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Lysyl oxidase initiates cross-linkage of collagen and elastin by catalysing the formation of a lysine-derived aldehyde. In order to study cross-linking in scleroderma, we used monoclonal antibodies to lysyl oxidase to determine the localization of this enzyme in systemic and localized scleroderma, and compared the distributions obtained with that in normal skin. Using an indirect immunofluorescent antibody method and an avidin-biotinylated enzyme complex method, 11 cases of diffuse type of systemic scleroderma and seven cases of localized scleroderma were studied. In the oedematous stage of systemic scleroderma, intracellular and extracellular lysyl oxidase were remarkably increased in the dermis, particularly in groups around blood vessels. In the sclerotic stage of systemic scleroderma, lysyl oxidase was detected intracellularly in fibroblasts and extracellularly among collagen bundles between the lower dermis and the subcutaneous fat tissue. In localized scleroderma, a marked increase in lysyl oxidase was observed in mononuclear cells and fibroblasts near blood vessels in the lower dermis and in the subcutaneous fat tissue, in addition to the extracellular deposits between collagen bundles. The increase in lysyl oxidase in localized scleroderma was much more common than in the oedematous stage of systemic scleroderma. These findings indicated that intracellular and extracellular expression of lysyl oxidase expression was greater in sclerodermatous skin than in normal skin.

A 64-year-old woman was transferred to the intensive care unit with dyspnea and palpitation on effort. Chest x-ray film showed cardiomegaly and pulmonary congestion. We carefully examined for sarcoidosis as a differential diagnosis of heart failure. Serum lysozyme was mildly high, but human atrial natriuretic peptide (HANP) and brain natriuretic peptide (BNP) were strikingly high. Angiotensin converting enzyme was within normal limit. Chest roentgenogram did not reveal bilateral hilar lymphadenopathy. Atrioventricular conduction block was not observed on electrocardiogram. Echocardiographic examination showed left ventricular global hypokinesis with septal thinning and enlargement. Mitral valve regurgitation was recognized by Doppler evaluation. Coronary arteriography showed normal coronary arteries. Endomyocardial biopsy revealed noncaseous epithelioid granulomas containing, Langhans type giant cell accompanied by fibrosis and lymphocyte infiltration. From these data cardiac sarcoidosis was diagnosed. Gallium scintigraphy showed diffuse uptake only in the heart. Treatment with oral prednisolone 20 mg/day was started. Her symptoms improved by several weeks after the medical treatment. In addition, both the value of HANP and BNP were markedly decreased and echocardiogram showed improvement of cardiac systolic function. In Japan, there is a higher incidence of cardiac sarcoidosis than in the West. The prognosis of this condition associated with cardiac dysfunction is reported to be very poor. When progressive heart failure in older patients is seen, cardiac sarcoidosis should also be kept in mind. Endomyocardial biopsy play an important role as the only accurate technique for the diagnosis of cardiac sarcoidosis.

We diagnosed phakomatosis pigmentovascularis type IIIb in an 11-month-old baby who had a giant nevus spilus, a nevus flammeus, and moyamoya disease. Development of the patient was normal until 6 months of age when he developed a sudden onset of focal seizures and left hemiparesis. This patient represents the sixth case of phakomatosis pigmentovascularis type IIIb, including three cases in the Japanese literature, reported thus far. However, to our knowledge, this is the first case with an association to moyamoya disease.

Cutaneous metastases from pancreatic carcinoma are uncommon. Eight cases have been described in the literature. The metastases display some common features such as umbilical nodules and the histologic pattern of adenocarcinoma. Our patient had erythematous infiltrated plaques in the left axilla and on the upper portion of the chest. A skin biopsy specimen disclosed many poorly differentiated atypical cells throughout the dermis. The diagnosis was confirmed by positive immunohistochemical staining for carbohydrate antigen 19-9.

We report a case of adult T-cell leukaemia/lymphoma (ATLL), in whom the polymerase chain reaction (PCR) on genomic DNA from saliva demonstrated the monoclonal integration of human T-cell lymphotropic virus type-1 (HTLV-1) proviral DNA in lymphocytes in the saliva. These results provided evidence of the possibility of saliva-borne transmission of HTLV-1.

We report a Japanese patient with congenital hypomelanosis with a segmental pattern on the lett abdomen, whorl-like pattern on the back; mosaic pattern on the chest, right abdomen, and proximal extremitiea; and with yellow hair on a portion of the scalp. Chemical analysis of the yallow hair revealed decreased eumetanin content, whereas the pheometanin content was normal.

