on CD34+ CD33high cells than on CD34+ CD33low cells, whereas IL-3R alpha was expressed more abundantly on CD34+ CD33low cells than on CD34+ CD33high and CD34+ CD33neg cells. Slight but significant expression of the beta c subunit was detected on CD34+ cells. Expression of not only GMR alpha and IL-3R alpha subunits but also c-kit was specifically downregulated by 48-hour incubation with their respective ligands. Receptor transmodulation between GM-CSF, IL-3, and stem cell factor (or kit ligand) was not detected on CD34+ cells in 48-hour cultures. We also detected upregulation of these alpha subunits by IL-1 alpha and interferon-gamma on leukemic cell lines. Our study showed expression levels for each receptor subunit--including GMR, IL-3R, and c-kit on human bone marrow and peripheral blood cells and leukemic cell lines--and revealed differential regulation of the expression of the receptor subunits.

3T3 Cells↗
PubMed1995

Roles of the cytoplasmic domains of the alpha and beta subunits of human granulocyte-macrophage colony-stimulating factor receptor.

The high-affinity and functional granulocyte-macrophage colony-stimulating factor receptor (GMR) is composed of two distinct subunits, alpha and beta; and the cytoplasmic domain of the beta subunit is essential to transduce growth-promoting signals. In contrast to the beta subunit, the role of the alpha subunit is not well characterized. We examined the requirement of the cytoplasmic domain of the alpha subunit and its functional region by deletion analyses. We demonstrated that the cytoplasmic domain of the alpha subunit, especially 29 amino acids residues near the transmembrane domain, was absolutely required for various signaling events including activation of immediate early genes, induction of tyrosine phosphorylation of cellular proteins, and cell growth. We further analyzed the role of the cytoplasmic domain of each subunit by constructing chimeric subunits, designated alpha/beta and beta/alpha, by exchanging cytoplasmic domains of the alpha and beta subunits of human (h) GMR. Reconstituted high-affinity chimeric hGMRs, hGMR(alpha/beta,beta/alpha) and hGMR(alpha/beta,beta), transduced signals at levels similar to the wild type hGMR(alpha,beta) in Ba/F3 cells and in NIH3T3 cells. These observations indicate that the original configuration between the extracellular and the cytoplasmic domains of the hGMR(alpha,beta) subunits is not required and that hGMR(alpha/beta,beta) transduced signals through the cytoplasmic domain of the beta subunit in an oligomeric form, without involvement of the cytoplasmic domain of the alpha subunit. Therefore human granulocyte-macrophage colony-stimulating factor signals are mainly transduced through the beta subunit, and the cytoplasmic domain of the alpha subunit is likely to activate the beta subunit in the normal hGMR.

3T3 Cells↗
PubMed1995

Gene structures of the alpha subunits of human IL-3 and granulocyte-macrophage colony-stimulating factor receptors: comparison with the cytokine receptor superfamily.

Recently, many genes encoding the members of the cytokine receptor superfamily (CRSF), which have common structural features, have been characterized. Analyses on the structures of the genes encoding the alpha subunits of human IL-3 (hIL-3R alpha) and granulocyte-macrophage colony-stimulating factor receptors (hGMR alpha) revealed that they have the structural features common to all members of the CRSF (i.e., conservation of the intron phase pattern as "1-2-1-0-1" rule in the fibronectin type III domains located in extracellular segments of type I cytokine receptor subunits. This finding led us to propose a possible model for gene evolution for the CRSF. We pointed out that the CRSF genes derived from a putative common ancestral gene. In addition to these common features, we found an additional intron that is unique to the IL-3R alpha and the GMR alpha genes. This additional intron suggests that the IL-3R alpha and the GMR alpha genes evolved closely in the evolution process of the CRSF genes. This evidence and results of recent studies on the evolution of mammalian X chromosome make it tempting to speculate that a putative common ancestral gene of the subfamily including IL-3R alpha, GMR alpha, and IL-5R alpha emerged in an autosome at least before the divergence of marsupials and eutherian mammals, early in the 200 million-year history of mammals.

Base Sequence↗
PubMed1995

Regulation of expression of the IL-2 and IL-5 genes and the role of proteins related to nuclear factor of activated T cells.

