Expression of smg p21A and ras p21 in epidermal neoplasms.
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GTP-Binding Proteins↗
Biomedical subjects
H Hachisuka
Publications and source records attributed to H Hachisuka.
Fractionation of Paget cells in extramammary Paget's disease.
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Aged↗
Purification and characterization of prolyl endopeptidase from rat skin.
An enzyme with the specificity of a prolyl endopeptidase was purified approximately 329-fold from rat skin. The enzyme has a molecular weight of 70,000 as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and a pH optimum of 5.8 as checked with 7-(Succinyl-Gly-Pro)-4- methylcoumarinamide (Suc-Gly-Pro-MCA) as the substrate. The optimal temperature for the enzyme activity was 40 degrees C. The Km and Vmax values for Suc-Gly-Pro-MCA were 0.7 mM and 68 nmol/min per mg protein, respectively. The enzyme activity was markedly inhibited by diisopropyl fluorophosphate, p-chloromercuribenzoic acid, N-ethylmaleimide, Zn2+ and Cu2+, while it was partially inhibited by phenylmethanesulphonyl fluoride. The purified enzyme was shown to release the N-terminal tetrapeptide, Arg-Pro-Lys-Pro, from substance P producing the C-terminal heptapeptide, Gln-Gln-Phe-Phe-Gly-Met- CONH2. In the skin, this enzyme might be related to the inactivation of substance P.
Amino Acid Sequence↗
Cell cycle analyses of extramammary Paget's disease.
The cell cycle of Paget cells in extramammary Paget's disease was investigated using microscopic analysis of the DNA content, and using immunohistochemistry for 5-bromodeoxyuridine (BrdU) incorporation. Stripped skin was treated with tetrasodium ethylendiamine tetraacetate dihydrate (EDTA) and trypsin, and epidermal cells suspensions were obtained. Microscopic analysis of the DNA content in the cells was performed. The resulting DNA histogram was close to the normal ploidy. Specimens from the involved skin were incubated with BrdU, and sections were immunostained with anti-BrdU monoclonal antibody. A few nuclei showed positive staining for BrdU. The present study showed that the proliferative potential of Paget cells in extramammary Paget's disease was low. This may explain why lesions of extramammary Paget's disease enlarge rather slowly.
Aged↗
Expression of ras proto-oncogene related protein p21 in normal human skin and cutaneous tumours.
The cellular proto-oncogene, ras, is known to play an important role in the regulation of cell growth and proliferation in normal and malignant conditions. The present study was undertaken to immunohistochemically examine the expression of ras protooncogene product p21 in normal human skin and some cutaneous tumours. In normal skin, the expression of p21 was found in sweat glands, sebaceous glands, capillary endothelium, and smooth muscles, while epidermis was devoid of reaction product. Keratoacanthoma and the granular cells of verruca vulgaris were immunoreactive to the antibody for p21. Bowen's disease and squamous cell carcinoma were positive for p21, but basal cell carcinoma and seborrheic keratosis were negative. In mammary and extramammary Paget's diseases, the immunoreactivity was inconsistent. The expression of p21 in malignant melanoma cells was intense, whereas normal melanocytes and nevus cells were devoid of the expression. These results suggest that the expression of p21 does not correlate with nuclear anaplasia and malignant behaviour of cutaneous tumours.
Bowen's Disease↗
Epidermal keratinocytes of bullous pemphigoid express intercellular adhesion molecule-1 (ICAM-1).
Intercellular adhesion molecule-1 (ICAM-1) is the ligand for lymphocyte function associated antigen-1 (LFA-1), mediating the adhesion of lymphocytes to vascular endothelium. Keratinocytes are known to express ICAM-1 in some inflammatory dermatoses. Using an indirect immunofluorescence method, we examined the patterns of ICAM-1 and LFA-1 staining in bullous pemphigoid (BP) lesions and compared them to pemphigus vulgaris (PV) cases. ICAM-1 was expressed on the cell surface and in the cytoplasm of epidermal keratinocytes at the sites of erythematous and bullous lesions of BP. LFA-1 molecules were expressed on T cells at the basement membrane zone. In addition, HLA-DR-positive keratinocytes were observed in the basal layer. ICAM-1 was not, however, expressed on epidermal keratinocytes in uninvolved skin from BP patients, PV or normal control skin. It is known that ICAM-1 is expressed on keratinocytes at the site of lymphoid infiltration in cutaneous dermatoses and that LFA-1-positive T cells can bind to interferon gamma-induced ICAM-1-positive keratinocytes. Our results suggest that cellular immunity involving ICAM-1 and LFA-1 may play a part in the pathogenesis of bullous pemphigoid.
