Hiromitsu Noguchi

Processing of cathepsins L, B and D in psoriatic epidermis.

Proteinase activity is increased in psoriatic epidermis. To elucidate the involvement of enzymes in psoriatic epidermis, the expression of cathepsins L, B and D was investigated by Western blotting and immunohistological studies. Normal epidermis contained abundant inactive precursors (39 kDa) of cathepsins L and B and an inactive intermediate form (45 kDa) of cathepsin D. Cathepsin L in psoriasis was processed to a variable extent from the precursor to a single-chain form (30 kDa) and a mixture of single- and heavy-chain (25 kDa) forms of the active mature enzyme, corresponding to the immunohistological staining patterns ‘diffuse dense’, ‘small granular’, and unevenly distributed ‘condensed granular’. Cathepsin B showed a mixture of precursor form (39 kDa) and single-chain (30 kDa) forms and was expressed as a ‘diffuse dense’ staining pattern in the mid-spinous layer and as a ‘condensed’ pattern in the upper spinous and granular layers. Cathepsin D was processed to the heavy-chain (31 kDa) form of activated mature enzyme with small granular staining and a mixture of heavy-chain and degraded protein (28 kDa) with larger and more condensed granular staining. The distribution patterns of ‘small granular’ cathepsin L, and of cathepsins B and D expression in suprabasal keratinocytes were very similar to that of involucrin. After complete clinical resolution of psoriasis by 8-methoxypsoralen plus UVA treatment, the expression of the three cathepsins was normalized. These results suggest that cathepsins L, B and D in forms activated to a variable extent may be involved in the pathology of psoriasis.

Granulocyte and macrophage colony-stimulating factors stimulate proliferation of human keratinocytes

Granulocyte/macrophage colony-stimulating factor (GMCSF), granulocyte CSF (G-CSF), and macrophage CSF (M-CSF), modulators of hematopoietic cells, are widely used for the treatment of neutropenia after chemotherapy for malignant disease and other neutropenic states [1, 2]. Some keratinocyte-derived cytokines, such as interleukin-1 (IL-1), IL-6, GM-CSF, transforming growth factor (TGF)-α, and tumor necrosis factor (TNF)-α [3–7] are known as autocrine factors. To elucidate possible autocrine mechanisms of these two CSFs, we have investigated stimulatory effects on keratinocyte growth by Gand M-CSFs. Neonatal human foreskin keratinocytes (more than 5 × 105 cells) in the Keratinocyte Growth Kit (Cascade Biologics, Portland, Ore.) were used. Thawed cells were seeded in modified MCDB 153 [8] (Nissui Seiyaku, Tokyo), that contained 150 μg/ml bovine pituitary extract, 5 μg/ml bovine insulin, 0.4 μg/ml hydrocortisone, 10 μg/ml bovine transferrin and 10 ng/ml human epidermal growth factor. Cells were used at the second or third passages and seeded in six wells at a density of 1.0 × 104 cells/cm2. The medium was changed to modified MCDB 153 without bovine pituitary extract, and with or without recombinant G-CSF (0.1–10 ng/ml) (Kirin Beer, Tokyo) or recombinant M-CSF (1-100 ng/ml) (Midori Jyuji, Osaka). Control experiments for specificity were performed by the addition of G-CSF(2 × 10–2 to 10–4 dilution) (the kind gift of Kirin Beer) or M-CSF-specific neutralizing antibody (1 × 10–2 to 10–4 dilution) (the generous gift of Morinaga Nyugyo, Kanagawa). After 4 and 7 days, cells were detached, collected and counted by trypan blue exclusion on a hemocytometer. [3H]Thymidine (0.5 μCi/well: Dupont/Nen Research Products, Boston, Mass.) was incubated in keratinocytes per well for 24 h. Cultured keratinocytes were detached and harvested with an automatic cell harvester (ACH-96, BioTec, Tokyo). Radioactivity was measured with a liquid scintillation counter (LKB 1216 Packbeta, Wallac Oy, Turk, Finland). The experiments were triplicated and were analyzed by Student’s t-test for unpaired values. Two CSFs studied showed dose-dependent stimulation of human keratinocyte growth. Cell number increased at G-CSF concentrations between 0.1 and 10 ng/ml. M-CSF also stimulated proliferation of keratinocytes with similar effects at higher concentrations (10–100 ng/ml) (Fig. 1). Gand M-CSFs induced the increase in [3H]thymidine uptake dose-dependently, indicating parallel results to cell number studies (Fig. 2). Control experiments with the addition of the specific neutralizing antibodies demonstrated no significant increase in cell number or [3H]thymidine uptake. This study demonstrated that Gand M-CSFs had stimulatory effects on keratinocyte growth in culture. These results indicate that they may be potential autocrine growth factors for keratinocytes, since these two CSFs are produced by keratinocytes as well as by immune cells [9, 10]. G-CSF exerted similar proliferative effects at lower concentrations than M-CSF, suggesting that G-CSF may have relevance to binding to keratinocytes. Receptors for MCSF are identified as the gene product of the c-fms-protooncogene, showing a possible role for M-CSF in nonhematopoietic tissue [11]. Each receptor is expressed in other cells [10], while little information is available for keratinocytes. An investigation of receptors of Gand MCSFs in human keratinocytes is currently under way. GM-CSF is synthesized by keratinocytes and has the ability to capacitate isolated murine Langerhans cells for Akira Kawada · Masataro Hiruma · Hiromitsu Noguchi · Akira Ishibashi · Kazuo Motoyoshi · Itaru Kawada

