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Biomedical subjects

T Taniguchi

Publications and source records attributed to T Taniguchi.

At least 631 records · Page 35Linked to original sources

The IL-2 receptor beta-chain (p70). Ligand binding ability of the cDNA-encoding membrane and secreted forms.

The high affinity IL-2R is composed of at least two distinct subunits; alpha (p55 or Tac)- and beta (p70)-chains. Recent cDNA expression studies with lymphoid cells unequivocally demonstrated that the IL-2R beta is an indispensable component for the ligand internalization and the signal transduction via the high affinity IL-2R. Furthermore, these studies confirmed that the IL-2R beta singly expressed on T lymphoid cells represents the intermediate affinity IL-2R. In the present study, however, we show 1) the IL-2R beta expressed on a mouse fibroblast line, NIH-3T3, did not bind IL-2 under the intermediate affinity conditions, 2) nevertheless, the IL-2R beta coexpressed with the IL-2R alpha not only formed the high affinity receptor but also internalized IL-2. By the use of competitive sandwich ELISA to study the IL-2 binding ability of the IL-2R beta molecule itself, we also show that the IL-2R beta present in the detergent-solubilized cell extracts and the secreted IL-2R beta (p37) produced by the truncated cDNA are actually capable of binding IL-2, but with a much reduced affinity compared with the intact IL-2R beta expressed on the lymphoid cells. These findings lead us to postulate a more complex feature of the IL-2R than ever speculated; the IL-2R beta molecule having on its own minimal IL-2 binding ability requires a putative gamma-chain specifically present on lymphoid cells to generate the functional intermediate affinity IL-2R. The gamma-chain, however, seems not to be required as far as the formation of the high affinity IL-2R by the alpha- and beta-chains and the ligand internalization through it are concerned.

Amino Acid Sequence↗

The human interleukin-2 receptor beta-chain gene: genomic organization, promoter analysis and chromosomal assignment.

The chromosomal gene for the human interleukin-2 receptor beta-chain (IL-2R beta) was isolated and characterized. The entire IL-2R beta gene is composed of ten exons spanning about 24.3 kilobases, in which the protein is encoded by the exons 2-10. The cysteine rich extracellular region which displays a significant evolutionary resemblance to other cytokine receptors, as well as growth hormone and prolactin receptors, is encoded primarily by exons 3 and 4, whereas the membrane proximal, cysteine poor domain showing a homology with type III modules of fibronectin is encoded by exon 7. Sequence analysis of the 5'-flanking region revealed the presence of potential binding sites for transcription factors such as Octamer binding factors, AP-1, AP-2 as well as the 'GC-clusters'. At least five potential cap sites were identified by S1 mapping analysis. The 850 bp DNA sequence of the 5'-flanking region exhibited constitutive promoter activity when it was linked upstream of the HSV-tk reporter gene and then transfected into YT cells, a human leukemic cell line. By applying the RFLP linkage analysis, the IL-2R beta gene has been assigned to chromosome 22q12-13.

Base Sequence↗

Purification and characterization of a cytosolic protein-tyrosine kinase from porcine spleen.

A cytosolic protein-tyrosine kinase has been highly purified from porcine spleen using [Val5]angiotensin II as a substrate. The purification procedure involves sequential column chromatographies on phosphocellulose, Sephacryl S-200, casein-Sepharose 4B, heparin-Sepharose CL-6B and anti-(4-aminobenzyl phosphonic acid)--Sepharose 4B. Analysis of the most highly purified preparation by SDS/PAGE revealed a major silver-stained band of molecular mass 40 kDa. The 40-kDa cytosolic protein-tyrosine kinase was purified approximately 10,000-fold with an overall yield of about 7%. It had autophosphorylation activity which was carried out by intramolecular catalysis. The stoichiometry of phosphate incorporation was about 1 mol phosphate/mol enzyme. In the autophosphorylation reaction, the apparent Km value for ATP was relatively low, 0.35 microM; Mn2+ was slightly preferred to Mg2+ as divalent cation. [Val5]Angiotensin II phosphorylation activity of the 40-kDa kinase increased with the amount of phosphate incorporated into the enzyme. A phosphate exchange reaction was observed during the autophosphorylation. These results suggest that the 40-kDa kinase described here is a different type of protein-tyrosine kinase than the enzymes so far reported.

