Ethanol, acetaldehyde and lactic acid stimulate type III and IV collagen synthesis of cultured Chang liver cells--immunoelectron microscope study.
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Acetaldehyde↗
Biomedical subjects
M Tsuchiya
Publications and source records attributed to M Tsuchiya.
Microcirculatory disturbance in indomethacin-induced intestinal ulcer.
Participation of microcirculatory disturbances, especially the role of 5-lipoxygenase products from neutrophils, was investigated in indomethacin (Indo)-induced ulcers of rat small intestine. After Indo treatment (20 mg/kg) in rats, small erosions appeared at 6 h and longitudinal ulcers developed 12 h later. At 6 and 12 h after Indo treatment, severe microcirculatory disturbances were observed under an intravital fluorescence microscope. Significant delay in the clearance and patchy pooling of injected fluorescein isothiocyanate-bovine serum albumin with sludge and stasis were observed in archade vessels of villi of Indo-treated rats. Increased numbers of sticking leukocytes were also detected along submucosal venules in these rats after the infusion of acridine orange. When regional venous blood was collected from the mesentery, a marked increase in neutrophil number and their increased production of oxygen-derived free radicals as determined by chemiluminescence assay were demonstrated at 6 h after Indo treatment. There was also a significant increase in myeloperoxidase activity of the intestinal mucosa at 6 and 12 h after Indo treatment, suggesting a significant neutrophil accumulation at this time. AA-861, a selective inhibitor of 5-lipoxygenase (80 mg/kg), attenuated these microcirculatory changes and neutrophil accumulation in the intestinal mucosa. AA-861 also significantly prevented the formation of intestinal ulcers induced by Indo. However, Indo-induced ulcer formation and leukocyte accumulation in submucosal venules were not attenuated by the treatment of Ono-1078, a potent antagonist of sulfidopeptide leukotrienes. From these observations, it is considered that microcirculatory disturbances, especially leukocyte accumulation and 5-lipoxygenase products, possibly leukotriene B4, may be involved in the development of Indo-induced intestinal ulcer.
Animals↗
Ischemic bowel necrosis induced by endothelin-1: an experimental model in rats.
Local intra-arterial administration of endothelin-1 induced small intestinal mucosal damage in rats in a dose-dependent manner. A remarkable decrease in mucosal blood flow (15% of control values) was observed by a laser Doppler flowmetry 10 min after injection of endothelin-1 (1 nmol/kg). Endothelin-1 at this dose induced significant hemorrhagic and necrotic lesions in the small intestinal mucosa 30 min after the injection. Decreased mucosal blood flow was attenuated to some extent by the pretreatment with platelet-activating factor (PAF) inhibitor, CV-6209, superoxide dismutase plus catalase or the calcium antagonist, nicorandil. All these inhibitors significantly prevented the endothelin-1-induced ischemic necrotic damage in the small intestine. These results suggest a potential role of endothelin-1 in the pathogenesis of ischemic bowel diseases in clinical situations and also the possibility that PAF and oxygen-derived free radicals may be involved as secondary mediators in endothelin-induced intestinal tissue damage.
Animals↗
Proto-oncogene expression in three human hepatoma cell lines, HCC-M, HCC-T and PLC/PRF/5.
Changes of nucleotide sequences and expressions of cellular oncogenes in human hepatoma cell lines, PLC/PRF/5, HCC-M and HCC-T cells, were examined by Southern and Northern blot analyses. The probes used are DNA fragment of myc, N-, H-, K-ras, fos, fms, raf, erb-A, erb-B, and erb-B2 genes and synthetic oligonucleotides corresponding to the part of N-, H-, K-ras genes. The results are as follows. DNA amplification and rearrangement were not detected in these three human hepatoma cell lines. Point mutations at codons 12, 13, and 61 in N- and K-ras genes were not demonstrated in these cell lines. N-, H-, K-ras and myc transcripts were detected in these three cell lines. However, fos gene transcript was detected only in PLC/PRF/5 and HCC-M cells which were derived from hepatitis B related hepatocellular carcinoma and having integrated hepatitis B virus (HBV) DNA. These data showed that there are no specific proto-oncogene expression into RNA except for myc and ras genes, nor DNA rearrangement in these 3 human hepatoma cell lines with regards to at least 10 different oncogenes examined and suggest the relationship between fos gene expression and integration of HBV DNA in host cell DNA.