Tinea pedis in members of the Japanese Self‐Defence Forces: relationships of its prevalence and its severity with length of military service and width of interdigital spaces

Summary. We examined the relationship between prevalence and severity of tinea pedis and the length of service and the width of the spaces between the toes in 74 members of the Japanese Self‐Defence Forces (SDF) undergoing special training. The subjects were divided according to the width of these spaces into: group I, wide; group II, fairly wide; and group III, closed. The severity of tinea pedis was determined by its duration and the extent of the lesions in the 49 subjects who had tinea pedis. The combined prevalence of tinea pedis and tinea unguium was 66%. There was a tendency for the prevalence to be higher in subjects who had served for 10 years or more in the SDF than in those with fewer than 10 years of service. Classified by the disposition of their toes, 10 subjects fell into group I, 34 into group II, and 30 into group III. The prevalence of 90% (27/30) in group III was significantly higher than in the other groups. A significant positive correlation was seen between length of SDF service and severity. Subjects with both a long service record and closed interdigital spaces showed both a high prevalence and marked severity.

Hyperthermic treatment of sporotrichosis: Experimental use of infrared and far infrared rays

Zusammenfassung. Bei 14 Sporotrichose‐Patienten (sieben Kinder und sieben Erwachsene) wurden Taschenwärmer, Infrarot‐und langwellige Infrarotstrahlung zur Behandlung eingesetzt. Neun Sporotrichose‐Läsionen gehörten zum Typ der fixierten Hautverletzung, fünf waren Lymphgewebsverletzungen; sechs waren im Gesicht und acht auf den Extremitäten lokalisiert. Vier Patienten wurden mit Taschenwärmern behandelt, fünf mit Infrarotstrahlung und weitere fünf mit Langwellen‐Infrarotstrahlung. Die mit Taschenwärmern behandelten Läsionen waren Gesichtsverletzungen bei Kindern. Infrarot‐und Langwellen‐Infrarotstrahlung wurde bei drei Kindern und sieben Erwachsenen eingesetzt. Dabei handelt es sich um zwei Gesichtsverletzungen und zwei Lasionen an den Extremitäten. Bei den Infrarotbehandlungen war die applizierte Wärmemenge groUßer als bei Taschenwärmern; eine tägliche Behandlung von 15 min war ausreichend zur Erzielung zufriedenstellender Resultate. Die Anwendung der Infrarotstrahlung ist jedoch bei Kindern und im Gesicht schwierig, und zwei bis drei Behandlungen von je 40 min/Woche führte nicht zu zufriedenstellenden Resultaten. Im Vergleich zur Taschenwärmer‐Behandlung konnte bei den Infrarotbehandlungen die Behandlungszeit um drei Viertel verkürzt werden.

A Pediatric Case of Atypical Mycobacterium avium Infection of the Skin

We report a case of cutaneous atypical mycobacteriosis in a 12‐year‐old healthy girl due to Mycobacterium avium. The cutaneous symptoms were three well‐defined subcutaneous nodules on both buttocks and on the posterior surface of the left thigh. One had a fistulous opening on the skin surface. Histopathological examination revealed epithelioid cell granulomas surrounded by dense lymphocytic infiltration and acid‐fast bacteria were seen with modified periodic acid‐carbol fuchsin staining. Using Ogawas medium at 37°C, acid‐fast bacteria were isolated from the biopsied specimen and identified by the DNA‐DNA hybridization method as Mycobacterium avium. In drug susceptibility test, these were resistant to all antituberculous drugs. Oral administration of minocycline 100 mg/day for two months had little effect on the two remaining lesions, which were therefore excised. Based upon reported cases of Mycobacterium avium complex, we considered that our pediatric patient with multiple intradermal or subcutaneous nodules on the buttocks and the thigh exhibited the characteristic symptoms of M. avium infection.

