Hiroshi Ueda

Immunohistochemical Localization of Nitric Oxide Synthase in Normal Human Skin: Expression of Endothelial-type and Inducible-type Nitric Oxide Synthase in Keratinocytes

Nitric oxide (NO) is a critical mediator of various biological functions. NO is generated from L‐arginine by nitric oxide synthase (NOS), which has three isoforms; endothelial‐type NOS (eNOS) and brain‐type NOS (bNOS) are constitutive enzymes, and inducible‐type NOS (iNOS) is expressed after stimulation. We investigated the expression of NOS in normal human skin by an immunohistochemical technique and western blotting analysis. In human skin, epidermal keratinocytes and the outer root sheath were labeled with not only eNOS antibody but also with iNOS antibody. Both eNOS and iNOS protein in epidermal keratinocytes were confirmed by western blotting. eNOS immunoreactivity was observed in endothelial cells, fibroblasts, the arrector pili muscle, apocrine secretory gland, eccrine coiled duct, and eccrine secretory gland. bNOS immunoreactivity was observed in mast cells. No staining with anti‐bNOS antibody was observed in any other cell type. Our present findings suggest that epidermal keratinocytes in normal human skin contain both eNOS and iNOS.

Identification of causative chemicals of allergic contact dermatitis using a combination of patch testing in patients and chemical analysis Application to cases from rubber gloves

5 cases of allergic contact dermatitis from rubber gloves were investigated by our recommended procedures using a combination of patch testing in patients and chemical analysis of causative rubber products by gas chromatography (GC) and high‐performance liquid chromatography (HPLC). We previously confirmed that zinc ethylphenyldithiocarbamate (ZEPC), a dithiocarbamae‐type accelerator (DTC), was causative in a case of allergic contact dermatitis from rubber work gloves. Subsequently, we have clarified that DTCs such as zinc dimethyldithiocarbamate (ZDMC), zinc diethyldithiocarbamate (ZDEC) and zinc dibutyldithiocarbamate (ZDBC) and amines such as dimethylamine (DMA), diethylamine (DEA) and piperidine (PIP) were also causative in cases from surgical rubber gloves. Thus, our investigative studies revealed that, although thiurams have been taken much more notice of as allergenic compounds than their corresponding DTCs and amines, not only DTCs such as ZDMC, ZDEC, ZDBC and ZEPC, but also amines such as DMA, DEA and PIP were noteworthy causative candidates of allergic contact dermatitis from rubber gloves.

Rapid contamination of the environments with varicella‐zoster virus DNA from a patient with herpes zoster

Patients with zoster are considered to be less contagious than varicella patients because their infection is localised. It is not known, however, when and for how long a spread of varicella‐zoster virus (VZV) from a zoster patient begins and continues and the extent of virus spread from the patient. The polymerase chain reaction (PCR) was used to detect VZV DNA in samples obtained from the hands and throat of a patient with zoster and from her room environments including surfaces of the back of a chair, the door handle, the table and the air conditioner filter. VZV DNA was detected on the surfaces of the back of the seat and the table and in peripheral blood mononuclear cells (PBMCs) and serum on Day 4 of the illness. VZV DNAemia persisted for 4 days until Day 7 of the illness. It was also detected in samples collected from throat and the air conditioner filter on Days 6 and 7 of the illness respectively. All of the surfaces, that were examined in her home environment, were contaminated with VZV DNA by Day 7 of the illness. The present study showed rapid and wide spread of VZV DNA in the environment even from a patient with zoster. J. Med. Virol. 63:64–66, 2001.

4 cases of photocontact dermatitis due to ketoprofen

We report 4 cases of photocontact dermatitis due to ketoprofen, a non‐steroidal anti‐inflammatory drug derived from propionic acid. We carried out a skin biopsy to examine the amount of ketoprofen in the eruptive skin. We investigated the cross‐reactions between tiaprofenic acid, suprofen and ketoprofen by patch and photopatch testing. In case no. 1, 17 days after the discontinuance of Mohrus poultice (containing ketoprofen as an active ingredient), we detected ketoprofen 312.5 ng/g in the area of skin where the poultice was applied. All 4 cases reacted positively to the causative medicaments containing ketoprofen and ketoprofen 1% pet. 3 out of 4 cases reacted positively to tiaprofenic acid 1% pet. Only 1 case out of the ketoprofen and tiaprofenic acid positive cases reacted positively to suprofen 1% pet. Vehicles of patients medicaments were negative in all 4 cases. We suspected that the key structure of the cross‐reaction between ketoprofen and tiaprofenic acid and suprofen was the benzoyl radical.

