Superoxide dismutase (SOD), which catalytically scavenges superoxide anion (O–2), constitutes an essential defense against the toxicity of oxygen. We investigated the enzyme activity of pig skin epidermis. SOD activity was determined by monitoring the inhibitory effect of SOD on red formazan formation from neotetrazolium, which depends on (O–2) generation. (O–2) was generated by the hypoxanthine‐xanthine oxidase reaction. Pig epidermis contained significant amounts of heat‐labile SOD activity which was proportional to the added epidermal homogenate. The optimal pH of the reaction was between pH 8.2 and 8.5. Metallochelating agents such as cyanide, sodium azide, and diethyldithiocarbamate (DDC) inhibited the epidermal SOD activity in a dose‐dependent manner. It has been known that two types of SOD, a Cu, Zn‐type and a Mn‐type, are present in eukaryotes; that the latter is insensitive to cyanide inhibition. Using this property, the main SOD present in the epidermis was hypothesized to be the Cu, Zn‐type (8.6 ±1.1 unit/mg protein; around 75%); the Mn‐type was a minor component (2.8 ± 0.2 unit/mg protein; around 25%). SOD staining following acrylamide disc gel electrophoresis revealed two epidermal SOD bands, one of which was abolished by the addition of cyanide. These results are consistent with the view that pig epidermis contains two types of SOD, a Cu, Zn‐type and a Mn‐type; the former appears to be predominant.
In order to elucidate a part of calmodulin actions in the hyperproliferative state in human epidermis, calmodulin activities in the psoriatic and in the normal human epidermis were determined using calmodulin-deficient phosphodiesterase from bovine heart and purified pig skin epidermal calmodulin as a standard. Skin samples were obtained from 11 normal healthy controls and from both the uninvolved and involved regions of 8 nonconsanguineous psoriatic patients. Pure epidermal samples, prepared by the microdissection method, were used for calmodulin assays. Normal human epidermis contained 270 +/- 13 ng/mg dry weight, whereas calmodulin activities were significantly increased in psoriatic epidermis, 412 +/- 29 ng/mg dry weight for the uninvolved epidermis and 747 +/- 46 ng/mg dry weight for the involved epidermis, respectively. These results suggest that calmodulin may play an important role in cell proliferation in human epidermis.
Although retinoids reveal various biologic and biochemical activities on epidermal keratinocytes, their effects on the epidermal cyclic AMP (cAMP) system has been less well characterized. In order to elucidate the relation between them, an in vitro pig skin-slice incubation system was employed. After a long-term (up to 24 h) incubation in vitro, control skin responded to epinephrine only slightly. The addition of Ro 10-1670, an active derivative of Ro 10-9359 (etretinate) in the incubation medium, resulted in an increase of the beta-adrenergic adenylate cyclase response of epidermis. On the other hand, histamine-induced cAMP accumulation was decreased by the retinoid treatment after long-term incubation. The augmentation of the beta-adrenergic response was observed at 1 microM concentration and the maximal effect was observed at 10 microM. There was no significant difference in cAMP phosphodiesterase activities between the control and retinoid-treated skin. The effect was also observed by the addition of all-trans-retinoic acid, retinol, and Ro 10-9359; the latter two compounds revealed much lesser effects. The addition of combinations of various drugs (Ro 10-1670 and hydrocortisone; Ro 10-1670 and colchicine) resulted in more marked (additive or synergistic) effects than the single addition of each chemical. On the other hand, the addition of Ro 10-1670 and all-trans-retinoic acid resulted in neither additive nor synergistic effect, suggesting that they probably work on the same site. Our data indicate that the epidermal beta-adrenergic adenylate cyclase response is modulated by retinoids probably as an independent mechanism stimulated by glucocorticoids or colchicine.
Psoriasis is a chronic inflammatory skin disease, which has been increasing during the last 50 years in Japan. The aim of the present study is to analyze psoriatic patients registered from 1983-2007 in Asahikawa Medical College Hospital, which is located in the northern part of Japan. A total of 607 cases were registered at the first inspection in the Department of Dermatology, Asahikawa Medical College. Men (403 cases, 66.4%) were predominant over women (204 cases, 33.6%). The clinical types of psoriasis were psoriasis vulgaris (91.5%), guttate psoriasis (4.2%), psoriasis arthropathica (2.8%), psoriatic erythroderma (0.6%), generalized pustular psoriasis (0.6%), localized pustular psoriasis (0.15%) and infantile psoriasis (0.15%). Topical corticosteroids (78.1%) and vitamin D3 (18.1%) products were the main previous topical agents. Previous systemic treatments included etretinate (7.7%), cyclosporine (1.5%) and methotrexate (0.3%). Use of topical vitamin D3 and cyclosporine therapies have been gradually increasing during the past 25 years. Regarding the previous phototherapy, topical psoralen and ultraviolet A therapy (PUVA) (4.9%) was predominant over ultraviolet B (0.9%), and systemic PUVA (0.7%). Use of ultraviolet B phototherapy has been increasing during the past 5 years. The results are essentially similar to those of a survey of psoriasis in Japan from 1982-2001. Although the incidence of psoriasis might be higher in Hokkaido Prefecture, there is essentially no variation in the disease profile of psoriatic patients.
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