A Statistical Study of Calcifying Epithelioma, Focusing on the Sites of Origin

We statistically investigated 396 lesions taken from 355 cases of calcifying epithelioma. The distribution of these lesions did not correlate with the density of the hair follicles, but it was in accord with the distribution of intermediate hairs, such as those in the hair border. This relationship may have etiologic significance.

Case Report. Sporotrichosis successfully treated with itraconazole in Japan

A case of lymphocutaneous sporotrichosis that had developed from the dorsum nasi to the left buccal region of a 65‐year‐old woman was treated with itraconazole 100 mg day−1. The lesion healed in 10 weeks after starting treatment leaving crusts, mild erythema and pigmentation, and the treatment was completed 16 weeks after the start of treatment. Neither adverse reactions nor abnormal clinical laboratory values were noted. Until the present time, 1 year and 2 months after the completion of treatment, no recurrence has been observed. In Japan, 43 cases of sporotrichosis have been treated with itraconazole, and 38 cases (88%) have been assessed as effective or better. The mean dose of itraconazole and the mean duration of administration are 100 mg day−1 and 11 weeks, respectively, in these reported Japanese cases. The use of itraconazole is expected to be one of the effective therapies for sporotrichosis.

Tinea nigra showing a parallel ridge pattern on dermoscopy.

An 18‐year‐old healthy female student noticed a brown macule measuring 21 mm in diameter on the left palm and visited our clinic concerned about a cancerous mole. Dermoscopic examination revealed a brown, fine‐dotted and granule‐like structure overlapping an amorphous light brown macule. However, unlike previous cases, analysis of the high dynamic range‐converted image revealed the parallel ridge pattern frequently observed in malignant melanomas. Brown mycelia were detected on direct microscopic examination; black colonies were isolated on fungal culture and the fungus was identified as Hortaea werneckii. The lesion was treated with topical ketoconazole cream, and it diminished 1 month later.

Effectiveness of treatment of severe tinea pedis with 1% terbinafine cream in members of the Japanese Self-Defense Forces

A clinical study was conducted to determine the efficacy of terbinafine cream in severe cases of tinea pedis. The subjects were 21 men, members of the Japanese Self‐Defense Forces who were judged to have severe symptoms of tinea pedis. The description ‘severe’ was defined as ‘considered to require the concurrent internal administration of antifungals for complete cure’, or as meeting criterion 5 or 6 for the severity of tinea pedis. A simple surface application of terbinafine cream was given once daily, the subjects’ clinical manifestations, mycological cure rates and safety‐related changes were observed, and a final assessment was made in the 12th week. In the final assessment, the improvement rate of the cutaneous symptoms was 95.2%, and the fungal eradication and efficacy rates were 81.0%. As for side‐effects, one patient complained of local irritation. These results suggested that terbinafine cream is a beneficial topical antifungal cream for severe tinea pedis.

Tinea pedis survey in members of the Japanese Self-Defense Forces undergoing ranger training

Summary. The conditions surrounding tinea pedis infection were surveyed—the degrees of infection and of aggravation attendant upon training, and the skin temperature within the leather boots of members of the Japanese Self‐Defense Forces undergoing winter and summer ranger training. Before winter training, tinea pedis was diagnosed in 8 of the 15 subjects. The mean atmospheric temperature was 7.1°C, the total precipitation, 23 mm, and the mean skin temperature of the feet during training, 33.7 °C. After training, 1 further subject had contracted tinea pedis, 5 of the 8 showed slight aggravation, and 3 showed no change. Before summer training, tinea pedis was found in 9 of another 15 subjects. The mean atmospheric temperature was 20.0 °C, total rainfall, 353 mm, and the mean skin temperature of the feet during training, 36.6 °C. After the training, aggravation had occurred in 3 of the 9, and slight aggravation in the other 6. In these 9, vesiculation, pustules and erosion were seen, so that summer training had a more severe effect than winter training. Investigation of the predisposing factors for tinea pedis due to the wearing of boots and shoes is considered extremely important for the prevention and treatment of this disease.