We report the case of a Japanese female infant with Chédiak-Higashi syndrome born to consanguineous parents. At birth she had fair skin but, when she was three months old, marked hyperpigmentation of the sun-exposed skin areas developed. Microscopic examination of blood and electron microscopic examination confirmed the diagnosis. She enjoyed good health until she was two years old when she had pneumonia with marked hepatosplenomegaly. It is important for dermatologists and pediatricians to be aware of the skin manifestations of this disease because hyperpigmentation after sun exposure may be a characteristic, initial feature of this condition.

Vogt-Koyanagi-Harada disease is a rare disease characterized by uveitis, meningitis, dysacusis, alopecia, poliosis, and vitiligo. We describe a 48-year-old patient with Vogt-Koyanagi-Harada disease associated with thin inflammatory raised erythema and plaque-type inflammatory erythema superimposed on vitiligo. Interestingly, inflammatory raised erythema was separated from the perfect vitiligo, and the incomplete vitiligo lay between them initially. Thereafter, incomplete vitiligo became completely depigmented with diminution of inflammatory raised erythema. This is the second case of vitiligo with inflammatory raised borders associated with Vogt-Koyanagi-Harada disease. Our results of immunohistochemical and electron microscopic studies suggested the involvement of T-cell-mediated cytotoxicity and apoptosis in the development of skin lesions.

Two patients with epidermolysis bullosa dystrophica recessiva who had squamous cell carcinoma are presented. Case 1 is a 40–year-old woman who had ulcers on her left lower leg. Case 2 is a 42–year-old man who had a tumor on his left first toe. Wide surgical excision with skin coverage by autograft was performed in case 1. Amputation of the toe in case 2 was performed. A review of the cases of epidermolysis bullosa dystrophica associated with cancer reported in Japan is also presented.

By using antibodies to type I, III, IV, V, and VI collagens and laminin, we examined the localization of interstitial collagens and basement membrane components with indirect immunofluorescence and immunoelectron microscopy (IEM). Furthermore, the morphological changes of these collagens in malignant transformation were investigated. In neurofibroma, IEM showed type I, III, and V collagens to be present diffusely on cross-striated collagen fibrils, whereas type VI collagen was present between the fibrils. Type IV collagen and laminin were observed to surround tumor cells. In neurofibrosarcoma, tumor collagen bundles that reacted with antibodies to type I, III, V, and VI collagens were irregularly arranged. Immunofluorescent deposits that reacted with anti-type IV collagen and anti-laminin antibodies were decreased in number, showing a thin and sparse arrangement.

Recently, we have reported two cases with endothelin (ET)-secreting tumor presenting with hypertension and elevated plasma endothelin-1 (ET-1) levels. The present study examines the histopathology of the ET-secreting tumor in one of these cases. The neoplasm was located in the skin and microscopically characterized as a malignant hemangioendothelioma by remarkable intravascular proliferations of atypical endothelium with the expression of Factor VIII-related antigen in the tumor cells. The tumor enlarged rapidly with rising ET-1 and blood pressure levels. The ET-1 content and its messenger RNA expression in the tumor extract were higher than those from normal parts of skin. ET-1 may play a pathophysiological role in patients with ET-secreting malignant hemangioendothelioma.

A primary adenoid cystic carcinoma of the skin is reported. Light microscopy revealed pseudocysts. PAS-positive basement membrane and true glandular lumen, which in aggregates are specific for adenoid cystic carcinoma. Perineural invasion was also observed. Ultrastructural examinations revealed three types of cystic spaces; pseudocysts, true glandular lumens and intercellular spaces. Enzyme histochemical examinations showed positive reactions for eccrine enzymes, including phosphorylase and succinic dehydrogenase and negative for apocrine enzymes. Immunolocalization of collagens and laminin revealed that basement membranes of the pseudocysts involve Type V collagen as well as Type IV collagen and laminin.

Three patients with subungual exostoses, two involving the great toe and one the fourth toe, are presented. Local excision was performed, and the specimens were processed for light and electron microscopy. Light-microscopic study revealed that the tumours consisted of a proliferative fibrocartilaginous cap that merged into mature trabecular bone at its base. Electron-microscopic examination revealed that the tumour was composed of two types of cells: one was rich in cell organelles including rough endoplasmic reticulum, well-developed Golgi apparatus, and glycogen granules; the other cell had few such cell organelles. The former cells seemed to be osteoblasts actively engaged in bone formation, and the latter to be osteocytes related to those situated deeper in bone matrix in normal bone. However, ossification or calcification in subungual exostosis is rather casual, and osteocytes in this disorder may lack the capacity to elaborate compact bone.