Blotting, Northern↗
Thioctic acid and dihydrolipoic acid are novel antioxidants which interact with reactive oxygen species.
Thioctic acid (TA) and its reduced form dihydrolipoic acid (DHLA) have recently gained some recognition as useful biological antioxidants. In particular, the ability of DHLA to inhibit lipid peroxidation has been reported. In the present study, the effects of TA and DHLA on reactive oxygen species (ROS) generated in the aqueous phase have been investigated. Xanthine plus xanthine oxidase-generated superoxide radicals (O2-), detected by electron spin resonance spectroscopy (ESR) using DMPO as a spin trap, were eliminated by DHLA but not by TA. The sulfhydryl content of DHLA, measured using Ellman's reagent decreased subsequent to the incubation with xanthine plus xanthine oxidase confirming the interaction between DHLA and O2-. An increase of hydrogen peroxide concentration accompanied the reaction between DHLA and O2-, suggesting the reduction of O2- by DHLA. Competition of O2- with epinephrine allowed us to estimate a second order kinetic constant of the reaction between O2- and DHLA, which was found to be a 3.3 x 10(5) M-1 s-1. On the other hand, the DMPO signal of hydroxyl radicals (HO.) generated by Fenton's reagent were eliminated by both TA and DHLA. Inhibition of the Fenton reaction by TA was confirmed by a chemiluminescence measurement using luminol as a probe for HO.. There was no electron transfer from Fe2+ to TA or from DHLA to Fe3+ detected by measuring the Fe(2+)-phenanthroline complex. DHLA did not potentiate the DMPO signal of HO. indicating no prooxidant activity of DHLA. These results suggest that both TA and DHLA possess antioxidant properties. In particular, DHLA is very effective as shown by its dual capability by eliminating both O2- and HO..
Antioxidants↗
Generation of probucol radicals and their reduction by ascorbate and dihydrolipoic acid in human low density lipoproteins.
Probucol, 4,4'-[(1-methylethylidene)bis(thio)]bis-[2,6-bis(1,1- dimethyl)phenol], is a lipid regulating drug whose therapeutic potential depends on its antioxidant properties. Probucol and alpha-tocopherol were quantitatively compared in their ability to scavenge peroxyl radicals generated by the thermal decomposition of the lipid-soluble azo-initiator 2,2'-azo-bis(2,4-dimethyl-valeronitrile), AMVN, in dioleoylphosphatidylcholine (DOPC) liposomes. Probucol showed 15-times lower peroxyl radical scavenging efficiency than alpha-tocopherol as measured by the effects on AMVN-induced luminol-dependent chemiluminescence. We suggest that probucol cannot protect alpha-tocopherol against its loss in the course of oxidation, although probucol is known to prevent lipid peroxidation in membranes and lipoproteins. In human low density lipoproteins (LDL) ESR signals of the probucol phenoxyl radical were detected upon incubation with lipoxygenase + linolenic acid or AMVN. Ascorbate was shown to reduce probucol radicals. Dihydrolipoic acid alone was not able to reduce the probucol radical but in the presence of both ascorbate and dihydrolipoic acid a synergistic effect of a stepwise reduction was observed. This resulted from ascorbate-dependent reduction of probucol radicals and dihydrolipoic acid-dependent reduction of ascorbyl radicals. The oxidized form of dihydrolipoic acid, thioctic acid, did not affect probucol radicals either in the presence or in the absence of ascorbate.
Antioxidants↗
Establishment of epithelial cell lines from human and mouse thymus immortalized by the 12S adenoviral E1a gene product.
To understand the role of thymic epithelial cells in the development of immature thymocytes, the establishment and cloning of thymic epithelial cell lines must be required. In the present study, we established human and mouse thymic epithelial cell lines through the immortalization by the transfection of cDNA sequences of adenoviral E1a 12S mRNA. This procedure resulted in the isolation of five stable cell lines (one human cell line and four mouse cell lines). These cell lines were positive in cytokeratin demonstrated by immunohistochemistry. Electron microscopic study revealed that they had bundles of tonofilaments and desmosome-like tight junctions. These findings indicate that the cell lines immortalized by E1a gene have retained the properties of epithelial cells. MHC class II antigens were not expressed on these cell lines. When interferon-gamma was added to the cultured epithelial cell lines, MHC class II antigens were induced in their cytoplasm and on their surface membrane, demonstrated by immunohistochemical and immunofluorescent studies. It is suggested that these stable cell lines from human and mouse thymus might serve a good tool for the further study of thymocytes differentiation and of unknown cytokines derived from thymus epithelium.
