Yuhsuke Suzuki

A Possible Pathogenesis for Blackfoot Disease

Blackfoot disease (BFD) is an endemic peripheral vascular occlusive disease found among the inhabitants of the southwest coast of Taiwan. The clinical features of BFD are similar to those of Buergers disease. Pathology shows arteriosclerosis obliterans and thromboangiitis obliterans. The high arsenic content of artesian well water in the area is regarded as the main causal factor of this disease. Therefore, the purpose of this study was to observe the toxic effects of various arsenic concentrations on cultured human umbilical vein endothelial cells (HUV‐EC). The methods of this study included cell growth assay, 51Cr‐release assay, and staining of Factor VIII related antigen (FVIII‐RAg) and Ulex europaeus agglutinin I (UEA‐I) binding sites of HUV‐EC. The following data were obtained: 1) no obvious cytotoxicity in 51Cr‐release assay; 2) inhibition of the synthesis of both FVIII‐RAg and UEA‐I binding sites when the arsenic concentration was above 100 ng/ml; 3) dose‐dependent inhibition of growth of HUV‐EC by any concentration of arsenic. At a higher concentration of more than 100 ng/ml, arsenic inhibited endothelial cell proliferation and glycoprotein synthesis, whereas it only inhibited the proliferation at a lower concentration of less than 50 ng/ml. It is suggested that arsenic, at both higher and lower concentrations, may damage endothelial cells. Such damage may play an important role in the pathogenesis of BFD.

A Case of Sneddon's Syndrome with Positive ANA and Anti‐cardiolipin Antibodies: Primary Anti‐phospholipid Syndrome?

A 22‐year‐old woman developed ulcerative lesions on the lower extremities which usually exacerbated during the summer. Histological analysis revealed a micro‐thrombotic lesion in the deep dermis without inflammatory cell infiltration or fibrinoid degeneration of blood vessels. Magnetic resonance imaging revealed multiple cerebral infarctions. Abnormal laboratory findings included an elevated anti‐cardiolipin antibody titer and positive speckled pattern ANA (x80), but without other manifestations or signs of SLE. FACS analysis revealed that the patients serum reacted with ethanol fixed endothelial cells in addition to keratinocytes and peripheral blood neutrophils. This case was thought to be livedo reticularis and cerebral thrombotic lesions (Sneddons syndrome) associated with atrophie blanche or livedo(id) vasculitis and may be one clinical subset of primary anti‐phospholipid syndrome.

A monoclonal antibody, SKH1, reacts with 40 Kd sweat gland-associated antigen.

Several monoclonal antibodies (MAB) have been produced using an eccrine carcinoma cell line as an immunogen. One such MAB, SKH1, reacted with both the secretory portion and coiled duct of the eccrine and with the secretory portion of apocrine gland. SKH1, however, did not react with myoepithelial cells, intradermal ducts of both types of sweat gland, or with other components of normal axillary skin including the epidermis and follicular apparatus. The reaction was strongest if the specimen was fixed with 80% methanol, and moderate on non‐fixed or acid‐alcohol‐fixed specimens. Only weak reaction was obtained on cold acetone‐fixed specimens, and reaction was negative with formalin‐fixed, paraffin‐embedded tissues. SKH1 reacted positively with the cytoskeleton of the eccrine carcinoma cell line, Colo‐16 and MCF‐7. Applied to pathological skin specimens, SKH1 reacted with the tumor cells of clear cell hidradenoma, syringocystadenoma papilliferum, and extramammary Pagets disease. SKH1 also reacted with the tumor cells of meta‐static adenocarcinomas arising from lung, breast and ovary. SKH1 did not react with the majority of tumor cells of eccrine poroma, but reacted with single–layered cells lining narrow ductal lumina. SKH1 did not react with epithelial cells lining cystic or ductal lumina of syringoma, but reacted moderately with the amorphous keratin–like substance filling the lumina. Immunoblot analysis revealed that SKH1 recognizes a 40 Kd sweat gland‐associated antigen, and can be an aid to identifying tumors arising from sweat gland structures.

A Case of Sclerosing Lymphangitis of the Lip

A 62‐year‐old woman noted a cord‐like swelling on the inner surface of her upper lip. A cross‐section of the lesion showed a radial‐shaped lumen surrounded by a thickened fibrous wall. Immunohistochemical staining with the anti‐factor VIII‐related antigen (FVIII‐RAg) antibody was negative on the luminal surface and swollen wall, although the vasa vasorum in the swollen wall were positive. The lesion was thus considered to be of lymphatic origin; that is, it derived from a lymphatic collecting vessel. For the present case, the term ‘lymphangiopathia obliterans’ is considered appropriate. This is the second report of such a lesion appearing on the lip in the literature.

GUINEA PIG ENDOTHELIAL CELLS IN CULTURE

In pursuit of a model for the studies of endothelial cells in a readily available experimental animal, culture methods and the in vitro characteristics of endothelial cells isolated from guinea pig aorta are described. Endothelial cells were harvested by trypsinization with perfusion techniques and cultured in Eagles minimal essential medium supplemented with 10% fetal calf serum. Their passage cultivation was possible for more than six months and they have maintained the in vitro morphological characteristics of endothelial cells.