Cyclic adenosine monophosphate (cAMP) inhibits phorbol 12-myristate 13-acetate (PMA)-induced IL-2 production while it inhibits IL-5 production at the transcriptional level in EL-4, a mouse lymphoma line. The -321 to +46 region of the mouse IL-2 promoter is required for activation by PMA and inhibition by cAMP. This promoter region contains several elements that interact with transcription factors, such as nuclear factor of activated T cells (NF-AT), NF-kappa B, AP-1, and octamer. With use of reporter plasmid carrying multiple copies of each element, we found that the construct that contained the NF-AT site was most effective for responding to PMA activation and cAMP inhibition. In electrophoretic mobility shift assay (EMSA), PMA-induced NF-AT binding complex was altered by cAMP. Furthermore, overexpression of the cytoplasmic component of NF-AT abrogated the inhibitory action of cAMP. These results indicate that the NF-AT site is a target of the inhibitory action of cAMP. We have previously reported that the -1200 to +33 region of the mouse IL-5 promoter can mediate transcriptional stimulation by PMA and cAMP in EL-4 cells. Here we identified the element IL-5P, which is required for maximal activation of the IL-5 promoter. We found that this element is homologous to the binding site for NF-AT and interacted with NF-AT-related factors induced by PMA and cAMP. Thus it appears that an NF-AT factor is involved in the regulation of IL-5 gene transcription.

Animals↗
PubMed1995

A human GM-CSF receptor expressed in transgenic mice stimulates proliferation and differentiation of hemopoietic progenitors to all lineages in response to human GM-CSF.

Granulocyte-macrophage colony-stimulating factor (GM-CSF) mainly stimulates proliferation and maturation of myeloid progenitor cells. Although the signal transduction pathways triggered by GM-CSF receptor (GMR) have been extensively characterized, the roles of GMR signals in differentiation have remained to be elucidated. To examine the relationship between receptor expression and differentiation of hemopoietic cells, we used transgenic mice (Tg-mice) that constitutively express human (h) GMR at almost all stages of hemopoietic cell development. Proliferation and differentiation of hemopoietic progenitors in bone marrow cells from these Tg-mice were analyzed by methylcellulose colony formation assay. High affinity GMR interacts with GM-CSF in a species-specific manner, therefore one can analyze the effects of hGMR signals on differentiation of mouse hemopoietic progenitors using hGM-CSF. Although mouse (m) GM-CSF yielded only GM colonies, hGM-CSF supported various types of colonies including GM, eosinophil, mast cell, erythrocyte, megakaryocyte, blast cell, and mixed hemopoietic colonies. Thus, the effects of hGM-CSF on colony formation more closely resembled mIL-3 than those of mGM-CSF. In addition, hGM-CSF generated a much larger number of blast cell colonies and mixed cell colonies than did mIL-3. hGM-CSF also generated erythrocyte colonies in the absence of erythropoietin. Therefore, GM-CSF apparently has the capacity to promote growth of cells of almost all hemopoietic cell lineages, if functional hGMR is present.

Animals↗
PubMed1995

Characterization of cis-regulatory elements of the c-myc promoter responding to human GM-CSF or mouse interleukin 3 in mouse proB cell line BA/F3 cells expressing the human GM-CSF receptor.

Interleukin 3 (IL-3) or granulocyte macrophage colony-stimulating factor (GM-CSF) activates c-fos, c-jun, and c-myc genes and proliferation in both hematopoietic and nonhematopoietic cells. Using a series of deletion mutants of the beta subunit of human GM-CSF receptor (hGMR) and inhibitors of tyrosine kinase, two distinct signaling pathways, one for activation of c-fos and c-jun genes, and the other for cell proliferation and activation of c-myc gene have been elucidated. In contrast to wealth of information on the pathway leading to activation of c-fos/c-jun genes, knowledge of the latter is scanty. To clarify the mechanisms of activation of c-myc gene by cytokines, we established a transient transfection assay in mouse proB cell line BA/F3 cells expressing hGMR. Analyses of hGMR beta subunit mutants revealed two cytoplasmic regions involved in activation of the c-myc promoter, one is essential and the other is dispensable but enhances the activity. These regions are located at the membrane proximal and the distal regions covering amino acid positions 455-544 and 544-589, respectively. Characterization of cis-acting regulatory elements of the c-myc gene showed that the region containing the P2 promoter initiation site is sufficient to mediate the response to mIL-3 or hGM-CSF. Electrophoretic mobility shift assay using an oligonucleotide corresponding to the distal putative E2F binding site revealed that p107/E2F complex, the negative regulator of E2F, decreased, and free E2F increased after mIL-3 stimulation. These results support the thesis that mIL-3 or hGM-CSF regulates the c-myc promoter by altering composition of the E2F complexes at E2F binding site.

Animals↗
PubMed1995

Intrathecal administration of N-methyl-D-aspartate receptor antagonist reduces the minimum alveolar anaesthetic concentration of isoflurane in rats.