Animals↗
[The CO2 absorption capacity and color indication of a newly developed soda lime (Wakolime)].
The CO2 absorption capacity and the color change of ethyl violet (color indicator) of Wakolime A (newer type). Wakolime (older type), and Sorb 800 were studied under the clinical anesthetic condition. A two chamber canister was filled with soda lime and anesthetic gas flowed upward through the canister at the rate of 4 ml.min-1. The wall temperature of each chamber was measured with surface temperature sensing device. After 30 hours of use, the colored soda lime in the first chamber was examined with a color analyzer. The change of wall temperature indicated that the CO2 absorption capacity of Sorb 800 was the largest and that of Wakolime was the smallest among the three. The indicator color of both Wakolime A and Sorb 800 was clearer and of more vivid violet than that of Wakolime, which was dull and grayish. These results indicated that Wakolime A is superior in the CO2 absorption capacity and color indication to Wakolime.
Absorption↗
Intralobular heterogeneity of carbon tetrachloride-induced oxidative stress in perfused rat liver visualized by digital imaging fluorescence microscopy.
Spatial and temporal alterations in carbon tetrachloride-induced oxidative stress were studied in perfused hepatic microcirculatory units of the rat by digital imaging microscopic fluorography using dichlorofluorescin diacetate, a hydroperoxide-sensitive fluorogenic probe. The surface of the liver loaded with dichlorofluorescin was microscopically visualized, and the fluorescence of dichlorofluorescin (DCF), a highly fluorescent molecule generated by hydroperoxide-mediated oxidation of dichlorofluorescin, was digitally processed. After completing the experiments, fluorescein-labeled albumin was injected into the perfusate to confirm the state of sinusoidal perfusion and the topography of hepatic microangioarchitecture in the area studied. During transportal infusion of carbon tetrachloride, DCF fluorescence was observed predominantly in the pericentral area and was attenuated by pretreatment with SKF-525A, suggesting the involvement of cytochrome P-450. After peaking at the maximum level, the pericentral DCF fluorescence gradually decreased in parallel to the loss of viability, implicating the causal role of intracellular hydroperoxide formation in hepatocellular injury. In retrogradely perfused liver, in which intralobular O2 gradient was reversed, no significant activation of DCF fluorescence was observed among hepatic lobules. It is therefore conceivable that the zonal heterogeneity of carbon tetrachloride-induced lipid peroxidation may depend on at least two factors, sinusoidal oxygenation and the intralobular distribution of cytochrome P-450. Furthermore, development of the current technique will provide a useful method to investigate the microtopography of oxidative stress in organ microcirculatory units.
Animals↗
[Convenient and useful computer software for the anesthesiologist].
Convenient and useful computer software for the anesthesiologist to draw figures for presentation, or to perform statistical analysis were presented. Each software is selected in the condition that it does not need special optional hardware, and processes the data quickly even with an old type computer, and also that even a computer beginner can easily operate it. The presented software are as follows: 1) Silhouette (drawing figures), 2) NEO (drawing figures), 3) Graph note (drawing figures), 4) Chart up science (drawing figures), 5) Z's word JG (making slide manuscripts) and 6) Biostat (doing statistical analysis).
Anesthesiology↗
Thymectomy in ulcerative colitis: a report of cases over a 13 year period.
Seventy eight patients with ulcerative colitis were treated by thymectomy combined with conventional therapy. An interim analysis was made after a median follow-up of 40 months in the thymectomized group and after 25 months in 173 from a non-thymectomized group. The percentage of remission periods in the thymectomized group was significantly higher than that in the non-thymectomized group as estimated by the "patient-month" method. Histological examination of the excised thymus revealed hyperplasia of the thymic epithelial cells and/or the formation of lymphoid follicles. Anticolon antibody activity of the serum from the thymectomized patients decreased gradually and disappeared in 5 years or more. "Thymectomy via the suprasternal notch with parasternal incision" which was applied in this study, is simple and not invasive. Therefore, it is recommended that thymectomy should be considered as one of the treatment for patients who are resistant to conventional therapy.
Adolescent↗
[The hemodynamic and metabolic changes following tourniquet release during halothane anesthesia].