Adenosine Triphosphate↗

Changes in signal transduction in Alzheimer's disease.

We studied the signal transduction system including the receptor and protein kinase C (PKC) in Alzheimer's disease (AD) brains. We used 3H-TCP as a ligand for the NMDA receptor-ion channel complex. The total concentrations of 3H-TCP binding sites were significantly reduced in AD frontal cortex. 3H-TCP binding sites spared in AD brains retained the affinity for the ligand and the reactivity to NMDA, L-glutamate, and glycine. We utilized antibodies to assess the degree of involvement of different PKC isoforms in AD. The concentration of PKC (beta II) was lower in AD particulate fractions and higher in AD cytosol fractions. Immunocytochemical studies revealed reduced numbers of anti-PKC (beta II)-immunopositive neurons. Anti-PKC (alpha) faintly stained entire plaques and surrounding glial cells. Anti-PKC (beta I) stained dystrophic plaque neurites. Anti-PKC (beta II) stained the amyloid-containing portions of plaques. These results suggest an involvement of second messenger cascades in the pathogenesis of AD in addition to neurotransmitters and their receptors.

Aged↗

Binding of Cu(II), Tb(III) and Fe(III) to chicken ovotransferrin. A kinetic study.

The kinetics of binding of Cu(II), Tb(III) and Fe(III) to ovotransferrin have been investigated using the stopped-flow technique. Rate constants for the second-order reaction, k+, were determined by monitoring the absorbance change upon formation of the metal-transferrin complex in time range of milliseconds to seconds. The N and C sites appeared to bind a particular metal ion with the same rate; thus, average formation rate constants k+ (average) were 2.4 x 10(4) M-1 s-1 and 8.3 x 10(4) M-1 s-1 for Cu(II) and Tb(III) respectively. Site preference (N site for Cu(II) and C site for Tb(III] is then mainly due to the difference in dissociation rate constant for the metals. Fe(III) binding from Fe-nitrilotriacetate complex to apo-ovotransferrin was found to be more rapid, giving an average formation rate constant k+ (average) of 5 x 10(5) M-1 s-1, which was followed by a slow increase in absorbance at 465 nm. This slow process has an apparent rate constant in the range 3 s-1 to 0.5 s-1, depending upon the degree of Fe(III) saturation. The variation in the rate of the second phase is thought to reflect the difference in the rate of a conformational change for monoferric and diferric ovotransferrins. Monoferric ovotransferrin changes its conformation more rapidly (3.4 s-1) than diferric ovotransferrin (0.52 s-1). A further absorbance decrease was observed over a period of several minutes; this could be assigned to release of NTA from the complex, as suggested by Honda et al. (1980).

Animals↗

Reovirus type 2-induced diabetes in mice prevented by immunosuppression and thymic hormone.

Reovirus type 2 that had been isolated from a cow with diarrhoea and passaged in bovine kidney cell culture produced a Type 1 (insulin-dependent) diabetes-like syndrome when inoculated into NC mice. The infection resulted in insulitis and destruction of islet cells. Viral antigens were found in islet cells by staining with fluorescein-labelled antibody to reovirus type 2. The destruction of islet cells resulted in abnormalities shown on glucose tolerance testing. Studies on the susceptibility of the host showed that only certain strains of mice had overtly abnormal glucose tolerance tests when infected with reovirus type 2. To assess the immunological role in the pathogenesis of reovirus type 2-induced diabetes, infected mice were subjected to immunosuppressive or thymic hormone treatment. The administration of either anti-thymocyte serum or serum thymic factor reduced or prevented the development of the diabetes-like syndrome, while Arg-Lys-Asp-Val-Try did not show any therapeutic effects.

Animals↗

Diverse effects of poly-basic amino acids, heparin and ionic strength on the phosphorylation of various substrates by cytosolic protein-tyrosine kinase from porcine spleen.

