Walter Gebhart

Enzyme histochemical analysis of cell viability after argon laser-induced coagulation necrosis of the skin.

Reduction of nitroblue tetrazolium chloride, a redox indicator, by nicotinamide adenine dinucleotide diaphorase produces in frozen tissue sections an intense blue cytoplasmic pigment. The activity of this enzyme has been shown to subside immediately upon cell death. Twelve patients with port-wine stains were treated with an argon laser. Frozen tissue sections from biopsy specimens obtained before and 10 minutes, 24 hours, and 48 hours after laser application were processed for nitroblue tetrazolium chloride staining. In normal skin all epidermal and dermal cells displayed dense cytoplasmic blue granular pigment that spared the nuclei. In port-wine stains the laser-induced coagulation necrosis was first seen as an arc-shaped, sharply demarcated, unstained, nitroblue tetrazolium chloride-negative area. Initiation of epidermal repair could be observed in all 48-hour sections. The nitroblue tetrazolium chloride method, when compared with hematoxylin and eosin staining, allowed an easier and more accurate definition of laser injury because of the color difference between damaged and normal tissue.

Hydroxyethylstarch deposits in human skin--a model for pruritus?

Severe itching for unknown reasons has been reported after administration of hydroxyethylstarch (HES) in haemodilution therapy of humans. After HES treatment, vacuoles in cells of various organs in humans have been shown, predominantly affecting the mononuclear phagocyte system. These vacuoles present indirect evidence for phagocytosis of HES particles. Since phagocytosis is also known to occur in the skin, this organ might represent a target for HES deposition, resulting in subsequent release of mediators responsible for the observed itching. The aim of the present investigation was to study skin biopsies of patients, who had received HES and suffered subsequently from itch. Skin sections were investigated for morphological impairment by means of light and electron microscopy, immunohistochemistry and immunoelectron microscopy using a polyclonal anti-HES antiserum. Storage of HES was demonstrated in the skin of all patients, mainly in dermal macrophages, endothelial cells of blood and lymph vessels, some perineural cells and endoneural macrophages of larger nerve fascicles, some keratinocytes and Langerhans cells. Treatment with antihistaminic agents proved ineffective in these patients; this fits with the observation that morphological signs of histamine release from mast cells were absent. These findings indicate that other mediators from HES-affected cells must be responsible for the development of the itching. Thus, investigation of HES storage may be a useful contribution to the elucidation of release of itch mediators and induction of pruritus.

“Lumpy‐Bumpy” Elastic Fibers in the Skin and Lungs of a Patient with a Penicillamine‐Induced Elastosis Perforans Serpiginosa

Penicillamine‐induced cutaneous elastosis perforans serpiginosa associated with a large air‐cyst in the right lung is described in a 29‐year‐old female patient with Wilson disease. Identical light and electron‐microscopic changes were present in both dermal and pulmonary elastic tissue, suggesting a disseminated drug‐induced cutaneo‐visceral elastosis. Lung cysts have not been previously reported in association with long term penicillamine treatment. The electron‐microscopic morphology of the elastic fibers was found to be “specific” enough to allow separation of penicillamine‐induced elastosis perforans serpiginosa from other forms of this disease.

Cyclosporin-A-induced hair growth in human renal allograft recipients and alopecia areata.

Cyclosporin A is now widely used as a potent immunosuppressive agent in organ transplantation. One of the most common dermatological side effects of the drug is dose-dependent hypertrichosis, which affects more than 80% of the patients with variable degrees of hirsutoid hair growth on different areas of the face and body [4, 7]. At present, the cause of cyclosporin-A-induced hypertrichosis is unclear; a multifactorial origin with both androgen-dependent and independent components has been suggested [2, 6]. Data obtained from experiments in nude mice indicate that cyclosporin A transforms lanugo hair producing follicles into vellus or even terminal hair follicles or stimulates dormant follicles by shifting them from a resting phase to an active one [8]. The aim of the present study was to investigate the ovarian, adrenal and gonadotropine hormone aspects of cyclosporin A therapy. Blood samples were obtained from 11 cyclosporinA-treated renal allograft recipients (4 females, 7 males) at 8:00 a.m. The mean age in this group was 32.8 years (18 54 years), mean duration of therapy 10.1 months (3 -29 months), mean dosage of cyclosporin A 8.6 mg/kg per day (3.5 13 rag), and mean prednisone dosage 15.2 rag/day. Blood samples from 11 azathioprine-treated renal allograft recipients (4 females, 7 males) served as control. In this group, the mean age was 37.5 years (26-59 years), the duration of therapy 56.8 months (36-164 months), the mean azathio-

Histological and immunohistochemical investigations of hydroxyethyl-starch deposits in rat tissues.

Tissue storage of hydroxyethyl starch (HES), a widely used artificial colloid, has been reported. In order to clarify whether storage of HES can be detected in tissues by immunohistochemical methods, use was made of a polyclonal rabbit anti-HES antiserum. Thirteen days after a single intravenous injection of HES rats were sacrificed and liver, spleen, lymph node, lung, kidney and skin were removed. On paraffin sections in all organs the anti-HES antiserum stained mainly cells which could be attributed to the mononuclear phagocyte system, as confirmed by the use of the antimacrophage monoclonal antibody ED1. The use of a polyclonal anti-HES antiserum may allow analysis of long-term storage and possible side effects in various tissues of man.