A patient with diabetes mellitus who developed the typical classic lesions of Kaposi's sarcoma is described. Our patient presented with a reddish-purple papulonodular lesion on the right foot of five months' duration. A skin biopsy specimen showed a proliferation of spindle cells forming numerous vascular slits and a diffuse extravasation of erythrocytes. The patient's sera was negative for human immunodeficiency virus (HIV) antibodies and cytomegalovirus (CMV) antibodies. Ultrastructural examination demonstrated fibroblast-like spindle cells phagocytosing and digesting red blood cells to form vascular spaces. The patient died, due to gastrointestinal hemorrhage, and the autopsy revealed an extensive visceral involvement of Kaposi's sarcoma.

The proliferation and cell cycle phase composition of human dermal fibroblasts cultured on or in type I collagen lattices (reconstituted dermis model) were examined. On collagen lattices, as compared with conventional cultures on plastic dishes, the proliferation of human dermal fibroblasts was suppressed, being arrested at about one-half the saturation density after 10 days of culture. In collagen lattices, proliferation was further suppressed, being nearly arrested within 4-7 days of culture. Cells were analyzed for cell cycle phases by two-color flow cytometry using DNA staining and S phase cell staining with FITC-conjugated antibromodeoxyuridine antibody. After 5 days of culture, the number of S phase cells on collagen lattices was 49.3% of that on plastic dishes, with an increase in G0G1 phase cells of 79.8%. In collagen lattices, the number of S phase cells was very small (4.3% of all cells), and most of the cells accumulated in G0G1 phase. These findings suggest that the cell cycle of fibroblasts is arrested at G0G1 phase by their interaction with collagen. On the basis of these results, the reconstituted dermis model using collagen lattice is considered to be analogous to the dermis in vivo with respect to cell growth and cell cycle phase composition.

Human dermal fibroblasts were cultured in a hydrated type I collagen lattice. When collagen fibers were arranged in one direction, fibroblasts were arranged in the same direction. Cell proliferation was markedly suppressed in the collagen lattice as compared with that on plastic, with growth being arrested after day 5. No differences in proliferation were observed between aligned cells and randomly oriented cells. Flow cytometry with DNA staining was performed to analyze each phase of the cell cycle of fibroblasts. Among the 10,000 cell population, S phase cells on day 2 of culture accounted for 43% on plastic but were markedly inhibited to 25% in the lattice. On day 4, S phase cells accounted for 33% on plastic but only for 10% in the lattice. These findings suggest that cell advancement to the S phase is markedly inhibited in the collagen lattice, resulting in accumulation of most of cells in the G0G1 phase. The present study clearly showed that culture in the collagen lattice allowed alignment of fibroblasts with a definite orientation as observed in vivo and produced a status resembling that in vivo in terms of proliferation and cell cycle phase composition.

A 7-year-old girl with alopecia universalis had dystrophy of all 20 nails. A nail biopsy specimen disclosed features of lichen planus. The patient also had ichthyosis vulgaris and hypogammaglobulinemia. We are not aware of any previous reports of these associations, which we believe to be noncoincidental.

A case of subcorneal pustular dermatosis (SPD) is reported in a 48-year-old man with a 2-year history of hyperthyroidism (Graves' disease). SPD has been reported in association with IgA monoclonal gammopathy, but to our knowledge it has not been associated with hyperthyroidism. The coexistence of hyperthyroidism and SPD suggests that immunologic factors may play a role in the pathogenesis of SPD.

A 14-year-old boy with linear localized scleroderma had a dramatic improvement in contractures after treatment with N-(3',4'-dimethoxycinnamoyl) anthranilic acid (tranilast, Rizaben). The observation that this anti-allergic drug was effective in localized scleroderma lends further support to the concept that mast cells play a role in increased collagen synthesis in this disease.

Summary Four cases of solitary glomus tumour are reported, three of the angiomatous type, and one of the epitheliomatous type. In all cases, mucinous degeneration was seen, the extent of which correlated with the number of glomus cells.

