Characterization of metal ion-implanted titanium oxide photocatalysts operating under visible light irradiation.
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Journal Article↗
Biomedical subjects
M Takeuchi
Publications and source records attributed to M Takeuchi.
An essential role for NF-kappa B in IL-18-induced IFN-gamma expression in KG-1 cells.
IL-18 is a multifunctional cytokine playing various regulatory roles in the immune system including induced cytokine production. As a part of our ongoing studies on the molecular mechanisms of IL-18-induced IFN-gamma production, we have examined the transcriptional regulation of the IFN-gamma gene by IL-18 in a human myelomonocytic cell line, KG-1. On the basis of DNA/protein binding, we have determined an IL-18-inducible NF-kappa B binding site located at -786 to -776 of the IFN-gamma gene regulatory region (designated KBBsite). Transient transfection of promoter-reporter gene constructs revealed that the KBBsite is required for full IL-18-induced activation of the IFN-gamma gene transcription induced by IL-18. In addition, stable transformants of a dominant-negative form of the I kappa B alpha showed an inhibition of IL-18-dependent I kappa B alpha degradation, NF-kappa B activation, and expression of IFN-gamma. These results are the first to show the actual significance of the NF-kappa B pathway in the regulation of IFN-gamma gene expression by IL-18.
Binding Sites↗
IkappaBalpha ubiquitination is catalyzed by an SCF-like complex containing Skp1, cullin-1, and two F-box/WD40-repeat proteins, betaTrCP1 and betaTrCP2.
Destruction of the transcriptional inhibitor IkappaB by the ubiquitin (Ub) system is required for signal-dependent activation of the multifunctional transcriptional factor NF-kappaB, but details of this ubiquitination are largely unknown. We report here that the IkappaBalpha-ubiquitin ligase (IkappaBalpha-E3) is an SCF-like complex containing Skp1, cullin-1, and two homologous F-box/WD40-repeat proteins, betaTrCP1 and betaTrCP2. Intriguingly, all these components are cooperatively recruited to bind to a phosphorylated IkappaBalpha (pIkappaBalpha) produced by tumor necrosis factor-alpha (TNF-alpha) stimulation. IkappaBalpha-E3 bound to pIkappaBalpha catalyzed in vitro ubiquitination of pIkappaBalpha in the presence of ATP, Ub, and E1-activating and E2-conjugating enzymes. Forced expression of betaTrCP1 and betaTrCP2 resulted in dramatic augmentation of the in vitro polyubiquitination activity of IkappaBalpha-E3. These results indicate that the long-sought IkappaBalpha-E3 is an SCF-like complex consisting of multiple proteins which are coordinately assembled during phosphorylation of IkappaBalpha in response to external signals.
Amino Acid Sequence↗
In vivo and in vitro recruitment of an IkappaBalpha-ubiquitin ligase to IkappaBalpha phosphorylated by IKK, leading to ubiquitination.
Activation of the transcriptional factor NF-kappaB is triggered by signal-dependent degradation of its inhibitor protein IkappaB through the ubiquitin (Ub)-proteasome pathway. We found here that a phosphorylated IkappaBalpha immunoprecipitated (IP-pIkappaBalpha) from the crude extract of HeLa cells which had been treated with tumor necrosis factor-alpha (TNFalpha) caused a dramatic ubiquitination of itself, termed autoubiquitination, when incubated with ATP, Ub, and E1-activating and E2-conjugating enzymes. IP-pIkappaBalpha also catalyzed ubiquitination of an in vitro synthesized 35S-IkappaBalpha previously phosphorylated by IkappaB-kinase (IKK) which is referred to as transubiquitination. No appreciable activity of auto- and transubiquitination was observed in an unphosphorylated IP-IkappaBalpha. Moreover, the putative IkappaBalpha-Ub ligase (IkappaBalpha-E3) present in HeLa cell cytosol associated in vitro with an IKK-phosphorylated recombinant IkappaBalpha, a process independent of NF-kappaB binding to IkappaBalpha or TNFalpha stimulation. Replacement of the two Ser residues at positions 32 and 36 corresponding to IKK phosphorylation sites by Ala resulted in almost complete prevention of binding of an IkappaBalpha-E3 to IkappaBalpha. These results indicate that phosphorylation of IkappaBalpha is necessary and sufficient for recruitment of this IkappaBalpha-E3 to associate with IkappaBalpha.
