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Biomedical subjects

K Fukuyama

Publications and source records attributed to K Fukuyama.

At least 145 records · Page 8Linked to original sources
PubMed1986

[A case of CSF (colony-stimulating factor)-producing lung cancer and a review of the literature].

A 36-year-old man with large-cell carcinoma of the lung showed marked leukocytosis of up to 109,300/mm3. As high CSF activity was detected in the pleural effusion and the supernatant of cell cultures originating from the effusion, a diagnosis of CSF-producing lung cancer was made. Twenty-one CSF-producing lung cancers have been reported in Japan. They comprised 13 large-cell carcinomas, six squamous cell carcinomas, one adenocarcinoma and one small cell carcinoma.

Adult↗
PubMed1985

Preliminary X-ray diffraction studies on a ferredoxin from the thermophilic archaebacterium, Thermoplasma acidophilum.

A ferredoxin from the thermophilic archaebacterium, Thermoplasma acidophilum, is supposed to contain two (4Fe-4S) active centers; one center could be linked by four cysteine residues to the protein and the other bonded with three cysteines and an unknown group. This ferredoxin has been crystallized by salting-out against 2.3 M-ammonium sulfate solution. The space group is P21212 with cell dimensions of a = 59.20 A, b = 52.77 A and c = 41.28 A. Four molecules pack in the unit cell with Vm = 2.03 A3/dalton.

Amino Acid Sequence↗
PubMed1985

Purification and characterization of major extracellular proteinases from Trichophyton rubrum.

Two extracellular proteinases that probably play a central role in the metabolism and pathogenesis of the most common dermatophyte of man, Trichophyton rubrum, were purified to homogeneity. Size-exclusion chromatography and Chromatofocusing were used to purify the major proteinases 42-fold from crude fungal culture filtrate. The major enzyme has pI 7.8 and subunit Mr 44 000, but forms a dimer of Mr approx. 90 000 in the absence of reducing agents. A second enzyme with pI 6.5 and subunit Mr 36 000, was also purified. It is very similar in substrate specificity to the major enzyme but has lower specific activity, and may be an autoproteolysis product. The major proteinase has pH optimum 8, a Ca2+-dependence maximum of 1 mM, and was inhibited by serine-proteinase inhibitors, especially tetrapeptidyl chloromethane derivatives with hydrophobic residues at the P-1 site. Kinetic studies also showed that tetrapeptides containing aromatic or hydrophobic residues at P-1 were the best substrates. A kcat./Km of 27 000 M-1 X S-1 was calculated for the peptide 3-carboxypropionyl-Ala-Ala-Pro-Phe-p-nitroanilide. The enzyme has significant activity against keratin, elastin and denatured type I collagen (Azocoll).

Chromatography, Gel↗
PubMed1985

Stimulation of pheomelanogenesis in cultured B16 melanoma cells by 4-tertiary butylcatechol.

Intermediates of pheomelanin in tissue cultured B16 melanoma cells were analyzed by high performance liquid chromatography, and reduced glutathione (GSH), L-dopa, 2-[(L)-S-cysteinyl]-L-dopa (2-SCD) and 5-[(L)-S-cysteinyl]-L-dopa (5-SCD) were quantified. The effects of 4-tertiary butylcatechol (TBC), an antioxidant which causes skin depigmentation, on the levels of the intermediate were then examined. A concentration of 10(-4) M TBC increased the intracellular levels of GSH, 2-SCD and 5-SCD, whereas the L-dopa level was unchanged. The time-course of the increased intermediates corresponded to the elevation of glutathione-metabolizing enzyme activities previously reported by Kawashima et al. [J. invest. Derm. 82, 53 (1984)] in the same cell line exposed to 10(-4) M TBC. The findings establish chemical evidence that TBC stimulates pheomelanogenesis in melanocytes.

Animals↗
PubMed1985

Immunological detection of a cysteine protease in the skin and other tissues.

Monospecific antibody directed to cysteine protease of 2-day-old rat epidermis recently characterized as being different from the proteases previously reported was produced in rabbits. By immunofluorescence microscopy and immunoperoxidase staining with an avidin-biotin-peroxidase method the protease was found to be present in the epidermis of rodents of different ages as well as that of humans, but not in the dermis. The staining in germinative cells was more intense than in cells in the superficial layers. It appeared as irregular patches in the nuclei and stained more diffusely in the cytoplasm where small granular components, strongly stained, were identified. The staining patterns in granular cells showed accumulation of the antigen in a granular form. The morphology and distribution of granules resembled those of keratohyalin-like granules in the nucleus and dense homogeneous deposits in the cytoplasm. In cornified cells the reaction product was localized by the plasma membrane where concentration of the dense homogeneous deposits occurred, suggesting that the cysteine protease is one component of the unique and characteristic structure of differentiated keratinocytes. In addition, the cysteine protease antigen having the same molecular weight as the epidermal enzyme was detected in liver, kidney and lung indicating a wider tissue distribution of the protease. The significance of the protease in regulation of cellular functions remains to be investigated.