Background. Rhinophyma is an end stage of acne rosacea. It results in a large nose due to a proliferation of sebaceous glands and fibrous tissue. Many cases of rhinophyma have been reported in the Western world; however, in Japan, rhinophyma has been an uncommon disease. Methods. We present two patients associated with rhinophyma who were treated by cross-shaped full-thickness excision followed by direct closure and compare the rhinophyma in Japan with that in the West. Results. To date, only 20 rhinophyma cases including our 2 cases have been reported in Japan. Epidemiologic factors of rhinophyma between the West and Japan do not differ except for location, malignancy and surgical treatment. Conclusions. In Japan, almost all cases are located on the lower half of the nose, treated by full-thickness excision followed by application of either skin grafts or direct closure. None have been malignant.

Summary Secondary cutaneous T-cell lymphoma may present with various types of skin lesions, hut rarely produces ulceration in contrast to primary cutaneous T-cell lymphoma. We report a case of malignant lymphoma of the tonsil that recurred as a giant cutaneous ulcer on the back of a 45-year-old man, Biopsy of the ulcer revealed a malignant lymphoma of diffuse, mixed-cell type. Surface marker analysis of the tumour cells showed suppressor/cytotoxic T-cell characteristics. The skin lesions were treated by electron beam therapy.

A 45-year-old woman presented with persistent headache which was of acute onset and occurred daily. Laboratory findings included thrombocytosis, elevated levels of antinuclear antibody, rheumatoid factor, and antibodies to Epstein-Barr virus (EBV). Magnetic resonance image of the head, left selective external carotid angiography, temporal arterial biopsy, and cerebrospinal fluid revealed no abnormal findings that could explain her headache. Her headache was compatible with NDPH, which has been reported by Vanast, and was thought to be related to EBV reactivation.

Summary Articular tissue was obtained at surgery for a femoral neck fracture in a patient with psoriasis without arthritis. The proteoglycan of the cartilage of the sample was analysed biochemically. Normal cartilage is known to produce two types of proteoglycan monomers (fast- and slow-sedimenting groups), which are distinguishable by density-gradient ultracentrifugation. In the psoriatic cartilage analysed in the present study, it was shown that the former group was absent and only the latter group remained.

Summary A case of a solitary pigmented neurofibrosarcoma involving the lateral side of the face in a 19-year-old female is described. Clinically it resembled a haemangioma, and angiography showed hypervascularity of the tumour. Light-microscopic study revealed that the major cellular element of the tumour was composed of bizarre neoplastic cells with large pleomorphic, hyperchromatic nuclei, conspicuous nucleoli and moderate to abundant amounts of conspicuous cytoplasm. Mitotic figures and multi-nucleate cells were present in most fields. Neurofibrosarcoma is one of the most highly malignant tumours; however, in the present case, there was no evidence of recurrence during a follow-up period of 5 years.

A 21-year-old male patient had localized papules on the chest and arms. Biopsy of a lesion showed it to be a malignant lymphoid neoplasm, and immunohistochemical studies confirmed a lymphoblastoma origin. Aberrant lymphoid cells were noted in the cutaneous blood vessels, but simultaneous examination of the peripheral blood showed no evidence of leukemia. One month elapsed before observable leukemic cells were found in the blood and sternal marrow, confirming the diagnosis of acute lymphocytic leukemia. Examination of a cutaneous biopsy specimen led to early diagnosis of the disease.

Tumor necrosis factor-alpha (TNF-alpha) has been demonstrated to selectively decrease the production of type I and type III collagens in human dermal fibroblasts. The effects of the commonly used food antioxidants, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), alpha-tocopherol, propyl gallate, superoxide dismutase (SOD), and catalase on TNF-alpha-induced growth enhancement and collagen metabolism were evaluated in the present study. BHA at concentrations of both 5 x 10(-5) and 10(-4) M inhibited cell proliferation and DNA synthesis induced by 10 ng/ml TNF-alpha in human dermal fibroblasts, while other antioxidants had minimal effects. Further, BHA (5 x 10(-5) M and 10(-4) M) significantly blocked TNF-alpha-induced decreases in collagen synthesis. These results suggest that antioxidants such as BHA may be involved in the modulation of collagen synthesis by TNF-alpha in human dermal fibroblasts.

S-carboxymethylated (SCM) fibrous proteins (FPs) from the scalp hairs of the three different hereditary hair abnormalities (trichorrhexis invaginata in Netherton's syndrome, pili trianguli et canaliculi in uncombable hair syndrome, and fine hair in anhidrotic ectodermal dysplasia) were analyzed by two-dimensional polyacrylamide gel electrophoresis. Comparison of the SCM FP compositions of the three hair abnormalities with that of the normal revealed that the SCM FP compositions of these three abnormal hairs were all electrophoretically different from each other and that of the normal.