Adenosine Triphosphate↗
Effects of opioids on neuronal survival in culture of embryonic chick dorsal root ganglion neurons.
We studied the effects of endogenous and synthetic opioids on survival of primary-cultured dorsal root ganglion neurons of 8-day chick embryos. In the basal growth medium containing nerve growth factor (NGF), [Leu5]enkephalin, [Met5]enkephalin or selective mu-agonist, [D-Ala2, N-Me-Phe4, Gly5-ol]enkephalin (DAMGO) produced significant increase of neuronal survival in a naloxone-reversible manner. On the other hand, dynorphin A (1-13), [D-Pen2, L-Pen5] enkephalin (DPLPE) (a delta-agonist) and U-50,488 (a kappa-agonist) showed no such effect. No viable neurons were observed in the absence of NGF in spite of the presence of any opioid peptides. These results suggest that [Leu5]enkephalin and [Met5]enkephalin might increase neuronal survival by stimulating mu-opioid receptors, and that these opioid peptides are working with other growth factors but, in and by themselves, do not promote survival.
Animals↗
Effect of astaxanthin rich red yeast (Phaffia rhodozyma) on oxidative stress in rainbow trout.
The antioxidative biological effect of dietary red yeast, Phaffia rhodozyma, which is rich in astaxanthin, on rainbow trout, Oncorhynchus mykiss, was examined. The levels of serum transaminase (glutamic-pyruvic transaminase and glutamic-oxaloacetic transaminase) activities and of lipid peroxides (LPO) of fish fed oxidized oil were significantly higher than those of the control fish fed non-oxidized oil. However, the supply of red yeast considerably decreased both enzyme activities and LPO level. Furthermore, the serum lipid (triglycerides, total cholesterol and phospholipids) concentrations were also significantly decreased. Especially, the serum triglyceride level of fish fed the red yeast was as low as that of the control. It was also observed that there were no significant differences in muscle LPO levels between the fish fed red yeast and the control. The present results suggest for the first time that dietary red yeast may effectively suppress the LPO generation of tissue and normalize liver function as well as improving muscle pigmentation of trout. Thus, red yeast should have a reducing effect on oxidized oil-induced oxidative stress in fish.
Animal Feed↗
Autoimmune diseases developed in athymic nude mice grafted with embryonic thymus of xenogeneic origin.
Reconstitution of T cell functions in athymic BALB / c nude mice was investigated after transplantation of embryonic thymus grafts under the renal capsule (TG nude mice). Thymi were obtained from mice, rats, rabbits, hamsters, guinea pigs, swine and cows. All of the grafted thymi grew and formed proper thymic structures with host CD90(+) cells. The TCRalpha beta expression pattern of the lymphocytes in the grafted thymi was quite similar to that of normal mouse thymocytes. Sufficient CD4(+) populations were observed in the peripheral lymphoid organs of all groups of TG nude mice. Plaque-forming cell assay revealed that the TG nude mice had acquired considerable helper T cell functions. Histological and serological studies, however, showed multiple organ-localized inflammatory diseases in TG nude mice of all xenogeneic thymus groups, but not with syngeneic or allogeneic thymi. The organs affected were the thyroid, the lacrimal, submandibular and sublingual glands, the eyes, stomach and ovaries. The incidence of lesions was variable depending on the species of origin of the grafted thymus. Lesions from TG nude mice were successfully transferred into naive nude mice by host CD4(+) cells. These results together suggested that the microenvironment of grafted thymi, even if xenogeneic, is able to educate host T cell precursors. However, this reconstitution of functions does not always induce tolerance to certain autoantigens, resulting in development of multiple autoimmune lesions.