1. Effects of poly-basic amino acids, heparin and ionic strength on the activity of cytosolic protein-tyrosine kinase from porcine spleen (CPTK-40) have been studied. 2. Both polylysine and polyarginine stimulated the phosphorylation of [Val5]angiotensin II and E11 G1 (synthetic peptide of EDAEYAARRRG), but could neither stimulate nor inhibit the phosphorylation of random copolymers; poly(EY)4:1 and poly(EAY)6:3:1. 3. Heparin stimulated the phosphorylation of poly(EY)4:1 by 2.5-fold, however, it inhibited those of E11G1, poly(EAY)6:3:1, casein and H2B histone. 4. Elevation of ionic strength of either NaCl, KCl or (NH4)2SO4 stimulated the phosphorylation of poly(EY)4:1 by greater than 5-fold, but inhibited those of casein, tubulin, H2B histone, E11G1 and poly(EAY)6:3:1. 5. These effectors did not change the Km for substrates but increased the Vmax. 6. These results suggest that the effects of poly-basic amino acids, heparin and ionic strength on the activity of CPTK-40 are mainly on the substrates employed rather than on the enzyme itself.

Amino Acids↗

Advantages and limitations in the use of porcelain veneer restorations.

Porcelain veneer restorations including preparations, impression materials, cast materials, refractory casts, handling of porcelain, the try-in, and the final luting are discussed. Techniques are described to increase marginal accuracy, establish predictable natural polychromatic color within the porcelain, establish good contours and surface textures, and shorten final placement time. Tooth preparation is routinely recommended, as is the combination of epoxy dies, refractory casts, and anatomic buildup techniques. Emphasis is placed on a close working relationship and communication among the dentist, dental technician, and patient.

Dental Bonding↗

Platelet size and function in septic rats: changes in the adenylate pool.

Cecal ligation and puncture (CLP) were performed in rats. After 4 hr (early sepsis) and 16 hr (late sepsis), platelet morphology and function were studied. At 16 hr, platelet counts for the CLP group were significantly lower than for the sham-operated control group. Low maximum aggregation rates (MAR) and decreased platelet counts were elicited in platelet-rich plasma with 4 M ADP and 2 micrograms/ml collagen. However, with platelet counts equalized, MAR for the CLP group increased significantly, especially after 16 hr. The platelet-large cell rate and platelet distribution width decreased temporarily at 4 hr, then rose significantly at 16 hr. No significant changes were observed in the mean platelet volume after 4 hr, but there were significant increases after 16 hr. Total adenine nucleotide (TAN) levels within the platelets rose significantly in the CLP group, suggesting the appearance during the late sepsis of large, heavy platelets or adenine nucleotide-rich platelets. The platelet adenylate pool was divided into granular and cytoplasmic fractions, respectively characterized by ADP and ATP increases. However, no septicemia-related differences were noted in the degree of binding between goat antirat fibrinogen and platelet surface glycoprotein IIb/IIIa complex. Internal environment changes in the platelets indicated that during septicemia hyperfunctional or hypersensitive platelets with a latent capacity for active aggregation and release appeared in the circulation. Hypercoagulability in septicemia involves activation of coagulation factors, stimulation of the coagulation cascade, volume changes accompanying increased platelet TAN content, and changes in AN distribution in the two pools. These findings significantly increase our understanding of the transition from the prethrombotic state to thrombosis in septicemia.

Adenine Nucleotides↗

Possible involvement of kinins in cardiovascular changes after alcohol intake.

Japanese healthy male subjects were divided into two groups, i.e., a normal aldehyde dehydrogenase (ALDH) group with a low Km isozyme of ALDH for acetaldehyde, and a deficient group without it. After intake of 0.4 g/kg alcohol, the deficient group showed high levels of blood acetaldehyde, facial flushing including an increased pulse rate and a fall in diastolic blood pressure, while the normal group did not manifest these changes. In the deficient group, the total kininogen concentration gradually decreased after alcohol intake due to a reduction in low molecular weight kininogen, and plasma prekallikrein remained unchanged. The normal group showed no significant changes in any of these values after alcohol intake. In an in vitro study with pooled plasma, the low concentrations of urinary kallikrein caused a decrease in the low molecular weight kininogen only. These results suggest that kinins released by acetaldehyde-induced activation of glandular kallikreins are associated with the changes in cardiovascular symptoms in deficient group which display flushing after alcohol intake.