We measured the hemodynamic and metabolic changes following the release of lower limb tourniquet during halothane anesthesia, and discussed the cause of reduction in the blood pressure and the effective method for prevention. Cardiac index decreased transiently following the release, and gradually increased afterward. Systemic vascular resistance index decreased continuously and progressively until 10 minutes after the release. These results indicate that the causative factors of reduction in the blood pressure are temporary decrease in cardiac output and subsequent decrease in systemic vascular resistance. In mixed venous blood, pH decreased, PCO2 increased and HCO3- remained unchanged after the release. It seems that pH reduction was chiefly caused by the increase in PO2. Thus, the preventive method for reduction in the blood pressure following the tourniquet release may be as follows; 1) intravenous fluid loading, 2) hyperventilation, 3) elevation of lower limb, 4) lightning the anesthesia, and 5) finally giving vasopressors.
Anesthesia, Inhalation↗
[Determination of fentanyl concentrations in the blood with the application of high performance liquid chromatography].
We developed a method to determine the blood concentration of fentanyl by using reverse phase partition liquid chromatography. The extract rate of fentanyl from blood 10 ml was 79 +/- 2%, and the minimum detective concentration of fentanyl was 158 pg.ml-1. These performances are sufficient to determine the blood concentration of fentanyl in clinical anesthesia.
Anesthesiology↗
Dynamics of resonant and nonresonant electron tunneling in double-quantum-well structures under electric fields.
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Journal Article↗
[Clinical relevance of oxidative stress--with special references to organ microcirculatory disturbance].
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Animals↗
Multiple forms of Go alpha mRNA: analysis of the 3'-untranslated regions.
Go, a guanine nucleotide binding protein found predominantly in neural tissues, interacts in vitro with rhodopsin, muscarinic, and other receptors and has been implicated in the regulation of ion channels. Despite the virtual identity of reported cDNA sequences for the alpha subunit of Go (Go alpha), multiple molecular weight forms of mRNA have been identified in tissues from all species examined. To investigate the molecular basis for the size heterogeneity of Go alpha mRNAs, four cDNA clones were isolated from the same retinal lambda gt10 cDNA library that was used earlier to isolate lambda GO9, a clone encompassing the complete coding region of Go alpha. These clones were identified as Go alpha clones based on nucleotide sequence identity with lambda GO9 in the coding region; they diverge, however, from lambda GO9 in the 3'-untranslated region 28 nucleotides past the stop codon. An oligonucleotide probe complementary to a portion of the 3'-untranslated region of lambda GO9 that differs from the newly isolated clones hybridized with 3.0- and 4.0-kb mRNAs present in bovine brain and retina whereas a similar probe for the unique region of the new clones hybridized with a 4.0-kb mRNA in both tissues and with a 2.0-kb mRNA found predominantly in retina. A similar hybridization pattern was observed when brain poly(A+) RNA from other species was hybridized with the different 3'-untranslated region probes. It appears that differences in the 3'-untranslated regions could, in part, be the basis for the observed heterogeneity in Go alpha mRNAs.
Animals↗
[A case of Weil's disease complicated with liver disorder, acute kidney failure and myocarditis treated by hemodialysis and plasma exchange].
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Acute Kidney Injury↗
Multiple elements in the promoter of granulocyte colony-stimulating factor gene regulate its constitutive expression in human carcinoma cells.
Some human carcinoma cells constitutively produce granulocyte colony-stimulating factor (G-CSF) which stimulates the proliferation and differentiation of the progenitor cells of neutrophilic granulocytes. By introducing mouse G-CSF chromosomal gene or its promoter DNA into human carcinoma cell lines of CHU-2, SK-HEP-1, and U-87MG, it was shown that the constitutive expression of G-CSF in these carcinoma cells was due to the intrinsic activation of nuclear factors which work on the promoter region of the G-CSF gene. A series of 5' deletion mutants, linker scanning mutants, and internal deletion mutants was constructed in the promoter of mouse G-CSF gene and was introduced into human CHU-2 cells to analyze their promoter activities. These studies demonstrated that at least three regulatory elements in the promoter of the G-CSF gene are essential for the constitutive expression of G-CSF in CHU-2 cells. These elements include the consensus decanucleotide "GAGRTTCCA/CC" present on G-CSF, granulocyte/macrophage colony-stimulating factor, and interleukin 3 genes and the "ATTTGCAT" octamer transcription factor binding site. Some point mutations in these consensus sequences significantly diminished the promoter activity in CHU-2 cells.
Adenocarcinoma↗