Disseminated superficial actinic porokeratosis: Experimental induction and exacerbation of skin lesions

A 55-year-old woman with disseminated superficial actinic porokeratosis (DSAP) had lesions on sun-exposed skin areas that were exacerbated during the summer months and involuted in winter. This is the third report in which induction and exacerbation of DSAP lesions were achieved by irradiation with artificial ultraviolet light sources. Our data show that UVB plus UVA is more effective in inducing new or exacerbating preexisting skin lesions than either wavelength alone. We believe that testing with the appropriate ultraviolet light sources is a practical means to differentiate between DSAP and disseminated superficial porokeratosis.

Colloid milium: light and electron microscopic investigations

A case of colloid milium (‘juvenile type’) is presented. Histochemically the colloid material showed the well known similarity to amyloid where especially in the upper portions of the colloid immunoglobulin and complement depositions could be found. Fibrin was demonstrated predominantly in the periphery of individual colloid islands.

Untersuchungen über die Blasenbildung beim Lichen ruber planus

SummaryTwo cases of bullous Lichen planus were examined electron-microscopically. The following results were obtained: 1.Blister formation takes place predominantly between the basal lamina and the cytomembranes of basal keratinocytes, that meansintrabasal.2.Basal cells show distinct degenerative changes, with may result in cytolysis. Frequently a fibrillar transformation of keratinocytes is observed. No evidence for cleft formation caused by a damage of the tonofilament-desmosome complex was found. Acytolytic origin of the blister is assumed.3.In the blister cavity fibrin masses and numerous inflammatory cells can be demonstrated besides cytolytic and “fibrillar” keratinocytes. The roof of the bulla is formed by the lower stratum spinosum. There the keratinocytes are swollen with intracellular edema; dyskeratotic cells and fibrillar bodies can be found. Similar alterations are observed in the surroundings of the bulla.ZusammenfassungDie elektronenmikroskopische Untersuchung von 2 Lichen ruber bullosus-Fällen ergab folgende Befunde: 1.Die Blasenbildung erfolgt vorwiegend zwischen Basallamina und Plasmalemm der basalen Keratinocyten, das heißtintrabasal.2.Die Basalzellen weisen ausgeprägte, bis zur Cytolyse gehende Degenerationszeichen auf. Auch eine fibrilläre Umwandlung von Keratinocyten ist häufig nachweisbar. Im vorliegenden Material fand sich kein Hinweis, daß eine spezielle Schädigung des Tonofilament-Desmosom-Komplexes die Hohlraumbildung verursacht. Einecytolytische Entstehung der Blase wird angenommen.3.Im Blaseninhalt finden sich neben cytolytischen und fibrillär umgewandelten Keratinocyten Fibrinmassen und zahlreiche, eingewanderte Infiltratzellen. Die Blasendecke wird vom unteren Stratum spinosum gebildet. Die Keratinocyten sind in diesem Bereiche durch ein intracelluläres Ödem geschwollen, dyskeratotische Zellen und fibrilläre Körper sind nachweisbar. Ähnliche Veränderungen lassen sich auch in der Blasenrandzone erkennen.

Some electron microscopic and autoradiographic results concerning cutaneous neurofibromas in von recklinghausen's disease

SummaryThe cutaneous tumors of 5 patients with clinical symptoms of Recklinghausens neurofibromatosis were investigated by light and electron microscopy. Special reference was given to the study of initial changes of the disease and the structure of the nerve fascicles which end in the tumor. In 2 patients the proliferating cells were determined by using an antoradiographic technique. It was discovered that all cellular elements of a peripheral nerve are involved by the tumor formation. The results were discussed and compared with the alterations during nerve regeneration after traumatic injury.ZusammenfassungDie Hautknoten von 5 Patienten mit klinisch diagnostizierter Neurofibromatose Recklinghausen wurden im Hinblick auf die initialen Veränderungen der Erkrankung licht- und elektronenmikroskopisch untersucht. Besonderes Augenmerk wurde auf die Struktur der Nervenfaszikel, die für die Bildung der Tumore verantwortlich sind, gerichtet. Bei 2 Patienten wurden mit Hilfe autoradiographischer Methoden die proliferierenden Zellen bestimmt. Die Untersuchungen ergaben, daß bei der Tumorbildung alle Zelltypen des peripheren Nerven beteiligt sind: Die Schwannschen Zellen, die Endoneuralfibroblasten und die Perineuralzellen. Die Veränderungen wurden mit dem normalen Ablauf der Nervenregeneration nach traumatischer Schädigung verglichen.

Epidermal Changes in Lichen Planus

Ultrastructural investigations in 15 patients with lichen planus (LP) revealed a multitude of epidermal changes. The most consistent findings were (1) multiplication, irregular folding or dislocation of the basal lamina, (2) fragmentation with degenerative changes of basal keratinocytes, (3) formation of numerous fibrillar Civatte bodies and, (4) presence of dyskeratotic elements. In addition, mitotic figures of keratinocytes and Langerhans cells were observed. These findings support the view that the primary event in LP represents an injury involving epidermal basal cells. Furthermore, the replacement of destroyed tissue seems to originate not only from the margin of the lesions or the skin appendages but also from unaffected cells within the LP papule.