Animals↗
PubMed1985

Autoradiographic studies of cell dynamics in immunogenic granulomas transplanted into the skin of athymic nude mice.

Schistosome egg granulomas in the livers of thymus-intact (nu/+) mice are large and contain eosinophils and mast cells, while those in nude athymic (nu/nu) mice are small and devoid of eosinophils or mast cells. To investigate the cell sources and cell kinetics of hepatic granulomas of nu/+ mice isolated and grafted into the skin of nu/nu mice, biopsies taken after grafting were examined by light and electron microscopy and autoradiography after 3H-thymidine (TdR) injection of either the donor or recipient mice. At 1 week, the grafted granulomas appeared to be amorphous and were surrounded by leukocytes, and the 3H-TdR-labeled donor cells had disappeared. After 2 weeks, repopulation with macrophages began and by 3-5 weeks, the granulomas morphologically resembled hepatic lesions of nu/+ mice. Injection of recipients with 3H-TdR before grafting, showed that labeled macrophages, eosinophils, and mast cells repopulated in granulomas. No granulomas were seen when nu/nu mice were grafted with schistosome eggs alone, and organ culture of nu/+ granulomas before grafting reduced the number of repopulated granulomas. These findings indicate that nu/+ cells in grafted granulomas are replaced by nu/nu cells. Granulomatous reaction of the grafted sites in nu/nu mice is influenced by a substance in nu/+ granulomas, and cells of nunu mice locally acquire a nu/+ type response.

Animals↗
PubMed1985

Detection of granuloma-associated plasminogen activator in experimental murine schistosomiasis.

Hypersensitivity granulomas induced by infection with Schistosoma mansoni were isolated from the livers of BALB/c mice after 7, 8, 10, and 12 weeks. The parasite egg-granulomas were sequentially extracted with a Tris-buffered saline (soluble fraction) and 2 M KSCN (bound fraction). Fibrinolytic enzyme activity measured with both synthetic substrates and fibrin plates demonstrated an elevated level of plasminogen activator activity in the bound fraction 7-8 weeks after infection when mature granulomas first began to appear, followed by a gradual decrease 10-12 weeks after infection. An electrophoretic enzymography technique revealed multiple molecular species of plasminogen activator at Mr = 95K, 74K, 60K, 45K, and 24K. The bands with Mr = 45K and 24K were found compatible with the electrophoretic pattern of macrophage-plasminogen activator. When the granulomas reached maximum size after 10 to 12 weeks, the plasminogen activator with 45K and 24K diminished, while plasminogen activator activity at Mr = 95K, 74K, and 60K remained unchanged suggesting the presence of both vascular and tissue types of plasminogen activators. There was no urokinase-type plasminogen activator detectable in granulomas at any time. In the soluble fraction no enzymatic activity was found, whereas regulating inhibitor activity for plasminogen activator was consistently detectable.

Animals↗
PubMed1985

Characterization of Fe2+-activated acid phosphatase in rat epidermis.

A particulate acid phosphatase (EC 3.1.3.2, orthophosphoric monoester phosphohydrolase (acid optimum)) was extracted in 1 M KCl, from 2-day rat epidermis. The enzyme has a Mr of 32,000, but two forms, F1 and F2 with pI values of 8.6 and 8.3, respectively, were identified while the pI values of other acid phosphatases soluble in sucrose and Triton X-100 were all acidic. F1 and F2 also differed from other epidermal acid phosphatases because they were (a) activated by Fe2+ and reducing agents, (b) showed immunological cross-reactivity with purple acid phosphatase of rat spleen and (c) dephosphorylated phosvitin and alpha-casein even though they had rather high Km values.

Acid Phosphatase↗
PubMed1985

Effect of ultraviolet B radiation on S-100 protein antigen in epidermal Langerhans cells.

Ultraviolet B (UVB) radiation has been shown to induce significant alterations in both function and surface antigen expression of epidermal Langerhans cells (ELC). In this study we investigated the effect of UVB radiation on ELC marker S-100 protein antigen (S-100 Ag) which is present in the nucleus and cytoplasm of human ELC. A total of 34 sites on 31 volunteers were exposed to 3 MED (minimal erythema dose) of UVB and biopsied at various times up to 7 days after irradiation. Skin from 9 noninjured and 7 slice-wounded subjects served as controls. The avidin-biotin-peroxidase staining technique was used to identify S-100 Ag in sections of formalin-fixed, paraffin-embedded tissue, and the numbers of stained suprabasal dendritic cells were then counted over a 200 basal cell length of interfollicular epidermis. Noninjured skin had 3.56 +/- 3.01 cells, whereas slice-wounded skin had elevated numbers (greater than 10.0 cells) at 1, 24, and 48 h after injury. Following UVB irradiation, a significant (p less than 0.001) increase in antigen-positive cells (14 +/- 3.46) was found at 1 h; this number declined to just below normal at 12 h, but by 48 h returned to and remained at preinjury levels. In contrast to previous observations of the depletion of ELC surface markers by UVB radiation, we demonstrate here that the numbers of S-100 Ag-positive ELC actually increase following comparable doses of radiation. Since this increase occurs so rapidly following both UVB irradiation and slice injury, S-100 Ag may be synthesized or unmasked within the ELC as a response to wounding of the epidermis.