We report a 22-year-old man with acquired dermal melanocytosis on the hand which developed without any history of previous inflammation. He had no history of contact with or oral administration of any drugs or metals which might have caused pigmentation. Histopathologically, spindle-shaped dermal melanocytes were observed running parallel to the collagen bundles and scattered in all portions of the dermis. The dopa reaction of these cells was positive. Ultrastructurally, dermal melanocytes containing numerous melanosomes (stages II to IV) which were surrounded by the extracellular sheath were observed. Differential diagnosis of other dermal melanocytosis was discussed.

Seven patients who had surgery for malignant skin tumors of the heel were followed up for several years. The reconstructive procedure performed was repair of the large skin defect after excision of a tumor, by using a flexor digitorum brevis muscle flap and an overlying free graft. Although this procedure is a good method without any serious complications, no long-term follow-up data supporting this opinion have been reported. In this study, plantar flexion of the toes and flattening of the plantar arch appeared 3 months after surgery and developed for 1 year, and the patients complained of easy fatigability in walking and difficulty in going up stairs. These sequelae were probably due to sacrifice of the flexor digitorum brevis muscle. We feel that this procedure should be replaced by another procedure that does not sacrifice any muscle, for example, that using a medial plantar flap.

The patient, a 34-year-old Japanese woman who noticed worsening of her rash after using topical corticosteroid preparations on her neck, was patch tested for both commercial preparations and corticosteroids themselves. The patch lest results revealed that she had a contact allergy to gold, oxytetracycline, and 2 types of corticosteroid (acetonides and esters) in 7 compounds (betamethasone valerate and dipropionate, hydrocortisone butyrate and hydrocortisone butyrate propionate, ameinonide, budesonide, and fluocinonide).

Summary Herpes simplex viral DNA was detected in biopsy specimens obtained from four patients with Kaposi's varicelliform eruption using an in situ hybridization technique with btotinylated complementary DNA probes. Herpes simplex viral DNA was consistently found almost exclusively in the nuclei of giant cells and balloon cells in vesicles during day 2, 5 and 7 of the disease. In the nuclei, the viral DNA staining pattern was granular or agglomerate. No positive staining was observed in the cytoplasm.

Endothelin-1, a potent vasoconstrictor peptide that has recently been isolated from the supernatant of cultured porcine aortic endothelial cells (1), is involved in some vascular disorders (2). Mal...

The effects of human recombinant tumor necrosis factor-alpha (TNF-alpha) on human keratinocytes cultured in a serum-free medium were investigated. TNF-alpha markedly suppressed cell growth. The growth-inhibitory effect was reversible and cytostatic at a concentration of 1-5 U/ml, but appeared to be irreversible and cytocidal at 10 U/ml. The growth suppressive effect was more marked when TNF-alpha was added in the late growth phase or preconfluent phase than when it was added in early or mid-growth phases. No effects of TNF-alpha on cell adhesion to the substrate were observed. These results indicate that TNF-alpha is a very potent anti-proliferative agent for human keratinocytes.

We report a case of microcystic adnexal carcinoma (MAC) occurring on the upper lip of an 82-year-old woman. Microscopically the tumor showed both pilar and sweat gland differentiation, involved the entire dermis and subcutaneous tissue, and invaded perineural spaces. Immunoperoxidase studies revealed carcinoembryonic antigen to be present in the ductal lining cells and in the amorphous content in the lumen, confirming sweat gland differentiation. The S-100 protein was positive in dendritic cells within the solid cell nests, but negative in cells lining cystic spaces. Ultrastructural study confirmed that the neoplasm was composed of two components, with pilar and eccrine differentiation. The former showed concentric layers of squamous epithelial cells with well-developed desmosomes and cytofilaments. The latter had ductal and alveolar structures; the ultrastructural features included: i) numerous villous folds of plasma membrane to interdigitate each other by focal desmosomes, ii) aggregates of cytofilaments, and iii) basally located myoepithelial cells which were separated from the surrounding stroma by rather thick basement membrane. In addition, distinct amyloid deposition was also observed on ultrastructural examination. To our knowledge, amyloid deposition has not been previously reported in MAC.

A case of generalized granuloma annulare associated with temporal arteritis is described. The patient, a 79-year-old man, noticed numerous asymptomatic lesions on his trunk and extremities for 3 months. Four months later, he suffered from headache and loss of vision. Both were successfully treated by oral administration of prednisolone.