Animals↗
Paradoxical increase in coronary flow velocity after termination of acetylcholine infusion is a marker of the impaired vasodilatation at coronary microvessels in patients with angina and normal coronary arteries.
Some patients with anginal chest pain and normal coronary arteries exhibit a paradoxical increase in coronary flow velocity after termination of acetylcholine infusion. The aim of this study was to investigate whether this paradoxical increase in the flow velocity is associated with the impaired microvascular dilatation to pharmacological stimuli. We infused graded doses of endothelium-dependent vasodilator acetylcholine (10, 50, and 100 microg/min for 2 min) and the endothelium-independent vasodilators, papaverine and nitroglycerin, into the left coronary artery in 15 patients with chest pain and normal coronary arteries. Coronary blood flow responses were evaluated by Doppler guidewire and quantitative angiography in the proximal left anterior descending coronary artery. Seven patients showed a paradoxical increase in coronary flow velocity after termination of acetylcholine infusion with the highest dose (100 microg/min), whereas eight showed no change or a decrease in the flow velocity. This was also observed in 50 microg/min of acetylcholine infusion. In patients with a paradoxical increase in the flow velocity, coronary flow velocity responses to acetylcholine during three graded doses were significantly blunted in comparison to those without a paradoxical increase. In contrast, coronary flow reserve to papaverine and nitroglycerin was similar in the two groups. Epicardial artery vasoreactivity to acetylcholine did not differ between the two groups. Papaverine and nitroglycerin also caused a similar degree of coronary dilatation in both groups. These results suggest that the paradoxical increase in coronary flow velocity observed immediately after termination of the intracoronary acetylcholine infusion is a marker of impaired nitric oxide-dependent dilatation of the coronary microvessels in patients with normal coronary arteries. Cathet. Cardiovasc. Intervent. 48:170-177, 1999.
Acetylcholine↗
Intracellular production of interleukin-18 in human epithelial-like cell lines is enhanced by hyperosmotic stress in vitro.
Interleukin-18 is a novel multifunctional cytokine, which enhances natural killer cell activity and promotes the induction of cytokine production, including that of interferon-gamma by T cells and antitumor effects. Interleukin-18 is produced by cells of several different tissues (e.g., macrophages, keratinocytes, osteoblasts, and intestinal epithelium); however, it is unclear what physiological conditions or stimuli induce interleukin-18 production. To determine physiological conditions for the production of interleukin-18, we have examined the effect of mannitol-induced hyperosmotic conditions on normal human umbilical vein endothelial cells (HUVEC) and eight established human epithelial-like cell lines (Intestine 407, Caco-2, A253, HeLa, SCC25, HT1197, ACHN, A549). Hyperosmotic conditions induced interleukin-18 immunoreactivity in all the human cell lines tested, as detected by immunocytochemistry. The enhanced interleukin-18 production was also observed when mannitol was replaced with NaCl as the inducer of hyperosmotic stress. Enzyme-linked immunosorbent assays revealed that interleukin-18 concentrations in cell extracts were significantly increased by hyperosmotic conditions. Reporter gene assays also revealed that hyperosmotic conditions stimulated transcriptional activity of the interleukin-18 promoter. These results show for the first time that hyperosmotic stress is a stimulator of interleukin-18 production in epithelial-like cells.
Cells, Cultured↗
A novel biphenotypic B-cell precursor leukemia cell line (NALM-29) carrying t(9;22)(q34;q11) established from a patient with acute leukemia.