We have studied the effect of intrathecal administration of N-methyl-D-aspartate (NMDA) receptor antagonists on the minimum alveolar anaesthetic concentration (MAC) of isoflurane in rats. In Wistar rats fitted with indwelling intrathecal catheters, we determined the MAC of isoflurane after administration of a competitive NMDA receptor antagonist, APV (0.01, 0.1, 1.0, 10, 30 micrograms), a non-competitive NMDA receptor antagonist, MK801 (0.1, 1.0, 10, 30 micrograms). NMDA (0.01, 0.1, 1.0, 10, 30 micrograms) and saline. APV at all doses except 0.01 micrograms decreased MAC by 17.1-32% (P < 0.001 and P < 0.0001). Although MK801 at 10 and 30 micrograms reduced MAC by 24.3-31.7% (P < 0.001 and P < 0.0001), lower doses did not affect MAC. Intrathecal administration of NMDA reversed these decreases in MAC, but not to control values with APV 10 and 30 micrograms and MK801 30 micrograms. We suspect that NMDA and NMDA receptor antagonists play important roles in the spinal cord in determining the MAC of isoflurane.

2-Amino-5-phosphonovalerate↗
PubMed1995

Immunoneutralization of inhibin and estradiol during the follicular phase of the estrous cycle in cows.

To investigate the physiological importance of inhibin and estradiol in the regulation of FSH secretion during the follicular phase of the estrous-cycle in cows, animals were passively immunized against the two hormones. Sixteen cows were divided into four equal groups and given injections of prostaglandin F2 alpha (PG) i.m. twice at 8-h intervals on Day 10 of the estrous cycle (Day 0 = day of estrus) to induce luteal regression. At 48 h after the first PG injection, each group of four cows received injections of one of the following: 100 ml castrated goat serum (control serum), 100 ml antiserum against inhibin, 100 ml antiserum against estradiol, or a combination of 100 ml inhibin antiserum and 100 ml estradiol antiserum. The LH surge occurred within 2 days after injection of the control serum or inhibin antiserum, whereas it was not detected in either of the groups passively immunized against estradiol, indicating that passive immunization against estradiol blocks a positive feedback effect of estradiol on LH secretion. There was no clear difference in basal concentrations of LH among the four groups. Injection of the inhibin antiserum resulted in a marked increase (p < 0.01) in plasma concentrations of FSH compared with values in the control group, while there were no significant changes in concentrations of plasma FSH after injection of the estradiol antiserum. Combined administration of the inhibin and estradiol antisera also produced a marked increase (p < 0.01) in plasma concentrations of FSH. The FSH response to the combined immunization was longer in duration than the FSH response to immunization with inhibin alone.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Use of recovery-enhanced thermography to localize cutaneous perforators.

Recovery-enhanced thermography implies the scanning, after cooling, of the skin surface with ice water for several seconds. Using this method, cutaneous perforators were clearly identified in constant distributions on the trunks of 12 healthy volunteers. Clinically, preoperative recovery-enhanced thermography is useful for the design of perforator-based flaps. We describe this method in detail and demonstrate its reliability with successful clinical cases.

Adult↗
PubMed1995

Simultaneous expression of type IX collagen and an inhibin-related antigen in proliferative myoepithelial cells with pleomorphic adenoma of canine mammary glands.

To identify an alcian blue-positive component expressed in the early stage of mammary mixed tumor and to pursue the possible involvement of a tumorigenic inducing factor, monoclonal antibodies to type IX collagen were generated and used to investigate the immunohistochemical kinetics of type IX collagen expression by the myoepithelial cell-derived chondrocyte-like cells during the development of chondrometaplasia. We also examined the expression of inhibin-related antigen using antibodies to a synthetic peptide spanning amino acids 1-30 of the inhibin alpha chain. At the earliest stage of chondrometaplasia, where myoepithelial cells began to proliferate inside the basement membrane, the cells expressed type IX collagen together with an inhibin-related antigen which was immunoreactive with the anti-inhibin peptide antibodies. The expression of the inhibin-related antigen was also demonstrated in normal embryonic chondrocytes and myoblasts, but was much less strong in mature chondrocytes and myotubes, strongly suggesting that the inhibin-related antigen is involved in the development of chondrocytes and myoblasts from undifferentiated mesenchymal cells as well as proliferating myoepithelial cells as a chondro-progenitor cell in the mammary mixed tumor. The pathophysiological significance of type IX collagen expression as a possible cell marker of the progenitor cell in myoepithelial cell-related chondrometaplasia is also discussed.

Adenoma, Pleomorphic↗
PubMed1995

Basic fibroblast growth factor as a candidate tumor marker for renal cell carcinoma.