We report a 19-year-old woman with the associated disorders of generalized pruritus, hypertrichosis and anorexia nervosa, all of which had been present for approximately 4 months. Psychotherapy induced both weight gain and resolution of the pruritus. We propose that anorexia-associated pruritus be considered one of the important cutaneous signs in anorexia nervosa.

A case of fixed drug eruption due to colchicine was reported. A 31-year-old man developed a dark purplish-red round macule on the glans penis 5 h after taking 2.5 mg of colchicine. An oral challenge test with 0.5 mg of colchicine provoked flare-up of the eruption.

Lysyl oxidase (EC 1.4.3.13), a copper-dependent enzyme which catalyses the formation of aldehyde cross-links, and acts primarily on collagen and elastin, is known to be increased during wound healing and in fibrotic disorders including liver cirrhosis and atherosclerosis, and to be decreased in some hereditary connective tissue diseases and in malignant cell lines. A recent study showed that lysyl oxidase might possess tumour suppressor activity as an antioncogene for ras. Little is known about the localization of this enzyme in human skin. In this study, we determined immunohistochemically the localization of lysyl oxidase in normal skin of young and elderly subjects obtained from sun-exposed and unexposed regions of the body. All skin samples tested had similar distributions of lysyl oxidase. The enzyme was present both extracellularly and intracellularly. Extracellularly, a few granular aggregates of immunoreactants were observed along collagen and elastic fibres. These granules were more common in the adventitial portion of the dermis than in the reticular portion. Of all sun-exposed and unexposed regions studied, the skin of the face displayed the greatest amount of extracellular immunoreactants. Immunopositive granules were observed intracellularly in fibroblasts, vascular endothelial cells, sweat glands, sebaceous glands, arrector pili muscles and some keratinocytes. These findings provide evidence that, as suggested in recent reports, lysyl oxidase may have a variety of intracellular functions.

An allergic drug eruption due to nitrazepam was reported in a patient who had been administered methotrexate and etretinate as concurrent therapies for severe pustular psoriasis. The clinical manifestation included a moon face-like appearance with edema, a diffuse erythematous rash over almost the entire body the day after taking nitrazepam and, two days later, eroded lesions developing on the cheeks, axillae and genitocrural regions. It was proved by provocation that this drug eruption had an allergic nature.

Unusual skin manifestations of a dermatophyte infection in a 30-year-old female were reported. These included a remarkable peripheral hyperkeratosis with central yellowish pus. The pinkish papillomatous lesions were covered with thick crusts. They gave off a foul smell. Abundant fungus elements were present in the crusts and keratotic layer. The trichophytin skin test was positive. Immunologic test results revealed that the patient had hypergammaglobulinemia with hypoalbuminemia. However, she had no edema or immunologic abnormalities. The skin lesions were successfully treated with topical miconazole cream, oral griseofulvin, and topical sulconazole cream. The etiologic agent was identified as Microsporum ferrugineum Ota 1922. It seemed to us that the unusual skin manifestation were induced by her immunological response to fungal and bacterial infection due to a long history of Candida granuloma.

20 patients with distal onychomycosis were given daily application of an ointment containing 2% tolnaftate and an ointment containing 20% urea under ODT. Following this, 17 of 20 patients developed onychomalacia and seven of these developed onycholysis 1 or 2 weeks later. The separated nails were cut as short as possible. Similarly to those patients with onychomalacia alone, occlusive dressing technique was continuously performed until the newly developed nails became macroscopically normal and no fungi were observed microscopically (responders). Following treatment, out of 20 patients, 14 responded. 5 patients who had a short course of treatment did not respond. Side-effects such as pain, hemorrhage and infection did not occur.

Samples of normal human skin were obtained from 48 sites in 26 subjects ranging in age from 2 to 85 years. The samples were examined by indirect immunofluorescence and immunoelectron microscopy using anti-human type IV collagen antibodies produced by immunizing rabbits with type IV collagen extracted from human placenta. Fluorescence was observed as granular or fine fibrous patterns, not only in the basement membrane at the dermo-epidermal junction, around the vessels, and the accessory organs of the skin, but also in the dermal regions in the vicinity of the basement membranes. This suggests the presence of type IV collagen in the dermis deep to the basement membrane. Ultrastructurally, the extrabasal lamina distribution of type IV collagen was noted as a partial distribution around the fibroblasts that existed close to the basal lamina. These findings are considered to be important in examining the function of this collagen in the dermis and the dynamics and metabolism of the basement membrane under normal and abnormal conditions.