Adult↗
PubMed1985

Effect of captopril on transplanted schistosome egg granulomas in skin.

Groups of athymic nude (nu/nu) and littermate (nu/+) mice received transplants into skin of hepatic egg granulomas isolated from Schistosoma mansoni-infected nu/+ mice. Some were fed captopril, an angiotensin-converting enzyme inhibitor, and other animals served as controls. Lesions were removed at 2, 4, and 6 weeks after implantation and prepared for light and electron microscopy. The number of new egg granulomas repopulated by recipient cells was compared by counting more than 100 eggs per specimen, randomly, double blind, and the effects of captopril on developing granulomas were determined. Results in nude mice indicated a trend for increasing granuloma takes with time in captopril-treated animals compared to controls, especially in mice who started therapy 2 weeks after transplantation. In nu/+ recipients the differences were more striking and statistically significant at 2 and 6 weeks after grafting. The granulomas in treated mice in both groups were larger than those in controls, but the cellular composition in the granulomas that developed were similar. New granulomas consisted mainly of eosinophils and mixed-function macrophages, although some purely secretory macrophages also were seen. The findings demonstrate that captopril ingestion enhanced granulomatous inflammation induced by skin transplantation of schistosome egg granulomas and significantly increased isograft takes. The results may have relevance for treating cutaneous diseases like sarcoidosis with captopril.

Animals↗
PubMed1984

An immunohistochemical study of nuclear proteins in differentiating epidermal cells.

Both DNA and RNA disappear from the nucleus during differentiation of granular cells into cornified cells but the fate of nuclear proteins remains unknown. We investigated localization of nuclear proteins in rat epidermis by light and electron microscopic immunoperoxidase techniques. As a probe, three sera that reacted, respectively, with the nucleoplasm, nucleolus, and nuclear envelope of basal cells of rat epidermis were used. In granular cells both the antinucleoplasm serum and antinucleolus serum increased intensity of the nuclear staining, but they reacted also with ribosomes, filaments, and periphery of keratohyalin granules in the cytoplasm. The staining appeared diffusely in cornified cells and identification of nuclear components became impossible. In contrast, the antinuclear envelope serum stained only the nuclear outline in granular cells and continued to stain the nuclear contour in cornified cells of the fourth and fifth proximal cell layers. The antigenic components surrounded amorphous but not filamentous materials in cornified cells. These findings suggest that some nuclear proteins become immunologically indistinguishable from cytoplasmic protein. However, the nuclear envelope protein maintains its localization even after nucleic acids are lost and the nuclear space is detectable in cornified cells by use of autoantibody directed to this protein(s).

Animals↗
PubMed1984

Enzymatic activity of metal-binding proteins in epidermal cells.

Enzymatic activity was investigated in metal-binding proteins from rat epidermal cells. Tris-HCl buffer soluble and KSCN solubilized proteins were extracted stepwise from granular and cornified cells of 2-day old rat epidermis. Each extract was separately applied to a Cu2+ or Zn2+ chelate Sepharose 6B column and the proteins were eluted with buffers of different pHs and finally with EDTA solution. Metal chelate-binding proteins were found in both soluble and solubilized proteins but there was a larger amount in the latter. Affinity of the proteins to bind with Cu2+ chelate was greater than that with Zn2+ chelate. In Tris-HCl buffer extract, histidase activity was detected in Cu2+ chelate-binding proteins, but not in Zn2+ chelate-binding proteins. Acid phosphatase, cysteine proteinase, dipeptidase, cathepsin D, beta-galactosidase, gelatin hydrolase, and superoxide dismutase did not bind to metal chelates although these enzymes, except acid phosphatase, were inhibited by Cu2+, but not by Zn2+. In contrast, KSCN solubilized metal chelate-binding proteins showed plasminogen activator, acid phosphatase, and gelatin and casein hydrolases while histone hydrolase did not bind to either chelate column. Since metal-binding proteins in rat epidermal cells have been shown previously to be histidine- and cysteine-rich proteins concentrated in keratohyalin granules, interaction of metals and the structural proteins with certain enzymes may be involved in the regulation of epidermal cell functions.