Immunohistochemical localization of NO synthases in normal human skin and psoriatic skin

Nitric oxide (NO), a diffusible free radical with a very short half-life, is a bioregulatory molecule with multiple physiological functions. For example, it is the messenger of macrophage-dependent cell-mediated cytotoxicity, it is an important mediator of vascular endothelium relaxation, and it functions as an intercellular messenger in neural transmission [1–3]. Recently, Heck et al. [4], using an in vitro assay of human keratinocytes in primary culture, have demonstrated that keratinocytes produce NO. However, little is known about the role of NO in the human skin. NO is generated from L-arginine by NO synthases (NOS). Three types of NOS are currently known: neuronal-type NOS (n-NOS), endothelial-type NOS (eNOS) and inducible-type (i-NOS). n-NOS and e-NOS are constitutive Ca2+/calmodulin-dependent enzymes. i-NOS is a Ca2+-independent enzyme induced in a wide range of tissues and cells by cytokines and bacterial products [5]. In this study, we focused on constitutive-type enzymes, and found that e-NOS was localized in normal human epidermis and that the immunoreactivity of e-NOS was reduced in epidermis from psoriasis vulgaris patients, while n-NOS was negative in normal skin and in psoriatic skin. Three normal skin specimens were obtained from patients undergoing elective plastic surgery. Skin biopsies were obtained from untreated lesions of three patients with psoriaris vulgaris. Tissue samples were fixed with 4% paraformaldehyde, and embedded in paraffin. Sections (3 μm) were cut and mounted on slides. After deparaffinization and hydration in phosphate buffered saline (PBS), specimens were incubated with anti-n-NOS antibody (Affinity Bioreagents, Neshanic Station, NJ, USA, 1/5000), or anti-e-NOS antibody (Affinity Bioreagents, Neshanic Station, NJ, USA, 1/5000) for 12 h. The avidinbiotin-peroxidase complex (ABC) technique was then used. The sections were washed in PBS, incubated with biotinylated goat antirabbit IgG (1/200) for 1 h, washed in PBS and incubated with avidin-biotin complex (1/100) for 1 h. The sections were then washed and developed in 0.02% 3,3′-diaminobenzidine tetrahydrochloride containing 0.003% H2O2 for 4 min. Normal rabbit IgG was used as a negative control. To evaluate the specificity of e-NOS Masao Sakai · Yoshinori Shimizu · Ikuko Nagatsu · Hiroshi Ueda

Contact sensitivity and cross‐reactivity of budesonide

5 cases of allergic contact dermatitis due to budesonide are described. We studied the antigen determinant structures in these cases by applying patch tests with several substances related to budesonide. 2 cases showed cross‐reactions to both amcinonide and prednisolone acetate. The antigen determinant structure is also discussed.

A Case of Idiopathic Vulvar Calcinosis: The First in Japan

Idiopathic calcinosis of the scrotum is a rare condition of unknown etiology. As counterparts of this male disorder, only two female cases have been reported (1, 2). We report a third case, the first of its kind, in Japan. Histochemical stains revealed acid mucopolysaccharide (acid‐MPS) within the calcinosis and many infiltrated mast cells near it; these might histogenese the calcium deposition. Immunohistochemically, polyclonal carcinoembryonic antigen (CEA) and keratin stains revealed no positive cells near the calcium depositions.

Immunocytochemical localization of copper-zinc superoxide dismutase in mouse olfactory mucosa.

Copper-zinc superoxide dismutase (Cu-Zn SOD) has been localized in mouse nasal mucosa. Immunocytochemical staining using polyclonal antibody against Cu-Zn SOD revealed endogenous Cu-Zn SOD in sustentacular cells in the olfactory area, and in ciliated epithelial cells in the respiratory area of the olfactory mucosa. Since these cells are located in the superficial portion of the olfactory mucosa, Cu-Zn SOD may serve as an intracellular antioxidant.