Cell Adhesion↗
Epidermolysis bullosa acquisita with oesophageal stenosis.
Epidermolysis bullosa acquisita (EBA) is a rare acquired subepidermal blistering disorder associated with autoimmunity to type VII collagen. Although the full clinical spectrum of EBA is still being defined, it is now known that EBA has greater clinical heterogeneity than previously suggested. We describe a patient with EBA which closely approximated the severity of the recessive form of dystrophic epidermolysis bullosa.
Autoantibodies↗
Sclerotic fibromas of the skin--reports of two cases and a review of the literature in Japan.
Two cases of sclerotic fibromas of the skin are presented. Clinical and light-microscopic findings are also reviewed from 8 sclerotic fibromas of the skin arising in other Japanese patients. The patients in this series had no manifestations of Cowden's disease.
Adult↗
Surface charge of fractionated guinea pig keratinocytes measured by free-flow cell electrophoresis.
Keratinocytes differentiate from basal cells to spinous, granular, and horny layer cells. It is known that alterations in the surface charge of cell membranes in most cases reflect the processes of differentiation. By using a continuous colloidal silica (Percoll) density gradient, keratinocytes may be separated into three fractions which correspond to their arrangement in vivo. Using a free-flow cell electrophoretic technique, we measured the electrophoretic mobility of guinea pig keratinocytes. Electrophoretic mobility histograms of basal and granular cells showed slow and fast monophasic patterns, respectively. In spinous cells, a biphasic pattern of slow and fast electrophoretic mobility was present. The electrophoretic mobility level of guinea pig keratinocytes was slightly reduced with neuraminidase digestion. Those of human red blood cells and lymphocytes, however, were markedly decreased. These results indicate that membrane charge density is lower in basal cells and higher in granular cells and that the membrane charge density of guinea pig keratinocytes involves not only neuraminic acid residues but also other substance(s). Our results illustrate the alterations of cell membrane charge properties during epidermal cell differentiation.
Animals↗
Densitometric determination of human papillomavirus DNA quantities by chromato-scanning in the fluorescence mode.
The quantitation of human papillomavirus DNA isolated from warts by chromato-scanning (fluorescence mode) photographs of ethidium bromide-stained agarose gels is described. Excitation at 200 nm (with a cutoff filter at 400 nm) generates fluorescence from the white portion of the printing paper. The fluorescent intensity correlated with the quantities of DNA in the band of interest. The amounts of DNA were determined using calibration curves of approximately the same size as lambda phage DNA fragments. This general method of quantification is applicable to photographs of other types of polynucleotides capable of being separated and stained in a gel medium.
Bacteriophage lambda↗
Bullous pemphigoid antigen expression in Pam 212 cells induced by the addition of platelet activating factor.
Platelet activating factor (Paf-acether) is a phospholipid which has various activities including platelet and neutrophil aggregation and eosinophil chemotaxis. We have previously reported that the blister fluids of bullous pemphigoid possess platelet aggregation activity and suggested that Paf-acether might be concerned with the accumulation and activation of neutrophils and eosinophils. In this study, we examined the influence of Paf-acether on BP antigen expression on Pam 212 cells. Paf-acether enhanced the expression of BP antigen on Pam 212 cells and this expression was blocked by Paf-acether antagonist. Our observations might suggest that Paf-acether contributes to the blister formation not only by the activation of inflammatory cells but the enhancement of BP antigen.
Animals↗
Monoclonal antibody KOLT-2 enhances DNA synthesis and expression of IL-2 receptors on activated T cells.
