Masataro Hiruma

Systemic phaeohyphomycosis caused by Exophiala dermatitidis

Summary. We report a case of systemic phaeohyphomycosis due to Exophiala dermatitidis in a 24‐year‐old man. At the age of 17, the patient had noticed cervical swellings. On palpation at his initial examination, more than a dozen firm lymph nodes between 1 and 5 cm in diameter were found on both sides of the neck and in both axillae. Examination of biopsy specimens of lymph nodes revealed pale brown hyphae in granulomatous lesions and Exophiala dermatitidis was isolated from mycological cultures of the local tissues. The clinical course was marked by an episode of jaundice six months later, and surgery disclosed a fist‐sized inflammatory mass in the region of the head of the pancreas and the duodenum. The patient was treated by providing external biliary drainage and by the administration of antifungal agents, but two years later complained of diplopia and a slight heaviness in the head. Computed tomographic scans were made of the head, revealing multiple plum‐sized masses in the brain. The patient died a year later. In Japan, 10 cases of systemic infection by this organism (including the present one) have been reported, all in patients of up to 30 years of age, with lesions appearing in the brain, lung, liver, digestive organs and lymph nodes. The prognosis is grave.

Stimulation of human keratinocyte growth by alginate oligosaccharides, a possible co-factor for epidermal growth factor in cell culture

Oligosaccharides, involved in regulation of plant developmental and defensive processes, were tested to determine their ability to enhance proliferation of human keratinocytes. A mixture of alginate oligosaccharides remarkably stimulated keratinocyte growth and [3H]thymidine uptake in the presence of epidermal growth factor (EGF). The activity was comparable to bovine pituitary extract, a common complement in keratinocyte culture, and additive on BPE‐induced stimulation. The most effective oligosaccharide in the mixture was identified and its chemical structure was determined. These findings demonstrate a novel activity of alginate oligosaccharide(s) in keratinocyte growth and suggest a possible co‐factor for EGF‐dependent stimulation in medium for keratinocytes.

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 chromomycosis with disposable chemical pocket warmers Report of a successfully treated case, with a review of the literature

A case of chromomycosis in which hyperthermia proved effective is reported. The patient was a 56-year-old male bean curd maker who, without any previous history of minor trauma, developed on the extensor side of the left upper arm an eczematous lesion that underwent gradual radial expansion. The lesion showed a well-defined, 7×10 cm infiltrated erythematous plaque with the central area healed and, at the upper and lower borders, adherent scales and crusts on the surface. Histological examination revealed granulomatous changes in the dermis, as well as sclerotic cells within giant cells and microabscesses. On culturing,Fonsecaea pedrosoi was isolated. The patient was treated with disposable chemical pocket warmers, which were secured over the lesion with a rather tight elastic bandage, so that they kept the affected area warm for 24 hours a day. After a month of such hyperthermic treatment, the erythema and infiltration had decreased considerably, and microscopic examination and culture of the crusts both yielded negative results. Examination of biopsy specimens of the lesion after the third month showed that it had cicatrized. The treatment was stopped after 4 months, and no relapse occurred. We also summarize the published results of local hyperthermic treatment of chromomycosis in Japan.

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.

Cathepsin B and D expression in squamous cell carcinoma

To elucidate involvement ol protcinascs in malignancy of keratinocytes. expression ol catbepsin B, a cysteine proteinase. and catbepsin D. an aspartic proteinase. was ascertained in lormalin‐lixed paraffin‐embedded specimens of nortnal skin, squamous cell carcinoma (SCC). Bowens disease, seborrboeic keratosis and basal cell carcinoma (BCC). Presence of procalbepsin B and an intermediate form of catbepsin D was coiilirmed by Westerti blolling and enzytne activity analysis. Cathepsin B stained more Intensely In SCC tumour cells tban in normal epidermis: staining patterns were diffuse, grantilar or botb. Diflusc and granular patterns (procatbcpsin B and mature enzyme, respectively) appeared in inner and outer parts of tumour islands, respectively. Five of 20 cases of Bowens disease sbowed diffuse enhanced catbepsin B expression: 20 cases of seborrhoeic keratosis or BCC did not. Cathepsin D stained intensely in tumour cells of half the SCC cases. The staining manner and distribution of cathepsins B and D was similar in the cytoplasm of cancer cells. No enhanced staining of cathepsin I) was seen in any cases of Bowens disease, seborrhoeic keratosis. or BCC. Coexistence and localization of active mature forms of cathepsins B and D suggests that cooperation between tbe two enzymes may play an important part in invasion of SCC.

Ultrastructure of asteroid bodies in sporotrichosis

Summary. Ultrathin sections were prepared from a paraffin‐embedded tissue specimen which had been obtained from a sporotrichosis patient and in which large numbers of asteroid bodies had been observed. Electronmicroscopic examination of the ultrastructure of the asteroid bodies revealed a central fungal cell with peripheral rays. These cells were almost structurally identical with fungal elements found elsewhere than in asteroid bodies; and in the rays, electron‐dense granular substance was arranged in layers marked by fissures and material resembling remnants of destroyed host cells. The outermost layer was formed of a granular substance of rather low electron density, surrounded by adherent cell membrane fragments. These findings suggest that an asteroid body might be composed of crystalline products of disintegrated host cells deposited around a fungal cell.

Prurigo pigmentosa, Ketonemia and Diabetes mellitus

The etiology of prurigo pigmentosa still remains unknown. We present a 16-year-old female patient with ketonemia caused by diabetes mellitus. The eruption subsided when blood glucose and total ketone levels were controlled by subcutaneous insulin injection. We propose that ketonemia caused by diabetes mellitus may play a role in the pathogenesis of prurigo pigmentosa.