Acetaldehyde↗

Heterogeneity of human islet cell antibodies in terms of cross-species reactivity.

There has been no consistent view about the cross-species reactivity of islet cell antibodies (ICA), and no report about their target antigens in the pancreatic islet cells of different species. Therefore, we examined the cross-reactivity of human ICA against rodent pancreatic islet cells, and found at least two types of ICA, one having a comparatively strong cross-reactivity and the other lacking it. Furthermore, using human as well as some rodent pancreatic tissues that had been modified chemically, we came to suspect that the target antigens of ICA were sialic acid residues of glycolipids. Therefore, we suggest that there are at least two types of ICA recognizing sialic acid residues of glycolipids, one reacting with antigen(s) commonly present in human and rat islets, and the other with antigen(s) only present in human islets.

Animals↗

Interleukin 2 receptor beta chain expressed in an oligodendroglioma line binds interleukin 2 and delivers growth signal.

Interleukin 2 (IL-2) is a potent growth factor for T lymphocytes, playing a crucial role in the immune response. In view of the considerable evidence that the immunoregulatory cytokines (or lymphokines) also play a role in the growth and differentiation of cells in the central nervous system (CNS), we examined the operation of the IL-2 system in a cell line of CNS origin by expressing a cDNA encoding the beta chain of the human IL-2 receptor (IL-2R beta, a 75-kDa protein). When the cDNA was expressed in a human oligodendroglioma cell line, ONS-21, the IL-2R beta bound IL-2 with an affinity similar to that in lymphoid cells (Kd, approximately 2 nM). Furthermore, cell proliferation ([3H]thymidine incorporation) was stimulated by IL-2. These results demonstrate that the same cytokine receptor is functional in cells of the immune system and CNS and point to a molecular mechanism that is similar for growth-signal transduction between lymphoid and neural cells but that may be different in other cells, such as fibroblasts.

Antibodies↗

Murine interleukin 2 receptor beta chain: dysregulated gene expression in lymphoma line EL-4 caused by a promoter insertion.

The functional, high-affinity interleukin 2 receptor (IL-2R) consists of at least two receptor components, IL-2R alpha (p55) and IL-2R beta (p70-75). The cDNA encoding the murine IL-2R beta has been isolated by using the previously cloned cDNA for human IL-2R beta as a probe. Analysis of the cDNA revealed that the murine IL-2R beta shows a marked homology with the human IL-2R beta and that it is also structurally related to other cytokine receptors such as erythropoietin receptor. The cDNA-directed murine IL-2R beta formed high-affinity IL-2R in conjunction with the endogenous IL-2R alpha in a murine pro-B-cell line and could transduce IL-2-induced growth signal. In mouse lymphoma line EL-4, the IL-2R beta gene was found to be rearranged by the insertion of the long terminal repeat sequence of an intracisternal A particle, giving rise to constitutive expression of the IL-2R beta mRNA.

Amino Acid Sequence↗

Characterization of an antigen defined by monoclonal antibody KMO1.

We have isolated an antigen defined by the monoclonal antibody KMO1 from tissue culture supernatant as a glycoprotein and from cancer cells as a glycolipid. The antigenic determinant was a carbohydrate. Antibody KMO1 inhibited the binding activity of 19-9 to CA19-9, but 19-9 antibody did not inhibited the binding activity of KMO1 to KMO1 antigen. The glycolipid antigen of KMO1 was a monosialoganglioside and was separated into 3 components on thin layer chromatography. The 19-9 antibody reacted with one of the components. These results suggest that the KMO1 antigen is similar to but not identical to CA19-9.

Animals↗

[3H]N-[1-(2-thienyl)cyclohexyl]-3,4-piperidine ([3H]TCP) binding in human frontal cortex: decreases in Alzheimer-type dementia.

