Acquired progressive lymphangioma as a flat erythematous patch on the abdominal wall of a child.

An erythematous patch was noted on the abdominal wall of an 8-year-old boy. The lesion showed a prolonged initial clinical course, followed by rapid later growth, finally reaching 3.7 X 7.0 cm in size over four years. Despite the harmless clinical appearance, the lesion was histologically characterized by tortuous vascular channels with some cellular atypia. Immunoperoxidase staining disclosed no factor VIII-related antigen or reaction to Ulex europaeus I lectin on tumor cells. There has been no recurrence three years after local excision. Although many features in our case resemble those reported in the literature under the term low-grade angiosarcoma, our preferred designation for such cases is acquired progressive lymphangioma, rather than angiosarcoma, because of their benign behavior.

Modification of three avian viruses passaged in Chinese hamster lung cells (Don) in pathogenicity to chicken embryo.

The Beaudette 42 strain of avian infectious bronchitis virus, Sato strain of Newcastle disease virus, and Uchida strain of avian reovirus were passaged in Chinese hamster lung cells (Don), and some properties were examined. The Don-passaged strains showed a difference in replication in Don and chicken embryo kidney cells in one-step growth curve examinations and a partial modification in pathogenicity to chicken embryos; nevertheless, neutralization tests revealed no serological alteration.

Serotypes of avian infectious bronchitis virus isolates from field cases in Japan.

Eight etiologic agents isolated from field cases in Japan were identified as isolates of infectious bronchitis virus by agar-gel diffusion, buoyant-density determination on sucrose-density centrifugation, and morphological study by electron microscope of the purified viruses. In studies of the antigenic relationships of the eight isolates and six known infectious bronchitis viruses, antigenic diversity of these viruses was recognized from the degrees of relatedness using a plaque reduction in the "constant-virus decreasing-serum" method.

Protection by vaccination against bovine leukemia virus infection in sheep.

Four preparations were tested as potential vaccines to protect sheep against bovine leukemia virus (BLV) infection. Purified glycoprotein (gp) 51 and protein (p) 24 antigens from the virus and glutaraldehyde-fixed fetal lamb kidney (FLK) cells chronically infected with BLV or sheep fibroblasts transformed with BLV (SF-28 cells) were used to inoculate 12 sheep. Each vaccine was given 3 times 2 and 4 weeks apart to 3 sheep. Six sheep vaccinated with gp51 antigen or fixed FLK cells developed complement-fixing antibody against gp51; the titers ranged from 1:8 to 1:128 at the time of virus challenge exposure at postinoculation week 9. Although the 6 sheep inoculated with p24 antigen or fixed SF-28 cells developed antibody against the respective inoculum, none of these sheep had gp51 antibody at the time of challenge exposure. All 12 vaccinated and 2 control sheep were challenge exposed with BLV-infected lymphocytes, and cells from the sheep subsequently were tested for infection by syncytium assay. Sheep inoculated with gp51 antigen or FLK cells were protected, but sheep inoculated with p24 antigen or SF-28 cells became infected. The cytotoxic activity of lymphocytes from protected and nonprotected sheep was not different from that of normal sheep. Seemingly, purified gp51 antigen or fixed FLK cells were capable of preventing BLV infection in sheep. Humoral immune responses to gp51 appears to have an important role in protection against BLV infection, whereas cytotoxic activity of lymphocytes does not.

Pigmentary incontinence in fixed drug eruptions. Histologic and electron microscopic findings.

Pigmentary incontinence is a phenomenon observed in some inflammatory skin disorders. Clinically it may be seen as a slate-colored pigmentation. Histologically it is seen as an accumulation of melanin in the upper dermis. The possible mechanism for development of pigmentary incontinence is discussed based on a review of the literature and electron microscopic studies of fixed drug eruption.

Electron microscopic studies on Civatte body in Riehl's melanosis.

So-called Civatte bodies were investigated with light and electron microscopes in six patients with Riehl's melanosis. They were present in the lower part of the epidermis and/or in the upper dermis of the lesions of all patients. In the epidermis they were composed of wavily arranged fine filaments and entangled melanosomes, desmosomes and other cell organelles. In the dermis Civatte bodies seemed to transform into net-like or more amorphous masses. Melanosomes, desmosomes and other cell organelles were also observed within them, athough in small numbers. Dermal components containing collagen fibrils were occasionally seen to have merged into the rims of Civatte bodies in the dermis. And these Civatte bodies appeared to incorporate the adjacent dermal components and further mix with them to transform into amyloid-like filament masses. The amyloid-like filament masses consisted of straight and nonbranching filaments and were observed in the lesions of four patients under the electron microscope. However, they could not be identified as amyloid itself with light microscope, because they were negative in both the thioflavin T and congo red stainings.

Immunofluorescence studies on civatte bodies and dyskeratotic cells with anti-keratin antibody.

So-called Civatte bodies and dyskeratotic cells were investigated in some skin disorders by using indirect immunofluorescence techniques with anti-human keratin antibody. In the disorders with lichenoid tissue reaction such as lichen planus and DLE, Civatte bodies were observed in the lower epidermis and upper dermis and they reacted distinctly to the anti-keratin antibody. In malignant skin tumors which show dyskeratotic cells in the epidermis, such as basal cell epithelioma and Bowen's disease, dyskeratotic cells were more clearly reacted to the antibody than other keratinocytes. These observations present additional new evidence for the hypothesis that Civatte bodies are derived from tonofilaments of keratinocytes.