Animals↗
PubMed1984

Biochemical characterization of arylsulfatases detected in granulomatous inflammation.

Arylsulfatases A and B were measured in the liver of mice infected with Schistosoma mansoni. The increase of total arylsulfatases paralleled enlargement of the granulomas. It began at 7 weeks after infection and reached a maximum at 10 to 14 weeks when the enzyme activity became about 2.5 times that of normal liver. The elevated enzyme activity was due to granulomatous tissue, because when granulomas were separated from hepatic cells, the former contained the increased activity but the latter did not. Arylsulfatase A, arylsulfatase B, and arylsulfatase Bv, in both normal liver and granulomas, were separated by anion-exchange column chromatography and differences in net charges of these enzymes were demonstrated by polyacrylamide gel electrophoresis. Biochemical properties were indistinguishable between arylsulfatase B and arylsulfatase Bv while they differed from arylsulfatase A. Granulomas at 8 weeks after infection showed 3.0-, 3.5-, and 5.0-fold increases in activity for arylsulfatase A, B, and Bv, respectively. As the granulomas enlarged, by 12 weeks, arylsulfatases B and Bv activities further increased but the arylsulfatase A value remained the same as that of 8 weeks. The finding suggests that arylsulfatases are involved in granuloma development and arylsulfatases B and Bv activities may reflect functions of macrophages and other cells including fibroblasts.

Animals↗
PubMed1984

Detection and characterization of epidermal proteinases by polyacrylamide gel electrophoresis.

Electrophoresis of cornified cell extracts of 2-day-old rats, using SDS polyacrylamide gels copolymerized with alpha-casein or gelatin with or without plasminogen, was performed. Both Tris-HCl buffer soluble protein and KSCN solubilized protein contained a number of hydrolases for alpha-casein and/or gelatin, but PA (mol. wts 57 and 50K) was found only in the KSCN extract. The pH dependency, substrate specificity and mol. wt of plasminogen-independent proteinases were comparatively determined and DFP inhibition tested. This simple technique helped to identify the presence of several proteinases and to characterize them in partially purified epidermal extracts.

Animals↗
PubMed1984

Immunohistochemical demonstration of S-100 protein antigen-containing cells in beryllium-induced, zirconium-induced and sarcoidosis granulomas.

S-100 protein has been detected in epidermal Langerhans cells, interdigitating reticulum cells, and large mononuclear cells of cutaneous T-cell lymphomas. Experimental data imply these cells are involved in the presentation of antigens and/or maturation of T-lymphocytes. The authors studied the morphology and distribution of S-100 protein antigen-containing cells in cutaneous sarcoidosis and metal-induced granulomas, both immunogenic and foreign-body types, with light and electron microscopic immunoperoxidase technics. Immunological reaction was seen in Langerhans cells, peripheral nerves, and granulomatous lesion dendritic cells. The latter showed a large, irregular nucleus and branching cytoplasm. They intermingled with other mononuclear cells in the granulomas but not with organized epithelioid cells. Morphometric quantification of dendritic cells in the three types of granulomas revealed statistically significant differences (P less than 0.005) and electron microscopy demonstrated their typical cytoplasmic appearance, without Birbeck granules. The increased number of dendritic cells in immunogenic granulomas, and their shared antigenic components with Langerhans cells suggest they act as accessory cells in eliciting the granulomatous response.

Beryllium↗
PubMed1984

Effects of 4-tertiary butyl catechol on glutathione-metabolizing enzymes in vivo and in vitro.

4-Tertiary butyl catechol (TBC) causes depigmentation in humans and animals and stimulates formation of pheomelanosomes. In this study, we investigated the effects of noncytotoxic doses of TBC on glutathione S-transferase (GST) activity in the skin of Uscd strain mice and B16 murine melanoma cells in culture, in relation to changes in activities of glutathione reductase (GR) and gamma-glutamyl transpeptidase (GGT) reported to be involved in pheomelanogenesis. Occurrence of pheomelanosomes in skin melanocytes was demonstrated by electron microscopy and reduction (25%) of eumelanin content in melanoma cells was shown by spectrophotometry. Topical application of 1 M TBC-DMSO-acetone solution on the ear skin elevated GST activity about 27%, and activities of GGT and GR to 35% and 19%, respectively, within 1 week. Melanoma cells cultured in 10(-4) M TBC-containing medium for 2 h showed no changes in GST and GGT activities, but 12% increase of GR activity during the first 12 h. Activities of all 3 enzymes was elevated (11-17%) 24 h later. The elevation detected by 48 h was 25% for GST, 26% for GGT, and 14% for GR. The findings were interpreted to show that depigmentation produced by the antioxidant results from stimulated pheomelanogenesis through activation of glutathione-metabolizing enzymes and suppressed oxidation of eumelanin intermediates.

Animals↗