A novel biphenotypic leukemia cell line, NALM-29, was established from a 46-year-old Japanese male patient with acute lymphoblastic leukemia (ALL). The primary leukemic blasts showed a common ALL phenotype with CD19+, CD10+, CD13-, HLA-DR+ and Igs-. NALM-29 cells display biphenotypic characteristics: expression of the intracellular enzyme myeloperoxidase at the mRNA and protein level and cell surface positivity for CD19, CD10, CD13, CD33 and HLA-DR. NALM-29 fulfills EGIL criteria as B-cell precursor (BCP) leukemia B-II type. NALM-29 cells have a lymphoblastic morphological appearance; the immunoglobulin heavy chain gene is rearranged. NALM-29 cells responded significantly to the proliferative stimuli of FLT-3 ligand and IL-7, but not to GM-CSF, IL-3, IL-6, PIXY-321 or SCF. Proliferation of cells was inhibited significantly by IL-4, TNF-alpha or TNF-beta treatment. Cytogenetic analysis revealed the characteristic t(9;22)(q34;q11); expression of the m-bcr e1-a2 BCR-ABL fusion gene (typically found in ALL) was determined by PCR amplification of cDNA. The immunological, cytogenetic and functional characterization of NALM-29 suggests that this cell line may represent a scientifically significant in vitro model for BCP-type leukemia cells with biphenotypic characteristics.
Base Sequence↗
Enhanced visualization of intravascular and left atrial appendage thrombus with the use of a thrombus-targeting ultrasonographic contrast agent (MRX-408A1): In vivo experimental echocardiographic studies.
Echocardiographic evaluation for the recognition of intravascular and left atrial appendage thrombus remains a difficult problem. A thrombus-specific ultrasonographic contrast agent has the potential for an alternative approach for their delineation. The aim of this study was to investigate the usefulness of thrombus-specific contrast agent MRX-408A1 for the detection of acute experimentally created intravascular and intracardiac thrombus. In the first study, we created inferior vena cava thrombus in 9 dogs. With the use of fundamental 2-dimensional echocardiography imaging, we recorded images of the inferior vena cava thrombus at baseline (n = 9), with the thrombus-specific contrast agent MRX-408A1 (n = 9), and with nonspecific contrast agent MRX-113 (n = 6). In the second study, we created a left atrial appendage thrombus in 8 dogs. We imaged left atrial appendage thrombus at baseline and during MRX-113 and MRX-408A1 infusion. Thrombus was successfully created in all dogs in study 1 and in 6 of 8 dogs in study 2. MRX-408A1 produced a visually apparent increase in ultrasonographic contrast enhancement of the thrombus in all cases in which thrombus was found on autopsy. In both studies, MRX-408A1 increased the videointensity of the thrombus significantly compared with baseline images and images obtained during MRX-113 infusion. The size of the visually detectable thrombus on the image was also significantly larger during MRX-408A1 infusion than at baseline and during MRX-113 infusion. These data provide in vivo demonstration of the efficacy of a thrombus-specific contrast agent, MRX-408A1, in the detection of acute intravascular and intracardiac thrombus. It has the potential to improve the diagnostic accuracy of ultrasonography for the detection of acute thrombi at various cardiovascular sites in the clinical setting.
Animals↗
Inhibitory effects of beer and other alcoholic beverages on mutagenesis and DNA adduct formation induced by several carcinogens.
The possibility that beer and other alcoholic beverages could be antimutagenic against the heterocyclic amines (HAs), a group of carcinogens produced on cooking proteinaceous foods, has been explored. In the Salmonella mutation assays, beer showed inhibitory effects against several HAs [preactivated Trp-P-1, Trp-P-2(NHOH), and Glu-P-1(NHOH)] that are directly mutagenic in bacteria. Japanese sake, red and white wines, and brandy were also effective. However, ethyl alcohol alone did not show these effects. The formation of O(6)-methylguanine by N-methyl-N'-nitro-N-nitrosoguanidine in the DNA of Salmonella YG7108 was also inhibited by beer. Nonvolatile beer components were administered orally to CDF(1) mice together with Trp-P-2. Adducts in the liver DNA were significantly decreased by the beer, as compared to those in controls fed Trp-P-2 only. Although several phenolic compounds known to be present in beer were antimutagenic toward these mutagens, their effects were very small. It was concluded that some yet to be identified component(s) of beer is (are) responsible for this antimutagenicity.