Amyloid in localized cutaneous amyloidosis: immunofluorescence studies with anti-keratin antiserum especially concerning the difference between systemic and localized cutaneous amyloidosis.

Amyloid of localized cutaneous amyloidosis and systemic amyloidosis were subjected to study with an indirect immunofluorescence technique using anti-keratin antiserum. Anti-keratin antiserum was prepared ad modum Sun & Green. Amyloid of localized cutaneous amyloidosis was positively stained for the antiserum, whereas amyloid of systemic amyloidosis (primary and multiple myeloma-associated) was negative. There was no difference between primary localized cutaneous amyloidosis (lichen amyloidosus and macular amyloidosis) and secondary localized cutaneous amyloidosis (amyloidosis associated with skin tumor). These results indicate that amyloid of localized cutaneous amyloidosis contains components derived from epidermal fibrous protein, probably tonofilaments of keratinocytes.

Immunofluorescence studies on epidermal keratinization in some skin disorders with keratin antisera.

The immunofluorescence technique using antisera against human plantar stratum corneum fibrous protein (total keratin) and 64K M.W. keratin subunit isolated from total keratin by SDS polyacrylamide gel electrophoresis (PAGE) was used to examine epidermal keratinization in some skin disorders. In normal skin, total keratin was distributed throughout the epidermis, while 64K keratin was localized at the suprabasal layers. In the case of squamous cell carcinoma (SCC), the tumor cells were uniformly stained positive with total keratin antiserum, whereas diminished or negatively stained cells were observed with 64K keratin antiserum (64K antiserum), suggesting that the tumor included cells in various stages of differentiation. In basal cell epithelioma (BCE), most of the tumor cells were negatively stained with 64K antiserum being consistent with the histologic observation that BCE is originated from the basal cells. However, some of the tumor cells were stained positive with 64K antiserum, indicating that individual cell keratinization might occur in BCE. In lichen planus, an inflammatory disease demonstrating so-called lichnoid tissue reaction, positively stained colloid bodies in the upper dermis were observed either with total keratin antiserum or with 64K antiserum. It was suggested that colloid bodies resulted from individual keratinization of damaged keratinocytes during inflammation.

Immunofluorescence studies on cutaneous amyloidosis with anti-keratin antibody.

Skin specimens obtained from two cases of primary localized cutaneous amyloidosis (PLCA) were subjected to study with an indirect immunofluorescence technique using anti-keratin antibody. Distinct fluorescence was observed in stratum corneum, and in spinous and basal cells of the epidermis and also in amyloid masses located in the upper dermis. These results appear to indicate that keratinous protein is one of the constituents of amyloid masses of cutaneous amyloidosis.

Quantitative autoradiographic studies in RNA and protein synthesis of dyskeratotic cells in morbus Darier.

Dyskeratotic cells in morbus Darier demonstrate various fine structures indicating that the keratinization process has proceeded to a certain extent in these cells; that is, there are Odland bodies, a marginal thickening, and keratohyalin granules. Thus, acantholytic cells in the lower epidermis are supposed to perform an incomplete and premature keratinization, and eventually transform to corps ronds and grains. The biosynthesis of RNA and proteins in these dyskeratotic cells was studied by electron microscopic autoradiography after in vitro incubation. Biopsy specimens taken from typical lesions of 3 patients with morbus Darier were incubated in one of the following media for 1-2 hours at 37 degrees C in air containing 5% CO2: (1) Earle's MEM containing 200 muCi/ml of [3H]uridine; (2) Earle's balanced salt solution containing 200 muCi/ml of [3H]leucine or 200 muCi/ml of [3H]histidine. After pulse labeling of [3H]-leucine or [3H]histidine, some specimens were fixed, dehydrated, embedded in Epon, and then prepared for electron microscopic autoradiography. These 3 labeled precursors were incorporated into early acantholytic cells in the base and wall of lacunae, but they did not accumulate in completely isolated acantholytic and dyskeratotic cells including corps ronds and grains. The nuclei of corps ronds in an early stage occasionally took up uridine. The accumulation of [3H]histidine was most remarkable in the granular cells which were not dyskeratotic, and silver grains showed a strong tendency to accumulate over keratohyalin granules by 3-6 hours of chase. [3H]histidine did not incorporate in dyskeratotic cells, even though they possessed a large number of keratohyalin granules. From these findings it seems difficult to assume that these acantholytic cells keratinize themselves further by synthesizing their structural proteins under the control of the RNA system during their upward movement. On the contrary, it appears more likely that most dyskeratotic cells result from neighboring cells which have already keratinized to some extent.

Immunoelectron microscopic examination of IgA deposition in dermatitis herpetiformis.

Ultrastructural localization of IgA in the skin of three Japanese patients with dermatitis herpetiformis (DH) was studied with the immunoelectron microscopic technique using periodate-lysine-paraformaldehyde fixation. In direct immunofluorescence studies, two of the three cases showed fine fibrillar deposition of IgA and the other case fine granules in the dermal papillae. In the former, the reaction products of IgA were present in the upper dermis forming various-sized aggregates which were occasionally arranged perpendicularly to the epidermis and appeared to be associated with microfibrillar bundles of the elastic tissue. Reaction products were also deposited to a lesser extent on the microfibrillar component of the elastic fibers at the lower part of the dermal papillae. However, in the latter, the reaction products were found to form smaller aggregates on and around the collagen fibrils rather than on the elastic tissue. Such a localization of IgA reaction products has not yet been reported in DH. The difference of the distribution patterns of IgA and the possible singularity of Japanese DH cases are discussed.