The functional properties of the KOLT-2 antigen, which was placed among the CD28 group in the 3rd International Workshop on Human Leukocyte Differentiation Antigens, are described. The method of production and a detailed characterization of the monoclonal antibody KOLT-2 including stain affinities and influence of T cell activation are presented. While KOLT-2 antigen was selectively expressed on human thymocytes, adult T-cell leukemia-lymphoma cells and activated T cells, the antigen was also spontaneously expressed by resting T cells after 24 hrs culture. Expression was transient, since it disappeared after 96 hrs. DNA synthesis and expression of Tac antigen on activated T cells was enhanced by stimulation with the KOLT-2. These observations suggest that the KOLT-2 antigen may have an important role in the early stages of T cell activation.
Antibodies, Monoclonal↗
Correlation between DNA quantities in agarose gel and chromatographic areas of photographed DNA bands.
To quantitate DNA from photographs, correlations between chromato-scanned (in the fluorescence mode) and virtual areas from white DNA bands on photographs were examined. A linear correlation between the two measurements with coefficients higher than 0.89 was observed. The dynamic range of the fluorescence detection yielded good linearity for a wide range of amounts of DNA (microgram-ng) on the photograph.
Chromatography↗
Low concentrations of interferon-gamma stimulate DNA synthesis in normal human keratinocytes during the initial stages of cultivation.
High concentrations of interferons can inhibit keratinocyte proliferation and induce the expression of HLA-DR antigen on keratinocytes. In the present study, the effects of low concentrations of recombinant human interferon alpha, beta and gamma were examined on DNA synthesis and the expression of HLA-DR and bullous pemphigoid (BP) antigens in normal human keratinocytes. Low concentrations of interferons induced DNA synthesis in normal human keratinocytes when epidermal cell growth factor and low Ca++ were used during the initial stages of culturing. This result indicates that physiological concentrations of interferons can induce keratinocyte activation.
Animals↗
Bialaphos poisoning with apnea and metabolic acidosis.
A 64-year-old man with ethanol intoxication, ingested a bottle of Herbiace (100 ml, 32 w/v% of bialaphos, CAS #35597-43-4, Meiji Seika Kaisha, Tokyo, Japan). He had severe metabolic acidosis and was treated with infusions of sodium bicarbonate and furosemide, plus gastric lavage and enema. The metabolic acidosis improved 15 hours after treatment but nystagmus, apnea and convulsions were progressive. Although his sensorium was clear, spontaneous respirations were not observed for 64 hours. The electroencephalographic findings of atypical triphasic waves and slow waves suggest a unique response to bialaphos poisoning. His clinical course indicates that the management of apnea is critically important to recovery from bialaphos poisoning.
Acidosis↗
Alterations in membrane fluidity during keratinocyte differentiation measured by fluorescence polarization.
The epidermis shows a distinctive pattern of differentiation wherein keratinocytes proliferate in the basal cell layer and mature into spinous and granular cells. Using a discontinuous density-gradient centrifugation method, guinea-pig keratinocytes were separated into high (HDF), intermediate (IDF), and low (LDF) density fractions. Morphological and flow cytometrical observations demonstrated that HDF, IDF, and LDF were basal, spinous, and granular cell-rich fractions, respectively. Membrane fluidity of the fractionated keratinocytes was measured by diphenylhexatriene fluorescence polarization. Polarization (p)-value of keratinocytes was negatively correlated with temperature. At each temperature, HDF cells showed a lower p-value than IDF or HDF cells except at 40 degrees C. Since a low p-value indicates a high degree of Brownian motion, membrane fluidity is higher in basal cells and lower in spinous and granular cells. Our results indicate that membrane fluidity of guinea-pig keratinocytes decreases during their maturation.
Animals↗
Human papillomavirus DNA from warts for typing by endonuclease restriction patterns: purification by alkaline plasmid methods.
The alkaline plasmid DNA extraction method of Birnboim and Doly was applied for the isolation of human papillomavirus (HPV) from warts. Tissue from common and plantar warts was digested with proteinase K, and the extrachromosomal circular covalently-closed form of HPV-DNA was rapidly extracted by alkaline sodium dodecyl sulphate and phenol-chloroform treatment. Recovery of HPV-DNA from the tissue was sufficient for determination of endonuclease restriction patterns by agarose gel electrophoresis.
Adolescent↗