Alcoholic Beverages↗
Asn-linked sugar chain structures of recombinant human thrombopoietin produced in Chinese hamster ovary cells.
Human thrombopoietin (TPO) that regulates the numbers of megakaryocytes and platelets is a heavily N- and O-glycosylated glycoprotein hormone with partial homology to human erythropoietin (EPO). We prepared recombinant human TPO produced in Chinese hamster ovary (CHO) cells and analyzed the sugar chain structures quantitatively using 2-aminobenzamide labeling, sequential glycosidase digestion and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF/MS). We found bi-, tri- and tetraantennary complex-type sugar chains with one or two N-acetyllactosamine repeats, which are common to recombinant human EPO produced in CHO cells. On the other hand, there were triantennary sugar chains with one or two N-acetyllactosamine repeats that were specific to the recombinant human TPO, and their distributions of branch structures were also different. These results suggested that proximal protein structure should determine the branch structure of Asn-linked sugar chains in addition to the glycosyltransferases subset.
Animals↗
Theophylline induces apoptosis of the IL-3 activated eosinophils of patients with bronchial asthma.
In bronchial asthma, eosinophils are upregulated and their survival is suggested to be prolonged by the action of some cytokines such as Interleukin (IL)-3, IL-5 and granulocyte-macrophage colony-stimulating factor (GM-CSF). We find here that the survival of eosinophils in the peripheral blood of patients with asthma is correlated with the serum levels of IL-3 but not of IL-5 and GM-CSF. Interestingly, theophylline is revealed to induce apoptosis of the prolonged survival eosinophils by IL-3, as judged by morphological changes and nucleosomal DNA fragmentation. During the apoptosis, caspase-3 in eosinophils stimulated by IL-3 is activated by theophylline. The substrate of caspase-3, poly (ADP-ribose) polymerase (PARP), is cleaved in the eosinophils after theophylline treatment. These results suggest that theophylline is able to induce apoptosis of the IL-3 activated eosinophils in patients with bronchial asthma, and that its clinical effectiveness may be due to the reduction of inflammatory cells in the airway.
Journal Article↗
Spontaneous remission of large granular lymphocyte T cell leukemia.
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Aged↗
Anterior chamber-associated immune deviation-inducing cells activate T cells, and rescue them from antigen-induced apoptosis.
Immune responses to antigens injected into the anterior chamber of the eye are devoid of T helper 1 (Th1)-type responses of the delayed hypersensitivity type, which has been termed anterior chamber-associated immune deviation (ACAID). Recently, it has been found that peritoneal exudate cells (PEC) from normal mice can be made to acquire the capacity to induce ACAID in vivo when the cells are pulsed with antigen in vitro in the presence of transforming growth factor-beta2 (TGF-beta2), a major cytokine in the ocular microenvironment. We now report that when ovalbumin (OVA)-specific T cells from DO11.10 transgenic mice, or from OVA-primed normal mice, were activated in vitro by normal (untreated) PEC pulsed with OVA, the responding T cells were induced to undergo apoptosis. However, when PEC were first treated with TGF-beta2 and then used to stimulate DO11.10 T cells in the presence of OVA, T-cell proliferation occurred without evidence of increased apoptosis. The ability of TGF-beta2 to rescue responding T cells from apoptosis rested with the capacity of this cytokine to inhibit interleukin-12 (IL-12) production by PEC. Untreated PEC produced large amounts of IL-12 upon interaction with responding T cells. Under these conditions, tumour necrosis factor-alpha (TNF-alpha) production was up-regulated, and this cytokine, in turn, triggered apoptosis among T cells stimulated with OVA-pulsed PEC. From these results, we conclude that TGF-beta2-treated APC promote ACAID by rescuing antigen-activated T cells from apoptosis, and by conferring upon these cells the capacity to down-regulate delayed hypersensitivity.