We studied [3H]N-[1-(2-thienyl)cyclohexyl]-3,4-piperidine [( 3H]TCP) binding to human frontal cortex obtained at autopsy from 10 histologically normal controls and eight histopathologically verified cases with Alzheimer-type dementia (ATD). Extensively washed membrane preparations were used to minimize the effects of endogenous substances. In ATD frontal cortex, the total concentration (Bmax) of [3H]TCP binding sites was significantly reduced by 40-50%. The apparent dissociation constant (KD) values showed no significant change. The reduction in binding capacity was also apparent in Triton X-100-treated membrane preparations, and there was a linear correlation between the number of [3H]TCP binding sites and that of N-methyl-D-aspartate (NMDA)-sensitive [3H]glutamate binding sites. [3H]TCP binding sites spared in ATD brains retained the affinity for the ligand and the reactivity to NMDA, L-glutamate, and glycine. These results suggest that the primary change in NMDA receptor-ion channel complex in ATD brains is the reduction of its number, possibly reflecting the loss of neurons bearing these receptor complexes, and that the functional linkage within the receptor complexes spared in ATD brains remains normal.

Alzheimer Disease↗

Phosphoinositide breakdown in rat hippocampal slices: sensitivity to glutamate induced by in vitro anoxia.

We examined the effects of in vitro anoxia on phosphoinositide (PI) breakdown in rat hippocampal slices stimulated by glutamate and quisqualate. In addition to assays of accumulations of 3H-inositol phosphates (3H-IPs) degraded from prelabeled PI, we adopted direct assay procedures of inositol 1,4,5-triphosphate (1,4,5-IP3) using 1,4,5-IP3-specific binding protein to determine the formation of 1,4,5-IP3. The first effect, observed with anoxic incubation by itself, was the diminished quisqualate (10(-5) M)-stimulated accumulation of 3H-IPs degraded from prelabeled PI under prolonged anoxia. Quisqualate caused a transient increase in 1,4,5-IP3 formation in the early phase of anoxia, similar to that under oxygenated conditions. Glutamate (10(-5) M), under normal conditions, influenced neither the accumulation of 3H-IPs nor the formation of 1,4,5-IP3. Also, the accumulation of 3H-IPs under prolonged anoxia was unaffected. The same concentration of glutamate, however, gave rise to a transient increase in 1,4,5-IP3 content in the early phase of anoxia, similar to that caused by quisqualate. The second effect, observed by oxygenation following anoxia, was the induction of glutamate-stimulated accumulation of 3H-IPs. When the hippocampal slices were oxygenated following a sufficiently long (greater than 30-min) exposure to anoxia, glutamate (10(-5) M) caused a significant increase in accumulation of 3H-IPs degraded from prelabeled PI. Quisqualate-stimulated accumulation of 3H-IPs under oxygenated incubations was also increased by prior exposure of slices to anoxia. These results support the hypothesis that an exposure of hippocampal slices to anoxia induces a sensitivity of the PI breakdown pathway to glutamate and that, given an oxygen supply following sufficiently long exposure to anoxia, the slices maintain their sensitivity to glutamate with an apparent increase in the accumulation of 3H-IPs.

Animals↗

Endotoxemia in patients on hemodialysis.

We examined endotoxins and limulus amebocyte lysate-reactive material (LAL-RM) in serum from 87 patients on hemodialysis, using the conventional chromogenic limulus test (CCLT) and a new specific endotoxin assay with factor G-free LAL (endotoxin-specific test: EST). All patients with regenerated cellulose dialyzers had increased CCLT values, whereas the EST values were not higher than in controls. This discrepancy can be explained by the LAL-RM which is cellulose-derived and cross-reacts with LAL via factor G. Using EST for measurements of endotoxin, 6 patients out of 87 had pathological endotoxemia and all of these patients were associated with either cirrhosis, infection, or malignancy. Some patients who had no complications showed fever before or during dialysis, but they did not have high endotoxin levels. EST is useful for the diagnosis of endotoxemia in patients on hemodialysis because of the presence of LAL-RM in serum. Endotoxemia is rare in patients on hemodialysis, if they are not associated with infection, cirrhosis, or other complications.

Adult↗