The present investigation was conducted to determine serum levels of basic fibroblast growth factor (FGF) by enzyme immunoassay in patients with various urogenital tumors. Renal cell carcinoma had a higher tendency (28 of 52, 53.8%) toward increased serum levels of basic FGF than any of the other urogenital tumors, and increased serum basic FGF was detected more frequently in patients with advanced renal cell carcinoma. Analysis of histological pattern indicated that renal cell carcinoma with a solid or tubular component is more likely to produce basic FGF. However, no significant difference was seen between the percentage of clear cell type tumor patients with increased serum basic FGF (50.0%) and the percentage of granular cell type tumor patients with increased serum basic FGF (66.7%). Five of 8 patients with renal cell carcinoma who underwent selective renal venous sampling before nephrectomy showed increased serum basic FGF in the renal vein from the affected kidney. After resection of the affected kidney to remove the cancer, serum basic FGF disappeared within 2 weeks. However, residual huge tumor or postoperative disease prolonged the increased levels of basic FGF in 2 patients, indicating that basic FGF is produced from and secreted by tumor tissue itself. These findings suggest that serum basic FGF can be useful in the diagnosis, and in evaluating the prognosis, of patients with renal cell carcinoma.

Biomarkers, Tumor↗
PubMed1995

Expression of erythroid-specific genes in acute megakaryoblastic leukaemia and transient myeloproliferative disorder in Down's syndrome.

Acute megakaryoblastic leukaemia (M7) and transient myeloproliferative disorder in Down's syndrome (TMD) are characterized by rapid growth of abnormal blast cells which express megakaryocytic markers. To clarify properties of the blast cells in M7 and TMD cases, we examined erythroid markers expression in blasts from six cases with M7 and seven cases with TMD in this study. Erythroid-specific mRNAs encoding gamma-globin and erythroid delta-aminolevulinate synthase were found to be expressed in blasts from most of these cases, indicating that majorities of the blasts in M7 and TMD cases have erythroid and megakaryocytic phenotypes. We also found that mRNAs encoding GATA-1 and GATA-2 are expressed in all these cases. These results suggest that M7 blasts and TMD blasts correspond to the erythroid/megakaryocytic bipotential progenitor cells.

5-Aminolevulinate Synthetase↗
PubMed1995

NFATx, a novel member of the nuclear factor of activated T cells family that is expressed predominantly in the thymus.

The nuclear factor of activated T cells (NFAT) regulates cytokine gene expression in T cells through cis-acting elements located in the promoters of cytokine genes. Here, we report the cDNA cloning, chromosomal localization, and initial characterization of a transcription factor related to NFATp and NFATc. The novel molecule, designated NFATx, exhibits in its middle a region very similar to the Rel homology domain in NFATc and NFATp. The amino-terminal region of NFATx also shows significant similarities to corresponding sequences in NFATc and NFATp and contains three copies of a conspicuous 17-residue motif of unknown function. We provide evidence showing that NFATx can reconstitute binding to the NFAT-binding site from the interleukin 2 promoter when combined with AP1 (c-Fos/c-Jun) polypeptides and that NFATx is capable of activating transcription of the interleukin 2 promoter in COS-7 cells when stimulated with phorbol ester and calcium ionophore. NFATx mRNA is preferentially and remarkably found in the thymus and at lower levels in peripheral blood leukocytes. The expression pattern of NFATx, together with its functional activity, strongly suggests that NFATx plays a role in the regulation of gene expression in T cells and immature thymocytes.

Amino Acid Sequence↗
PubMed1995

No apparent mineralocorticoid receptor defect in a series of sporadic cases of pseudohypoaldosteronism.

Pseudohypoaldosteronism (PHA) is characterized by congenital resistance of the kidney and/or other mineralocorticoid target tissues to aldosterone, resulting in excessive salt wasting. Although the mineralocorticoid receptor (MR) was suggested as a potential locus of the defect in this disease, no such abnormality was found in 3 recently reported cases, one of whom belongs to this series of 5 patients. Molecular studies of the MR complementary DNA and gene in this series of sporadic cases of pseudohypoaldosteronism are reported. Four of these patients had multiple mineralocorticoid target tissue resistance, whereas 1 had transient isolated resistance in the kidney. A nonconservative homozygous mutation (C944-->T944, Ala241-->Val241) was identified in the complementary DNA of 4 of the patients but was also found in 62 of 100 normal alleles. One of these 4 patients had an additional conservative heterozygous mutation (A760-->G760, Ileu180-->Val180), which was also present in 11 of 100 normal alleles. None of the patients had any abnormalities in the first untranslated exon and 0.9 kilobases of the 5'-regulatory region of the MR gene, which were fully sequenced and compared with the normal sequence. It is concluded that the mutations identified in 4 of these 5 patients with PHA are polymorphisms, which on their own have no apparent pathophysiological significance. It is hypothesized that the defect causing PHA might be in a post-MR step of aldosterone action or in an unsuspected nonclassic receptor for this hormone.

Adolescent↗