Animals↗
Nitric oxide inhalation as a chemical assist for circulation in patients after cardiovascular surgery.
The objective of this study was to investigate whether nitric oxide (NO) inhalation might be an alternative strategy as a chemical assist for the circulation in patients showing a deterioration in oxygen delivery. Twelve adult patients whose oxygen delivery indices (DO2I) were less than 400 ml/min/m2 after cardiovascular surgery were included in this study. NO was administered via a premixing system or a side stream system at doses between 1 and 10 (5.1+/-2.4) ppm. Data obtained before and during a 120 min NO inhalation were compared using the paired Student's t-test. The increase in PaO2/FiO2 resulting from NO inhalation was significant (from 162 to 251 mm Hg). DO2I increased significantly from 326 to 417 ml/min/m2 concomitantly with significant increases in both arterial oxygen content (CaO2) and cardiac index (CI) (from 14.1 to 15.4 vol% and from 2.31 to 2.71 L/min/m2 , respectively). The increase in SvO2 during NO inhalation was significant (from 55.2 to 62.6%). Among the other hemodynamic parameters, both total pulmonary resistance and systolic pulmonary arterial pressure (SPAP) showed significant decreases during NO inhalation, but right atrial pressure did not change significantly. There was a close relationship between the baseline SPAP level (bSPAP) and the decrease in SPAP during NO inhalation (dSPAP) (r = -0.88). However, negative correlations were observed between bSPAP and percentage increase in CI (%CI) (r = -0.61) and between bSPAP and percentage increase in DO2I (%DO2I) (r = -0.48). Moreover, positive relationships were observed between dSPAP and %CI (r = 0.62) and between dSPAP and %DO2I (r = 0.45). Hemoglobin (Hb) increased significantly from 11.0 to 11.4 g/dl. There were no significant changes in Fio2, pH, PacO2, or base excess (BE) during NO inhalation. The level of methemoglobin measured during the study period remained within the normal range (0.86+/-0.23%). In conclusion, NO inhalation could be an efficient and alternative assist for the circulation in patients whose oxygen delivery deteriorates after cardiovascular surgery.
Administration, Inhalation↗
Novel B-cell precursor leukemia sister cell lines, NALM-33 and NALM-34, established from a patient with acute lymphoblastic leukemia.
Two novel B-cell precursor (BCP) acute lymphoblastic leukemia (ALL) cell lines, designated NALM-33 and NALM-34, were established from a 72 year-old male patient with ALL at relapse. Subcultures of each initial flask were first made after eight weeks of continuous incubation; thus, the two cell lines are simultaneous sister cell lines. The cells proliferate consistently singly and free-floating in suspension. They are negative for Epstein-Barr virus (EBV) infection by polymerase chain reaction (PCR) and are negative for mycoplasma infection. They have the morphological appearance of lymphoblasts with a scanty rim of cytoplasm, fine nuclear chromatin and distinct nucleoli. The primary leukemic blasts showed a common ALL phenotype with CD19+, CD10+, CD13-, HLA-DR+ and Igs-; the cell lines NALM-33/-34 display an identical immunophenotype. They fulfill "European Group for the Immunological Characterization of Leukemias (EGIL)" criteria as BCP leukemia B-II type. While the immunoglobulin heavy chain genes were found uniquely to be in their germline configuration, rearrangement of both kappa and lambda light chain genes was noted by Southern blot analysis. CDR-II detection by reverse transcriptase-PCR was also not detected. NALM-33/-34 did not respond significantly to the proliferative stimuli of various hematopoietic cytokines. In the cytogenetic analysis, they revealed the t(8;14)(q24.1;q32) with additional numerical and structural chromosomal abnormalities. The extensive immunological, cytogenetic and functional characterization of NALM-33/-34 suggests that these two novel cell lines may represent unique and relevant in vitro model systems for BCP-type leukemia cells.
Aged↗